It's Still in the Water! [Archive] - Reef Central Online Community Archives

Habib

08/13/2002, 08:20 AM

Thanks for entering the discussion, Habib.

You are welcome!

I know that you posted a huge amount of data and links to papers on this topic in my forum a few minutes ago, and it would be great if you could bring it over here (a single cut and paste looses the links).

OK I will give it a try. If links are missing then I will try to correct it ASAP.

Here is one of my posts:

Tatu, Randy & others,

Sorry, it has become a tiny bit loooooooong:D

Toxicity of (trace) elements depends on the speciation, that is how it is present in the water column.

Free ionic forms will be far more toxic when present in high concentrations than organically bound forms.

The method used for measurement of the elements by Ron also measures besides the free ionic and organically bound forms also particulate forms. E.g. strongly adsorbed on other particles or as the solid oxide or carbonate forms. They will have no direct toxic effect.

Also the data in Ron's study regarding antimony and arsenic is IMO highly suspect. The values are high and are (virtually) the same for all tanks.

If it were known that reef tank inhabinants intentionally release species that control metal concentrations, and that these are not just random happenings but controlled events, then it brings validity that perhaps reef tank inhabitants, either intentionally or unintentionally, detoxify their own environment by releasing metal chelating agents.

O.K. here are some to start with.

The first one is easy to read and is very informative for a large crowd:

Chelation, uptake, and binding of trace metals (http://www.princeton.edu/~cebic/chelbindintro.html)

The following is the same as above but is the more scientific version:

Extra-cellular iron siderophores: structure and regulation (http://www.princeton.edu/~cebic/chelbindadvanced.html)

Another one:

Acquisition and Utilization of Transition Metal (http://www.chem.ucsb.edu/~butlergroup/Articles/080201a6.jrn.pdf)
In the above links you will find also other interesting pages on other topics.

The second parts deals with algae:
Intracellular binding: details (http://www.princeton.edu/~cebic/bind-detail.html)

The following is from:

http://www.liv.ac.uk/~sn35/Documents/Research_Statement.html

Marine chemical findings
Our measurements are among the first to demonstrate that the biogenic metals (copper, zinc, iron, and cobalt) occur organically complexed in the oceans. Our measurements have shown that organic complexation reactions control the transport of copper, nickel and zinc through estuaries, and that dissolved metal concentrations in estuarine and coastal waters follow dynamic patterns. Unexpectedly anionic elements such as antimony, molybdenum, and uranium were also shown (by CSV) to occur to a significant extent as non-labile species in estuarine waters. A voltammetric study of the geochemistry of platinum in the Indian Ocean conclusively showed that this element has a geochemical behaviour reminiscent to that of manganese in those waters. Our investigations have shown that titanium and aluminium occur as unknown non-labile species, either colloidal or organically complexed, in the oceanic water column. We used our methods to determine the complex stability of sulfide with several metals in seawater, and found that the complex with copper is much more stable than expected.

Probably the most important finding of the last decade in oceanography has been that lack of iron limits oceanic productivity (the Iron Hypothesis). Our finding that iron is organically complexed provides the explanation for its apparent poor availability, and moderates the Iron Hypothesis to a combination of lack of iron and unavailability.

Photochemical effects and the existence of transient species were investigated in the field, showing that the redox chemistry of chromium appears to be controlled both by photochemical and biological processes in the upper water column, causing the presence of significant amounts of chromium(III) where it is thermodynamically unexpected.

We demonstrated that the biologically important folic acid and glutathione occur dissolved in the oceanic water column of the NE Atlantic. Further work has now shown the importance of thiols like glutathione on the chemical speciation of copper suggesting that thiols may well account for at least part of the ligands we see in natural waters.

The complexation reactions likely regulate the rate at which these metals are transferred through the membranes of microorganisms. For this reason we looked at interactions with microorganisms and found that marine algae (the important bloom forming coccolithophore Emiliania huxleyi) release iron-binding ligands in response to iron additions. This work stimulated the use of seawater cultures without modifier additions

Another one:

Nature 400, 858 - 861 (1999)

Competition among marine phytoplankton for different chelated iron species

DAVID A. HUTCHINS*, AMY E. WITTER*, ALISON BUTLERâ€* & GEORGE W. LUTHER III*

* College of Marine Studies, University of Delaware, Lewes, Delaware 19958, USA
â€* Department of Chemistry, University of California, Santa Barbara, California 93106, USA

Correspondence and requests for materials should be addressed to D.A.H. (e-mail: dahutch@udel.edu).

Dissolved-iron availability plays a critical role in controlling phytoplankton growth in the oceans,. The dissolved iron is overwhelmingly (99%) bound to organic ligands with a very high affinity for iron, but the origin, chemical identity and biological availability of this organically complexed Fe is largely unknown. The release into sea water of complexes that strongly chelate iron could result from the inducible iron-uptake systems of prokaryotes (siderophore complexes) or by processes such as zooplankton-mediated degradation and release of intracellular material (porphyrin complexes). Here we compare the uptake of siderophore- and porphyrin-complexed 55Fe by phytoplankton, using both cultured organisms and natural assemblages. Eukaryotic phytoplankton efficiently assimilate porphyrin-complexed iron, but this iron source is relatively unavailable to prokaryotic picoplankton (cyanobacteria). In contrast, iron bound to a variety of siderophores is relatively more available to cyanobacteria than to eukaryotes, suggesting that the two plankton groups exhibit fundamentally different iron-uptake strategies. Prokaryotes utilize iron complexed to either endogenous or exogenous siderophores, whereas eukaryotes may rely on a ferrireductase system, that preferentially accesses iron chelated by tetradentate porphyrins, rather than by hexadentate siderophores. Competition between prokaryotes and eukaryotes for organically-bound iron may therefore depend on the chemical nature of available iron complexes, with consequences for ecological niche separation, plankton community size-structure and carbon export in low-iron waters.

Citing a few:

Ahner, B. A.. Cornell University, baa7@cornell.edu
Oleson, J. A.. Cornell University, jro5@cornell.edu
Slinski, K. M.. Cornell University, kms32@cornell.edu

GLUTATHIONE CONCENTRATIONS IN FRESHWATER AND MARINE PHYTOPLANKTON

Small organic sulfur compounds play a large role in the intracellular speciation of trace metals in marine and freshwater algae. Glutathione, the principle free thiol in most algae, can complex metals directly or is polymerized enzymatically into metal-binding ligands called phytochelatins. In Emiliana huxleyi, concentrations of g-glutamyl cysteine (a precursor of glutathione) are significantly higher than glutathione which has implications with respect to phytochelatin synthesis. We have examined the effect of prolonged metal exposure on glutathione synthesis in both freshwater and marine algae. We found that deviation from control concentrations of glutathione is highly variable among species, though short-term exposure to Cd stimulates glutathione synthesis in some organisms. Utilizing published and experimentally determined binding constants for glutathione and phytochelatin it is possible to evaluate the probable intracellular speciation of various trace metals such as Cd and Hg. In addition, we are evaluating the use of various metal-specific fluorescent probes to quantify intracellular metal speciation.

From:

http://www.science.plymouth.ac.uk/DEPARTMENTS/Environmental/marchem/marine_chem.htm#Interactions%20between%20Trace%20metal%20and

The uptake and availability of metal species to macrophytes (seaweeds) is currently under investigation within the group. Sensitive electrochemical techniques are used to determine the speciation of metals within culture media containing juvenile stages of seaweeds. These studies have shown that seaweeds release organic material which alters the metal speciation in the culture media. The concentration of this organic material (complexing ligand) released is dependant on both the growth of the the macrophyte and on the concentration of metal, perhaps indicating that these complexing ligands are released in a direct response to the presence of a toxic metal. In parrallel to this, investigations are being undertaken into the intracellular production by algae of specific binding ligands (termed phytochelatins) in response to metals. The distribution and occurrence of phytochelatins is being investigated under laboratory conditions in order to assess the variation in response between different algal species and under variations in temperature and salinity. In addition, concentrations of phytochelatins in natural assemblages of phytoplankton in contaminated and uncontaminated estuaries are being measured. The aim of these studies is to relate phytochelatin production with metal toxicity and to develop a useful biomarker for metal contamination.

Other links:

Lack of phlorotannin induction in the brown seaweed Ascophyllum nodosum in response to increased copper concentrations (http://www.int-res.com/abstracts/meps/v192/p119-126.html)

Bioavailability of biologically sequestered cadmium and the implications of metal detoxification (http://www.int-res.com/abstracts/meps/v147/p149-157.html)

From the following link:

ABILITY OF IMMOBILIZED CYANOBACTERIA TO REMOVE (http://www.engg.ksu.edu/HSRC/JHSR/v1_no2.PDF)

Reports also indicate that carboxyl groups on algal cell biomass are
responsible for binding to various ions (Gardea-Torresdey et al.,1990). Live algae possess
intracellular polyphosphates which participate in metal sequestration, as well as algal extracellular
polysaccharides that serve to chelate or bind metal ions (Zhang and Majidi,1994; Kaplan et
al.,1987; Van Eykelenburg,1978). Strains of Synechocystis spp. have been shown to develop a
thickened calyx when exposed to copper-stressed growth conditions (Gardea-Torresdey et
al.,1996a). Synechococcus sp. PCC 7942 was found to possess a copper-transporting P-type
ATPase in the thylakoid membrane (Bonilla et al.,1995). Synechococcus cedrorum 1191 was
shown to be tolerant to heavy metals and pesticides (Gothalwal and Bisen,1993). Other
investigators have studied the biosorption of heavy metals by algal biomass (Volesky and Holan,
1994; Volesky and Holan, 1995; Volesky and Schiewer, 1997). Such findings show the
possibility of manipulating or overexpressing existing resistance mechanisms and the use of such
organisms to remove harmful metals from the environment.

Other links:

The effect of Fe and Cu on growth and domoic acid production by Pseudo-nitzschia (http://aslo.org/lo/toc/vol_47/issue_2/0515.pdf)

See page #2 of:

Phytoplankton Physiology and Ecology of Metals (http://shrimp.ccfhrb.noaa.gov/research/pep2000.pdf)

rshimek

08/13/2002, 09:24 AM

Originally posted by Randy Holmes-Farley

Hi Randy,

You'll of course need a control

Gee, will I really? Sage advice coming from some one, who has to the best of his published aquarium record, never run an experiment, or done a test.

I think you need to show that any organics are having any effect.

Until you do that you are simply "blowing smoke."

So, boy, spend some time and spend some money doing the work.

I would be glad to see a way of detoxifying these metallic poisons, and I would be more than happy to see you demonstrate it. But, you haven't done it....

Which corals? Any that people keep?

Try reading the article.

If so, I take away just the opposite

ROFL - It is truly fun to argue when you have absolutely no data to work from isn't it.

Do some experiments, collect some data. Then your arguments will have some credibility.

To suggest it is magircal because you are unaware of it is only natural, but not very scientific, or accurate.

To say that it is happening in a system when you have not ONE shred of evidence to support it, is absurd.

rshimek

08/13/2002, 09:32 AM

Originally posted by Habib

Habib,

Toxicity of an element depends besides the concentration also very strongly on speciation.

So when you say that there are toxic concentrations in our tank are you just guessing or did you actually determine the speciation?

These tests were run exactly as standard pollution monitoring tests are run. The data are compared in exactly the same manner with the same conclusions being reached.

If the tests show that levels of a certain metal kill animals, and if the tests show that the levels are above those levels, and animals do die then it is reasonable to suggest that the metal is responsible. It really doesn't matter the type of ion that is involved.

Mythical chelators do not exist. Natural chelators excreted by marine microorganisms, algae,....do exist.

I will grant you that they do. But there are no data that they are found in any concentrations in our aquaria, nor that they have any effect there.

We not only did the same type of tests but we went even much further.

Sure you did. Yup. Somewher you have those data....

Now lets see them published somewhere.

I can back this.

Publish the data.

It is really easy to criticize someone else without a stitch of published data to support your arguments.

Adam

08/13/2002, 09:39 AM

Hi all,

No that I have stopped laughing, I had to chime in....

Dr. Ron,

More than likely, you - and most of us - have morphologies or species of corals that are pollution tolerant growing in our systems; in effect, they would have to be to survive. Such corals are found in nature, as well - but are not normally dominant as they generally grow slower and compete less well than their non-tolerant cousins. These animals likely be able to detoxify additional amounts of the toxic chemicals.

How lucky we are that the collectors target such pollution tolerant variants for the coral trade! How is that more rational than chelation as an explanation?

Did you poll the participants in your study to gather some meaningful guage of the health and survival of the animals in their aquariums?

Such data would be very telling in terms of the real toxicity of these agents in our tanks.

To say that it is happening in a system when you have not ONE shred of evidence to support it, is absurd

I think the posted abstracts represent far more than mere shreds!

Lastly, I hope that your response to the need for a control was only professional insult over the reminder, and not denial that it is necessary.

Please understand, I am not asserting that one hypothesis is more correct than the other. I believe that our corals probably to build tolerance, and I do believe that heavy metals are chelated. I mostly want to make the point that both hypotheseses must be tested (with controls!)

In a thread in Randy's forum, someone mentioned testing for heavy metals in skimmate. Elevated concentrations would be pretty compelling evidence for Randys theory.

Adam
;) ;)

rshimek

08/13/2002, 09:44 AM

Originally posted by Randy Holmes-Farley

From the Marine Chemistry Group at the University of Plymouth (the bolding is mine):

"The uptake and availability of metal species to macrophytes (seaweeds) is currently under investigation within the group.

Gee, I am so glad.

Sensitive electrochemical techniques are used to determine the speciation of metals within culture media containing juvenile stages of seaweeds. These studies have shown that seaweeds release organic material which alters the metal speciation in the culture media. The concentration of this organic material (complexing ligand) released is dependant on both the growth of the the macrophyte and on the concentration of metal, perhaps indicating that these complexing ligands are released in a direct response to the presence of a toxic metal.

Very nice, what algae or maybe what other plants... sea weeds, don' t they know which ones? More to the point does this happen with algae that occur in aquaria?

. In parrallel to this, investigations are being undertaken into the intracellular production by algae of specific binding ligands (termed phytochelatins) in response to metals. The distribution and occurrence of phytochelatins is being investigated

Lots of things are being investigated. To show that it works in a lab is nice. Now does it work anywhere else.

The aim of these studies is to relate phytochelatin production with metal toxicity and to develop a useful biomarker for metal contamination."

Hey, great, they will have a biomarker. So we can test our tanks and find that biomarker and know "fur shure" that metals toxicitiy is indeed occurring. LOL!

Basically what they are saying is that the metals are being toxic and they are finding a way in which some algae try to detoxify them. Well, I hope they find their marker, and I hope they can convince somebody to market the commercial version. It might be useful.

And what happens when the concentration rises yet futher?

This is nice post but it is simply irrelevent to our systems.

rshimek

08/13/2002, 10:02 AM

Originally posted by Habib

Also the data in Ron's study regarding antimony and arsenic is IMO highly suspect. The values are high and are (virtually) the same for all tanks.

Well, let's see... Arsenic was found only in one tank. Antimony in many. So, run tests on 20 random aquaria and see what you found.

You guys are missing the point....
The tests are the same ones that are used to detect high metals concentrations in environmental monitoring. These tests are used to assess environmental conditions around polluted localities. They work. Positive high values are correlated with signficiant environmental degradation. Such degradation has also been shown with corals under the same conditions. Aquarists have problems keeping corals alive for extended periods, many other corals die immediately after they are added to tanks. Every environmental scientist EVERY SINGLE ONE who works with these animals and who considers metals considers these materials as poisons.

We have concentrations of these materials, specifically Copper, Zinc, Nickel, and Vanadium from 10 to 500 x more concentrated in our tanks that in nature.

I suspect is it very reasonable to suggest these poisons are having an effect.

If they are not having an effect then it is incumbent upon Habib and Randy to demonstrate that they are not.

Sure there are data that in very low concentrations in natural environments, the metals may be - operative words - may be detoxified. So... show that it works in aquaria.

Get your hands wet, boys.....

Randy Holmes-Farley

08/13/2002, 10:04 AM

Gee, will I really? Sage advice coming from some one, who has to the best of his published aquarium record, never run an experiment, or done a test.

So, boy, spend some time and spend some money doing the work.

Let's not let this degrade into name calling. As the Vice President of Chemical Research at a pharmaceutical company, as the inventor of approved drugs that you may take yourself, as the holder of many dozen chemical patents, and as a very intelligent scientist, I am quite aware of what it takes to do quality science as opposed to junk science. Have you not read any of my published papers and the experimets therein? Ahh, such a pity.

Since neither you nor anyone else knows what organics are in your tank water, to attribute any found toxicities to metals is simply ignoring the other possibilities. Nearly all organics are toxic at some concentration. Some at very low levels, and some at high levels. Since you don't know what you've got, or what the concentrations are, how can you possibly say that this isn't a real possibility?

To show that it works in a lab is nice. Now does it work anywhere else.

Isn't that what an experiment is? Isn't that what you plan to do? Isn't that even what a reef tank is?

Besides interfering with your current pet theory, why could it not happen in reef tanks if it happens in tanks in labs?

rshimek

08/13/2002, 10:15 AM

Originally posted by Adam

Adam,

How lucky we are that the collectors target such pollution tolerant variants for the coral trade! How is that more rational than chelation as an explanation?

Pollution tolerant animals abound. We are lucky they do, elstwise our coasts would be barren.

We can keep alive a small subset of all the animals imported, and relatively few species are collected at all. In a lot cases, there are only certain clones (frags) that may be kept alive and well. Hmmm.....

Chelation may work. Prove it. I have given a reasonable explanation of much of the mortality in aquaria, and related it to demonstrable testable source.

If it is not the cause, then let those who suggest that it isn't show that it isn't.

Did you poll the participants in your study to gather some meaningful guage of the health and survival of the animals in their aquariums?

No. I don't think this can be done observation. For example, stress in corals is often measured by increasing respriation rate, and this is impossible for most hobbyists to do.

Such data would be very telling in terms of the real toxicity of these agents in our tanks.

Yes, but it can't be done.

I think the posted abstracts represent far more than mere shreds!

No, they didn't. The have no evidence that such materials are even found in our aquaria, let alone that they are working as suggestted there.

Lastly, I hope that your response to the need for a control was only professional insult over the reminder, and not denial that it is necessary.

Actually it was a comment to the poster, who seems to have remembered that one needs controls in experiments, but seems not to know how to do the experiments necessary to validate his assertion.

someone mentioned testing for heavy metals in skimmate. Elevated concentrations would be pretty compelling evidence for Randys theory.

I have such data and will be publishing it soon... There is not a lot actually going out with skimmates. I suspect most heavy metals are ending up in the tank sediments.

andy-hipkiss

08/13/2002, 10:22 AM

Guys,

Before descending into the sticks and stones diatribes can we actually take something useful out of Ron's work?

OTS exists IME. Now is it as Bob Stark/Richard Harker suggests related to excessively high bacterial populations or is it in relation to elevated micronutrient levels, or indeed both or none.

As I posted above, measuring Cu or Pb (Zn was also high in my tank but only a paltry :lol: x200 above NSW), I "know" that simple ICP measurements alone do not give the whole story. Chelation certainly would appear applicable as to the reason that I can have x100,000 NSW Cu and still have spawning Acropora. (note to self: I really must submit a Micrpore filtered sample to ICP testing, as we might just be measuring the fact that microalgae contains hugely elevated "trace elements")

However, since as Habib's references allude, some cyano/algae can break the chelation bonds, how "leaky" is this process? Is there an event or trigger that can (in the longer term) lead to excessively high "trace elements" available within the water column?

Perhaps Bob Stark was right all those many years ago on CI$ (fishnet) to suggest replacing 10% (IIRC) of the sand bed per month whether it be in terms of bacteria or "trace elements" ... dunno.

So, for me at least, the question remains, micronutrient levels above NSW are pragmatically not a problem until maybe point X is reached. What is "point X", and how do we avoid reaching that point?

rshimek

08/13/2002, 10:24 AM

[QUOTE]Originally posted by Randy Holmes-Farley

Hi,

Have you not read any of my published papers and the experimets therein? Ahh, such a pity.

And have you done anything that supports what you assert for aquaria?

Nada, I'd guess.

Since neither you nor anyone else knows what organics are in your tank water, to attribute any found toxicities to metals is simply ignoring the other possibilities. Nearly all organics are toxic at some concentration. Some at very low levels, and some at high levels. Since you don't know what you've got, or what the concentrations are, how can you possibly say that this isn't a real possibility?

So why don't you try find out what organics are in tank water. If I don't know them - neither do you and your assertions are valueless.

Oh, I would say it is a possibility. Just an unlikely one. We have known poisons in our systems at known high levels. Pretty good smoking gun, in my opinion.

Isn't that what an experiment is? Isn't that what you plan to do? Isn't that even what a reef tank is?

So... do you have any data that support your suppositions, in reef tanks...

You have no data for types of organic materials in your tanks, and you have no data for their effects on organisms or if they temporarily detoxify metals.

Time to get out of your vaunted Vice Presidente's chair and do some experiments.

I would love to be proven wrong on this, it would be nice to say we can add heavy metals to our tanks and call them additives instead of poisons, and it would be nice to by cheap, bottom of the barrel salt and know it isn't poisoning our systems. Unfortunately, it ain't so.

It is not about a pet theory. The toxic trace metal explanation is reasonable and supported by data, if you - or anyone else - can provide data from aquaria that support your suppositions, let's see them.

rshimek

08/13/2002, 10:45 AM

Originally posted by andy-hipkiss

Andy,

Chelation certainly would appear applicable as to the reason that I can have x100,000 NSW Cu and still have spawning Acropora.

A more likely explanation is that you simply have a tolerant morph of Acropora.

Chelation might be a factor. Let's see the proof...

That shouldn't be much a problem, if anybody would care to gather the data... I suspect.

However, since as Habib's references allude, some cyano/algae can break the chelation bonds, how "leaky" is this process? Is there an event or trigger that can (in the longer term) lead to excessively high "trace elements" available within the water column?

Of course there is. Chelation of these metals simply provides food for some bacteria, and in the process the metals are back in the water.

So, for me at least, the question remains, micronutrient levels above NSW are pragmatically not a problem until maybe point X is reached. What is "point X", and how do we avoid reaching that point?

According to all physiologists who study corals, and other inverts. "X" is natural sea water levels. Anything above it is dangerous.

sven

08/13/2002, 11:12 AM

First things first, all the given, given. I do not want or intend to offend or upset anyone with this post.

I believe that all of the contributors are so far over my head on this subject it is like a space-shuttle joke. (So far overhead I don't even know it went by). To illustrate this I would like to suggest to Habib, that even though the first article was "very informative for a large crowd", maybe I should see the "picture book" version. These discussions are extremely informative for most of us that have no means nor the education to perform the research that you all do. Each of you have helped me greatly with answers to my questions in the past and I certainly hope that it will continue. My goal is to be able to help you someday too. No, I won't be able to answer any chemistry questions or identify a mystery invertebrate for you. But someday you may have a question that I can answer. In the meantime. Debate and discussion is a cornerstone of the scientific process. Those who can't listen to or take criticism well are doomed to failure. Obviously you all know what you are talking about. Obviously there is a difference of opinions too. This is all good. I would just like to recommend that you don't get so mad at each other and spend your time sharing the ideas rather than barbs. I believe that there is actually a lot of common ground in what you are saying, just some differences in what it all means. Each piece of research will help to answer that. Just remember, the more personally involved you become, the less you see. And if, heaven forbid, you are proven to be wrong, it is a whole lot easier to show yourself in public in the future.
This is a great topic, those of us on the sidelines can learn a lot. Teach us.

andy-hipkiss

08/13/2002, 11:35 AM

Ron,

Chelation might be a factor. Let's see the proof...

OK, tell me how I'd go about doing this. Sadly the lab at the place I work is only geared towards elemental analysis of a limited subset of metals (they make high precision wiring and connectors), but if you can tell me how I can take this forward, I'll endeavour to do so.

At the end of the day 211ppm (not ppB!) Cu should mean all my corals are dead ... but they are not. With one eye on the consequences of an event that may make this high Cu level available within the water column, I am keen to contribute.

JohnL

08/13/2002, 11:52 AM

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rshimek

08/13/2002, 12:03 PM

[QUOTE]Originally posted by andy-hipkiss

Andy,

OK, tell me how I'd go about doing this.

Frankly Andy, this chemistry is nothing I know how to do.

Now I presume Randy and Habib can do such work, being as how familiar they are with the literature.

I suggest you ask them, or better yet, they do it.

At the end of the day 211ppm (not ppB!) Cu should mean all my corals are dead ... but they are not.

Actually, if my subcontractor who has done chemical analyses for me on environmental projects had come up with such a value, I would suggest that there is an error in the test.

221 ppm Cu will - should - kill everything, chelation or not.

andy-hipkiss

08/13/2002, 02:13 PM

221 ppm Cu will - should - kill everything, chelation or not.

Indeed. The lab even asked me how come the value was so high. I guess I'll see on Monday when I go back to the office in Belgium and pick up the results of the most recent tests (I had my tank water and a freshly mixed sample of IO tested). I do so hope that they were wrong!

Regarding chelation, Randy/Habib, any suggestions?

wade

08/13/2002, 03:16 PM

I think a number of valid points have come up, so for my own benefit, and for the benefit of those watching this conversation, I would like to try to outline them again:

- Chelation (biological/chemical? active/passive?) is possibly a major factor in reducing toxicity of various metals.

-Speciation/VALENCE is of specific importance, especially as regards toxicity and ICP tests giving total metals do not consider this.

-Precipitation/complexation is another factor (did that get mentioned in this thread?) that probably plays a real role. Especially as regards copper. CuS isn't toxic until you choke on it (and has a very small ability to return to ionic copper). And yes, in small amounts its likely to float around in our highly kinetic reef tanks.

I believe everyone has made valid points and from thinking about it in the past, I believe that metals are a real concern in 'old tank syndrome'. However, the levels you see in most analyses don't take all avenues of detoxication into account and those must be mentioned. So, before making broad pronouncements and scaring those unable to follow the science, think a little more and dig a little deeper.

Testing for general organics in a reef tank would be beyond even a full length PhD. Its known that many many saponins, terpenes, etc etc are released and look at the volume of slime produced by acroporids.... that alone may play a major role in "resistance" as its shed repeatedly. So why not test the released slime for increased metal concentrations?

I'd suggest the following as far as future posts/articles on the subject... Make certain not to pronounce absolutes. There are too many factors and too many unknowns, especially when someone reaches beyond their immediate education. Bring up the possibility of other mechanisms or avenues. Science is a process of questioning data that others derive, nothing more. Without the questioning its meaningless. When someone asks a question and you have no immediate answer, acknowledge it and allow it as a possibilty unless you can prove otherwise. *steps down from the soapbox*

Randy Holmes-Farley

08/13/2002, 03:58 PM

So... do you have any data that support your suppositions, in reef tanks...

I have data that there are organic chelators in my tank far in exess of the copper concentration. That's because I add them in the form of large doses of iron citrate.

Beyond that, the problem is analysis. There are literally tens of millions of known organic compounds. Even in well studied natural seawater, there is very little information available on more than the basics of these molecules. People have perhaps studied and quantified a few hundred in seawater.

However, it is generally becoming accepted that metal ions (such as iron) are mostly chelated even in natural seawater. I don't see why that would not be the case in reef tanks, and see real reasons why there would be more chelation based on what we add and what we grow.

Wade:

I think that's a nice summary :)

Fredfish

08/13/2002, 04:10 PM

Ahh, its so hard to resist getting involved in a good argu.. er... discussion.

If I understand correctly, nobody disputes that most of these metals provide no usefull biological function. In fact, they are, in the correct form, toxins/polutants/nasty-bad-stuff.

Also, for those that are required to some extent - such as cu - they are found in our tanks in absurdly high concentrations that do not provide any additional value.

If we agree that the above two are correct, does it not make sense to make an attempt to remove or minimize them as much as possible as they provide the potential for problems.

Surely it is reasonable to use a source of salt that minimizes the content of these metals so that we entirely eliminate the potential for problems (or at liest minimize potential risk).

Out of curiosity, how easily do metals change speciation (hope I am using this term correctly)? Is it the same for all metals?

Another question. Will something like the Poly-Filter adsorb chelated forms of these metals (can we really remove them all from our tanks even if we wanted to?)?

Politely and respectfully :D

Fred.

rshimek

08/13/2002, 05:38 PM

Originally posted by Randy Holmes-Farley

I have data that there are organic chelators in my tank far in exess of the copper concentration. That's because I add them in the form of large doses of iron citrate.

As an aside, iron seems to be a major algal nutrient and the suggestion that has been made in algology literature, is that the algae don't really use it much, but rather sequester it to prevent bacterial use, as the bacteria that really require iron are primarily cyanobacters which are, in turn, of course, competitors of the algae. So, the question - in I hope a non-confrontational mode - is why add iron? At best, you drive algae, at worst, you drive cyanobacters.

Of course iron hydroxides do detox a lot of the trace metals in at least some natural situations (hot vents) where these are added naturally. So... adding iron might be a good way of counteracting trace element over additions.

However, it is generally becoming accepted that metal ions (such as iron) are mostly chelated even in natural seawater. I don't see why that would not be the case in reef tanks, and see real reasons why there would be more chelation based on what we add and what we grow.

Randy, I agree that at the metal concentrations found in natural situations chelation occurs. That isn't my issue, my issue is that we are dealing with situations where the trace metals are much more highly concentrated.

And additionally, where there is not one shread of evidence that such pathway is occurring in our aquaria, or that if it is, that the chelated byproducts themselves are non-toxic. For you to categorically dismiss the possibility of trace element poisoning without any support for such a position seems, well, absurd.

I think we have a complex soup in our aquaria, where at times both organic and inorganic toxic materials come into play. I have chosen to examine heavy metals simply because I have the seen environment degredation caused by them, and have spent many years in the employ of various consulting firms examing the effects of copper, arsenic, lead, zinc and so forth.

Using methodology that is identical to that found in environmental consulting, I have demonstrated that the heavy metal concentrations in aquaria are sufficiently high to be considered severely polluted.

Bioassays done by other researchers show that metals (again measured the same way) at lower concentrations than in our tanks caused coral death and disruption of coral reproduction, as well as effects in other invertebrates. This means to me, that this could, and probably should be happening in aquaria.

I have provided an explanations for several types of aquarium "maladies" which are consistent with, and supported by the data I have collected. To say that these explanations are wrong is your right, but you have no evidence to support what you are claiming, other than the fact that chelation occurs in natural situations.

In any case, your explanation, still lacks support from aquaria.

rshimek

08/13/2002, 05:42 PM

Originally posted by Fredfish

Hi Fred,

If I understand correctly, nobody disputes that most of these metals provide no usefull biological function. In fact, they are, in the correct form, toxins/polutants/nasty-bad-stuff.

At any concentrations above those found in natural sea water, they are at best null and at worst poisons. Most of them are considered to be poisons.

Surely it is reasonable to use a source of salt that minimizes the content of these metals so that we entirely eliminate the potential for problems (or at liest minimize potential risk).

Indeed, but no such salts are marketed presently for the aquarium trade. I am trying to change that.

Another question. Will something like the Poly-Filter adsorb chelated forms of these metals (can we really remove them all from our tanks even if we wanted to?)?

I don't know, and will ask.

Sardonic Wit

08/14/2002, 12:26 AM

Mr. Holmes-Farley:

If there is so much organic chelation going on why should we worry about heavy metals pollution of the ocean and the world's reefs?

It sounds like the metals should be quickly bound up and not pose a threat in any environmental system.

Habib

08/14/2002, 06:52 AM

The attached table shows that approx 0.1 % of the copper is present as free or inorganically complexed copper. The remaining copper is bound to organics or is particulate.

cfockler

08/14/2002, 09:19 AM

Originally posted by Fredfish
If I understand correctly, nobody disputes that most of these metals provide no usefull biological function. In fact, they are, in the correct form, toxins/polutants/nasty-bad-stuff.

Also, for those that are required to some extent - such as cu - they are found in our tanks in absurdly high concentrations that do not provide any additional value.

If we agree that the above two are correct, does it not make sense to make an attempt to remove or minimize them as much as possible as they provide the potential for problems.

Fred, you beat me to it! I was getting ready to post the exact same opinion! :thumbsup:

Nobody has tried to claim that elevated levels of these metals is a benefit to anything. And there is data showing that they are toxins at least in some forms. Chelation and such may or may not be there, but why rely on it to detoxify our polluted water? Why would anybody argue against correcting a potential (and very likely) problem?

I want my tank water to match natural ocean water where feasible. I eagerly await more information on alternative salt mixes and ways of detoxifying my current setup.

And hopefully Ron and Randy will give peas a chance. ;)

rshimek

08/14/2002, 09:19 AM

Hi,

The point remains regarding chelation in tanks and its relative importance or lack thereof.

Randy has pointed out he cannot test for this. Interestingly enough, then he cannot propose a testable or falsifiable hypothesis. Given that testable hypothesis cannot be formed, he (we, they) have to take this explanation without any sort of verification by the scientific method, which requires testable hypotheses.

So, we have an explanation of events based on documentation in dissimilar systems which must be taken on faith, and on the word of a true believer.

Sounds to me like you guys are creating a religion; The Cult Of
The Totipotent Chelator.

When and if you can use the scientific method to show that this has any effect in tanks I will be glad to hear about it. 'Till then...

Another point as well, if these chelators are being produced by the all of these algae and bacteria, do you not start to wonder why? Simply put it is because the trace metals are damnably toxic to them, too and this is there way of getting rid of them.

Which bears on my point, that if any of these chemicals are in the water in our systems, it is bad news.

rshimek

08/14/2002, 09:22 AM

Originally posted by Habib

The attached table shows that approx 0.1 % of the copper is present as free or inorganically complexed copper. The remaining copper is bound to organics or is particulate.

Which simply means it gets eaten by something and kills them during digestion rather than by epidermal absorption.

Six of one, half a dozen of another.

Sardonic Wit

08/14/2002, 09:31 AM

Habib:

So are you saying that we only have a need to be concerned with free copper or free metals? Don't you think there is reason for concern about the other 99.9%?

Here is an article specifically regarding pollution in San Diego Bay.

http://www.tsrtp.ucdavis.edu/newsletters/fall_2000/IndicatorsofPollution.html

It opens as follows:

Ocean pollution is a serious environmental concern, especially in developed coastal areas where shoreline and bay sediments may become reservoirs of urban and industrial pollutants, including heavy metals, As a consequence, organisms living in or on the sediment, or those that prey upon bottom-living organisms, face heavy-metal exposure.

I have a DSB and animals that eat those critters that wallow in those sediments. It looks like oceanographers are concerned. It looks like invert. biologists are concerned. Why shouldn't I be concerned, especially if the concentrations in my tank are much higher than in the San Diego Bay?

Just curious,

andy-hipkiss

08/14/2002, 10:15 AM

Ron,

Previously in this thread you suggested that water changes are not effective in controlling "trace element" levels due to the food we add increase the levels faster than any sane water change regime could cover for (I think that's what you meant by referring me to your April article ??).

If this is the case then the starting point, no matter whether we use this trace element free salt or any of the normal artificial salts, is equally irrelevant? Or have I missed your point?

rshimek

08/14/2002, 10:43 AM

Hi Andy,

I probably didn't explain it clearly.

As long as we are using salt mixes that have exceptionally high levels of trace metals, water changes are futile, the best we can do is get back to the original very level from an even higher level (after feeding).

If we can use salt with low trace metal concentrations, we can lower the metal concentration significantly and keep it under control. The key is that we need salt that is not loaded with these materials because, among other things, we add them each time we feed.

At the time I wrote the April article, I didn't know of any salt that had low trace metal concentrations or I surely would have tried to explain it better.

Sorry for the garble, and hope this helps.

:D

andy-hipkiss

08/14/2002, 11:37 AM

Thanks Ron,

However I think you were right first time (even if you didn't mean to be :) ). In your article you wrote:

These concentrations are sufficiently high that if these materials were all converted to soluble form, the water volume of the tank would receive enough of them to be raised from zero to around NSW levels, EACH DAY, EVERY DAY.

So what if "Mr Salty's Poison UR Tank salt (TM)" is x7 NSW on day 1? Give it a month and the difference between the "pure" and old Mr. Salty's will be trivial. The feeding input is such a massive influence vs. the starting point that I really can't see it matters greatly. Change 100% of the water and you've only bought yourself potentially 2 days grace before you are once again double NSW (ok not all of the elements will enter the water column for some reason, but the principle remains).

I still believe the method of control lies in tank water filtration (if indeed we are looking in the right area for the causes of OTS, which to a greater or lesser degree I do, certainly my corals have a growth spurt after a 50% water change). If "pure" salt is available for an equivalent cost as the normal stuff then why not I guess. It is certainly of some benefit. Probably the most important factor (to my mind) is designing the system such that large (>= 50% water changes) can be performed with ease (therefore done!), anything above and beyond that is a bonus.

YMMV :D

cfockler

08/14/2002, 12:54 PM

Originally posted by rshimek
Another point as well, if these chelators are being produced by the all of these algae and bacteria, do you not start to wonder why? Simply put it is because the trace metals are damnably toxic to them, too and this is there way of getting rid of them.

You go Doc! Hammer it home. :hammer:

IF these chelators are being produced then it is a defensive response to the toxins.

Humans have been poisoning themselves by smoking tobacco for years and survived and some have even managed to reproduce (although with some birth defects), but you wouldn't tell your children today that there's nothing wrong with smoking would you? "Hey, go ahead junior, light up! It won't kill you (cough, cough, hack) and your body will naturally cough up the phlegm in the morning to protect itself! No problem!"

Now that I know the toxins are there I feel a moral obligation to remove them for the sake of my captive organisms.

Instant Karma's gonna get you... :dance:

Randy Holmes-Farley

08/14/2002, 01:32 PM

So, the question - in I hope a non-confrontational mode - is why add iron? At best, you drive algae, at worst, you drive cyanobacters.

It has helped algae in many tanks. This effect can be seen instantly (well, in a day or so) after adding iron to tanks with certain macroalgae getting pale and not growing well. THey quickly green up, grow faster, and hence remove more nutrients like phosphorus.

Also, my survey shows that it may help in prevent what people refer to the sexual reproduction phase of caulerpa. The statistical significance was high enough to make this worth trying for people who've had the problem.

FWIW, my article on iron detailing all of these things shold post any minute now.

IF these chelators are being produced then it is a defensive response to the toxins.

In some cases, yes. In other cases, it is because the organisms WANT the metals, and are scavenging for them. This is especially true for iron, but may hold for many other metals. If they are not getting enough metal, they relase more chelators. There is a long thread about this and a paper that Tatu posted in my forum.

In other cases, it is entirely coincindental that the organics bind metals, as their intent was something entirely different (for example, bacteria surface themselves bind heavy metals). So dead bacteria parts will bind metals.

Now that I know the toxins are there I feel a moral obligation to remove them for the sake of my captive organisms.

Unfortunately, there is no good way to do that (that I've heard). Polyfilters are a sledgehammer that bind many metals, including some that I don't want bound (like iron). They also will not bind chelated metals to any significant extent. And they will not bind sediments that are either microscopic or macroscopic.

Salts with lower trace metals? I'll believe that when I see a tank running for a few months with such a salt and that maintains the low levels. Remember, it's not just the salt mixes to worry about. It's the calcium supplements, alkalinity supplements, metal devices in the tank, etc. etc.

If "pure" salt is available for an equivalent cost as the normal stuff then why not I guess.

That if seems pretty large to me. I don't see how it is even remotely possible. Were talking about ppb concentrations for some things.

I have a DSB and animals that eat those critters that wallow in those sediments. It looks like oceanographers are concerned. It looks like invert. biologists are concerned. Why shouldn't I be concerned, especially if the concentrations in my tank are much higher than in the San Diego Bay?

But Ron is comparing tank water to ocean water, and comparing tank water to known toxic levels for metals in water. Not tank sediments to Sand Diego bay sediments. Habib's concern is that Ron's test may have included sediments in the tank water test. Microfiltration only gets down so far.

Randy Holmes-Farley

08/14/2002, 01:36 PM

Randy has pointed out he cannot test for this. Interestingly enough, then he cannot propose a testable or falsifiable hypothesis.Given that testable hypothesis cannot be formed, he (we, they) have to take this explanation without any sort of verification by the scientific method, which requires testable hypotheses.

So, we have an explanation of events based on documentation in dissimilar systems which must be taken on faith, and on the word of a true believer.

Sounds to me like you guys are creating a religion; The Cult Of
The Totipotent Chelator.

When and if you can use the scientific method to show that this has any effect in tanks I will be glad to hear about it. 'Till then...

Please, I'll speak for myself. It is not hard to design a proper experiment in this case. If you spent more time thinking about the issue and less thinking up witty ways to make fun of people's ideas, you might have thought of it yourself.

Put free metals in artificial seawater (or whatever is preferred for toxicology tests) to the total metal concentrations found in tanks. If both waters (tank and artificial + metals) are equally toxic and substantially more toxic than the artificial seawater alone, then the binding to organics is unimportant for toxicity.

OTOH, if the toxicity of the tank water is lower than the artificial seawater plus the metals, then orgnaics may well be the explanation (though there are others, such as particulates being measured in tank water that are not toxic). In any case, it will show whether ICP measurements are appropriate for determining the metal toxicity of tank water samples.

To be perfectly honest, I'm not claiming that metals are toxic in our tanks, or not. I've said so several times. They might be. They might not be at the levels found in most tanks. As per the scientific method, however, I'm pointing out other possibilities that had not been considered in coming up with the conclusion that we are "killing corals". FWIW, you make it very hard for people to keep a middle ground position because you present your opinions as conclusive facts, and that leaves some of us to point out other opinions that may be true, or may not be true, but that have not been ruled out by any existing data.

Randy Holmes-Farley

08/14/2002, 01:44 PM

If there is so much organic chelation going on why should we worry about heavy metals pollution of the ocean and the world's reefs?

It sounds like the metals should be quickly bound up and not pose a threat in any environmental system.

Obviously not, or the various organisms would not spend the energy to make organic compounds to detoxify their environments.

Two concerns:

1. The organics in our tanks may be far higher than in the ocean, so the toxicity may be much lower. After all, the relative concentration of the creatures that release organics is higher in our tanks, so it makes sense that their products would also be higher. I have no data, however, on the concentrations of any specific organic in our tanks.

2. The metals in the ocean may be far less toxic because of binding to organics. Maybe the problem would be 10,000 times worse if not for the organics. So the fact that there is or is not a problem says nothing about the role (if any) that organics play.

If we want to get into talking about contaminants in sediments (where the environmental problem often lies), that is well worth discussion. We have, however, no data on sediment pollutants in reef tanks.

I'd also point out that pollution with organics is just as pervasive and problematic as pollution with metals. GE is being forced to dredge the Hudson to take PCB's out of the sediments. PCB's are organic! So by all rights we should be just as concerned about organics as metals. Unfortuantely, analysis of organics is far, far more complicated than ICP.

SciGuy2

08/14/2002, 02:46 PM

Habib

08/14/2002, 02:54 PM

Ron,

These tests were run exactly as standard pollution monitoring tests are run. The data are compared in exactly the same manner with the same conclusions being reached.

If the tests show that levels of a certain metal kill animals, and if the tests show that the levels are above those levels, and animals do die then it is reasonable to suggest that the metal is responsible. It really doesn't matter the type of ion that is involved

Many hobbyists are running tests by having one or more aquarium, keeping corals, growing them, multiplying them,...

They are alive. With they I also mean the corals and not only the aquarists in case that is not clear enough.
Now if the elements present in the average aquarium are toxic at the levels you measured of these average (random) aquaria within which period of time should the corals die? a few days, a few weeks.

If you answer this remember what you have said in previous posts.

Sure you did. Yup. Somewher you have those data....

Now lets see them published somewhere.

As a matter of fact I have recently been invited for an article........

From your respons to Randy :

We do know that concentrations of the chemicals seen our systems can cause toxic effects. We do know that they even if such metals are rendered temporarily insoluble that minor changes of condition can make them soluble and toxic.

So what you said is that metals are rendered temporarily insoluble then change of condition can make them soluble and toxic.

So according to you if it can not be made soluble it is not toxic.
That is being not bio-available. Is this what you are saying or do you diasagree with the scientific community?

BTW EPA recognizes/accepts that toxicity of an element depends on the speciation and how it is present.
It is generally accepted that speciation etc. determines the toxicity, bioavailability , bioaccumulation,...

With respect to the data of copper speciation in san diego bay posted by me you said:

Which simply means it gets eaten by something and kills them during digestion rather than by epidermal absorption.

If this is no speculation then show us the data for the above concentrations and speciation.

Sounds to me like you guys are creating a religion; The Cult Of The Totipotent Chelator.

Sounds to me as an insult. Or is it because my knowledge of the English language is so poor. Please explain.

Bomber

08/14/2002, 03:51 PM

So what you said is that metals are rendered temporarily insoluble then change of condition can make them soluble and toxic.

Wouldn't acid meet that requirement?

Habib

08/14/2002, 04:21 PM

Bomber:

Wouldn't acid meet that requirement?

Considering the very low solubilty products of heavy metal carbonates and oxides it would require a pH far below necessary to dissolve calciumcarbonate. So the system will resist low enough pH concentrations being reached as long as calciumcarbonate is present.

Furthermore natural seawater and aquariumwater have a pH buffering effect thanks to the bicarbonate/carbonate (carbonate alkalinity) system.

Furthermore the solubilty products for inorganic particulate matter will be even much lower than the values obtained in seawater free from organics.

These particles are covered rapidly by many organics, decreasing the solubility product tremendously.

Also thanks to the above mentioned at least some aquatic organisms are able to secrete a skeleton (bio-mineralization) which resists dissolution or atleast transformation of their crystalstructure. Without these organics they might dissolve.

Habib

08/15/2002, 06:27 AM

Ron,

After you are done with replying to the above posts here is something more.

It is a description of EPA Grant Number: R825220 ;
Biogeochemical Control of Heavy Metal Speciation and Bioavailability in Contaminated Marine Sediments

EPA Grant Number: R825220
Title: Biogeochemical Control of Heavy Metal Speciation and Bioavailability in Contaminated Marine Sediments
Investigators: Shine, James P.
Institution: Harvard University
EPA Project Officer: Manty, Dale
Project Period: December 2, 1996 through December 1, 2001
Project Amount: $453,630
Research Category: Early Career Awards

Description:
The total concentrations of contaminants in an environmental sample are not indicative of the potential for adverse ecological effects. Contaminant speciation and its effect on bioavailability are critical to understanding ecotoxicology. This information is also crucial for development of policies concerning the use and disposal of toxic material in the environment.
An initiative is currently under way at the national level to understand the factors controlling the toxicity of heavy metals in aquatic sediments. Heavy metals discharged into aquatic ecosystems are likely to be scavenged by particles and removed to the sediments, perhaps leading to a situation where the water is ?clean' yet the underlying sediments have accumulated toxic levels of heavy metals with resultant adverse effects on ecosystem health. The presence of sulfides and particulate organic carbon have been appropriately identified as factors buffering the availability of heavy metals in contaminated sediments. However, even when the ability of these constituents to buffer the toxicity of metals is exhausted, toxicity is not always observed. This implies other binding phases which may also contribute to reduction of metal availability in sediments. These other binding phases may include dissolved and colloidal organic matter in porewaters which can form stable complexes with heavy metals, thus reducing their bioavailability.

The proposed work will examine the role of dissolved and colloidal organic matter on metal speciation and bioavailability in marine sediments. Speciation will be measured at two contaminated locations in New Bedford Harbor, USA, and at a comparison ?clean' location in Buzzards Bay. Observations will be made over multiple seasons to observe temporal and spatial variability these ligands have on metal speciation and bioavailability.

The specific objectives of this proposal are as follows:

1. Develop methods to determine the role of dissolved and colloidal organic ligands on the partitioning of metals between particles and pore water in marine sediments and observe how partitioning varies in space and time.

2. Determine, in conjunction with acid volatile sulfides, how adsorbed (particle bound) and porewater ligands control the availability of heavy metals to transplanted and native benthic organisms.

3. Develop methods to quantify the thermodynamic characteristics of adsorbed and porewater metal binding ligands.

4. Develop the study findings in a framework that assists development of criteria for protection of aquatic ecosystems.

Please explain to me if a certain concentration of metal is determined without discriminating the speciation etc. , that is in the way your measurements were conducted, then how you could deduct the speciation from just these data?

If that can not be deducted , and it can not be deducted, then nothing can be said about the toxicity or any inhibition effects.

Or do you disagree that speciation etc. of a metal affects toxicity? If so show us any as recent as possible reference or any data which would support your disagreement.

But on the other hand if you agree then how can you say anything about the toxicity of the metals found in the measured concentrations without knowing speciation etc ?

From your last Reefkeeping magazine article:

The literature on the effects of these chemicals, specifically, Cadmium, Copper, Chromium, Lead, Mercury, Nickel, Vanadium, and Zinc, is rather depressing, frankly. It deals with nothing more than mortality factors or how the organisms deal, perhaps more importantly, how they cannot deal, with enhanced levels of the chemicals.

I suggest you do a better literature search. Many enzymes will not function without heavy metals. Without them no corals, no coralline algae, no bacteria, no..........

Another thing:
If toxicity takes place then it automatically means that the metal which is causing toxicity is taken in by whatever pathway by that organism.
If that happens the metal is bio-available. In such situations relatively huge amounts of that metal will be taken in until the organism dies or the pathways of toxic metal intake have ceased to function (almost dead).

Now if the metal is that much bio-available to be toxic then how come there is still that metal present in the water column (limited volume)?

It should be used up quickly and become undetectable by your methods if it were toxic and therefore bio-available. Isn't it???

And why there was no iron detectable in the aquariums you tested while all sea salts tested have a large amount of iron in them and there were other metals such as copper (which you consider to be toxic) still present in a concentration similar to what is present in saltmixes?

rshimek

08/15/2002, 08:41 AM

Originally posted by Randy Holmes-Farley

Please, I'll speak for myself. It is not hard to design a proper experiment in this case. If you spent more time thinking about the issue and less thinking up witty ways to make fun of people's ideas, you might have thought of it yourself.

I did, but I think the issue is unimportant. You seem to think it is important, but seem totally unable to present any data from aquaria supporting your position. Basically, you are "jumping up and down and waving your arms, " but don't seem to be willing or able to do the work.

I have pulled together a short reseach project over the last few years, showing the absurd levels of poisons in our tanks and the paths they take through our systems. I started with the sea water composition work by Atkinson and Bingman and Working through the foods added to our systems, the amounts of poisonous trace metals in our tanks water what is being exported (the data are on hand and will be published this autumn) and the toxicity of various waters to standard test.

It seems to me that if you thought that organic materials had some real effect in our systems, you could do the work to show those effects.

Basically, Randy, put up or shut up; all you have said is simple conjecture.

If we want to get into talking about contaminants in sediments (where the environmental problem often lies), that is well worth discussion. We have, however, no data on sediment pollutants in reef tanks.

I will - if I can get the $$ - be examining the metals in tank sediments late this autumn as one of the upcoming components of the this project of mine.

Perhaps you should undertake some research about the organic components, if you think they are important.

Habib

Considering the very low solubilty products of heavy metal carbonates and oxides it would require a pH far below necessary to dissolve calciumcarbonate. So the system will resist low enough pH concentrations being reached as long as calciumcarbonate is present.

The system will resist this, true enough, but these materials will still be toxic to organisms as they will encounter sufficiently low pH values during digestion. Many aquarium animals, including corals ingest and digest particulate material, additionally much fine particulate material - including potentially toxic material- is moved from tank sediments or the surface of live rock into the water where suspension-feeding animals will encounter it. The animals will convert this insoluble - and harmless - material into a toxic material within their body.

If that can not be deducted , and it can not be deducted, then nothing can be said about the toxicity or any inhibition effects.

Certainly, it can. In the same way, observational/correlative research is done. If the level of a certain chemical is measured in the same way it has been measured elsewhere, and if mortality has been associated with those measurements elsewhere, then the mortality may correlated with the measured value.

This does not imply direct cause. However, if such a correlation exists then there is certainly reason to suspect the measured factor is having an effect.

The "species" of the metal is unimportant. What is important is that every time the metal is measured at, or exceeds certain values, mortality occurs. It may well be due to the form of the metal in the solution, or to the form that the organism converts it to. However, it happens, the metal is toxic.

suggest you do a better literature search. Many enzymes will not function without heavy metals. Without them no corals, no coralline algae, no bacteria, no..........

Read the statement in the article again, and don't take it out of context. In the context of the article I was referring to the environmental physiology of these materials, and the statement is correct.

Certainly, in natural trace amounts these materials are necessary. Neither in the environmental literature nor in our aquaria are these materials at anything near natural trace amounts.

If toxicity takes place then it automatically means that the metal which is causing toxicity is taken in by whatever pathway by that organism.
If that happens the metal is bio-available. In such situations relatively huge amounts of that metal will be taken in until the organism dies or the pathways of toxic metal intake have ceased to function (almost dead).

This is absolutely incorrect. The material may be ingested in an non-toxic form and converted to a toxic form within the animal. Happens all the time....

And why there was no iron detectable in the aquariums you tested while all sea salts tested have a large amount of iron in them and there were other metals such as copper (which you consider to be toxic) still present in a concentration similar to what is present in saltmixes?

The test method is rather insensitive to iron. But more to the point, iron hydroxides have been shown to complex with some of the trace metals, and I suspect it is bound that way and removed from solution. Also, iron is preferentially taken up by cyanobacteria and some algae, so the concentration of iron is probably lowered biologically.

Copper is not taken up preferentially by any animal. I do consider it toxic, copper kills animals... and algae.... and bacteria. Maybe this isn't toxic by your definition, but it surely is by mine.

I am glad you will finally publish some results. Regardless of the result, at least you seem to have been doing some research, unlike other posters.

wade

08/15/2002, 10:29 AM

Do you think testing "substrate" for metals will yeild any information that conclusions will be drawn from?

In general, toxicologists consider what is in sediment as an indicator of overall health of a system, nothing more. And, in light of that, would you be looking at pore water samples, overall water, sediment? I think each has relevance and in order to draw any real conclusions, I think we would need to see each piece of that puzzle. I believe this is especially important in light of the fact that the only mechanism in our tanks of moving a toxicant from one region to another is via biology. If the organisms are consuming the sedimentary organics and re-releasing those as larvae.

rshimek

08/15/2002, 10:50 AM

Hi Wade,

If I get around to doing this I will look at the sediment proper, basically to find out the metals levels are. I don't want to get into sampling pore water, etc. Simply put, at this time it isn't worth the money for me to do this, as all of this work has been either self-funded or funded with the help of some aquarists, money really is a driving force.
:D

cfockler

08/15/2002, 12:12 PM

Originally posted by andy-hipkiss
Ron,

Previously in this thread you suggested that water changes are not effective in controlling "trace element" levels due to the food we add increase the levels faster than any sane water change regime could cover for (I think that's what you meant by referring me to your April article ??).

If this is the case then the starting point, no matter whether we use this trace element free salt or any of the normal artificial salts, is equally irrelevant? Or have I missed your point?

I don't think you missed the point at all. And after finding a better salt mix the focus will likely turn to finding or culturing cleaner foods. There are encouraging possibilities...

Check out this thread...
http://www.reefcentral.com/vbulletin/showthread.php?s=&postid=726769#post726769 (http://archive.reefcentral.com/vbulletin/showthread.php?s=&postid=726769#post726769)

I've gotta admit I'm surprised this thread has dragged on the way it has. The metals are present in our aquariums in amounts far above natural levels. Getting our tanks closer to natural levels where feasible is a worthwhile and logical goal.

Randy Holmes-Farley

08/15/2002, 08:21 PM

Basically, Randy, put up or shut up; all you have said is simple conjecture.

Conjecture? That's a laugh. Doesn't the fact that thousands of people have reef tanks growing corals at rates similar to natural growth rates in "absurd levels of poisons" seem to suggest that it is your theory about toxicity, and not my theory about why people have been successful that is most lacking in evidence? Actually, I won't ask you the question. I'll state it: yes, it does suggest that there is a problem with the theory that we are killing corals. It is not conjecture that these corals do fine. It is conjecture that some are being killed.

Can you show even one coral from anywhere in the world that died in a reef tank from metal poisoning? I thought not.

Remember, you have absolutely zero real toxicity data in reef tanks. You ridicule me because I don't have data on organic levels, but you don't have any tox data.

Perhaps you should undertake some research about the organic components, if you think they are important.

That doesn't sound like a scientific suggestion, but more like a way to make me sound uninformed or uninterested in helping the hobby. What if I told you the levels of 3,215 different organics, 256 of which are bound to 25 different metals in my reef tank? What would that do for anyone? Total squat, that's what. Then you'd simply say, OK you've shown nothing that says anything about toxicity of metals. It all boils down to toxicity tests if toxicity is the claim. Running such a test in reef tank water is as easy as running one in low metal seawater. Most scientists would run such a test before claiming that we are killing corals. Since you made the claim, it is incumbent on you to back up the assertion, not on your critics to "prove" you wrong.

I've gotta admit I'm surprised this thread has dragged on the way it has. The metals are present in our aquariums in amounts far above natural levels. Getting our tanks closer to natural levels where feasible is a worthwhile and logical goal.

I'm sure we'd all like to see what happens when that is done. Can it be done? That remains to be established for anything short of a reef tank for Bill Gates. Even so, let's see what happens. Then we can really see the merits.

The animals will convert this insoluble - and harmless - material into a toxic material within their body.

Yes, that could very well be, to some extent, but it isn't appropriate to extrapolate the toxicity of such materials from the toxicity of free metal ions. Suppose the real toxicity level of such particulates was slightly, or greatly, higher than the tank concentration? Aluminum ions are quite toxic, aluminum oxide is not, even when passed through the low pH stomach of humans. One cannot quantitatively equate them.

Neither in the environmental literature nor in our aquaria are these materials at anything near natural trace amounts.

Of course, that may not be true of all ions that we are concerned with. Did you detect any iron? In fact, I recall many ions that you did not detect, and some are necessary nutrients. Don't tell me that you want to lower those too, even without knowing what the levels are?

Copper is not taken up preferentially by any animal.

Another incorrect generalization. It is preferentially taken up in humans. Ever hear of Wilson's disease? Since you didn't apparently know that even about yourself, can I assume that you aren't sure about every single one of the 5,234 organisms living in my reef tank?

I am glad you will finally publish some results. Regardless of the result, at least you seem to have been doing some research, unlike other posters.

Ahhh, such a cutting blow. How can I even hope to have a scientific discussion when I'm burdened with such guilt over leaving toxicity studies to you? Oh, I forgot, you haven't run any either.

rshimek

08/15/2002, 08:44 PM

Originally posted by Randy Holmes-Farley

Oh, I forgot, you haven't run any either.

Ah, yes... but they are scheduled.

And as for your efforts - - - have you done any studies at all, Randy? Have you generated any aquarium data, at all, that support your ramblings? Any supporting data whatsoever for your conjectures? Or, perhaps better put, any supporting data for your "unsupported guesses?"

.

.

.

.

I thought not.

SDBDRZ

08/15/2002, 11:22 PM

I am sure that Ron and Randy are having fun with this manhood swinging contest but I have a somewhat off the immediate topic question. I will first say that I am not a Biologist or Chemist, I am a CPA and therefore a complete idoit when it comes to biology or chemistry (I had to read the article and thread several times to follow it). My question for Dr. Ron has to do with sample size. I have read the article (and previous Reefkeeping magazine articles you have written) and as far as I can see your sample size for these experiments is 23 aquarium water samples. Is a statistical sample size of 23 adequate to draw conclusions about the population as a whole? I am not exactly sure how many reeftanks there are in the world but I would dare say that your sample is less than 1/10th of 1% at best. If I am incorrect about your sample size please accept my appology in advance.

As my disclaimer I am not trying to throw my manhood in on this I am just curious about your sample size as it relates to the total number of reeftanks. My remarks are not intended to insult, insinuate, irridate, annoy, etc. There that should keep me from getting flamed :D

saltshop

08/16/2002, 12:09 AM

I think you guys just need to spend a day in each others shoes! I have a father who is a chemistry professor, while I am rather interested in biology and ecology when it comes to my glass box (dare I say I was a business/law major :D ). From the chemist point of view, one usually knows what is going to happen beforehand and then you perform simple experiments to prove what you had already formulted on paper and from what is already known. In this case organics+life+metals=chelators and detoxification of various elements. From the biology point of view, you usually have an effect and then set about determining the cause through experimentation, observation, and previous works. In this case we have the mysterious "old tank syndrome" combined with what is known about heavy metals in "real life" ecosystems.

I am really not sure why the two fields have clashed so much here. :confused: FWIW, from someone who is "rather familiar" with you both through postings I am actually quite surprised! I know Randy prefers to keep things, if at all possible, as close to NSW be it good or bad. Iodine for instance is added to keep NSW values even if there is no conclusive evidence of what it may do...but there really is no arguing with what is the natural environment, correct? Now if metals are measured through ICP in the wild and shown to be a problem then, even if our "organic soup" tanks most likely are somewhat different, why not try to keep NSW values if at all possible just in case. Now for Doc, I know you are not real fond of someone making conclusions without thoroughly investigating all other possibilties and are quick to point it out when someone else does it. To be honest I was quite surprised when I read the last column stating that LR and sand should be discarded every four years or so. I saw nothing to support that LR and sand is releasing large amounts of these metals and that there is a demonstrated chain of events leading to death of organisms...heck it could be some build-up of some type of organic that cause old tanks to crumble or even a combination of the two You have mentioned here that it may be looked in to as far as what the levels are in the sand. I am really unsure of how you came to the conclusion you did already when there is still more to be explored??? :confused:

Now, you both can banish me from your forums or whatever you want to do (I can still hang out with Eric ;) ), but as an outsider this is how it looks to me. You are arguing about the forest but can't see through the trees. If you were to put your heads together you could probably come up with some really good "experiments/trials" that would lead to a much better understanding than either of you could provide by yourselves. Peace....

jayo

08/16/2002, 12:17 AM

What bothers me about the toxicity argument is that people that use natural seawater (with "normal" metal levels) don't seem to have systems that are any healthier than those that use artificial "toxic metal levels" salt. I've never heard anyone claim that they could keep delicate animals better with natural sea water (unless it was an open system, and then it is more likely plankton that is making the difference). Even Ron is working hard to get more "natural" water (with a salt mix lower in metals) - why bother when using water with ideal concentrations (natural sea water) doesn't seem to work any better than artificial for any reef tank I've ever seen?

jayo

saltshop

08/16/2002, 12:18 AM

LOL...hey Scott at least us business guys see things clearly! What are the odds of tax law coming up in a reef forum? :D I have mentioned this elsewhere, but in some cases the sample size was less than N=23, for the case of Arsenic N=1, Iodide N=14, etc. as the results that were below detection limits were thrown out and considered non-consequencial. I am sure that would cause much distress among any statisticians out there.

Habib

08/16/2002, 06:20 AM

Ron,

I will be running some sea urchin bioassays on tank water and sea water mixes within the next month. Provided that I can complete the tests in time for MACNA, I will report on them there.

I assume you will be counting the number of fertilized eggs.
Is this correct?

If so which percentage range of fertilized eggs would you consider to be acceptable?

And if the results are within the acceptable range what would your conclusion be?

And what would your conclusion be if it is not within the acceptable range?

TIA

Randy Holmes-Farley

08/16/2002, 09:24 AM

Ah, yes... but they are scheduled.

Wonderful:) When you finish testing on corals and find them suffering, I'll help you promote the concern far and wide that we are harming corals .

wade

08/16/2002, 10:29 AM

Sorry to keep this going, but I'm still a bit confused...
If I get around to doing this I will look at the sediment proper, basically to find out the metals levels are. I don't want to get into sampling pore water, etc. Simply put, at this time it isn't worth the money for me to do this, as all of this work has been either self-funded or funded with the help of some aquarists, money really is a driving force.
When you say sediment proper, what do you mean? Typically, sediment is tested by pouring off any residual water and then dessicating it, which excludes any water soluble (non-bound) chemicals. Perhaps, the misunderstanding is mine about ICP methodology. Can you run a sample that is equal parts water and araganite/sediment?

Also, from a scientific standpoint/methodology issue, I would like to see your water column tests performed on samples that have had a bit more in the way of control/explanation. I realize that you had to draw from aquarists who don't know too much about science (generally speaking, no offense meant to the readers), but were things such as PTFE solvent rinsed bottles used? were samples microwaved or microfiltered prior to use? Questions like that make me nervous about the data you presented. The reason being, as an example, what if the water from a given tank had bacteria/algae counts on the order of 10^3 per ml? This would indicate that many of the elements you analyzed for were likely a part of that biological system and then we get into issues of coral/fish feeding and selectivity.

In all, this is a very complex issue and a few simple tests will tell us little. True tox testing (urchin bioassay, et al ) will give us an indicator of overall health, but only if statistically sigificant sampling is used (with every effort made to control and get random samples). I guess my point to this is that it would be nice to have the data you are striving for, but I question its worth in a standalone fashion. And as you say, money drives everything, especially science, so the cost of doing this 'right' would be noticable.

That make sense?

Wade

Habib

08/16/2002, 11:16 AM

Wade,

but were things such as PTFE solvent rinsed bottles used? were samples microwaved or microfiltered prior to use? Questions like that make me nervous about the data you presented. The reason being, as an example, what if the water from a given tank had bacteria/algae counts on the order of 10^3 per ml?

I also had concerns regarding this and also some other related matters.

A discussion with Ron about this can be found here:

What we test in the water (http://archive.reefcentral.com/vbulletin/showthread.php?s=&threadid=85249)

rshimek

08/16/2002, 11:45 AM

Originally posted by SDBDRZ

Hi,

IIs a statistical sample size of 23 adequate to draw conclusions about the population as a whole?

No, probably not. And I hope that other studies will be done. I think reef tanks are relatively diverse. However, that having been said....

This series of tanks was taken from across America (NY to AZ to TX to MT, etc). The data are surprisingly consistent.

As I have said, there is no compelling reason to regard these few tanks as representative. However, there is likewise no compelling reason not to either.

Presently, they are all we have. I would love to see more studies done testing for these materials in a comparable manner. Such tests are unlikely. I advertise on both Reef Central and Reefs.Org for volunteers (who had to help pay for the the tests) and this is the best I got. The tests are spendy - the base costs are over $200 per tank (although we got a volume discount :D)

There that should keep me from getting flamed :D

:thumbsup:

rshimek

08/16/2002, 11:57 AM

Originally posted by Habib

Habib,

I assume you will be counting the number of fertilized eggs.
Is this correct?

No, this will be a developmental test, running from fertilization up to the end of the prism stage.

If so which percentage range of fertilized eggs would you consider to be acceptable?

And if the results are within the acceptable range what would your conclusion be?

And what would your conclusion be if it is not within the acceptable range?

The test will be a modified EPA protocol (I can send it to you if you wish). The modification will primarily be the use of a tropical urchin, as the test was designed for a temperate species.

Anyway the endpoint will be a comparison between the number of normal embryos versus the number of deformities.

I will be testing development in:

Catalina Salt Water (NSW) being shipped to me;
Several salt mixes newly made up;
Several tank waters.

The working hypothesis is that that there is no difference in the waters from all the tanks.

If salt mixes contain toxic materials, these will have an effect on the tests.

If tank wates are modified (by any means - unfortunately I will not be able to determine how they are modified) so that the embryos survive that will show up as well in the tank waters.

I have not yet decided whether I wish to do other modifications - such as dosing the NSW with various levels of metals to mimic salt levels, etc.

rshimek

08/16/2002, 12:06 PM

Originally posted by Randy Holmes-Farley

Wonderful:) When you finish testing on corals and find them suffering, I'll help you promote the concern far and wide that we are harming corals .

There is a voluminous literature on the basis of using bioassays as a way of determining toxicity of waters. If you are unfamiliar with such methods, I can provide you with some literature citations to get you started.

It is a standard methodology, and has been used in coral reef environments as well as elsewhere. The results are accepted by effectively every govenmental agency examining environmental degradation in the world, as well as being considered definitive by environmental scientists world wide.

The presumption is that injury to the the bioassay organism reflects injury to all organisms in the community. Coral bioassays exist, and they are cited in my recent article, and that metals concentrations resulting in coral mortality are lower than those found in our tanks. I would love to use these tests, but unfortunately, I cannot afford to purchase the corals to run them. Fortunately, the urchin developmental test, which I can afford to do, is widely accepted throughout the environmental assessment scientific community.

rshimek

08/16/2002, 12:20 PM

Originally posted by wade

When you say sediment proper, what do you mean? Typically, sediment is tested by pouring off any residual water and then dessicating it, which excludes any water soluble (non-bound) chemicals.

Yes. And this is what would be done.

, but were things such as PTFE solvent rinsed bottles used?

The bottles were provided by the laboratory.

were samples microwaved or microfiltered prior to use?

No.

Questions like that make me nervous about the data you presented.

Yes, they should. On the other hand, these data are all we have. So... Accept them or reject them, unless someone else wants to do something else, they are all we have.

In all, this is a very complex issue and a few simple tests will tell us little.

With the amazingly high concentations found, they certainly give some indications.

but I question its worth in a standalone fashion.

I certainly welcome any effort to duplicate or replicate the sample analysis in any other form, so go to it...

And as you say, money drives everything, especially science, so the cost of doing this 'right' would be noticable.

Well, I don't think there is any evidence that the results are "wrong."

To invalidate these data would require additional testing and analysis, and I would love to see the results of such tests.

saltshop

08/16/2002, 02:02 PM

[QUOTE]Originally posted by rshimek
IIs a statistical sample size of 23 adequate to draw conclusions about the population as a whole?

No, probably not. And I hope that other studies will be done. I think reef tanks are relatively diverse. However, that having been said....
[QUOTE]

I have a few questions along these lines, but first I have to say "holy cow, don't get on the computer after coming home from a Nascar party!" My apologies for the rambling post!! :D

I was looking through the Feb. article that lists the tank conc. v. IO and for the most part most of the elements seem to be held in tow relative to the starting point. Are you at any point going to put up the data from each tank in some sort of spreadsheet format? The reason I ask, is that in some cases the values fluctuate by significant amounts resulting in too large of a standard deviation to have it make any sort of sense, partly because of the limited sampling. An example of this would be for Lithium which has a standard deviation which is larger than the mean itself. Some of the others that would fall in line as being too random would include Cobalt, Iodide, Molybdenum, Phosphorus, and Silicon. I also have a question about Vanadium which has a Min. of 0.030, a Max of 0.037, and a mean of .023 +/- .047? I think there has to be something wrong with that one to get a mean larger than the minimum and a crazy SSTD. I am not sure if it was corrected in the later issues? I think in these cases it would be really nice to see what the data sets actually were to get an idea of the representation both above and below mean. If done, it would also be nice to note what salt mix was used to begin with. In some cases a single tank or 2 that uses Sea Chem products might throw off the whole curve for something like Boron, likewise certain salts have really high Lithium levels.

Lastly, have you done (or will do) any statistical runs on tank age versus the elements listed? If the premise is that these elements are building up over time there should be a statistically meaningful measure showing the older the tank the higher the build-up. I could not find such a thing in any of the articles.

From just looking at the means while taking into account the large SSTD and range it appears that we start with a crappy mix to begin with and it doesn't get any better over time and in a few cases a little worse. In some cases people probably started with an even worse mix ... I am hopeful that someone will PLEASE find a good source for the raw materials to make adequate salt at home to start with and if the elements "hold the line" like they did against IO most of this thread would become moot. I would have no problem paying a dollar plus per gallon to make up water that was even close to NSW to start with, as I am pretty sure water changes could be held to something like a semi-annual event. Heck we might even end up saving money in the long run. Thanks!

El-ahrairah

08/16/2002, 02:14 PM

Ron,

Just thought I'd chime in on this interesting discusson. I have a couple of questions regarding your urchin experiment. What tropical urchin species are you planning to use? Will it be in season when you perform your experiments?

Actually, I can pretty much tell you what the results of your urchin experiment are going to be, since urchins don't like ASW. The tropical urchin will most likely give pretty much the same results as temperate species.

Your cultures using the Catalina sea water should have 90% or survivourship and high synchronicity or your doing something wrong. Whereas your cultures using ASW regardless of the brand will show much higher mortality and non-sychronous embryos. The water from your sample tanks will likely match the ASW samples, especially if the tanks use ASW.

The embryos and larvae of many other species will show the same results as well. This is not entirely due to the higher amounts of various metals (though it does play a large part) in the ASW but also the lack of many other compounds mostly organic that are found in NSW. Larvae are very intolerant to ASW this has been known for the past thirty years. I've actually tested this with L. pictus, S. purpuratus, S. neumayeri (antarctic species), C. gigas, and R. pachyptila (hydrothermal vent worm)myself.

I have to agree with you that the majority of tanks contain concentrations of metals that far exceed that of NSW, and that for most invertebrates these are poisons and that many(but not all) of the unexplained deaths that hobbyists see can be attributed to this. However how can you honestly say that the thousands of people with tanks that are capable of keeping healthy corals and other sensitive invertebrates is simply conjecture and that it is in itself not sufficient evidence that elevated levels of various elements will always poison the tank. Please do not argue with "those specimens that thrive are pollutant tolerant" I've been force to sit through enough toxicity seminars that that argument just doesn't wash (since even the tolerant species can't handle the levels of pollutants you say are in tank water).
I don't intend to come off harsh and apologis if I do.
-Michael

Randy Holmes-Farley

08/16/2002, 02:40 PM

For those thinking about sediment toxicity, here's an article where truly polluted sediments are being tested for toxicity, with clean sediments in Instant Ocean Salt mix used as the nontoxic control sample:

http://www.wes.army.mil/el/dots/pdfs/eedp03-1.pdf

Habib

08/16/2002, 02:48 PM

Ron,

No, this will be a developmental test, running from fertilization up to the end of the prism stage.

I don't know the EPA procedure for this test. I however know that the EPA procedure to say something about toxicity by determination of the percentage of fertilised eggs allows the use of NSW and synthetic seasalts such as Forty Fantoms, IO, HW-marinemix,...

The control should have a value of above 70% (IIRC).

The results of the controls that I have seen for some saltmixes which we use in our hobby are typically 80 - 90%!!!-----> Non-Toxic?!?

Spiking artificial seawater with IIRC 0.0025 ppm (2.5 ug/L) Cu --> 70%, 0.02 ppm Cu --> 40% (IIRC).

I also wonder what the EPA procedure allows to be used in the the test you intend to do? NSW and also artificial seawater??

In your last article you mention various results obtained for various coral stages. Abstracts which I have read for some of such studies mention the use of artificial seawater as control!

The studies you mention in your last article, has any of these used artificial seawater as control or was NSW used??

TIA

El-ahrairah

08/16/2002, 03:09 PM

I don't know the EPA procedure for this test. I however know that the EPA procedure to say something about toxicity by determination of the percentage of fertilised eggs allows the use of NSW and synthetic seasalts such as Forty Fantoms, IO, HW-marinemix,...

The control should have a value of above 70% (IIRC).

The results of the controls that I have seen for some saltmixes which we use in our hobby are typically 80 - 90%!!!-----> Non-Toxic?!?

Ferilization tests are basically of no value. Sea urchins are basically foolproof when it comes to fertilizing them. If you don't get better than 90% fertilization then there is something wrong with the eggs. The real test for toxicity is if they can make it past gastrulation. In our lab we can get around 90% fertilization everytime, even in conditions that kill them by the time they hit blastula. I'd like to see ASW mixes that can regularly get urchins to settle. ASW is a poor substitute when trying to culture invertebrate larvae.
-Michael

rshimek

08/16/2002, 04:56 PM

Originally posted by El-ahrairah

Michael,

ASW is a poor substitute when trying to culture invertebrate larvae.

As you have indicated this has been the case for some time, but it may not necessarily be so with all ASW. I have now recieved a mix that claims to have the trace element concentrations of NSW, and it is apparently marketed to labs working with EPA standardized bioassays. I will test this during the tests. I am interested to see if the larvae fare better in it than in other ASW mixes.

Actually, I can pretty much tell you what the results of your urchin experiment are going to be, since urchins don't like ASW

Maybe so, and maybe no. The last time I cultured urchins in Instant Ocean was for an invert class I taught 3 years ago, and we got in an open class room situation, larvae that lived for 5 weeks and had good juvenile rudiments in them prior to the termination of the class.

The urchins I will use are Lytechinus and they will be obtained from Carolina Biological Supply, and generally some individuals are ripe all year round.

As far as raising urchins through to competancy, I have raised:
Strongylocentrotus purpuratus, S. franciscanus, S. droebachiensis, S. pallidus, S pallidus x S. droebachiensis (I was the first to do this), and D. excentricus. Other animals I have spawned and raised include:numerous echinoderms of all classes except concentricycloideans, Serpula vermicularis, several anemones (no-brainers, they don't feed), sea pens (Ptilosarcus gurneyi) numerous gastropods with feeding planktotropic larvae, but specifically turrid gastropods such as Oenopota levidensis, and really a whole slug of others.

The water from your sample tanks will likely match the ASW samples, especially if the tanks use ASW.

I doubt this very much. After short periods in reef tanks, the salt water often seems quite fine for many larvae (I have collected gastropod, polychaete and flatworm larvae from my tanks and others have done the same). As much as I have been giving Randy a hard time about him not having any data to support his chelation myth, I actually think it or something like it is occurring, and that ASW may be effectively detoxified in a tank. However, the price that is paid for such detoxification is that the chelated or bound toxins end up in the sediments or live rock and will come back to haunt us.

(since even the tolerant species can't handle the levels of pollutants you say are in tank water).

Well, Michael, the pollutants are in the tank. And I have certainly seen pollution-tolerant species that handle far greater loads. See the comments below regarding bioassay labs.

As you indicate the problems with ASW have been know for 30 years or more - and actually I was part of the group (as a grad student at the UW Friday Harbor Labs) that first did some experimentation with such mixes.

Our presumption at the time was the the mixes lacked something. After the tests I have run, I suspect instead that they have too much of the trace metals.

Additionally, Michael, may I add....

Do you have Meg Strathmann's book on "Invert Reproduction and Development?" If so, you will find I am chapter advisor for the asteroid and prosobranch chapters. I have had significant experience with invertebrate embryology, development, and larval ecology.

One of the larger engineer-consulting firms in the Pacific NW is Parametrix, Inc. From 1988 until 1993 I was the head of their natural resources section, and set up and was the head of their bioassay lab from 1988 until 1991. During that time we ran literally thousands of urchin bioassays on Superfund site work (given the potential for litigation, these tests were very closely monitored for procedural compliance). I think I know how do the procedure :D.

Additionally, most of these tests concerned excess heavy metals concentrations (although quite a number also dealt with excess toxic organic compounds - so Randy, some folks can figure something to test for. :D ).

Habib

Yes, you will find tests done using several ASW mixes as controls. I think these tests are flawed, but if they show increased mortality above and beyond what is caused by the salt mix due to metals concentrations, it must mean the metals are REALLY nasty.

I will try to find the URL for the test protocol and you can down load it if you want.

El-ahrairah

08/16/2002, 05:57 PM

Ron,

First off I am not challenging your ability to raise larvae or your knowledge on them. I posted the species that I had worked with to back up my claim that I had some experience with invertebrate larvae (that I didn't just walk in off the street - since you don't know me). Also the larval biology world isn't that big so naturally I know the Strathmann book quite well. You must have noticed then the fact that larvae - while they may reach settlement the numbers are not as high as when using NSW. When I first came into my lab I had already had a decades experience with marine aquaria and was quite shocked when my advisor told me ASW was a no-no, so naturally I had to check for myself. Consistantly the mortality with ASW is much greater than with NSW.

Our presumption at the time was the the mixes lacked something. After the tests I have run, I suspect instead that they have too much of the trace metals.

I still believe they are missing something. I agree that the heavy metals that are present are highly toxic to urchins (again this has already been done to death), but even if you make up seawater without the metals, I would say they don't do as well as with NSW (by "do" I mean their physiology not just thier mortality - ie respiration, protein synthesis, lipid metabolism...). I would argue that the seawater needs the natural DOC (animo acids and the like) for the urchins to survive as in NSW. I agree that the "toxins" present are part of the problem but only part of the problem with ASW - if you are trying to raise larvae.

As much as I have been giving Randy a hard time about him not having any data to support his chelation myth, I actually think it or something like it is occurring, and that ASW may be effectively detoxified in a tank. However, the price that is paid for such detoxification is that the chelated or bound toxins end up in the sediments or live rock and will come back to haunt us.

Okay then, from reading the earlier posts it wasn't apparent to me that you even considered accepting the idea. As far as coming back to haunt us, it might not be the case. The diversity of a reef tank will greatly differ from that in the wild - most likely from the bacterial point of view. Bacteria have a wonderful way of using everything and anything as an energy source. It may be (yes I know it's just specualtion) that the microbial community of a closed reef tank is able to utilize and "permenantly" detoxify many of the toxins in the water, thus the need for better toxin analysis of the sediments and rock of aged tanks. Naturally a monetary and timely undertaking for someone. Since I have never had it come back to haunt me personally and I have had tanks run (and still running) for a lot longer than four years, I am not yet convinced that the elevated levels of compounds in the ASW are that problematic.
-Michael

El-ahrairah

08/16/2002, 07:39 PM

Ron,

One other thing, you didn't quite answer my first question as to which species you plan to use, Lytechinus... ??

Since you covered an entire genus I'm not surprised their are some ripe all year round. :D
-Michael

Jabrams

08/16/2002, 09:25 PM

Originally posted by rshimek

The urchins I will use are Lytechinus and they will be obtained from Carolina Biological Supply, and generally some individuals are ripe all year round.

Ninong

08/16/2002, 09:49 PM

Oops! Double post.

:D

Ninong

08/16/2002, 09:49 PM

Joe, Michael knows the genus, he wants to know which species of Lytechinus. :D

Maybe it's Lytechinus variagatus?

:D

rshimek

08/17/2002, 07:08 AM

Originally posted by El-ahrairah

Michael,

One other thing, you didn't quite answer my first question as to which species you plan to use, Lytechinus... ??

Ah... frankly, what difference does it make? Do some looking around, I told you where I was getting them, if you are that interested, you can find the information from there.

Okay then, from reading the earlier posts it wasn't apparent to me that you even considered accepting the idea.

Oh, I considered it, but there are no data whatsoever confirming that it occurs, and I think taking it on faith is idiotic, as it seems to me that many other things could be occurring.

rshimek

08/17/2002, 07:17 AM

Folks,

We have beaten this topic to a pulp, and gone off on a number tangents, most of which have been unproductive.

The amount of time I have invested in this thread has been unfortunately, significant, and it has detracted from other things I need to be doing.

Consequently, I am closing the thread. If you wish to continue the discussion with me via email, we can do that, but the discussion here is at an end.

Randy Holmes-Farley

08/19/2002, 03:44 PM

Ron may choose not to post any more if he elects. Since the thread itself doesn't violate any reefcentral rules, however, it seems to me that it should remain open in case anyone else does.

Oh, I considered it, but there are no data whatsoever confirming that it occurs, and I think taking it on faith is idiotic, as it seems to me that many other things could be occurring.

FWIW, I find it unfortunate that Ron thinks the thread has been unproductive and the views of reefcentral members "idiotic".

Luckily, I don't think any people in this thread have taken anything on faith (certainly not me). I've mentioned several times that metal toxicity might be an issue in our tanks, and alternatively suggested reasons that it also may not be so critical that we are killing large numbers of corals in normal tanks. That's the whole point that several of use were trying to make: there may be alternative explanations that are worth considering.

Now that Ron is considering them too, as he suggests, then perhaps our time here has been well spent, and we can productively proceed to understand what is taking place in our tanks.

Fredfish

08/19/2002, 04:25 PM

Ron:

Though this thread has gone on a long time, I, for one, have learned a lot.

I thank you for your patience in all the questions that you have answered.

Randy:

How exactly can we proceed to determine what is going on in our tanks? It takes properly designed experiments, time, and, as Ron has pointed out several times, money.

How do you determine or eliminate organics as the source of toxicity in our tanks.

Its enough to drive me to buy a lottery ticket.:rolleyes:

Re: Old Tank Syndrome

This label is a bit of a red herring in my opinion. It has not in any way been defined let alone defined in a manner that one can conclude that it is the effect of one specific set of toxins.

Fred.

rshimek

08/19/2002, 05:10 PM

Originally posted by Randy Holmes-Farley

Ron Luckily, I don't think any people in this thread have taken anything on faith (certainly not me).

You have no data to support anything that you have suggested occurs in aquaria at all, cannot figure out a way to test for any of it Surely looks like you are taking it on faith, to me.

And, I don't think you have followed through on what would happen to anything that is "chelated" or bound to any organic material. This material would likely go to form flocculant material in the water or small particulate organic material and be eaten, by many organisms. In effect this "organic" binding, if it did occur would likely be a rather well designed "posion delivery system" using the particulate organic material that so many animals, including corals, eat. So, the question of whether or not the materials I have found are either simply ionic or bound to something is moot. They are still in the water, still bioavailable, and still toxic.

That's the whole point that several of use were trying to make: there may be alternative explanations that are worth considering.

When will you mention some of them?

Randy Holmes-Farley

08/19/2002, 06:28 PM

When will you mention some of them?

I don't know how you could have missed these POSSIBILITIES:

1. Organics chelate the metals, and so the metals are not at toxic concentrations to the corals that we choose to keep in most reef tanks.

2. That the metal concentrations you measured represent values that include solid, particulate, and colloidal materials, and are thereby less toxic then shown in studies that test (by addition) only freely dissolved metals.

3. That organics in our reef tanks are quite a bit more toxic than the heavy metals. Many of those known to be in seawater are mutagens and otherwise highly toxic. If I were to spend time studying toxicity, organics is where I'd put my money.

4. That our tankis can benefit from certain heavy metal additions. They may or may not be lower than NSW (iron, manganese, etc.), but tanks still clearly benefit.

It is lucky for you that my iron article posted yesterday. Now you don't have to spend time claiming that I am not addressing the issue of heavy metals, and that I have not run experiments.

http://www.advancedaquarist.com/issues/aug2002/chem.htm

rshimek

08/19/2002, 06:33 PM

Originally posted by Randy Holmes-Farley

I don't know how you could have missed these POSSIBILITIES:

And no data to support any of them.

It is lucky for you that my iron article posted yesterday. Now you don't have to spend time claiming that I am not addressing the issue of heavy metals, and that I have not run experiments.

Well, after fighting my way through the advertisements...

Randy, the only data you have in your poll are the results of an unscientific poll of reef hobbyists raising Caulerpa. woo woo....:eek2:

Do you have any measurements on the effects of the iron?
Did you test any hypothesis?
Do you have a schedule of dosing?
Did you measure iron uptake by anything?

You at one time mentioned controls? Where is your control?

How about a table of data? How about one datum?

You are right. I don't have to spend any time claiming that you don't run experiments; you have conclusively proven that you can't run one.

You sound like the old farmer who, after drinking some beer, added some self-generated liquid fertilizer to his garden and thinks his tomatoes are bigger. :bounce2:

:D:D:D

Fredfish

08/19/2002, 07:35 PM

Sigh...

OK, so we have all these metals in the water and a bunch (deliberately non acurate term since we don't know how much) of them are tightly bound by organics.

Habib, you talked about these bonds "holding tight" in nsw. Will these bonds be broken in the digestive tracts of the filter feeders we have in our aquariums?

Randy, how do we determine exactly what is killing organisms in our tanks?

Also, what organics in our tanks are more toxic than heavy metals?

Fred.

Randy Holmes-Farley

08/19/2002, 07:50 PM

And no data to support any of them.

You say this over and over, but of course it is completely untrue. Perhaps it is my fault for not doing your literature searching for you.

In "Captive Seawater Fishes" by Stephen Spotte, there is a multipage literature review with many references to copper toxicity and how the speciation of copper relates to toxicity in aquarium settings.

Here's a summary:

1. Copper is KNOWN to be CHELATED to ORGANIC compounds in aquaria.

He states "Immediately on dissociation the Cu(II) in copper salts forms very strong and complete nonlabile complexes with humic acids."

2. Copper chelation by organics causes a decrease in toxicity of copper to phytoplankton by reducing the free copper concentration.

He states "Toxicity in phytoplankton is reduced substantially when Cu(II) is complexed with inorganic and organic substances in solution and becomes unavailable for uptake by the cell walls." and "THE TOTAL CONCENTRATION OF TOTAL COPPER IS NOT A FACTOR".

3. To be pefectly fair, in orther organisms, organic copper compounds are still toxic. Notably things like certain polychaetes that absorb the organics as well as they do free metals.

Consequently, there is more than enough published literature data to support the HYPOTHESIS that organics may reduce the toxicity of copper and other metals towards corals in reef tanks. It is not a pure fantasy, as you suggest, but a possibility that flows naturally from the published literature.

rshimek

08/19/2002, 07:58 PM

Randy,

You make broad generalizations and treat them as known facts.

The fact remains that you have demonstrated no data supporting your suppositions in any of your systems or any others.

You now, finally, have a hypothesis to test. :D

Go ahead and test it or simply discuss it, but don't treat it as if it were anything but an asked question until you have the data in hand.

beerguy

08/19/2002, 08:02 PM

Originally posted by rshimek
Randy,

You make broad generalizations and treat them as known facts.

Maybe it's just me, but ..... oh nevermind.

SDBDRZ

08/19/2002, 08:09 PM

I think that we are getting caught up in the details here. As the saying goes we are looking at a tree instead of the forest. With all due respect to Dr. Ron and the wonderful work he is doing for this hobby I still think the sample size is inadequate to draw the conclusions that were drawn. What Dr. Ron says very well may be true but then again maybe not. There are just too many variables and without larger sampling we are not going to be able to draw conclusions about reeftanks in general.

There are too many things that can throw off such a small sample. Maybe most of the tanks contained Fiji LR and the LR is the problem and contains a high level of certain metals before it gets to the tank. Maybe 50% of the tanks sampled used trace element additives which throw off some readings. Maybe in one of the tanks someones child threw a penny in the tank when Dad was not looking. I could go on and on but you get my point. In such a small sample these errors do not get cancelled out.

I think that both sides have some interesting and valid points but we are argueing over the findings in 23 tanks. We are arguing about the tree not the forest.

Now someone pony up a ton of cash so Dr. Ron can test 2,000 tanks, then we can discuss findings :D

Randy Holmes-Farley

08/19/2002, 08:10 PM

Randy, the only data you have in your poll are the results of an unscientific poll of reef hobbyists raising Caulerpa. woo woo....

No, not exactly, but I do consider that an interesting and potentially useful piece of data as well. What's your beef with it? Do you believe it to be incorrect, and that adding iron has no effect whatsoever on sexual reproduction of caulerpa in reef tanks? Do you have any data for reef aquaria? Or is it just that you don't like information gathered from hobbyists? Or do you not accept standard stastical analysis?

As to experiments, maybe you missed that I personally dose iron at hundreds of millions of the NSW level and have seen only good effects. Hmmm, you'd think that heavy metal poisoning would have kicked in a little before that if it were an issue for iron.

FWIW, a great many people have asked me for help with caulerpa problems (fading, dieing, not growing much). In many of those cases I recommended iron, and what happened? Iron often IMMEDIATELY solved the problem (it greened up and began to thrive). What's you're beef with that? Do you not believe that it happens? These people have all lied?

Do you have any measurements on the effects of the iron?

Yep. Same as above.

Did you test any hypothesis?

Yep. Same as above.

Plus "Adding iron will not prevent sexual reproduction of caulerpa racemosa in my tank". That null hypothesis failed, and sexual reproduction has been prevented for years.

Plus "Adding iron will not encourage microalgae in a mixed tank of microalgae and macroalgae". That null hypothesis failed to, as the macroalgae thrived and the microalgae quickly lost out and disappeared, even where there was no predation. This effect was suggested to me years ago, and has been reproduced in many tanks.

Do you have a schedule of dosing?

Absolutely. Every day. Same amount. If you want details to try it in your own system (beyond what is in the paper, of course), I'll be glad to suggest some good supplements and schedules for you.

Did you measure iron uptake by anything?

And what would be the purpose of that? It is well established in the scientific literature that it is taken up. Again, I'll refer you to the many page literature review in "Captive Seawater Fishes" by Stephen Spotte. That review goes into great detail on where the iron is going and how it gets there.

My hypotheses did not involve uptake. It involved microalgae growth, macroalgae growth, color, and sexual reproduction. Since I do not know how much iron would be necessary to attain those effects, nor whether currently available techniques would see the uptake (recall you detected no iron), it seemed of little value. Besides, if I showed uptake, that doesn't imply benefit, does it? Copper uptake, after all, is what can kill things. So what would you or anyone else in the hobby do with iron uptake numbers for my reef tank? Such a criticism again looks more like an attempt to discredit someones work rather than to be asking for a legitimate piece of informnation that they would use to formulate some type of conclusion. Or am I missing some insight that you have about iron uptake rates?

Randy Holmes-Farley

08/19/2002, 08:20 PM

You at one time mentioned controls? Where is your control?

My tank is it's own control (before and after dosing; a well established technique in both biological and medical research). In the hobbyist survey, the control is people who dose no iron against those who dose iron. I'm surprised that you didn't understand these issues.

How about a table of data? How about one datum?

Not sure what you want. Some types of data are suited to a table, and some a stastical result. Have you ever seen epidemiological data that gave a table of every response? Of course not. They give a stastical analysis of the data, and set aside the primary data for review off line. The priamry data for my stastical experiment was carried out here at reefcentral, so you can look it up yourself and reanalyze it if you wish.

You are right. I don't have to spend any time claiming that you don't run experiments; you have conclusively proven that you can't run one.

OOOOOH. Struck through the heart again. You'll find I'm fairly immune to such lame criticism. :lolspin:

You sound like the old farmer who, after drinking some beer, added some self-generated liquid fertilizer to his garden and thinks his tomatoes are bigger.

Maybe you'be better explain this insult a little better. It doesn't seem to make sense. I though fertilization was a proven fact.

Randy Holmes-Farley

08/19/2002, 08:26 PM

Fred:

How exactly can we proceed to determine what is going on in our tanks? It takes properly designed experiments, time, and, as Ron has pointed out several times, money.

Absolutely. It is a very tough problem. If metals are toxic in our tanks, it is very hard to show what animals are suffering from it. Good old tox tests on each animal of interest would be necessary. Showing it for any coral or other animal that we wish to keep, however, would probably generate substantial interest and possibly money.

Randy Holmes-Farley

08/19/2002, 08:30 PM

Go ahead and test it or simply discuss it, but don't treat it as if it were anything but an asked question until you have the data in hand.

I need to repeat literature experiments that are discussed in Spotte? What on earth for? Do you not believe them?

Would you be more inclined to believe it if I found and reported the same thing 10-30 years later than the original research? I appreciate the confidence in my skills, but I think the original researchers did an OK job.

Randy Holmes-Farley

08/19/2002, 08:31 PM

Randy, how do we determine exactly what is killing organisms in our tanks?

What organisms?

Randy Holmes-Farley

08/19/2002, 08:33 PM

Ron:

You make broad generalizations and treat them as known facts.

Name one that I made, please.

Randy Holmes-Farley

08/19/2002, 08:38 PM

Also, what organics in our tanks are more toxic than heavy metals?

I'd have to know the tank concentrations of many organics to know the answer to that.

Still, some that are well studied in seawater that are made by algae include methyl iodide. It is very dangerous to people because it methylates DNA. I don't have any idea what it does to corals, but maybe that's where some of the unusual color morphs come from (that's a joke, by the way).

rshimek

08/19/2002, 08:46 PM

Originally posted by Randy Holmes-Farley

No, not exactly, but I do consider that an interesting and potentially useful piece of data as well. What's your beef with it?

No beef at all. It is a 2 x 2 contingency table and the statistics of these are rather well known. Following the formulations in Zar, Biostatistical Analysis, if my math is correct, the calculated chi squared value of such a table is 1.38, which puts the probability of obtaining a table with values such as that drawn randomly from a population as between 25% and 50%. So, there is no statistical difference between the dosing and non-dosing responses.

Do you believe it to be incorrect, and that adding iron has no effect whatsoever on sexual reproduction of caulerpa in reef tanks?

That is exactly what your data show. The adding of iron has absolutely no discernable effect.

As to experiments, maybe you missed that I personally dose iron at hundreds of millions of the NSW level and have seen only good effects.

Where are the data? Did you do anything like take a measurement of Caulerpa growth, time between reproductive events, any data that could be quantified.

Do you have any measurements on the effects of the iron?

Yep. Same as above.

Did you test any hypothesis?

Yep. Same as above.

So... NO DATA. NO HYPOTHESIS TESTING.

No replicates, no controls, no... science. Lotta arm waving though.

rshimek

08/19/2002, 08:52 PM

Originally posted by Randy Holmes-Farley

My tank is it's own control (before and after dosing; a well established technique in both biological and medical research).

That statement would certainly get you laughed out of any biology department that I know of. You have an N of 1, and no control. Ridiculous.

How about a table of data? How about one datum?

Not sure what you want.

Oh, something measurable... actually anything measureable. All you have is anecdote.

Some types of data are suited to a table, and some a stastical result. Have you ever seen epidemiological data that gave a table of every response? Of course not. They give a stastical analysis of the data, and set aside the primary data for review off line. The priamry data for my stastical experiment was carried out here at reefcentral, so you can look it up yourself and reanalyze it if you wish.

You have NO experiments, and the data published in your contingency table, which you were apparently unwilling or unable to analyze do not support your conclusions.

rshimek

08/19/2002, 08:59 PM

Originally posted by Randy Holmes-Farley

I'd have to know the tank concentrations of many organics to know the answer to that.

Well.... Duh... My point exactly.

How about taking some measurements?

Then you might have some data instead of supposition.

SDBDRZ

08/19/2002, 10:00 PM

That statement would certainly get you laughed out of any biology department that I know of. You have an N of 1, and no control. Ridiculous.

Most statisticians probabaly would not think much more of an N of 23. :rolleyes:

I really don't mean to distract from you and Randy's entertaining discussion (tomorrow night I think I am gonna pop some popcorn, pull up this thread and sit back and enjoy the show :D ) I keep bringing sample size up because I do not agree with your definitive findings based on such a small sample. Hey maybe your findings are right but you will have to sample more than 23 tanks to convince me.

EricHugo

08/19/2002, 11:02 PM

I couldn't bear to read the whole thread, so maybe this has been covered.

I will add my two cents. I sent a small box of papers to Ron, including a text, on Aquatic Toxicology, to add to what he had already unearthed in his own research. Included were what I feel are about the sum total of references on the effects of metals on corals.

I see it being argued that becasue we don't know certain things or the effects on specific animals, as scientists you know that science is a foundation. If a metal is found to be toxic to Hydra, to Drosophila, to C. elegans, then the assumption is that is is toxic to invertebrates and probably highly likely that it is toxic to closely related invertebrates, unless there is something that wouldindicate for a particular species that it is not.

Ron is totally correct about the research on invertebrates, and since sending him the information, I delved here and there and uncovered some other somewhat related and somewhat unrelted material. There is no question that levels above what are found in non-impacted areas are considered pollutants, posions and toxins...there is question and many unanswered questions as to the effects, and this is even more true with marine invertebrates.

There are studies investigating and that have investigated effects of polluted areas - many of the best happen to be in Kaneohe Bay and I happen to share a lab with someone who did their research there. There does indeed seem to be tolernace. There is little question that reproduction is heavily to totally affected by elevated levels of metals. I have listened to my fair share of talks at conferences on thsi very subject, and living in texas where chemical plants and oil industry makes our coastlines look like Miami Beach only with industry, there are no shortage of investigations, especially to the Flower Gardens Banks. Elevated levels of metals, pesticides, PAH's, and undetectable by standard EPA testing were found to affect settlement of coral planulae at levels in the parts per billion. How's that for a number?

Or how bout this regarding settlement studies:

In laboratory tests, planula recruitment bioassays were more sensitive than
standard toxicity tests performed on either adult corals or their larvae. Larval-substratum interactions are highly sensitive to the presence of certain pollutants.

I also see references being thrown around to support various alternative hypotheses - and from what I gleaned, they are algal and cyanobacterial studies....totally irrelevent. I mean, duh! Bacterial and algal uptake of toxins is old old news. In fact, the uptake of such things by algae is being propsed as one of the factors in some phase shifts from coral to algal dominated communities in impacted areas since algae tolerate it and invertebrates do not. Furthermore, metal studies on corals indicate that the zooxanthellae save the animal from much toxicity by taking up such pollutants and binding them. Corals also remove metals quickly from their tissue, releasing them into the water or binding them in the skeleton. In the very few studies on metals and zooxanthellae, elevated metal levels caused reduced mitotic indices and bleaching.

Irrespective of all this, and if corals can survive in the presence of all this garbage, why even postulate on why unless it is an area of research interest? Why even thinkabout adding more? What is this long-stading and bizarre fascination with trace elements and corals? Why do you think there is so little research going forth on effects of molybdenum on coral growth and coloration, as an example? Why, because no one would fund it becasue there is no scientific basis for it, no precedent, and every indication on a scientific foundation would indicate there is no need for it, and is probably toxic. If there is a release of it into the water by accident, be assured research would follow quickly to investiagte the potential harm to the reef community, not the potential benefits. Should it not be evident in the lack of sexual activity in our tanks?

Anyway, one last question - to Randy. Do you publish under the name Randy Holmes-Farley, because I searched PubMed, PubScience, Science Citation Index, WorldCat, Academic Search Premiere, ArticleFirst, MedLine, and SciFinder Scholar, and even my university search engine under pharmacology, and could not find any articles with you as the author or researcher. I would very much like to see what you have published. I like to read the works and interests of peers.

clkohly

08/20/2002, 01:11 AM

Eric: Well this is about the only part of the thread that I really understand. Common sense would tell me that Randy would probably go by something a little more professional such as Randall. Perhaps if you enter that name into your search Engines you might find something. I sure did.

Chris

Habib

08/20/2002, 09:18 AM

Eric,

Welcome to the club:D

If a metal is found to be toxic to Hydra, to Drosophila, to C. elegans, then the assumption is that is is toxic to invertebrates and probably highly likely that it is toxic to closely related invertebrates, unless there is something that wouldindicate for a particular species that it is not.

If e.g. sea urchin larve and Daphnia magna's tolerance to copper is significantly increased if the copper speciation is changed then would it not be unlikely that other invertebrates would show a similar increased tolerance to copper if the speciation is changed?

And is toxicity not proportional to bioavailability?
And does speciation not affect bioavailability?

Also it is well known that to treat fish with copper that the copper "disappears" and loses it's toxicity. The speciation of copper has changed.

Abstract:

Environ Toxicol Chem 2002 Feb;21(2):275-80

Speciation of copper in sewage effluents and its toxicity to Daphnia magna.

van VE, Burton N, Comber S, Gardner M.

Imperial College of Science, Technology and Medicine, Ascot, Berkshire, United Kingdom.

Copper complexation capacity was determined in a range of sewage treatment works final effluents and receiving waters, upstream and downstream of the discharge point. Forty-eight-hour immobilization tests on Daphnia magna were used to assess the toxicity of copper in the effluent matrix. Complexation capacities in effluents were typically in the range 50 to 100 microg Cu/L, with higher values being found in the poorer-quality effluents with higher dissolved organic carbon (DOC) concentrations. The tolerance of Daphnia to dissolved copper concentrations was more than quadrupled in a 50% effluent matrix, with the increase in tolerance being related to complexation capacity. Ligand concentrations in effluents were found to correlate strongly with effluent DOC. No such relationship was observed in surface waters. On mixing with river water, sewage-derived ligands behaved conservatively and were relatively stable over time scales of up to 10 d.

Abstract:

Aquat Toxicol 2002 Jul;58(1-2):27-41

Effect of humic acids on speciation and toxicity of copper to Paracentrotus lividus larvae in seawater.

Lorenzo JI, Nieto O, Beiras R.

Departamento de Ecoloxia e Bioloxia Animal, Universidade de Vigo, E-36200, Vigo, Galicia, Spain. lorenzo@uvigo.es

The effects of humic acid (HA) on the toxicity of copper to sea urchin Paracentrotus lividus larvae were studied in chemically defined seawater. Square Wave Anodic Stripping Voltammetry (SWASV) was employed to study the complexation of copper in seawater medium. A simple complexation model assuming one ligand type and a 1:1 reaction stoichiometry successfully explained the inverse titration experiments. A conditional stability constant of 6.53+/-0.05 and a complexating capacity of 230+/-7 micromol Cu/g HA were obtained. Sea urchin bioassay tests with two endpoints, embryogenesis success and larval growth were carried out in order to study the toxicity of dissolved copper in both the presence and absence of HA. The toxicity data obtained fitted well into a logistic model, and the high sensitivity of both endpoints (EC(50) were 41.1 microg Cu/l and 32.9 microg Cu/l, respectively) encourages their use for biomonitoring. The HA had a clearly protective effect, reducing the toxicity of Cu to the sea urchin larvae. The labile copper, rather than the total copper concentrations, explained the toxicity of the Cu-HA solutions, and the Cu-HA complexes appeared as non-toxic forms. These results are in agreement with the Free Ion Activity Model, because the labile Cu concentrations in this buffered and chemically defined medium covary with the free ion activity of the Cu, validating the model to naturally occurring HA in the marine environment.

EricHugo

08/20/2002, 10:15 AM

Habib:

Thanks for reminding me. Yes, I did mean to comment on that, too, and I do agree that speciation is very important...and also agree with you guys that complexing with organics can make a world of a difference in both biological effects and in testable results. And with Ron, I also agree that no matter how likely or pragmatic the argument, even less information is available and relevant to this subject that with the direct effects of compound x on species y in the marine environment. We are *all* skating on thin ice in such debate, and urge tempering of statements and careful throwing about of quotes and references so as not to appear to be a final word or gospel thats supports our views.

I'll turn to a non-scientific but darn applicable analogy of personal experience for a second. A long time ago, I did something out of desperation and something I repeated not too long ago. Also something I don;t ever want to hear someone else did cause Eric did it and it turned out ok....it is most certainly not something I would want anyone to do. By coincidence, it invovled copper.

I almost left the hobby in the mid 90's because of Aiptasia - I had tried everything and failed. I eventually put all my live rock in a bin and went out and bought several bottles of Copper-safe - a chelated copper treatment....and poured it in the bin. The anemones had turned dark within the day, but very few seemed to die. Knowing how much I had put in, I bought more bottles and added more a bit at a time. After pouring about five bottles over two weeks into about fifty gallons of water, I realized the anemones were not dying, and had even recovered - they seemed tolerant of this product, or the product was not availabel to affect them. I used a copper test kit, and while there was testable copper in the water, it wasn't off the scale as I would have expected.

I then went and bought another copper treatment that was supposedly a free ionic solution of copper. I added the bottle - and the bottle was only about one third the volume - of course, I don't know the concentration of either of the products, can't trust the manufacturer anyway, but the receommended treatment level for fishes, assuming that their recommendations are at all valid, suggested that the free ionic was roughly twice as strong based on dosage level per water volume. Thats a heck of an assumption, but let's go with it for the purposes of this analogy. The anemones started turnig dark again, and this time they died. In fact, bristle worms died like flies, I had peanut worms crawling out of the rock and dying, and the whole water turned blackish brown with decomposition of all the anemones and worms in a few days. It basically killed everything....and I felt I had won - I guess, if you consider mass mortality of all living invertebrates a victory. Now, none of the filamentous algae, coralline algae, or macroalgae on the rock was affected at all - at least visually or long-term in terms of survival and future growth. I then started using water changes and metal sponges to remove the copper, and eventually was able to replace the rock into the tank.

My second experience was similar - I used Copper Safe to try and kill snails in a rfeshwater planted tank. Snails weren't even phased and the plants did fine, too. Total waste of effort.

Now, the two most reasonable explanations for this are that 1) coppersafe is a lame and extraordinarily weak product that can't even kill a two cell layer thick tissue sac like an anemone or that 2) the chelation significantly affects the relative toxicity and/or testable amounts of copper in solution.

But, (and I do ask that you bring me up to speed on this thread if I repeat what's been said, fail to cover issues addressed, miss a point, etc., because as I said, I couldn't/wouldn't/shouldn't read it all), and excuse me for being presumptuous, but so what? Chelators themselves may have their own set of effects....as I pointed out in a thread on my forum, the standard method for decalcifying corals for electron microsocopy involves slow chelation with EDTA. It slowly chelates the calcium and dissolves the skeleton. That's just an example. Second, if things are taken up by plants, algae, microbes, coral skeletons, or bound by chelators, or other organics, and they die or are released by other means, we're in a world of trouble. The fact that they exist at levels way in excess of NSW and are likely to have toxic effects in at least one chemcial species, makes me sweat - especially when we are likely manipulating the biological and chemical environment far more in tanks than is done in the wild.

Now, I don't know what was said regarding toxic effects, and believe I tried to temper at least some conclusions in Ron's article when I reviewed it. The information he has discovered - and I might add in a scientific fashion using quality materials and methods, even if not ideal (as can be said for most of thepapers in speer reviewed science, I might add) and being one of the very very few truly available pieces of such information in the entirety of private aquarium science (term used loosely) - is pretty astounding and certainly has the very real potential of needing to taken with as much shock value as is obviously the case by this thread's interest. Debating the fine points and possible alternatives to explain survival in a toxic slurry sort of takes away from the importance of the finding. Believe me, I also know all too well how my friend can get when he gets fired up.

I have also watched many times as scientists I greatly respect begin to take opposite sides of a fence by pushing their own points to their logical extremes and wind up almost diametrically opposed from a fence they once stood nearly upon together. This can be highly counterproductive, too. Another example: Ove Hoegh-Guldberg and Clive Wilkinson are two scientists in Australia who agreed on the potential impacts to coral reefs by climate change, the increasing and warnings of mass bleachings, the somewhat dismal future for the world's reefs, etc. One of them sort of felt that corals will not be able to adapt, the other felt they might be able to. The press (lay persons, like most of those reading this thread), came to the conclusion, since both were well respected, that they had diametrically opposed views of the future of coral reefs and mass bleaching events, and so "since no one knew the answer yet" no longer felt either reefs or bleaching were probably an imminently important issue.

So, I would urge those of "well-heeled intellect" in this thread to bear this in mind for the sake of and in consideration of the hobby for whom this thread, board, site, entire world's aquarists, and information are being given.

cfockler

08/20/2002, 11:07 AM

Originally posted by EricHugo
Irrespective of all this, and if corals can survive in the presence of all this garbage, why even postulate on why unless it is an area of research interest? Why even thinkabout adding more?

Welcome to the show Eric. I'm reading you loud and clear! :wave:

I personally feel all of this energy would be better spent finding ways of getting our tanks closer to natural seawater conditions.

Ron, hows the search for a better salt progressing?

08/20/2002, 11:43 AM

Eric,

I notice you are quick to discredit your "peers" credentials, but didn't reply when someone suggested you were mistaken.

Was this due to you feeling the poster was incorrect, or do you feel it's acceptible to smear someone and not bother to see if your accusations are correct?

This kind of mud slinging is not helpful, and IMO is more indicative of acting on agendas than on helping the hobby.

I have also watched many times as scientists I greatly respect begin to
take opposite sides of a fence by pushing their own points to their logical
extremes and wind up almost diametrically opposed from a fence they once
stood nearly upon together.

It seems to me a bit of "Kettle, you are black, sincerely, Pot." is going on here.

SDBDRZ

08/20/2002, 12:14 PM

08/20/2002, 12:23 PM

I wasn't aware there was a post count limit to asking a question. I don't see my post as an attack, unlike Eric's post to Randy, which was most certainly an attack on his credentials.

I do feel that since Eric chose to make an attack on Randy's credentials, he is open to criticism for not responding when a rebuttal is made. Otherwise, it appears that he is ignoring any points that don't fit his agenda.

PS> If you look at my registration date, you'll see that I didn't just sign up today in order to post this message, thanks for the welcome.

SDBDRZ

08/20/2002, 12:33 PM

There is not and I apologize. I will delete my last post.

Threads like these get a little heated and it is easy to get pulled into the accussation mode, you have my apologies.

EricHugo

08/20/2002, 12:41 PM

F4, you are totally mistaken. I like Randy's advice and feel he offers a great deal to the aquarium hobby with his knowledge. I have sent Randy, on my dime, hundreds of pages of articles for his columns from my collections when he asked, and I reguarly read and commend his work. It was his post about all his papers that made me go and try to read some of his field of work, and I couldn't find any - which is why I asked. So save your presumptions, ok? Got anything to add to the discussion or are you just going to try and take me on...cause if you are, I'm ready...hope you are! :)

SciGuy2

08/20/2002, 12:45 PM

:D Good to have a peace keeper or two around.

08/20/2002, 01:01 PM

are you just
going to try and take me on

Unlike some here, I am not posting for any reason other than to clear up one tiny part of this mess. I have no agenda other than to understand whether you are still suggesting that Randy's claims of publication are false. You made what I consider a pretty serious attempt to discredit him, and I think it's important to understand exactly where you stand on the issue when it has been brought up that he DOES in fact use names other than "Randy".

Why is asking Randy for the name that he publishes under an attack?

What other purpose could be assumed by asking someone why you can't find any published works by them? The insinuation is VERY strong that there are none to be found.

Trying to be a peace keeper,

That's great, but I find it interesting that I am attacked for asking why Eric didn't respond to the additional information regarding Randy's name. Especially when it is not even important to the discussion. Eric felt it important enough to bring it up in the first place. I'm just wondering why, if not to attempt to discredit.

Eric, I'm sorry you feel that my wanting to understand your motives isn't productive, but I think it's important.

Randy Holmes-Farley

08/20/2002, 02:09 PM

No beef at all. It is a 2 x 2 contingency table and the statistics of these are rather well known.

I did a 2 x 2 chi square test and got the result posted. If you don't like the result, then I suggest you run your own survey.

Randy Holmes-Farley

08/20/2002, 02:18 PM

That statement would certainly get you laughed out of any biology department that I know of.

Ron, I'll go toe to toe with you on scientific credentials any day. Including where you got your biology degree and where I got my biology degree (leaving aside chemistry for a tiny second; Oh, did you not know that I too have a biology degree? It is easy to ridicule people when you know little about them). Still, that isn't the point of science, is it? Is it?

Sure, an N of one requires replication to be generally accepted, but that doesn't mean that it is wrong. That replication involved the tanks of many reefcentral members, including several of the moderators here. Though, they could all be lieing....

If I'd been suggestintg someting odd, I'd expect a bit of skepticism, but since I'm only recommending something that many people have been doing successfully for many years, I'm not sure I understand it, except again that it seems to not fit with your theory that addition of metals is undesirable.

rshimek

08/20/2002, 02:25 PM

Some points to start with.

F4: Start a new thread some place for your argument with Eric. Better yet take it off line. If it continues in this thread, I will close and delete the thread.

Habib and Eric have both discussed the ability of chelating agents to bind copper. Great.

Now ----

How abundant are these things in our tanks?
How available are they? sometimes? always? And which ones?

More importantly, what happens to them and their bound metallic poisons when they start to accumulate?

If they form any kind of particulate organic material, they will be eaten.

If they are eaten, in the acid or basic phases that virtually all animals have, some or all of the poison (copper, nickel, zinc, vanadium, ad nauseum) will be released.

When I tested aquarium waters, these chemicals were found in the waters in lethal amounts. If they are chelated, fine. They are likely also chelated in the natural systems when the test mortality occurs. It simply doesn't really matter whether they are ionic or chelated; sooner or later unless they are removed, they will be effecting the animals in our systems.

ASW is universally thought to be toxic to invertebrate embryos invertebrate embryologists. If ASW gets detoxified as it is added to our tanks by chelating organic compounds, what about those tanks without the appropriate organisms to produce the appropriate compounds? and what can be done to ensure their productivity?

Etc, etc, etc....

But most importantly of all.....

WHY ARE THE SALT WATER MIXES MADE WITH TOXIC CHEMICALS IN THEM?

In other words, why are we having this discussion in the first place?

Randy Holmes-Farley

08/20/2002, 02:35 PM

Eric:

I greatly appreciate all of the papers that you have sent me, and I also greatly welcome your input in this thread.

The primary reason that I am here in this thread is to try to provide some counterbalance to Ron's assertions that "we are killing corals" and that I am personally doing so. It could be true. We may be killing corals with metal toxicity. But we also may not be. An ICP measurement is not definitive enough, IMO, to start telling people that it is a demonstrated fact, as Ron has done.

I've provided several reasons why his test may not demonstrate that we are killing corals. Which one, if any, is true? I don't know. There is just no allowance in his articles or posts that there is any possibility that we are not killing corals. I'm allowing for tha possibility, and in the case of some corals, it really has to be a likely probability since many of us have kept many corals thriving for many years. Are these the ones that are especially resistant? Maybe. Or maybe they even exemplify the majority of corals.

ut, (and I do ask that you bring me up to speed on this thread if I repeat what's been said, fail to cover issues addressed, miss a point, etc., because as I said, I couldn't/wouldn't/shouldn't read it all), and excuse me for being presumptuous, but so what? Chelators themselves may have their own set of effects....

Hmmm. I seem to have suggested this as well. That organics themselves may be more toxic than metals in our tanks. Even beyond chelators, many organsims release organics specifically to KILL other things. Ron tossed out this hypothesis as ridiculous since I have not personally measured the levels of these organics in my tank.

08/20/2002, 02:46 PM

F4: Start a new thread some place for your argument with Eric. Better yet
take it off line. If it continues in this thread, I will close and delete the
thread.

I find it interesting that you allow Eric to bring the subject up, but threaten me for asking him for clarification of his comments.

I had hoped that the response would not be more attacks, but rather an acknowledgement of the real facts. I'm dissapointed in the reaction. My real hope was that the attempts to discredit would stop, but I see that is not going to happen.

But, never fear there is no more I can accomplish in this vein, there is no need to close the thread on my account.

Randy Holmes-Farley

08/20/2002, 02:49 PM

Eric:

No offense taken on the publications. It would be safe to look for most of my publications with just the last name Holmes-Farley, since there are only 4 of us, and my wife has only two searchable publications on ethics (my 2 and 5 year old daughters have yet to publish (well, maybe they published in China before I adopted them, but those don't show up in normal searches :D )).

My papers end up with all combinations of first and middle names and initials, as my full name is Stephen Randall Holmes-Farley.

The majority of my recent publications have been patents. Here's a link to 44 of them for those that don't have a good database available, but Patent Office documents are hard to read this way:

http://patft.uspto.gov/netacgi/nph-Parser?Sect1=PTO2&Sect2=HITOFF&p=1&u=%2Fnetahtml%2Fsearch-bool.html&r=0&f=S&l=50&TERM1=holmes-farley&FIELD1=INZZ&co1=AND&TERM2=&FIELD2=&d=pall

I am especially proud of my work on organic polymers that bind inorganic and organic materials (like iron, phosphate, bile salts, and fat), hence my keen interest in metal binding in this thread. My inventions have led to two FDA-approved pharmaceuticals (Renagel and WelChol) and have won several American Chemical Society awards for those research efforts. These awards were presented to me at symposia in my honor (co-honor with one other inventor in most cases) at regional and national ACS meetings.

Randy Holmes-Farley

08/20/2002, 03:02 PM

Eric:

Irrespective of all this, and if corals can survive in the presence of all this garbage, why even postulate on why unless it is an area of research interest? Why even thinkabout adding more?

Were only postulating on why because Ron tells us that they aren't surviving. It just seems to us that many corals are, and we'd like to understand how Ron's data and assertions about toxicity can be compatible with the experience of hordes of hobbyists.

I don't advise people to add any heavy metals except iron, and Ron detected none. If anyone has evidence that there is "enough" iron around in our tanks, I'd gladly back off on that opinion. But I've seen many people benefit from additions. They just don't typically post such in Ron's forum.:D I've also NEVER seen anyone who had a toxicity problem that they attributed, rightly or wrongly, to iron additions.

Still, I'd have to say that unless one knows the bioavailability of the metals present in reef tanks, they could be in too short of supply EVEN IF the total metal concentration were higher than NSW.

That's where the tox tests come in. I think it would be great to see some good quality experiments run on corals that we keep (or want to keep) to know if this is an issue or not.

Likewise, we should keep our minds open enough that if Habib or someone else publish results that show metal additions to be beneficial, that we should at least read it and see if it is good science, or see if it is easily refuted (not by whining about stastics, but by showing a genuine flaw in the test or showing it to not be reproducible). If the stastical numbers are believed to not be high enough, then the response is to test more tanks, not to state that it can't be happening because the author only tested 65 tanks, and not the 154 necessary to attain a p of 0.05.

rshimek

08/20/2002, 03:03 PM

Originally posted by Randy Holmes-Farley

Hi

I did a 2 x 2 chi square test and got the result posted. If you don't like the result, then I suggest you run your own survey.

I don't have to. Your data are conclusive and show that you are incorrect.

A 2 x 2 test, is a chi-squared test with 1 df. And such a test shows non-significant difference.

Well, I guess the contingency tests aren't as well known as they should be. :D

So...

Your data:

....................dose...................no dose.......total
spores............1...........................20.............21
no spores.......9............................31............40
Total..............10............................51............61

Chi-square statistitic = (61( abs val((1 x 31) - (9-20)) - 61/2)^2)/(10 x 51 x 40 x 21)

or ..... = (61 (abs val (31-180) -30.5 )^2)/(510 x 840)

or ..... = (61( 149-30.5)^2)/428400

or ..... = (61 (118.5)^2)/428400

or ...... = (61 x 14042.25)428400

or ...... 856577.52/428400 = 1.999

From tabled values of the chi squared statistic, with 1 df, such a value or greater one has a probability of occurring randomly in a sample of your size with a probability between 10 and 25 percent of the time.

Standard signficance in biological tests is a probablility of 5 percent or less.

Your data are therefore not statistically significant.

Test formulation from pp 64-66, and tabulated chi squared value from p. 479 of:

Zar, J. H. 1984. Biostatistical Analysis. Prentice-Hall, N. Y. 718 pp.

Randy Holmes-Farley

08/20/2002, 03:06 PM

How abundant are these things in our tanks?

Ron, this isn't rocket science. They are very adundent in many tanks. Far higher than in seawater. That's why tanks are more yellow than seawater: because humic substances have built up. Spotte discusses these facts in detail. I'd suggest you read his discussion and those references before continuing to assert that this is an imaginary process or not seen in reef tanks.

rshimek

08/20/2002, 03:12 PM

Originally posted by Randy Holmes-Farley

(not by whining about stastics, but by showing a genuine flaw in the test or showing it to not be reproducible).

Statistical analyses allow us to distinguish valid results from "noise." The only people who whine about statistics are those who can't understand them, or those whose data are not supported by them.

Your data, charitably, might be said to show a "tendancy." If you had done some actual experimentation with replication, used controls, and had a decent sized number of samples, you might be able to say something. As you didn't, didn't, and didn't, you really can't.

rshimek

08/20/2002, 03:19 PM

Originally posted by Randy Holmes-Farley

They are very adundent in many tanks. Far higher than in seawater.

So you claim. Why not show me some numbers?

That's why tanks are more yellow than seawater: because humic substances have built up.

That's nice, Randy, you are a true master of generalizations.

Now what humic substances?

Which ones are there? Are they toxic in their own right? Or if they bind metals are they more-or-less toxic then?

Which ones bind toxic metals?

What is their fate in tanks?

What are their concentrations with time? If they vary signficantly, should we not be adding some of these to our systems to detoxify our salt mixes?

dchisenh

08/20/2002, 03:28 PM

Hey Everyone :) , I've been following this thread for quite some time (along with half of the reefing community it seems) and something has caught my attention. Why is it the question is always about "tolerance" of metals and organics? Don't we want our inverts (and fish, and everything else alive in our tanks) to thrive and be "happy"? ;)

I definately think the "chemistry" side of the debate has it's points on the ability of organisms to detoxify metals, sequester them, etc. and it's definitely good science for someone to play devil's advocate in order to continue the research in the area to find out definitively (well, as best as we can) the effects of these chemicals along with many others on all aspects of marine life. As valuable as it is for us keeping aquaria on a hobbyist level, it may end up doing the world a favor as our reefs face more and more problems in the years to come.

Also, the "biology" side of the debate makes a great point on the fact that these metals aren't naturally found in these concentrations in NSW and they most likely (no one can be 100% sure all the time) that they are a major suspect in "mysterious" coral and invert deaths that are all too common in the aquarium supply chain. I can't remember the exact percentage of species harvested which end up dead in the process even before they get to the consumer (which the consumer himself is most often responsible for a LOT of the subsequent deaths), but I know that if we were dealing with dogs, cats, birds, ferrets, marmosets, pigmy elephants (just kidding) or any other cute and cuddly animal, all hell would break loose with animal rights activists and the entire public in general. Funding for the prevention of all of these causes of mortality would sky-rocket and the world could sleep at night thinking they've done their good for the day. Unfortunately, corals and inverts aren't exactly cute and cuddly (have you ever tried to hug a frag of fire coral or kiss a mantis shrimp?) and so their mass deaths go un-noticed and un-investigated. The biologists, and everyone on this planet, should be rightfully inflamed over this and should be actively looking for and investigating as many causes as possible.

Perhaps these metals are the killers they're made out to be, perhaps it's something else AND the metals, and maybe something else and the metals and something else..and so on, and so on. Either way, I think we're on the right track and in just this thread alone I see reason to continue research with metals, and organics, and chelators, etc. Hopefully, we'll figure this thing out and greatly reduce the mortality of corals coming into this trade, and perhaps help out the natural reefs in the process. But, in the meantime, I think we should be continuing to look for ways to allow our corals that have made it through the gauntlet and end up in our tanks to thrive and "enjoy" life, not just "tolerate" it. If reducing metals helps, great. If reducing organics helps, great. If ANYTHING helps, great, let's try it. I'm sure if the critters could talk, they'd thank us if we try.

That's just my thoughts on the subject,

Dan

Also, as a senior biology major working on options in biotechnology and ecology, it's an honor to engage in a conversation with researchers with the credentials we have here on this website. I think all of us reefers, professional and hobbyist alike, owe you all a great deal of thanks for sharing your knowledge and expertise with us. Thank you once again.

Randy Holmes-Farley

08/20/2002, 03:52 PM

Ron:

"Statistical analyses allow us to distinguish valid results from "noise." The only people who whine about statistics are those who can't understand them, or those whose data are not supported by them.

This (below) was the conclusion that I made in the paper. If you'll note, the conclusion was that the results were intriguing and worthy of additional study. If I thought that the result was a definitive fact, I would not have written that. Please don't exaggerate what I said and make it seem as if I stated that I had found and demonstrated something beyond all doubt.

"In other words, there is a 96% chance that there is a real difference between the iron dose group and the no iron group with respect to Caulerpa undergoing a sexual event. However, we must recognize that those who dose iron may be the same folks who dose other things (like iodine, which also was statistically significant against no dosing), or do something else in common, potentially confounding a statistical test. Nevertheless, the difference is intriguing, and worthy of additional study. In my opinion, it is also good enough evidence for those plagued with such events to try iron dosing. "

Habib

08/20/2002, 04:14 PM

Ron,

ASW is universally thought to be toxic to invertebrate embryos invertebrate embryologists.

Don't you know that various invertebrate stages require nutrients which they derive from the surrounding water? That these nutrients are a.o amino acids or other nitrogen compounds.

ASW will normally be devoid of such compounds. So toxicity could be in fact malnutrition?????

Does anybody know the exact nutrient requirement for corals not to mention the early stages of corals?

WHY ARE THE SALT WATER MIXES MADE WITH TOXIC CHEMICALS IN THEM?

Just look at the NSW given by you in the febr article. You corrected it later thanks to IIRC Tatu and Randy.

GP2 is perhaps crap and Crystalsea Marinemix can be perhaps and perhaps after some slight modifications mimic NSW.
But I want to have it confirmed by analyses. I am about ready to have them done and I am paying for them. And if somebody want to make donations that is fine; I only accept beer or californian pistache nuts;)

When I tested aquarium waters, these chemicals were found in the waters in lethal amounts. If they are chelated, fine. They are likely also chelated in the natural systems when the test mortality occurs. It simply doesn't really matter whether they are ionic or chelated; sooner or later unless they are removed, they will be effecting the animals in our systems.

They are, if present in elevated concentration, more likely to be chelated in the aquarium then in NSW. Especially when food is added in the aquarium. For example copper can bind strongly to the highly abundant amide groups in polypetides (proteins).

If they form any kind of particulate organic material, they will be eaten.

If the particles are very small and there is sufficient water movement then they will not be eaten. This is plain physics and rheology. If the particle size is larger it might be eaten; ever looked at metal concentration in naturally ocurring algae. They greatly magnify the metals concentration of NSW; upto 100,000x ?!?

You have asked a lot more questions:)

So lets say that if the heavy metals measured in the concentrations as presented in Reefkeeping etc etc when present as free ions have the potential to be (highly) toxic to at least some marine organisms.

It still remains to be answered if they are present as free ions or are in any way bound to organics or other inorganics and present as e.g. ion-pairs or as colloids.

Toxicity to atleast some marine organisms depends on the (meta)stability of these compounds in the water column or in the organism if ingested directly or indirectly.

Further research on the speciation of these metals in aquaria should be conducted and if possible simple methods of monitoring be devised. The search towards better synthetic seasalts should be continued in the meantime.

Can you agree on this as a further basis for discussions?

Randy Holmes-Farley

08/20/2002, 04:16 PM

That's nice, Randy, you are a true master of generalizations.

Actually, for the immediate section that you are so up in arms about, I'm simply the master of copy and paste. All of the discussion of humic substances is directly from the detailed description in Spotte and references included therein.

I haven't yet seen you name a specific instance where I incorrectly genralized something into a fact that wasn't, and I'm still waiting. Simply because you don't know the fact, doesn't mean that it doesn't exist.

Now what humic substances?

I'm afraid that even asking that question shows that you know little about seawater chemistry and humic substances.

According to references given in Spotte, humic materials are "amorphous, brown or black, hydrophilic, acidic, polydisperse substances of molecular weight ranging from hundreds to tens of thousands". It is a big class of many different compounds. So it is not a list of 1 or 34 or even 23,456 compounds. It is a continuum of different products that evolve chemically with time in each tank. They are typically studied as a class or at least in sub groups (like humic acids and fulvic acids).

Are they toxic in their own right?

Yes, in some stages of their formation they are (the polyphenol stage has an LC50 of 0.3 mg/L to plaice larvae). In some chemical stages they are not toxic to these same larvae.

Or if they bind metals are they more-or-less toxic then?

I already addressed this, but in the case of certain organsims the toxicity of copper goes down. In others, it stays the same as the free copper.

Which ones bind toxic metals?

All are polyphenolic and all polyphenols bind metals at the pH of seawater. Some bind more strongly than others.

What is their fate in tanks?

Some humic acids stay in solution forever. That's why they build up in both tanks and in seawater. Some have been floating around in seawater for thousands of years. These are termed the refractory substances. In a reef tank with certain types of filtration (skimming or carbon) they may be removed that way, which would not happen in the ocean. This is likely a great export mechanism for toxic metals, whether we want them exported or not.

What are their concentrations with time? If they vary signficantly, should we not be adding some of these to our systems to detoxify our salt mixes?

If you didn't mind a yellow tank, knew where to buy some, and believed that you had excess metals, then I think that is a wonderful idea:) Now were getting somewhere!

Of course, it isn't really the salt mix that is at fault, is it? The natural seawater samples in your study were among the worst. Perhaps if metal toxicity is a real issue, we ought to look to other sources, like calcium and alkalinity additives.

Randy Holmes-Farley

08/20/2002, 04:30 PM

I can't track them all down right now, but several times in this thread it has been asked: Why have all these metals in the salt mixes?

I can think of one critical reason (cost) but that aside, I think before beating the war drum for low metal salt mix, we need to ask whether it will help.

In Ron's test the answer really appears to be: Not a teeny tiny bit.

The natural seawater samples are among the worst in terms of metals content. In some instances, they are the absolute worst.

Here's the link:

http://reefkeeping.com/issues/2002-02/rs/feature/index.htm

and here's Ron's conclusion:

"but even aquarists using NSW as the initial medium don't have tanks where the chemical composition bears much similarity to Natural Sea Water. "

In fact, the copper concentration only varied by a factor of two between the lowest tank and the highest tank. That seems odd to me, but still it is what we have to work with. Is there some other metal that we think is a problem in salt mixes that would be "cured" if only we used natural seawater types of mixes?

So why the tremendous angst about salt mix?

If metal toxicity really is an issue, we must look deeper into the problem Since the media used for reactors have been studied by many people, we can see how much metal is there (lots, in some cases):

http://www.animalnetwork.com/fish/library/articleview2.asp?Section=Aquarium+Frontiers+--+Biochemistry+of+Aquaria&RecordNo=1571

http://www.animalnetwork.com/fish2/aqfm/1997/aug/bio/default.asp

Craig first reported this problem years ago, but few seemed to care.

dchisenh

08/20/2002, 04:35 PM

Hey Randy or Habib, do you think the sodium EDTA found in Kent's Marine Coral-vite additive has any appreciable chelating effect for a reef tank? It's listed as the fifth ingredient on the list on the bottle, but it doesn't mention a concentration. Just wondering. :)

Dan

Randy Holmes-Farley

08/20/2002, 04:37 PM

Sure, but it may be already chelated to something besides sodium in the mix, and that something may not come off until the EDTA is photochemically degraded in the tank. Still, if added with just sodium, it will undoubtedly chelate metals. That's why it is added to the product to begin with (I expect).

dchisenh

08/20/2002, 04:51 PM

So...if the EDTA is in the tank in any appreciable amount with normal dosing of that particular product (I'm guessing it's in others too, but that's the only one I have on my desk in front of me right now ;) ), would that create a "pool" of EDTA available in the tank that combined with the typically intense lighting reefkeepers employ, be routinely available via photochemical degradation? If so, would that help explain the seemingly successful tanks some people have with high metal concentrations, or has that already been taken into consideration when doing assays? I've taken my fair share of O. Chem through my academic career, but I'm not sure how it would react in a typical aquarium setting and I apologize if this has already been addressed earlier, I must have missed it.

Thanks,

Dan

Just realized this, are you saying that the metal the EDTA may be bound with initially won't be released until the bond with the EDTA is degraded or the EDTA itself is degraded?

And Ron, if this isn't in keeping with the theme of your main discussion and is too much of a chemistry related side-issue, just let me know and I'll address any other questions about it directly to Randy in the chemistry forum, I just thought it sounded like chelators were part of the original discussion.

Randy Holmes-Farley

08/20/2002, 06:24 PM

If so, would that help explain the seemingly successful tanks some people have with high metal concentrations,

It could, yes. It could effectively lower the free (and possibly more toxic) copper (II) ions, and hold it in a less toxic form.

At the same time, the EDTA may help maintain the solubility of iron that is typically very, very insoluble (it may be the least soluble metal ion in seawater).

Yes, it is when the EDTA is degraded photochemically that iron gets loose and is available to animals and plants for consumption. So in that sense, EDTA helps maintain a steady but low concentration of bioavailable iron.

rshimek

08/20/2002, 08:02 PM

Originally posted by Habib

Habib,

Don't you know that various invertebrate stages require nutrients which they derive from the surrounding water? That these nutrients are a.o amino acids or other nitrogen compounds.

No, sir, they don't.

In early development any nutrition is dependent upon yolk. They may absorb some compounds from the water, but they don't absorb meaningful amount of amino acids, or any other food.

If they feed, they do it by suspension-feeding.

They may require and absorb certain other things, including, in small amounts trace elements. In larger amounts, these elements kill and and this is the basis for bioassays of non-feeding larvae.

ASW will normally be devoid of such compounds. So toxicity could be in fact malnutrition?????

Not a chance. The larvae die before needing any food.

Does anybody know the exact nutrient requirement for corals not to mention the early stages of corals?

Yeah, for some corals some parts of the puzzle are known; it is out there in the literature; at least for a few. However, most corals are "black boxes" nutrient of some source goes in, coral tissue comes out. Pocillopora has been raised as "lab rat" since 1975 or there abouts and there is a lot written about it. You may be able to track down some of these data if you need it.

But....regarding the larvae and embryos...

Cnidarians, by and large, have large yolky eggs and do not feed or take nutrients from the water until after they settle and metamorphose into juveniles.

If the particles are very small and there is sufficient water movement then they will not be eaten. This is plain physics and rheology.

The operative part of your statement is very small. Once they aggregate to anything over about 0.01 micrometer they will be eaten. I suspect that happens fairly rapidly.

Also our tanks have an enormously high surface area to volume ratio compared to the ocean. With all of that suface these particles will find their way there, be adsorbed and then be eaten.

If the particle size is larger it might be eaten; ever looked at metal concentration in naturally ocurring algae. They greatly magnify the metals concentration of NSW; upto 100,000x ?!?

Yes, I know. Substance like iodine in particular (probably because it makes a great defensive factor against herbivores) and I have data from aquaria, on Caulerpa which I hope to publish soon. They don't concentrate the levels of tank water metals much.

You have asked a lot more questions:)

Yes, and most of them are not answerable with the data at hand.

It still remains to be answered if they are present as free ions or are in any way bound to organics or other inorganics and present as e.g. ion-pairs or as colloids.

Yes, agreed. It would be interesting find out what way they are found, and how stable the systems are.

Toxicity to atleast some marine organisms depends on the (meta)stability of these compounds in the water column or in the organism if ingested directly or indirectly.

Yes.

Further research on the speciation of these metals in aquaria should be conducted and if possible simple methods of monitoring be devised. The search towards better synthetic seasalts should be continued in the meantime.

Can you agree on this as a further basis for discussions?

Absolutely. :D

Randy Holmes-Farley

08/20/2002, 08:29 PM

The search towards better synthetic seasalts should be continued in the meantime.

Can you agree on this as a further basis for discussions?

Absolutely.

What will that do for us? Tanks using such seawater (natural seawater) had just about as much copper and other metals in your test.

Wouldn't we still be having all these same concerns if every tank used natural seawater or low metal salt mixes? This is, if every tank looked like the natural seawater ones in your study?

rshimek

08/20/2002, 08:30 PM

Originally posted by Randy Holmes-Farley

Randy,

"In other words, there is a 96% chance that there is a real difference between the iron dose group and the no iron group with respect to Caulerpa undergoing a sexual event.

And this should have been,

In other words, there is between a 75% and a 90% chance that there is a real difference between the iron dose group and the no iron group....

As Regards the NSW tank in the original study...

Yes, it had high levels of heavy metals. But that is immaterial to the study comments at hand. The salt water in the tank was polluted from the accumulation of heavy metals in foods, more than likely.

Toxic metals enter our tanks by a number of path ways.
Apparently, they don't exit the tanks much, although I will have more to say about that.

The net result is accumulation.

If we can develop some way of filtering the toxic material out, or otherwise exporting it, or rendering it non-toxic we will have animals that will live. I suggest that some folks are able to do this some of the time, maybe a few most of the time.

Nonetheless, we start out the game at a decided disadvantage; we start out with salt that is toxic.

We pay for a medium to keep animals alive. It makes no sense whatsoever that it should be so poorly and cheaply made as to be potentially toxic from the get-go.

Now... maybe, if I run these bioassays I intend to run I will find that not all salts are created equally bad. Maybe some will be pretty good. Then it makes sense to use those salts to start with in our tanks, all things being equal.

On the other hand, if they are all bad = significant larval mortality, done in an experiment with replication and controls, then what does that mean?

Well, it means that surrogate animals died in the salts. Not corals. I don't have either the money nor the space to do this experiment with corals. However, the use of delicate animal stages as surrogates is well-documented and well-supported in the ecological literature as well as in the environmental regulatory community. It is an understood and useful technique.

If there is mortality in these bioassays, will it prove corals will die? No. Most assuredly not. However, it will say that the water in the container kills animals in very short periods (4 days or less). The assumption is that any animals in such toxic soups for any long period of time will show stress and that many of them will die.

The fact that my bioassays will be directly comparable to bioassays run with coral larvae which already show that corals die at metals concentrations far lower than what we find in our tanks will allow a rather good assessment, I think, of the liklihood of long-term survival of many corals and other reef animals in our tanks.

rshimek

08/20/2002, 08:33 PM

[QUOTE]Originally posted by Randy Holmes-Farley [quote]

You truly [b]DO miss the forest for the trees, don't you. I am not saying perfecting salt mixes would be the end all. Just the first step. Contamination comes from many sources, salt mixes are just the first thing to fix.

If we can do that, we would at least be starting with sea water that didn't kill animal embryos on contact.

If we can then figure out ways to remove the excess metals, perhaps along with excess organic snot, then we can have tanks where animals can survive to their potential.

rshimek

08/20/2002, 08:52 PM

[QUOTE]Originally posted by Randy Holmes-Farley

I'm afraid that even asking that question shows that you know little about seawater chemistry and humic substances.

Well, gosh, golly, gee, Randy. That's why I asked you to discuss them.

Unfortunately, you passed the buck and gave generalizations, all of which boiled down to...

Simply put...

That you don't have a clue what is in our tanks. It could be any thing from a to zed, and you have no idea what is in the tank.

In other words, saying that humic acids are in tanks is lot like saying, "Salt is found in sea water," except for one thing. We really do know that salt is there. You are guessing about the organic compounds.

Strictly guessing.

You say the yellow color is indicative of them. I say, it could just as well be saffron yellow in coral pee.

You don't have tests, you make broad generalizations about them, and you lack any specific data about them. Truly magical, these chemicals. Sorta like a chemist's cosmic fudge factor, or shall we call them "Randy's variable constant." The are constantly there (maybe, some of them) and they vary a lot.

This general kind of data is patently useless.

If these things are detoxifying metals, then we need to know which ones are present. If nothing else, in one tank. We need to know when they are toxic in there own right and how and when it is necessary to remove them.

We need to know even if we can remove them.

Or if we should.

rshimek

08/20/2002, 09:03 PM

Hi,

At this time, I would like to summarize a bit, and close the thread. If anybody wants to continue the discussion, then do it elsewhere; every time somebody posts here it sends an email to me and that is getting to be significantly annoying.

Randy has made the point that organic materials are present in tanks and likely detoxifying metals. However, he cannot give any specifics about this process, nor has any work on this been done.
That notwithstanding, such detoxification seems likely, at least at times.

Habib has mentioned some of the tests showing detoxification.

I have tested and found high overabundances of metals in tank waters, whether or not they are bound in organic materials or free as ions. Similar tests on sea water or in aquaria dosed with chemicals to levels lower than in that what I have found have resulted in death of corals, snails, and sea anemones.

Presently, the result of this metals overabundance is unclear. I think it is cause for concern. Randy doesn't. He likes 'em.

I think it is unlikely that any further discussion in this thread will resolve anything, or change any opinions, so I am closing it. As I said above if you wish to carry on the discussions elsewhere open a new thread.

Randy Holmes-Farley

08/21/2002, 09:23 AM

Well, gosh, golly, gee, Randy. That's why I asked you to discuss them.

No, actually it's not. You asked because you were accusing me of overgeneralizing, thinking that I was generalizing about humic acids and did not have any specific info (reread your own post if you doubt that). I was being as specific as is possible with a class of substances. Hence my response. If I thought you were genuinely interested in knowing more about what humic substance that I was referring to, I would certainly not have responded that way.

Randy Holmes-Farley

08/21/2002, 09:31 AM

However, he cannot give any specifics about this process, nor has any work on this been done.

I thought that both Eric and I pointed out that a great deal of published work has been done on the toxicity of copper and it's speciation. On corals in reef tanks? No. Do you have any tox data on corals in reef tanks? No.

So we both fall back on what is known: that metals can be toxic to many organsims in many situations, and that binding by organics has been demonstrated to play a role in some of those situations. Hence, my assertion that organics MAY play a role in reef tanks, and that the situation may not be as dire as you claim when you flatly state that we are killing corals.

Randy Holmes-Farley

08/21/2002, 09:36 AM

You say the yellow color is indicative of them. I say, it could just as well be saffron yellow in coral pee.

Actually, if you feed saffron to your tank, it probably is part of the problem. For the remainder of us, I'll accept the comments by marine chemists and biochemists, as referenced in Spotte that

"The main refractory component consists of humus, a portion of which imparts yellow coloration to old aquarium seawater".

Do you think that these scientists are wrong? Or do you just find it easy and convenient to say that these ideas are my imagination, and forget that there is published literature to support it?

rshimek

08/21/2002, 09:56 AM

Well Randy,

The reason I asked if you had any specific data, to which you showed that you didn't, is that I was wondering if you personally actually had any numerical data to draw on, or if you were simply blowing in the wind.

Perhaps, I should explain that I didn't quite fall off the turnip truck with regard to these chemicals; I have had to use data on them in a number of projects, and some of them, at least, are easily tested for. Incidently, in really high concentrations, they are no longer yellow; they can be purple, black and ochre.

You see, there is a significant body of data regarding the some of these organics in both aquaria and the real world. And there is actually some information about them in coral reef situations, in areas downstream of logging sites.

In specific, in the Pacific NW, and probably elsewhere in the US, some of the major pollutants are pulp mills, and signficant amount of their pollution would be classified as humic acids, and similar kinds of organics. The EPA maintains a list of "Priority Pollutants" to be tested for in pulp mill remediation, etc. Included in this list are a reasonably large number of these types of organics. Data do exist for them with regard to their toxicity toward all sorts of organisms.

Now, are these chemicals the same ones found in our tanks? Maybe, but more likely, maybe not. However, certainly some chemicals characteristic of the various classes of them are. And then, contrarty to your assertions that they can't be examined and must be treated as a "black box," it appears that they can be examined and analysed, at least as first step.

If some of the chemicals on the EPA's priority pollutent list could be considered representative of some of the classes of these organics, then we could test for them in aquaria, and we could look at their effects. Analytical labs, such as the one I used for my metals studies, also routinely examine these pollutants. I am not sure of the costs, but I suspect one wouldn't have to look at the complete list of these organics (one could leave out some of the chlorinated hydrocarbons that are made as byproduct of the Kraft pulping process for example).

You say,

Hence, my assertion that organics MAY play a role in reef tanks, and that the situation may not be as dire as you claim when you flatly state that we are killing corals.

Yes, the organics may be killing corals and other animals just as much or more so than the metals. So, in fact the situation may more dire.

Incidentally, correct me if I am wrong. You added iron to tanks and got a reduction in sporulation of algae, right? Sporulation of this algae is a normal part of the life cycle, right? So, by adding iron, you effectively altered the life cycle of the algae to prevent reproduction, right? In other words, you poisoned the algae by the addition of the iron, right?

andy-hipkiss

08/21/2002, 10:08 AM

Minor (ok major) correction to the Cu result I posted earlier .... having checked with the lab it was 0.211 not 211 ppm :o

Duh, I'm used to "." as the decimal separator but here in Belgium they use ",". So "Cu ,211ppm" isn't as I originally posted. Still excitingly high though :eek:

rshimek

08/21/2002, 10:21 AM

Hi Andy,

Yup, that is enough to give everybody a cheap thrill. :D

Randy Holmes-Farley

08/21/2002, 10:42 AM

Incidentally, correct me if I am wrong.

Always glad to help out whenever necessary.

Sporulation and other forms of reproduction are a part of the normal life cycle of nearly every organism, yes. They are also often the response to stress. Such stresses are readily observed with the large numbers of organsims that desperately try to reproduce immediately when moved from the ocean to a tank. I've seen many organsims do that in my own tank (I can supply you with genus and species, if that will help you out).

Is that because they are really feeling good about being boxed up? Probably not. More likely it is the well known response to stress.

Likewise, it is also well known that many organisms attempt to flee in whatever way they can when a situation no longer meets their needs (many examples of which are seen even in our tanks). Sporulation is one way of doing so for an otherwise immobile organism.

Incidently, in really high concentrations, they are no longer yellow; they can be purple, black and ochre.

I seem to recall quoting colors to you earlier in this thread in the definition of humic substances. But thanks anyway for the info. If I see any purple or ochre tanks, or if any reefcentral members ask about such a situation, I'll remember this post.

The reason I asked if you had any specific data, to which you showed that you didn't, is that I was wondering if you personally actually had any numerical data to draw on, or if you were simply blowing in the wind.

Since you believe that I cannot carry out scientific experiments, then it would be smarter for me to rely on published works of other scientists, would it not? If I measured it in my tank, would that impress you somehow? What difference would it make? My tank wasn't in your metal study, and is not udergoing any tox testing at the moment, so its seems like unnecessary data. Suffice that other aquaria have been measured.

I have had to use data on them in a number of projects, and some of them, at least, are easily tested for.

Ahhh, I see. In your own words, which one?

Now, are these chemicals the same ones found in our tanks? Maybe, but more likely, maybe not. \

I agree. Spotte has an extensive discussion of the differences between terrestial and marine humic acids, if you want to understand the situation better. Here's a quote :

"Harvey et al (1983) remarked that reports of humus formation and composition in freshwater and terrestial environments are "irrelevant to seawater humus." "

And then, contrarty to your assertions that they can't be examined and must be treated as a "black box," it appears that they can be examined and analysed, at least as first step.

I don't believe that I said black box, so the quotes seem rather inappropriate. I said that they had to be treated in classes. And that is what I provided to you: quantitative tox data on one class (the polyphenolic exudates released from the seaweed Ascophyllum nodosum and then exposed to seawater prior to the test).

It has also been studied in the synthesis lab:

"Harvey et al (1983, 1984) reported that humic substances in seawater are water-soluble, aliphatic organic acids formed by the autoxidative crosslinking of two or more polyunsaturated faty acids...Humus allowed to form in the laboratory under controlled conditions was similar to humic substances that form naturally in seawater"

IN NO CASE, HOWEVER, DOES ANYONE CLAIM THAT A SINGLE SPECIES IS REPRESENTITATIVE. When a single species is not representative, monitoring the concentration of a class of compounds in the presence of many other compounds of similar molecular weight and solubility is very difficult and fraught with uncertainties. To suggest that I have missed the fact that many researchers regularly quatitate individual seawater humic acids and use those numbers to represent the whole class is simply nonsense. If you have any such references to SEAWATER humic substances, please post them.

Yes, the organics may be killing corals and other animals just as much or more so than the metals. So, in fact the situation may more dire.

I quite agree. I said as much earlier in this thread. According to Spotte "So few studies have been done on the toxicity of humus that no general conclusions can be made".

If some of the chemicals on the EPA's priority pollutent list could be considered representative of some of the classes of these organics, then we could test for them in aquaria, and we could look at their effects.

Good start. Most of the known marine toxins aren't man made, so won't be on this list, but it is a good place to begin to see if things like methyl iodide are a problem or not. You'll need a good setup to study such a highly toxic compound, however.

Randy Holmes-Farley

08/21/2002, 10:52 AM

So, by adding iron, you effectively altered the life cycle of the algae to prevent reproduction, right? In other words, you poisoned the algae by the addition of the iron, right?

One other comment on this. In my iron paper there is a picture of one of my refugia taken about 1 month ago. It shows a bit of Caulerpa racemosa growing around the edges.

Today I had to go in and remove about 2 gallons of the Caulerpa. It had grown to solid packing across the entire surface of the water, and through most of the water column down to the bottom (about 8 inches) preventing light from getting to the corals underneath.

That doesn't sound like poisoned macroalgae barely struggling to survive against a toxic onslaught of 800 million times the normal iron concentration every day. It sounds like a very active export mechanim for nitrogen and phosphorus (and potentially other things:D). Would that have happened without the iron additions? Maybe. I don't post it to claim the iron is responsible. But it did happen in the presence of the iron, so the iron is clearly not causing a substantial problem with using macroalgae as an export mechanism.

Randy Holmes-Farley

08/21/2002, 10:57 AM

Presently, the result of this metals overabundance is unclear. I think it is cause for concern. Randy doesn't. He likes 'em.

The only one that I dose, and the only one that I recommend others dose is iron. That's the one that I like.:love2:

Is there any evidence whatsoever that there is an overabundance of iron in any operating reef tank?

If so, please post it.

Otherwise, I'd appreciate your not generalizing your concerns about certain metals that you measured to others that you were unable to detect.

Randy Holmes-Farley

08/21/2002, 11:13 AM

You truly DO miss the forest for the trees, don't you. I am not saying perfecting salt mixes would be the end all. Just the first step. Contamination comes from many sources, salt mixes are just the first thing to fix.

But Ron, it doesn't help even a tiny bit. Why wage war against something that won't help even a TINY bit?

In your study, you write:

"Sample GD was from a tank that uses NSW as a medium, and as shown in the dendrograms, no matter how the data are manipulated, that particular sample is never very similar to NSW. "

Let's examine tank GD.

It is the highest or second highest for:

arsenic, boron, cobalt, copper, nickel, and vanadium

A list that incorporates most of your most-mentioned metal toxins.

This data suuggests that lowering the metals in the salt mixes will do NOTHING to help.

Do you have some other data that suggests that it will actually help something? Even a little bit?

Were there any other tanks in your study that claimed to use NSW? Which ones, so we can look at the metal concentrations in them as well.

rshimek

08/21/2002, 11:28 AM

Originally posted by Randy Holmes-Farley

Randy,

[That doesn't sound like poisoned macroalgae barely struggling to survive against a toxic onslaught of 800 million times the normal iron concentration every day. It sounds like a very active export mechanim for nitrogen and phosphorus (and potentially other things:D).

No, it simply sounds actually like you are exporting enough algae to keep the algae below the size necessary for reproduction. However, if the iron does shut off the reproductive process, that is still a poison, and vegetative growth can still occur.

The point being that poisons don't have to kill, simply altering the target metabolism in a way that destroys reproductive fitness also works.

Is that because they are really feeling good about being boxed up? Probably not. More likely it is the well known response to stress.

For the alga, it is more likely and simply a size thing. Although the release of the spores is rapid, the interal cell division that leads to it generally takes several days. This is not a process that occurs in the algae in response to immediate stress, unless the species was already primed to go.

If you have had other algae and animals respond to placement in your tanks by spawning, I would say that something is pretty drastically wrong. From my most recent article, I would suggest that they are being put into a situation where the toxic chemical load is far beyond their capabilty to compensate for, and that causes the spawning.

IN NO CASE, HOWEVER, DOES ANYONE CLAIM THAT A SINGLE SPECIES IS REPRESENTITATIVE. When a single species is not representative, monitoring the concentration of a class of compounds in the presence of many other compounds of similar molecular weight and solubility is very difficult and fraught with uncertainties.

Oh, I agree. However, it is a start. One war at a time...

To suggest that I have missed the fact that many researchers regularly quatitate individual seawater humic acids and use those numbers to represent the whole class is simply nonsense. If you have any such references to SEAWATER humic substances, please post them.

No, sir, it is not nonsense to suggest that you have missed the fact that researchers have regularly sampled these substances. You appear to be using only one reference, Spotte, and the references therein. Nothing more recent that the mid-1980's. A lot has happened since then.

Much of this is in the so-called gray literature, however, there is a lot, I think, in Marine Pollution Bulletin. I have not searched the literature recently for these data specifically, as I haven't been interested in addressing any problems revolving around organics in our systems. And, from where I am at, I really can't get into the data bases I need either - my local university can barely keep the library doors open, let alone subscribe to marine journals. It is also likely that it will be several months before I can make a trip to Seattle to get to the UW library which has these data. So... if you want some of this, you're going to find it on your own.

I will try to dig out my lists of the priority pollutents over the next few days (these are in boxes in my stacks of these publications in my garage) and if I can find a complete one, I will send it to you. Until then, if you want, you can do a search under "EPA priority pollutents" and you may find them. In case they vary from EPA region to region, most of my work was in Region 10.

I was going to suggest that we might be able to work up a collaborative project on this, one that might well result in publication in the professional literature as well as in the hobbyist press, but in light of your contentious response, I think such a project would have little chance of success.

rshimek

08/21/2002, 11:35 AM

Originally posted by Randy Holmes-Farley

This data suuggests that lowering the metals in the salt mixes will do NOTHING to help.

And you miss the point, again.

The poison levels are cumulative. Most of them initially come in ultrahigh doese with the water in most tanks. They are replenished with the water.

If we can remove them from the water the levels will be lower. Maybe well below toxic levels.

In the particular tank in question, they are undoubtedly coming in with foods, and maybe other additives. Accumulation continues through time as export of the damnable substances is inefficient. That they accumulate, simply means export is not occurring.

Habib

08/21/2002, 01:56 PM

Dear Ron, Dear Sir:D and others,

Humic Acids (HA):

Based on UV-VIS spectra which I had recorded of several aquaria I estimate the HA content to be approx. 0.5 - 10 ppm. It can be much higher but most probably not lower. I had to make some estimates regarding molecular extinction coefficient and have used a value which very likely undeestimates the actual HA concentration. I have also taken a low molecular weight which also can lead to an underestimation.

I am also including a few cuttings:

C-Functional Group Chemistry of Humic Substances and
Their Spatial Variation in Soils
S. C. B. Myneni 1 , T. A. Warwick 2 , G. A. Martinez 3 , G. Meigs 2
1 Earth Sciences Division, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
2 Advanced Light Source, Lawrence Berkeley National Laboratory, Berkeley, CA 94720, USA
3 Agriculture Experiment Station, University of Puerto Rico, San Juan, PR 00936-4984, USA
Organic molecules derived from biological processes and the biochemical alteration of plant and
animal residue are common in soils and natural aquatic systems and their concentration ranges
from <1 ppm to as high as 4x10 5 ppm. Their composition varies widely with location and origin
(e.g. soil, marine), and consists of small chain molecules (e.g. acetate, citrate), organic
macromolecules (e.g. proteins), and polyfunctional humic substances (HS)1 . Of these, humic
substances exist at high concentrations, and are stable to biochemical alteration with long
lifetime. In addition, HS can form strong complexes with both inorganic and organic
contaminants and mineral surfaces, and thus play a major role in geochemical processes 2 . At least
for a century, research has been focused on understanding the HS functional group chemistry and
the macromolecular structure - the properties of HS that control their behavior in the
environment.

The following is from an online aquariumfrontiers article by Randy:

Organics

The nature of organic molecules is certainly the most complicated aspect of seawater chemistry. Organics comprise about 2 ppm of seawater. Of this 2 ppm, the majority is in the form of dissolved organic carbon (DOC). DOC includes all fully dissolved organic compounds and any particulates that are small enough to pass through a 0.45-micron (Âµm) glass fiber filter. Strictly speaking then, it is not all fully dissolved. Any organic particles greater than 0.45 Âµm are called particulate organic carbon (POC). The POC is about a factor of 10 lower in concentration than DOC and is composed of living and dead organisms, as well as assemblies of organic molecules.

DOC is an incredibly complicated mixture of molecules that represents billions of years of biological waste products from uncounted numbers of different organisms, combined with reactions catalyzed by light, heat, inorganic catalysts (metals), biological processes, and many other factors. It includes carbohydrates (20 to 35 percent of the total), humic substances (10 to 30 percent of the total), amino acids and proteins (2 to 3 percent), hydrocarbons (less than 1 percent), carboxylic acids (1 percent) and steroids (trace).

There is also a great deal of uncharacterized organic material. In fact, the study of seawater organics is an active area of research. Additionally, the summation of all dissolved organics in the ocean is a pool of carbon larger than carbon dioxide in the atmosphere, so it cannot be ignored by those looking at the planetary carbon cycle. In addition to carbon, these organics contain significant amounts of oxygen, nitrogen, phosphorus, and sulfur.

It is probably also safe to say that most, if not all, closed marine systems have higher organic levels than the ocean, although hard numbers are difficult to come by. The desire to reduce these organic levels is one of the reasons for the popularity of skimmers with marine aquaria.

Anothe one:

........Humic and fulvic acids have strong affinities for most metals and therefore influence metal mobilization and bioavailability in the environment. [5,1]. Characterizing these complex multivariate interactions is fundamental to the prediction and control of metal speciation [6,1,5].
The nature of humic-metal complexation is neither understood nor agreed upon [1,6]. However, because HFAs have numerous binding sites with a continuum of binding strengths it is agreed that they can not be treated as simple, isolated molecules [7]. Because of the complexity of humic-metal interactions, their stability constants and complexation capacities depend on pH, ionic strength, metal concentration, presence of interferents, and the nature of the particular HFA [7, 8]. .........

Randy Holmes-Farley

08/21/2002, 03:22 PM

If you have had other algae and animals respond to placement in your tanks by spawning, I would say that something is pretty drastically wrong.

Absolutely, they responded to stress by trying to reproduce. I think it is a reasonable hypothesis that Caulerpa racemosa responds the same way when given a shortage of iron, but as I noted in my article, such a hypothesis requires much further testing, and is not something that I believe to be more than a working hypothesis.

I actually wasn't referring to algae at all in my tank, but spawning of bivalves, limpets, urchins, and hermit crabs.

Randy Holmes-Farley

08/21/2002, 03:29 PM

Based on UV-VIS spectra which I had recorded of several aquaria I estimate the HA content to be approx. 0.5 - 10 ppm.

Come on, Habib, since I haven't measured it personally in my tank, it can't be known.:D

For what it's worth, that puts it into the potential toxic range (0.32 mg/L for the LC50 with plaice larvae) IF what you measured has the same toxicity as that measured by Sieberth and Johnson. Of course, it likely does not because that toxic fraction is only a part of the total, and the toxicity is lower after complete humification.

Randy Holmes-Farley

08/21/2002, 03:35 PM

No, it simply sounds actually like you are exporting enough algae to keep the algae below the size necessary for reproduction.

You may have hit on something here, Ron. Adding iron may make it grow robustly enough that people do need to prune it, and the pruning is important in preventing the sexual phase.

Many people have suggested that pruning helps. Then in the absence of added iron, growth is slow enough that people don't need or bother to prune, and the algae can proceed to an unwanted sexual phase.

Thanks for the suggestion. If I put it into a followup article, I'll be sure and credit you with the idea:)

Randy Holmes-Farley

08/21/2002, 03:47 PM

Habib

08/21/2002, 04:01 PM

Randy,

For what it's worth, that puts it into the potential toxic range (0.32 mg/L for the LC50 with plaice larvae) IF what you measured has the same toxicity as that measured by Sieberth and Johnson.

You wanted to impress me with the 0.32 mg/L value?;)

This is nothing compared to the 0.03 mg/L PO4-P which is sufficient to decrease the ferilization of an Acropora eggs drastically. I will post it when I am back in the office (it's almost midnight here).

And if you consider the high PO4-P values..........

Randy Holmes-Farley

08/21/2002, 06:18 PM

Yes, I agree, there are loads of things that may be toxic in our tanks. Anions (phosphate, arsenate, etc) are certainly among them.

Inhibition of calcification of corals happens at fairly low phosphate levels (<0.2 ppm) and even organic phosphate levels. The levels are in my calcification article (the one that Eric kindly supplied many references for: http://www.advancedaquarist.com/issues/apr2002/chem.htm

Midnight? You sleep? What about all those people wanting your test kits? They have to wait while you sleep?

Habib

08/22/2002, 08:55 AM

Here is what I said about highly reduced fertilisation at a PO4 concentration of 1 microM (approx. 0.1 ppm as phosphate or 0.03 ppm as PO4-P).

Elevated levels of nitrogen and phosphorus reduce fertilisation success of gametes from scleractinian reef corals
P. L. Harrison1, and S. Ward1

(1) Centre for Coastal Management, Southern Cross University, P.O. Box 157, Lismore, NSW 2480, Australia

Communicated by G.F. Humphrey, Sydney

Abstract. Spawned gametes were collected from colonies of Acropora longicyathus at One Tree Island and Goniastrea aspera at Magnetic Island, Great Barrier Reef, Australia, for use in fertilisation trials. Mean fertilisation rates were significantly reduced compared with controls (P<0.003), when gametes from the branching coral A. longicyathus were exposed to elevated ammonium concentrations at 1 ÂµM and above in one cross (60-64% reduction), and at 100 ÂµM in another cross (16% reduction). Mean fertilisation success of A. longicyathus gametes was also significantly reduced compared with controls in both crosses (P=0.000) at concentrations of 1 ÂµM phosphate and above (35-75% reduction), and at 1 ÂµM ammonium plus 1 ÂµM phosphate and all higher concentrations (68-74% reduction). Similarly, the mean percentage of regular embryos that were developing normally was significantly reduced in most nutrient treatments compared with controls (P=0.000). Fertilisation trials using gametes from the brain coral G. aspera resulted in a significantly lower percentage of regular embryos (P=0.001) and a significantly higher percentage of deformed embryos (P=0.001) developing after exposure to elevated nutrient treatments compared with controls. Mean fertilisation rates for this species were only significantly reduced (P=0.034) in the 50 ÂµM ammonium plus phosphate treatment in one cross (8% reduction), compared with the control. Therefore, ammonium and phosphate enrichment significantly impairs fertilisation success and embryo development in scleractinian reef corals.

Diatom2

08/22/2002, 11:36 AM

I was going to suggest that we might be able to work up a collaborative project on this, one that might well result in publication in the professional literature as well as in the hobbyist press, but in light of your contentious response, I think such a project would have little chance of success.

Hmmm, proposed that way, I'd have to agree

I must say that after following this thread from the beginning and even printing it out so I could make some notes and keep it for future reference, this seems an excellent idea, to me anyway.

Each of you would bring an excellent balance and welth of knowledge to the project. I know things can get a bit heated in the scientific world, however this seems like potentially very important work at least in our small world of reefkeepers. I for one would like to see you two work together, for my selfish benefit if for no other reason, and I'm sure I'm not alone.

EricHugo

08/22/2002, 10:18 PM

Sorry for my absence but I've been busy. I spent the day in the library today, and not specifically for you guys...but in the process, well, my nature got the best of me again.

OK, I'm not finished yet, but here's what I have found so far in addition to all theprevious articles/books, etc..

Randy: Thanks for the links and I'll be sure and read them and any others as soon as I can. I also agree that if Habib or anyone would further our knoweldge of these things with real science, I would be more than open minded enough to read them. I also wanted to throw in, as an aside to others reading this thread the following before getting to the meat of my post: A lot of people tend to think because someone has written something in the aquarium literature, it is true. That's a very scary way to think.

Similarly, a lot of people think that if something apppears in the scientific literaure, it is true. Equally scary, on a different level. Yes, it is peer-reviewed and it helps, but is absolutely no guarantee of correctness or applicability to similar subjects. In fact, unless one is familiar with the materials and methods and can be assured, even if those were chosen properly, that the experiment was conducted properly, that the resutls are valid, that statistics were done correctly and using an appropriate test, and mostly that any conclusions based on thee results have any merit at all.

Now...

1. Glynn, Peter W., Szmant, Alina M., Corocoran, Eugene F. Cofer-Shabica, Stephen V. 1989. Condition of coral reef cnidarians from the North Flordia Reef Tract: pesticides, heavy metals, and histopathological examination. Mar Poll Bull 20(11): 568-576.

summary: found high levels of As, Cu, Pb in corals at low ppm levels in coral tissue, cadmium and mercury at less than 0.6 ppm. Histopathological abnormalities were present including disease, bite parks, bleaching, galls, algae, epidermal erosion, neoplasia and necrosis and presence of numerous foreign organisms - was not possible to determine effects of metals directly compared to all stresses. Trace metals applied at concentrations comparable to tissue levels here resulted in bleaching and mortality of reef building corals (Evans 1977), but no unusual morbidity or mortality at these sites. Questions exist as to how florida corals with a high "tissue burden" of pesticides and metals will respond to additional stresses.

2. Benson, A.A., Summons, R. E. 1981. Arsenic accumulation in Great Barrier Reef invertebrates. Science. 211 (4481): 482-3.

summary: massive accumulation of arsenic in Tridacna clams even though low levels in water. thought to be related to metabolsim of arsenic in low phosphate waters. Accumulation results by deposition into host tissue after accumulation by zooxanthellae. Massive accumulation also occuyrs in other symbiotic inverebrates, greatest in mollusks and ascidiians

3. Harland, A.D., Brown, B.E. 1989l. Metal tolerance in the scleractinian coral Porites lutea. Mar Poll Bull 20(7): 353-357.

summary (Randy take note!!): Exposure to elevated iron led to loss of zooxanthellae from tissues - repsonse is very marked in corals not regualrly exposed to high environmental concentrations of iron. Tolerance is suggested here, but other studies conflict as well as support tolerance theory. Some evidence exists to suggest effective metal regulation of exclusion in coelenterates (Brown and Howard 1985, Howard and Brown 1987). Skeletal studies suggest uptake and sequestration in skeleton relative to env. conditions.

(Ron, take note!!) The elevated seawater levels mg/l recorded around a tin smelter and ore washing effluent were Fe: 0.61-29 Zn 0.02-0.22 Cu <0.005-0.06 Cd 0.01-0.04 Mn 0.02-0.09 Pb <0.005 - 0.71

This particular article has an abundance of references I will be getting on my next afternoon there. Probably next week. the Brown and Howard papers look critical to this discussion.

Back to work...

Habib

08/23/2002, 06:06 AM

Similarly, a lot of people think that if something apppears in the scientific literaure, it is true. Equally scary, on a different level. Yes, it is peer-reviewed and it helps, but is absolutely no guarantee of correctness or applicability to similar subjects. In fact, unless one is familiar with the materials and methods and can be assured, even if those were chosen properly, that the experiment was conducted properly, that the resutls are valid, that statistics were done correctly and using an appropriate test, and mostly that any conclusions based on thee results have any merit at all.

I fully agree! And especially if a scientsist has to publish a minimum number of publications per year then it is more suspect:D

My professor once told me that if experiments are carried out properly etc the data will remain but conclusions and theories can change in time.

I have been used to being shot at or to shoot at others during presentations of research results for select groups. This not to denigrate or insult some one but only to improve the quality of the research, the quality of the conclusions, getting (better)aware of the weak points, to formulate further experiments,.......

Having said that a few notes on Eric's last post.

to be related to metabolsim of arsenic in low phosphate waters.

Phosphate has a strong tendency to complex a.o. with arsenate, molybdate and vanadate. Phosphate is an inorganic ligand for such species of metals.

A study conducted on vanadate uptake by tunicates showed that elevated phosphate inhibits vanadate uptake by tunicates.

Elevated phosphate showed decreasd Sr/Ca ratios in PO4 polluted areas in Australia.

Regarding the Harland and Brown's iron paper I would be interested to read at least the Experimental part of the paper. What sort of light was used? How high were the iron levels?

Some iron compexes will generate reactive oxygen species if UV light is present and might cause bleaching.

I would like to add one more which shows if iron is added an increase in zooxanthellae density and reduction of growth rate (I don't know if they have measured calcification rates!!):

Journal of Experimental Marine Biology and Ecology, Vol. 259 (2) (2001) pp. 249-261

Response of a scleractinian coral, Stylophora pistillata, to iron and nitrate enrichment
Christine Ferrier-PagÃ¨s * a , Vanessa Schoelzke a, Jean Jaubert a, Len Muscatine b and Ove Hoegh-Guldberg c
a Observatoire OcÃ©anologique EuropÃ©en, Centre Scientifique de Monaco Av. Saint-Martin, MC-98000 Monaco
b Department of Biology, University of California, Los Angeles, CA 90095, USA
c Center for Marine Studies, University of Queensland St. Lucia, 4069 QLD, Australia
Received 4 August 2000; received in revised form 31 January 2001; accepted 12 March 2001

Abstract
The purpose of this study was to determine whether the addition of iron alone or in combination with nitrate affects growth and photosynthesis of the scleractinian coral, Stylophora pistillata, and its symbiotic dinoflagellates. For this purpose, we used three series of two tanks for a 3-week enrichment with iron (Fe), nitrate (N) and nitrate+iron (NFe). Two other tanks were kept as a control (C). Stock solutions of FeCl3 and NaNO3 were diluted to final concentrations of 6 nM Fe and 2 ÂµM N and continuously pumped from batch tanks into the experimental tanks with a peristaltic pump. Results obtained showed that iron addition induced a significant increase in the areal density of zooxanthellae (ANOVA, p=0.0013; change from 6.3Â±0.7Ã—105 in the control to 8.5Â±0.6Ã—105 with iron). Maximal gross photosynthetic rates normalized per surface area also significantly increased following iron enrichment (ANOVA, p=0.02; change from 1.23Â±0.08 for the control colonies to 1.81Â±0.24 Âµmol O2 cm2 h1 for the iron-enriched colonies). There was, however, no significant difference in the photosynthesis normalized on a per cell basis. Nitrate enrichment alone (2 ÂµM) did not significantly change the zooxanthellae density or the rates of photosynthesis. Nutrient addition (both iron and nitrogen) increased the cell-specific density of the algae (CSD) compared to the control (G-test, p=0.3Ã—109), with an increase in the number of doublets and triplets. CSD was equal to 1.70Â±0.04 in the Fe-enriched colonies, 1.54Â±0.12 in the N- and NFe-enriched colonies and 1.37Â±0.02 in the control. Growth rates measured after 3 weeks in colonies enriched with Fe, N and NFe were 23%, 34% and 40% lower than those obtained in control colonies (ANOVA, p=0.011).

This one shows incorporation of iron by a totally different mechanism and could perhaps also be be applicable for other metals besides iron:

Brown, B.E., A.W. Tudhope, M.D.A. Le Tissier & T.P. Scoffin, 1991.

A novel mechanism for iron incorporation into coral skeletons. Coral Reefs, 10: 211-215.

Intertidal corals living in seawater with high concentrations of iron incorporate the metal into their skeletons. Cross-sections of the coral skeleton reveal orange-stained banding patterns reflecting periods of high availability of iron. The mechanism of metal incorporation involves deposition of iron compounds on to skeletal spines that are exposed as a result of temporary tissue retraction during periods of extreme stress. Subsequent tissue recovery and calcification trap the iron compounds which provide a visible environmental signature in the coral skeleton. This previously unrecognised mechanism has significant implications for the reconstruction of past environments from chemical analysis of annually-banded massive coral skeletons.

I have 7 year old measurements of metals in different genetically identical corals grown in NSW (original habitat) and in various aquariums and of course also the water analyses.
I hope to have them published soon after I am ready with the article.
These measurements were done to measure the effect of element concentration (total metal concentration) on metal incorporation in the skeletal material.;)

Besides that I have far more results and because most of them were meant for our own use they have nor been published.

I know that besides my research and Ron's there have been more studies done on aquariums and I am working to obtain them.

To all who read this thread I would like to say that I think that we all are here for further advancement of our hobby and if there are conflicting ideas, theories or whatever, it will only be of advantage :)

Bomber

08/23/2002, 06:46 AM

3. Harland, A.D., Brown, B.E. 1989l

received in revised form 31 January 2001

Guys as Habib mentioned, be very careful quoting references pre late 90's. A lot has changed.

Randy Holmes-Farley

08/23/2002, 08:03 AM

Thanks for the info, Eric!

Regarding the Harland and Brown's iron paper I would be interested to read at least the Experimental part of the paper. What sort of light was used? How high were the iron levels?

I don't have ready access to it today since I'm on my lkeave still. I'd also be interested in knowing what type of iron. The idea (not mine but the literature consensus, at least at the time of Spotte's publication in the 90's) with chelated iron (like with EDTA) is that the bulk of it is unavailable until UV light degrades the chelate and the iron falls out, available for uptake. Consequently, adding a large amount of chelated iron may only result in a low, steady concentration of free iron.

If the study in question added free iron itself, the situation may be very different.

I would have though that adding hundreds of millions of times the NSW amount of iron EVERY DAY would have shown a problem in my tank if there were one. Of course, I've not measured zoox densities, and that may be a critical difference, but the corals and anemones have certainly not bleached and are growing nicely.

Randy Holmes-Farley

08/23/2002, 08:05 AM

Bomber:

Do you know what the revised version says?

I'm keen to do some literature searching of more recent works myself, but that will have to wait until I'm back at work.

Bomber

08/23/2002, 08:20 AM

Randy
More than likely all you need to do it pick up the phone and call Mote.
Several years ago they had almost a complete wipe out because of Red Tide. As a result, they now have everything they could find on iron. What would more than likely apply for you is what was used as bioindicators.
Jerel

EricHugo

08/23/2002, 08:55 AM

Jerel:

Totally agree - papers on iron abound now. The reason I used this particular one was that it was investigating metal uptake in general and happened to use iron as the indicator metal. Personally, I was disappointed to see that it, of all metals, was used in this study.

I wouldn't go so far as to say anything older than late 90's should be approached cautiously, though, except in certain fields of study. I think all papers should be approached cautiously, and do agree sometimes, though, as I read and older paper (sometimes not even all that old) with something so totally known as wrong today... In contrast, the sometimes profound ignorance of young researchers of studies already done and totally ignored in their background search not only compromises their own study, but I keep running across more and more papers that "miraculously" discover something that was known thrity years ago and subsequently disproved or fell out of favor in the interim period by "Then new" research.

Randy: they used ferric nitrate.

I'm off to to work in a few minutes, but forgot another one with major references and was a good study.

Nipper, Marion, Carr, R. Scott. 2001. Porewater toxicity testing : a novel approach for assessing contaminant impacts in the vicintity of corals reefs. Bull Mar Sci 69(2): 407-420.

interesting points:

carbon is considered a major factor in reducing contaminant bioavailability because it can both sorb organic chemicals and complex metalsl.

Mobilization of metals caused by dredgingwas analyzed by Reichelt and Jones (1994).High uranium found in sediments corals, algae and seagrass of polluted sites in Gulf of Aqaba (Abu-Hilal 1994)

Sandy sediments believed not to retain contaminants...Clays do - but high carbon of clay ameliorates

Use of sea urchin gametes and embryos lauded as ideal and useful measure for toxicity tests for many reasons

Metals consistently referred to as pollutants, contaminants and potential sources of detriment to corals, coral reefs, etc.

Bomber

08/23/2002, 09:12 AM

It's just that the money was there for iron. Once iron was identified as a marker for African/Saharan dust - everyone's knee jerk reaction was to blame it for everything.

For other metals I would be contacting The South Florida Water Management, IFAS, and (of all things) The Florida Farm Bureau. The farmers have to spend untold amounts of money every year defending their right to farm South Florida (produces over 90% of your winter produce). They spray (copper), fertilize, etc. and every bit of that ends up in the aquifer (oolitic limestone, live rock) or as runoff. From there it's non-stop to 18 national parks and reserves. Or work it backwards from the parks and reserves.
At one time or the other all these "aquarium" critters have been used as bioindicators.

Good luck, you all have your work cut out for you
Jerel

EricHugo

08/23/2002, 09:36 AM

Jerel: LOL

Not really...once I get these last few refs, read them out of sheer morbid curiosity and post them, I'm outta here and back to my own real areas of interest....the other guys with the real expertise in the area can finish hashing this out...I'm just throwing some more fuel in the fire ;-)

Randy Holmes-Farley

08/23/2002, 02:54 PM

Habib:

Growth rates measured after 3 weeks in colonies enriched with Fe, N and NFe were 23%, 34% and 40% lower than those obtained in control colonies (ANOVA, p=0.011).

How do you interpret this last sentence? Are they talking about growth of the coral or something else? Things sound good for iron up to that point in the abstract. Is "lower" a typo?

rshimek

08/23/2002, 03:32 PM

Well, it is pretty straight forward...

They had controls...

The growth rate with the factors indicated was lower than in the controls. The probability of the coral growth with the factors added and in the corals without the factors added being the same was 0.011, or 1.1%. Basically it says Iron decreased the growth rate by 23%.

This is the advantage of using proper experimental design with replication, controls and the proper interpretation of statistics.

Habib

08/23/2002, 03:43 PM

Randy,

How do you interpret this last sentence? Are they talking about growth of the coral or something else? Things sound good for iron up to that point in the abstract. Is "lower" a typo?

It is not my typo. It could be a typo in the abstract but the chance that it is is minimal.

I doubt (but I can be wrong) if they have measured calcification rates (45Ca uptake rate). If they have just measured e.g. just extension rate then there is even the possibility that calcification rate was increased (with increased phtosynthesis rate) but that the growth was just more denser.

Getting hold of the article would give more information.

You might perhaps have also noted that the experiment with just addition of only 1 micromol/L of NO3-N (0.12 ppm NO3) gave a decrease of 40% in growth rate!

Also increased photosynthesis rate could have caused depletion of other nutrients resulting in a different calcium deposition.

The abstract however shows the opposite of coral bleaching when iron is added. Namely an increase in zooxanthellae.

One more thing I can add is that several manufactures sell liquid phosphate removers. All of them are iron based. They are used in several tens of thousands of tanks and the dosages are far more then what you add as a trace element.

I have never seen or heard of any bleaching events using such products.

I personally doubt that dosing of significant amounts of iron causes bleaching.

El-ahrairah

08/23/2002, 04:42 PM

Ron,

Don't you know that various invertebrate stages require nutrients which they derive from the surrounding water? That these nutrients are a.o amino acids or other nitrogen compounds.

No, sir, they don't.

In early development any nutrition is dependent upon yolk. They may absorb some compounds from the water, but they don't absorb meaningful amount of amino acids, or any other food.

I'm sorry Ron, actually larvae do uptake meaningful about of amino acids. Here are just a few papers demonstrating this fact.

Jaeckle WB, Manahan DT (1989) Feeding by a "nonfeeding"
larva: uptake of dissolved amino acids from seawater by lecithot-
rophic larvae of the gastropod Haliotis rufescens.
Mar Biol 103 :87-94.

Jaeckle WB, Manahan DT (1989) Amino acid uptake and metabol-
ism by larvae of the marine worm Urechis cuupo (Echiura), a newspecies in axenic culture. Biol Bull 176: 317-32.

Manahan DT (1983) The uptake and metabolism of dissolved
amino acids by bivalve larvae. Biol Bull 164: 236-250.

Manahan DT (1990) Adaptations by invertebrate larvae for nutrient acquisition from seawater. Am Zoo1 30: 147-160.

-Michael

Randy Holmes-Farley

08/24/2002, 06:23 AM

The probability of the coral growth with the factors added and in the corals without the factors added being the same was 0.011, or 1.1%. Basically it says Iron decreased the growth rate by 23%.

I guess that you then ought to be on a nitrate toxicity bandwagon as well, since it is more toxic than the iron at the tested levels, and is known to be present in most reef tanks at levels exceeding those tested in the paper. Many tanks exceed that level by several orders of magnitude. But the corals still grow well, including the exact species listed. Somehow, I think your simplistic interpretation is missing something. Perhaps if any of us had read the actual paper, we'd know better:D

EricHugo

08/24/2002, 09:26 AM

Last post to the topic:

I now have the following articles and the ones with stars by them I would suggest those invovled in this discussion should obtain - there are definitely some key works here and while still not answering many questions, definitely answer many of the issues posed here - I'll list them, make a general summary statement of all of them, and if Ron, Randy, Habib or other major contributors to this thread want a copy, I'll be happy to send them snail mail (cause its about a ream of paper).

1. Dalliner, Reinhard and Rainbow, Philip. 1993. Ecotoxicology of metals in invertebrates. Lewis Publishers, Boca Raton. 445+ pages.

2. **Howard, L.S., and Brown, B.E. 1984. Heavy metals and reef corals. Oceanogr Mar Biol Ann Rev 22: 195-210.

3. Deslarzes, Kenneth J. P., et al. 1995. Historical incorporation of barum in the reef-building coral Montastrea annularis at the Flower Garden bansk, North-West Gulf of Mexico. Mar Poll Bull 30(11) 718-722

4. **Brown, B.E.,and Holley, M.C. 1982. Metal levels associated with tin dredging and smelting and their effect upon intertidal reef flats at Ko Phuket Thailand. Coral Reefs 1(2): 131-137.

5. Pilson, Michael E. Q. 1974. Arsenate uptake and reduction by Pocillopora verrucosa. Limnology and Oceanography. 19(2): 339-341.

6. Meehan, William J., and Ostrander, Gary K. 1997. Coral bleaching: a potential biomarker of environmental stress. J Tox Env Health 50(6): 529-552

7. Readman, J.W. et al. 1996. Discrete bands of petroleum hydrocarbons and molecular organic markers identified within massive coral skeletons. Mar Poll Bull 32(5): 437-443.

8. ** Recihelt, A.J. and Jones, B.R. 1994. Trace metals as tracers of dredging activity in Cleveland Bay - field and laboratory studies. Aus J Mar Freshwater Res 45: 1237-1257.

9. **Howard, L.S., and Borwn B.E. 1987. Metals in Pocillopora damicornis exposed to tin smelter effluent. Mar Poll Bull 18(8): 451-454

10. **Denton, G.R.W., and Burdon-Joes, C. 1986. Trace metals in corals from the Great Barrier Reef. Mar Poll Bull 17(5): 209-213.

11. Brown, B.E., and Howard, L.S. 1985. Assessing the effects of "stress" on reef corals. Adv mar Biol 22: 1-63
(this is a paper I have used many many times and has less to do with metals, although some, but is a tremendously seminal paper in general)

There are a couple more papers that look like they are definitely worth looking into, but I'm finished.

1. St. John, B.E. 1973. Trace elements in corals of the Coral Sea: their relationship to oceanographic factors. In: Proc Int Symp on Oceanogr of the South Pacific (Wellington, R. Fraser, ed.)pp: 149-158 UNESCO

2. St. John, B.E. 1974. Heavy metals in the skeletal carbonate of sclearctinian corals In: Proc. 2nd Int Coral Reef Symp 2: 461-469 (note: this proceedings is hard to find)

3. Brown, B.E. and Howard, L.S. 1985. Responses of coelenterates to trace metals - a field and laboratory evaluation. Proc 5th Int Coral Reef Cong

Eric's overall summary:

Various trace metals are necessary in varying amounts for varying organisms at normal environmental levels through either food (primary) or absorption (secondary). Levels above normal environmental levels quickly become toxic at varying rates to varying organisms and must be dealt with through accumulation, sequestration, detoxification, or excretion. Some organisms can accumulate metals at very high amounts. The overall effect of these metals is variable, with some highly tolerant and some highly sensitive. In no case does it appear that anything above normal environmental levels is beneficial, and can cause mortality, reproductive failure, diminished growth rate, abnormalities, etc.

Trace metals are available as free ions (rarest) or complexed (most common) to various organics. It is likely that most of the trace metal excess found in both tissue and skeletons of marine organisms comes from trace metals at elevated concentrations in sediments and particulates. These particulates and free ions can be directly absorbed chemcially onto substrate or directly incorporated biologically. They can also be incorporated by physical deposition. Bioavailability is a key issue and either varies substantially
according to metal/metal complex/ and organisms, or is not known.

The effects of trace metals on corals is hard to characterize. There is pretty strong evidence for tolerance in areas of greater inputs or availability, but the effects of tolerance are not known and tolerance implies survival.... and equally strong evidence to support diminishing reproductive output and stunted growth rates, reduced diversity, fragile skeletons, different forms of carbonate, and direct histopathologic events. Bleaching seems like a common finding (and I do not necessarily mean a stark white coral, although maybe). I estimate there is fairly strong evidence to support the fact the zooxanthellae play a key role in uptake and accumulation of metals with subsequent release of zooxanthellae by the coral (bleaching) - has been shown for lead, copper, iron, barium, and others. Levels of metals in coral tissue seem surprisingly low in several studies, indicating a fairly effective means of dealing with them. Levels in skeleton seem to depend largely on if chitin is present in the skeleton (Pocilloporids, etc.), and the form of metal present. If based from particulates, it appears to be higher. If free, appears to be present in correlation with levels in water. Only uranium appears higher than expected.

Overall, I'd say any levels above NSW are probably deleterious - from minimally to lethally, depending on the organism and the metal. Given the information presented by Ron, irrespective of bioavailability and means of dealing with the excess, I'd say this is a very major area of concern for at least several of the metals in the study.

rshimek

08/24/2002, 10:35 AM

Here are a few more references with abstracts.

2177. Dallinger,R (1994): Invertebrate organisms as biological indicators of heavy metal pollution. Applied Biochemistry and Biotechnology 48, 27-31.
<Some species of invertebrate animals are known to be efficient accumulators of trace elements. Generally, metal accumulation by such organisms is based on efficient detoxification mechanisms, such as intracellular compartmentalization, or metal inactivation by binding to metallothioneins. Metal accumulators have often been used as accumulation indicators of environmental metal pollution. This means that, ideally, metal concentrations in the animal's body reflect quantitatively or semiquantitatively environmental pollution levels. In reality, however, many factors, such as the animal's weight and age, can disturb such quantitative relationships. These factors have, therefore, to be considered carefully before an invertebrate is utilized as accumulation indicator for metal pollution. Apart from accumulation, many invertebrates exposed to elevated metal concentrations respond to this stress by metal-induced synthesis of metallothioneins. Additionally, metallothionein in metal-loaded organisms can be present in different isoforms that are specifically synthesized in response to different metals. These facts make metallothionein a potential biomarker for metal stress in invertebrates. One possibility may be to assess parameters of metallothionein synthesis at the molecular or biochemical level. Moreover, metallothionein isoform patterns could provide information on different isoforms synthesized in response to different metals or chemicals. In any case, however, care must be taken to consider intrinsic physiological parameters, such as nutritional or developmental factors, which could also interfere with metallothionein synthesis>

2213. Wang,WX; Fisher,NS (1999): Delineating metal accumulation pathways for marine invertebrates. Sci. Tot. Environ. 237-238, 459-472.
<Delineating the routes of metal uptake in marine invertebrates is important for understanding metal bioaccumulation and toxicity and for setting appropriate water and sediment quality criteria. Trace element biogeochemical cycling can also be affected if the rates of metal uptake and regeneration by marine animals are dependent on the routes of metal accumulation. In this paper we review recent studies on the pathways of metal accumulation in marine invertebrates. Both food and water can dominate metal accumulation, depending on the species, metal and food sources. Trace elements which exist in seawater primarily in anionic forms (e.g. As and Se) are mainly accumulated from food. For metals that tend to associate with protein, uptake from water can be an important source. Kinetic modeling has recently been used to quantitatively separate the pathways of metal uptake in a few marine invertebrates. This approach requires measurements of several physiological parameters, including metal assimilation efficiencies (AE) from ingested food, metal uptake rates from the dissolved phase, and metal efflux rates (physiological turnover rates) in animals. For suspension feeders such as mussels and copepods, uptake from the dissolved phase and food ingestion can be equally important to metal accumulation. Metal AE and partition coefficients for suspended particles, which are dependent on many environmental conditions, can critically affect the exposure pathways of metals. For marine surface deposit feeding polychaetes such as Nereis succinea, nearly all metals are obtained from ingestion of sediments, largely because of their high ingestion rates and low uptake from solution. The bioavailability of metals from food and the trophic transfer of metals must be considered in establishing water and sediment quality.>

Note the following study was done using "mesocosms" = aquaria.

2214. Breitburg,DL; Sanders,JG; Gilmour,CC; Hatfield,CA; Osman,RW; Riedel,GF; Seitzinger,SP; Sellner,KG (1999): Variability in responses to nutrients and trace elements, and transmission of stressor effects through an estuarine food web. Limnology and Oceanography 44, 837-863.
<Aquatic systems are increasingly exposed to multiple stressors from anthropogenic sources. These stressors can vary in the consistency and magnitude of responses they elicit in biota and in how the presence of additional stressors modifies their effects. Understanding how the biological environment and temporal dynamics influence responses to stressors, and how stressors interact, is important to predicting their effects in the natural environment. We examined temporal variability in responses of an experimental estuarine food web to elevated trace elements and nutrients, as well as non-additive effects of the combination of these two stressors. Experiments were conducted four times during spring through autumn 1996 in 20 l-m3 mesocosms. We measured a range of system-, population-, and individual-level parameters to quantify responses of phytoplankton, bacterioplankton, heterotrophic nanoflagellates, copepods, fish, and benthic invertebrates to trace element and nutrient additions. The response to trace element additions was more variable both temporally and among phytoplankton and higher trophic level taxa than was the response to nutrient additions. Most taxa increased, either significantly or showed a trend toward increasing, in response to nutrient additions in all four mesocosm runs. In contrast, the direction as well as the magnitude of responses to trace element additions varied considerably among taxa and experimental runs. Two distinct types of nutrientXtrace element interactions were important. First, temporal dynamics of nutrient ratios appeared to affect the temporal pattern of toxicity of trace elements to phytoplankton. Second, in the June mesocosm run when trace element additions reduced production, abundance, or growth of many organisms, these reductions were often proportionately greater in nutrient addition tanks than where no nutrients were added. Our results suggest that considerable temporal and taxonomic variation in responses to trace element loadings are likely to be seen in field settings even under constant loadings to the system and that trace elements may mask the magnitude of the response to high nutrient loadings in eutrophic systems. More generally, the presence of multiple stressors may either increase or dampen the temporal and spatial variability seen in aquatic systems, depending on the interactions among stressors and the influence of background environmental conditions and sensitive species on the expression of stressor effects.>

2227. Wang,WX; Stupakoff,I; Fisher,NS (1999): Bioavailability of dissolved and sediment-bound metals to a marine deposit-feeding polychaete. Mar. Ecol. Prog. Ser. 178, 281-293.
<Assimilation efficiencies (AEs) of trace elements (Ag, Cd, Co, Se and Zn) in a marine deposit-feeding polychaete, Nereis succinea, from ingested sediments were measured using a pulse-chase radiotracer feeding technique. Radiolabeled sediments were encapsulated and fed to the worms for 1 h, after which the worms were allowed to depurate their ingested materials for 3 d. The ranges of AEs were 12 to 36% for Ag, 5 to 44% for Cd, 35 to 96% for Co, 29 to 60% for Se and 21 to 59% for Zn. Trace metal assimilation was little affected by sediment source and sediment grain size. Metals (Ag, Cd, Co and Zn) associated with anoxic sediments were assimilated with a significantly lower efficiency than metals from oxic sediments. The AE of Cd decreased with the duration of sediment radiolabeling; AEs of Ag, Co, Se and Zn were weakly affected by sediment aging. Metal uptake in worms from the dissolved phase was proportional to metal concentration in the dissolved phase, although there was some evidence of Cd and Zn regulation in response to an increase in ambient concentrations. Uptake rate constants were highest for Ag, followed by Zn > Co > Cd > Se. By incorporating metal influx from both the dissolved and particulate (sediment) phases, a bioenergetic-based kinetic model indicates that most (>98%) of the Cd, Co, Se and Zn in polychaetes arises from sediment ingestion due to the high ingestion rates of these animals and the low uptake rate of metals from the dissolved phase (porewater or overlying water). For Ag, approximately 5 to 35% is due to uptake from the dissolved phase. Our study suggests that the establishment of sediment quality criteria must consider sediment as a potentially important source for metal uptake in benthic invertebrates.>

4783. Nystrom,M; Nordemar,I; Tedengren,M (2001): Simultaneous and sequential stress from increased temperature and copper on the metabolism of the hermatypic coral Porites cylindrica. Marine Biology (Berlin) 138, 1225-1231.
<Stressors arising from human activities may interact not only with each other, but also with natural disturbances. However, experimental studies on disturbance complexity and physiological responses of corals to sublethal stresses, especially those due to human activities, are surprisingly few. In this study we investigated the stress response of the scleractinian coral Porites cylindrica after 24 h of exposure to copper (11 mug Cu l-1) and increased temperature (following a 4degreeC above-ambient curve), separately and in combination. We also investigated the effect of sequential stress where corals pre-exposed to increased temperature for 24 h were exposed to copper (for 24 h) after a 5-day recovery period. Changes in gross primary production (Pg: per milligram chlorophyll a per hour) and respiration (R:per square centimeter per hour) in terms of dissolved oxygen were used as indicators of stress. The results show that heat and the combination of heat and copper significantly reduced production rate. However, corals exposed to elevated temperature displayed a significantly higher production rate following the 5-day recovery period. The combination of the two stressors showed no additive or synergistic effects. Copper alone had no effect on the production rate. However, corals that were pre-exposed to increased temperature and again exposed to copper after 5 days displayed a significant reduction in production rate. The respiration rate was significantly reduced by all treatments, although no significant differences between treatments were detected. The results presented here illustrate how a stressor that does not affect corals when acting in isolation may do so in sequential combination with other stressors>

4929. Reichelt-Brushett,AJ; Harrison,PL (2000): The effect of copper on the settlement success of larvae from the scleractinian coral Acropora tenuis. Marine Pollution Bulletin 41, 385-391.
<This study examined the effect of copper on the settlement success of planula larvae of the reef-building coral Acropora tenuis during 1994 and 1996 at Magnetic Island, Great Barrier Reef. Copper concentrations of 2, 10, 20 mug l-1 did not inhibit larval settlement after 48-h exposure. However, copper concentrations of 42 mug l-1 and 81 mug l-1 significantly reduced settlement success of A. tenuis larvae after 48-h exposure compared with controls using normal seawater. At 200 mug l-1 copper, all larvae died. EC50 values for the effect of copper on A. tennis larval settlement were calculated from the 1996 results using measured copper concentrations. The 48-h EC50 was 35 mug l-1 with an upper and lower 95% confidence limit of 37 mug l-1 and 32 mug l-1, respectively. The 48-h NOEC value for both experiments was 20 mug l-1 copper. These experiments provide some of the first data on sub-lethal effects of trace metals on tropical marine organisms, and demonstrate that relatively low copper concentrations impair or inhibit settlement of coral larvae.>

rshimek

08/24/2002, 10:40 AM

Originally posted by Randy Holmes-Farley

Somehow, I think your simplistic interpretation is missing something. Perhaps if any of us had read the actual paper, we'd know better

Actually, Randy, I have read the paper. I suppose that it is possible that if you had read the paper, you might indeed know better.

My interpretation, however simplistic, is correct.

This is my last post in this thread.

Randy Holmes-Farley

08/25/2002, 08:03 PM

Actually, Randy, I have read the paper. I suppose that it is possible that if you had read the paper, you might indeed know better. My interpretation, however simplistic, is correct.

This is my last post in this thread.

Oh, that's disappointing that you won't tell us some of the details. I'll have to go about ordering it myself. Since it shows both positive and negative effects of both nitrate and iron to a coral and it's zoox that we actually keep, it would seem to be of general interest to many reefkeepers. There are lots of considerations on how this result might relate to our aquaria, and without knowing the details, it is hard to assess what it means.

For example, did increased iron and nitrate encourage growth of something else in the system (from algae to the mentioned zoox growth and photosynthesis increase) that used up phosphate as a limiting nutrient? If so, that wouldn't seem to relate to most aquaria, but would be as we expect for increased growth in a low phosphate system like natural or synthetic seawater (but not most tanks).

Without knowing details like that, we are unable to asses whether the iron and nitrate are "toxic' or are actually growth stimulants, as we generally recognize them.

After I get a copy (which may take me a while), I'll be glad to provide any necessary details to those who are interested in knowing what was tested, how it was tested, and what the endpoints were (from the abstract we can't even tell if that was coral growth rate or something else). If you change your mind and provide it in the meantime, I'm sure we would appreciate it.

FWIW, I'm glad that you are so impressed with the stastics of using 2 tanks for each test in this paper. Running 2 tests has apparently taken the test from being laughable (as you described mine) to one that you quite like. So if one other person duplicated my result with iron additions, you'd be happy with my results? Perhaps you should check out my forum. There are posts there from people with the same results. Should I say QED? Naw, I 'd hate for you to have to respond.

FWIW, I too am basically uninterested in pursuing heavy metal toxicity in reef tanks until someone shows there to be some toxicity in reef tanks. I am very interested in papers discussing the effects of iron additions to corals and other organisms in our tanks, since I supplement with it, and will strive to find out as much as is available on the subject, and then provide it to the reefkeeping public as warranted.

Randy Holmes-Farley

08/26/2002, 05:56 PM

OK, I bit the bullet and forked over the $30 for the full text and links associated with the paper (the Ferrier-Pages et al paper).

Since it involves a coral that we keep (Stylophora pistillata) and a supplement that we add (iron, either alone or in food), it is worthy of extensive discussion (not here, but likely in an upcoming AAOM article of mine).

FWIW, however, while the authors do use the word "toxicity" of the iron with respect to calcification (making Ron's assertion technically correct), they go on to state that it may be as simple as the demonstrated faster zoox growth using up nutrients or photosynthetic byproducts that the coral would otherwise have gotten, and hence calcification may be slowed in a nutritent poor system like theirs.

Habib

08/27/2002, 01:46 AM

Randy,
OK, I bit the bullet and forked over the $30 for the full text and links associated with the paper (the Ferrier-Pages et al paper).

Great! I phoned here yesterday but she is still on vacation.
If there is need to ask her still some things please let me know.

they go on to state that it may be as simple as the demonstrated faster zoox growth using up nutrients or photosynthetic byproducts that the coral would otherwise have gotten, and hence calcification may be slowed in a nutritent poor system like theirs.

This sounds very obvious.

BTW how did you get the paper. I did not came further then being only available to subscribers to that magazine.

Thanks

Randy Holmes-Farley

08/27/2002, 07:28 AM

Habib:

You can buy it on this page:

http://www.sciencedirect.com/science?_ob=GatewayURL&_origin=biology&_urlversion=4&_method=citationSearch&_version=1&_piikey=S0022098101002416&_volkey=00220981%23259%23249&_refkey=FerrierPages%232001%23249%23261&md5=c2352b1964e2b848916bd15a91321ba9

Habib

08/27/2002, 07:40 AM

Randy,

When I follow the link I see:

The article you have requested could not be found within the system or is not within your institution's current entitlements; access to journal entries is based upon current journal subscriptions

Probably buying articles is only allowed for the poor Americans and the rich Europeans have to take a subscription.:D

Perhaps Ron is willing to give me a subscription as a gift:D

Randy Holmes-Farley

08/27/2002, 12:28 PM

It could be that it is only a US thing. At that page, I see lots of negative stuff about needing a subscription, etc., but also:

"If you do not have a User Name and
Password click the "register to purchase"
button below to purchase this article.

Price: US $30.00 "

Maybe they don't trust euros:D

Tafabub

08/27/2002, 11:05 PM

Well, since I am a bit too lazy to read through 6+ pages of the chemist vs. Biologist battle, one claiming that nothing done in a lab is feasible in nature, the other claiming that the other one doesn't have a good enough control, I am going to propose a possible experiment that one could do to determine the toxicity of Cu in seawater.
It has been established that certain bacteria secrete siderophores to complex metals (usually Fe, but others are known), which either allow uptake of these metals to aid in biological processes or to protect themselves against them.

One could have a tank with a known concentration of a metal that at that concentration will kill 50% the animals used (whether it be a mollusk, coral, whatever) in X time. This of course is the control. In another tank, the metal and food supply (be it nitrate or ammonia) is added and to it a known concentration of a bacteria is added, allowed to multiply, and then the test subject is added. The concentration of the metal in the water won't change and since the kinetics of most siderophores is so fast, the only barrier is how much chelating ligand is produced.
This of course would have many trials, each with different concentrations of the metal and bacteria as well as time allowed for the bacteria to reproduce.

This could also be done by just adding straight siderophores or another natural chelating ligand to the metal spiked water but this wouldn't be as natural as what may happen in ones tank.

Argue away.

Matt

Oh ya, since I proposed this first (I think), if someone does do this, please cite me (email me for info :-)).

Randy Holmes-Farley

08/28/2002, 01:10 PM

Matt:

That would be a fine experiment for showing what COULD happen, but it wouldn't show if it does happen in a real reef tank.

IMO, the best first experiment would be to show that something (anything) in real reef tank water (taken as a whole) is toxic to a coral compared to natural seawater.

Then, one could begin the long task to find out what that agent(s) are (if any). There are many things different between tank water and seawater, and nailing down one or more culprits may be difficult (or may be easy if you find it on the first couple of trys).

Assuming that an artificial seawater can be found that is just as good as natural seawater for whatever tox test is used, then one can add back certain compounds to see what effect they have, both individually, and as part of a larger collection of things. This is where the experiment becomes tricky, as whatever is added needs to be similar to the form that it takes in reef tank water (for metals, this would include oxidation state and organic binding). You could do your siderophore experiment here, but the issue may be that the one you picked may not reflect what is really happening, and may give a misleading result, one way or the other.

Tafabub

08/28/2002, 01:29 PM

True true, but i was assuming that Dr. Ron's cites were accurate and that it has been proven that certain metals in concentration are toxic to corals.

Just think of the size and PITA it would be to do a HPLC of tank water to find all the organics. It makes me shudder just to think of it.

Matt

Habib

08/28/2002, 01:47 PM

Matt,

Biologist battle, one claiming that nothing done in a lab is feasible in nature,

If the questionon or a part of it is if ligands are present in aquariums then I can say yes they are.

It is relatively easy to measure them in aquariumwater.
I have done this already.

And since ligands are present in aquariums they can alter the toxicity of metals such as copper.

Randy Holmes-Farley

08/28/2002, 07:40 PM

Ron:

I noticed that there is a paper by Norris and Fenical (reference below) that tests a bunch of organic toxins from various seaweeds. One that may be of interest to your upcoming test of reef tank water on sea urchin eggs was from the tropical red algae Laurencia caraibica. It was found to be toxic to the fertilized eggs of the temperate sea urchin Strongylocentrotus purpuratus.

You might consider this type of information in drawing conclusions about what may be toxic in your tests.

"Chemical Defense in Tropical Marine Algae; in "The atlantic barrier reef ecosystem at Carrie Bow Cay, Belieze, I Structure and Communities" Rutzler and Macintyre (eds); Smithsonian Institute Press, pp. 417-431; 1982.

EricHugo

08/28/2002, 11:45 PM

LOL -

Randy, that book is sitting behind me as I type. Its a signed edition from Rutzler and MacIntyre. I had, in fact, just pulled that article for an article Anthony Calfo is writing on Caulerpa. How's that for unbelievable because that is an obscure book!

It also has one of the only sources of information around on the zoanthid Isaurus. The chapters are amazing - covers all the Caribbean corals, zooplankton, corallines, inverts, geomorphology, water currents and exchnage, and on and on - and completely, too, I might add.

Just another reference available from the library of Borneman :)

Habib

08/29/2002, 03:10 AM

Ron,

I will - if I can get the $$ - be examining the metals in tank sediments late this autumn as one of the upcoming components of the this project of mine.

Perhaps you should undertake some research about the organic components, if you think they are important.

Here is an abstract which is worth considering before drawing any conclusions on heavy metals in the sediment and their toxicity.

Marine Pollution Bulletin, Vol. 44 (4) (2002) pp. 286-293

An overview of toxicant identification in sediments and dredged materials
Kay T. Ho a * ho.kay@epa.gov , Robert M. Burgess a, Marguerite C. Pelletier a, Jonathan R. Serbst a, Steve A. Ryba a, Mark G. Cantwell a, Anne Kuhn a and Pamela Raczelowski b
a Atlantic Ecology Division, US Environmental Protection Agency, 27 Tarzwell Drive, Narragansett, RI 02882, USA
b University of Rhode Island, Kingston, RI 02892, USA

Abstract
The identification of toxicants affecting aquatic benthic systems is critical to sound assessment and management of our nation's waterways. Identification of toxicants can be useful in designing effective sediment remediation plans and reasonable options for sediment disposal. Knowledge of which contaminants affect benthic systems allows managers to link pollution to specific dischargers and prevent further release of toxicant(s). In addition, identification of major causes of toxicity in sediments may guide programs such as those developing environmental sediment guidelines and registering pesticides, while knowledge of the causes of toxicity which drive ecological changes such as shifts in benthic community structure would be useful in performing ecological risk assessments. To this end, the US Environmental Protection Agency has developed tools (toxicity identification and evaluation (TIE) methods) that allow investigators to characterize and identify chemicals causing acute toxicity in sediments and dredged materials. To date, most sediment TIEs have been performed on interstitial waters. Preliminary evidence from the use of interstitial water TIEs reveals certain patterns in causes of sediment toxicity. First, among all sediments tested, there is no one predominant cause of toxicity; metals, organics, and ammonia play approximately equal roles in causing toxicity. Second, within a single sediment there are multiple causes of toxicity detected; not just one chemical class is active. Third, the role of ammonia is very prominent in these interstitial waters. Finally, if sediments are divided into marine or freshwater, TIEs performed on interstitial waters from freshwater sediments indicate a variety of toxicants in fairly equal proportions, while TIEs performed on interstitial waters from marine sediments have identified only ammonia and organics as toxicants, with metals playing a minor role. Preliminary evidence from whole sediment TIEs indicates that organic compounds play a major role in the toxicity of marine sediments, with almost no evidence for either metal or ammonia toxicity. However, interpretation of these results may be skewed because only a small number of interstitial water (n=13) and whole sediment (n=5) TIEs have been completed. These trends may change as more data are collected.

Also worth considering is the heavy metal speciation and pollution in Cleveland Bay (Australia) , the nearby coral reefs (a.o. magnetic island) and the advice not to dredge only around mass spawning periods.

Randy Holmes-Farley

08/29/2002, 07:04 AM

Just another reference available from the library of Borneman

:thumbsup:

Bomber

08/29/2002, 08:53 AM

Its a signed edition from Rutzler and MacIntyre.

You're not old enough! LOL

EricHugo

08/29/2002, 09:28 AM

Hi Bomber

also LOL - yeah, you're right...I forgot

Bomber

08/29/2002, 09:56 AM

LOL

That book came out in the late 70's early 80's something like that, You're not a old man like me.

I meant you're not old enough to have been there and had it signed. ;)

Habib

08/29/2002, 10:02 AM

Eric,

You have got something like this:

rshimek

08/29/2002, 11:09 AM

Originally posted by Habib

Habib,

Here is an abstract which is worth considering before drawing any conclusions on heavy metals in the sediment and their toxicity.

I don't need you to tell me what is important in discussing heavy metals in sediments and their toxiciity; the following are some of my publications on the topic. I was senior or sole author in the Parametrix reports and they are available from the Region X EPA library in Seattle. Although most of this is in the grey literature, much of it has been the basis of TIE development in the areas considered. Publication in the peer-reviewed literature was not an option because the funding was corporate, or municipal and they wouldn't pay for my time.

When you gather some experience in the practical applications of this, we can discuss it further.

Shimek, R. L., T. Thompson, and D. Weitkamp. 1991. Slag, benthos, and bioassays: poor correlation of bioassay predictions and the benthos. In: Chapman, P., F. Bishay, E. Power, K. Hall, L. Harding, D. McLeay, M. Nassichuk and W. Knapp (Eds.) Proceedings of the seventeenth annual aquatic toxicity workshopL November 5-7, 1990, Vancouver, B. C. Canadian Technical Report of Fisheries and Aquatic Sciences. 2:1046.

Shimek, R. L., T. A. Thompson, T. H. Schadt, and D. E. Weitkamp. 1992. Interpreting conflicting biological and chemical results from a Puget Sound sediment data set. Puget Sound Water Quality Authority. Puget Sound Research '91, Proceedings. 2:546-552.

Sole or senior contributing author for the following major project reports, Parametrix, Inc.

Asarco-Tacoma = Arsenic refinery and the studies concern off shore heavy metal deposition.

Parametrix. 1989a. Asarco Tacoma remedial investigation. Prepared for Asarco, Incorporated, Salt Lake City, Utah.

Parametrix. 1989b. Asarco Tacoma smelter offshore marine sediments feasibility study. Prepared for Asarco, Incorporated, Salt Lake City, Utah.

Parametrix. 1990a. Asarco Tacoma smelter offshore feasibility study. Supplementary marine sediment survey. Prepared for Asarco, Inc., P. O. Box 1677, Tacoma, Washington.

Parametrix. 1991c. Asarco Tacoma Smelter Offshore Feasibility Study - Supplement Number 2. Prepared for Asarco, Inc., P. O. Box 1677, Tacoma, Washington

Parametrix, 2000. Descriptive analyses of the benthic infaunal communities of two 24 month Asarco Pilot cap stations with a brief comparison to two selected reference stations.

St. Paul Water Way - heavy metals and organics contamination

Parametrix. 1990b. St. Paul Waterway Remedial Action and Habitat Restoration Monitoring Report. 1988-1989. Unpublished report to Simpson Tacoma Kraft Company, Tacoma, Washington. 115 p.

Parametrix 1991a. St. Paul Waterway Remedial Action and Habitat Restoration Monitoring Report. 1990. Unpublished report to Simpson Tacoma Kraft Company, Tacoma, Washington.

Parametrix. 1992b. St. Paul Waterway Remedial Action and Habitat Restoration Project. 1991 Monitoring Report. Unpublished report to Simpson Tacoma Kraft Company, Tacoma, WA, and Champion International, Stamford, CT.

Parametrix. 1993a. St. Paul Waterway Remedial Action and Habitat Restoration Project. 1992 Monitoring Report. Unpublished report to Simpson Tacoma Kraft Company, Tacoma, WA, and Champion International, Stamford, CT.

Parametrix. 1994. St. Paul Waterway Remedial Action and Habitat Restoration Project. 1993 Monitoring Report. Unpublished report to Simpson Tacoma Kraft Company, Tacoma, WA, and Champion International, Stamford, CT.

Parametrix, and R. L. Shimek. 1994. Biological indicators - St. Paul Waterway area remedial action and habitat restoration project. Unpublished report to Champion International, Stamford CT., and Simpson Tacoma Kraft Company, Tacoma, WA.

Simpson-Tacoma Kraft mill - We fixed this one. It was the first and I think still is only remediated marine Superfund site. Organics here mostly.

Parametrix. 1990c. Outfall benthic survey results - Simpson Tacoma Kraft mill. Prepared for: Simpson Tacoma Kraft Company, P. O. Box 2133, Tacoma, Washington.

City of Tacoma - Organics, Heavy metals.

Parametrix 1991b. City of Tacoma Central Waste Water Treatment Plant. Draft 1990 Outfall Monitoring Report. Benthic Macro-Infaunal Invertebrate Analysis. Unpublished draft report to the City of Tacoma, Washington. 46p + appendices.

Parametrix. 1992a. Central Wastewater treatment plant, 1991 Outfall monitoring report. Benthic macro-Infaunal invertebrate analysis. Unpublished report to the City of Tacoma.

Sitcum Waterway - Habitat restoration. Heavy metals and organic contamination.

Parametrix, 1997. Principal Coordinate Analyses of the 1996 Benthic Infaunal and Epibenthic Zooplankton Data from The Sitcum Waterway area. Prepared for: City of Tacoma

Parametrix, 1999. Principal Coordinate Analyses of the 1998 Benthic Infaunal and Epibenthic Zooplankton Data from The Sitcum Waterway area. Prepared for: City of Tacoma

EricHugo

08/29/2002, 11:42 AM

Habib. That's hilarious. Did you come steal my book? Something happened with your title, though...it seems to have some text missing. ;-)

Randy Holmes-Farley

08/29/2002, 12:08 PM

When you gather some experience in the practical applications of this, we can discuss it further.

:lolspin:

Since I have extensive practical experience in binding metals with organic molecules (and you do not), you'd think the same would have applied to that discussion. Somehow you were able to comment on metal binding organics without any published practical experience, so why would you think Habib less able to participate in scientific discussions without having published sediment toxicity tests himself?

FWIW, all he did was point out a paper to you. If you've already read it, that's great. If you think it's nonsense, that's fine too, but it would be nice if you based your comments on the science of the paper in question and not the background of the person that pointed it out to you. That's what being an expert involves: people often, and hopefully frequently, point out things that you already know. It's not a reason to criticize them.

So, with your extensive background in sediment toxiciology, do you disagree with these conclusions in the abstract?

"while TIEs performed on interstitial waters from marine sediments have identified only ammonia and organics as toxicants, with metals playing a minor role. Preliminary evidence from whole sediment TIEs indicates that organic compounds play a major role in the toxicity of marine sediments, with almost no evidence for either metal or ammonia toxicity. "

Randy Holmes-Farley

08/29/2002, 12:15 PM

Something happened with your title, though...it seems to have some text missing. ;-)

Boy, I got sucked in with that one. I guess I'll have to be a little more skeptical of things you guys post. ;)

Good one!

Habib

08/30/2002, 07:32 AM

Eric,

I can't remember you responding that fast on a post of mine:D

though...it seems to have some text missing

Yes, I did not like to keep you wondering too long;)

Habib

08/30/2002, 08:06 AM

Ron,

much of it has been the basis of TIE development in the areas considered.

So others did or try to do TIE's but not you?

If the tests show that levels of a certain metal kill animals, and if the tests show that the levels are above those levels, and animals do die then it is reasonable to suggest that the metal is responsible.

Is this a TIE?

When you gather some experience in the practical applications of this, we can discuss it further.

You mean reading and writing?

Randy Holmes-Farley

08/30/2002, 03:33 PM

FWIW, Ron has initiated a new discussion of iron in my forum:

http://archive.reefcentral.com/vbulletin/showthread.php?threadid=110366

Enjoy!

rshimek

08/31/2002, 09:32 AM

Originally posted by Habib

Also worth considering is the heavy metal speciation and pollution in Cleveland Bay (Australia) , the nearby coral reefs (a.o. magnetic island) and the advice not to dredge only around mass spawning periods.

In almost no cases that I know of are dredging restrictions based solely on the relevent scientific findings. Most of the time, in fact, the science takes a back seat to politics/economics. As an example, dredging may have to be done during certain periods due to tidal cycles (In the Puget Sound region of Washington, USA; it is most frequently done around the equinoxes as the tidal currents are far less). Dredge spoils may be stored for some periods, but generally are dumped soon after dredging occurs. This will happen whether or not it is advantageous for the adjacent fauna. I tried - several times - to regulate dredging times, but was only successful once or twice. Nobody argued with us that the dredging would damage the adjacent fauna, but that was simply considered "a cost" of the project.

So others did or try to do TIE's but not you?

TIEs were being developed during the 1990's when I did most of the work indicated, and presently have only begun to be investigated, and mostly on the eastern seaboard of the US, not on the west coast where I work. We did do pore water toxicity testing (which amounts to much the same thing - but it was not as standardized as the present TIEs), but it is/was difficult to do. In some sediments, it is effectively impossible to extract, particularly since it may have be to be done undewater by divers.

If the tests show that levels of a certain metal kill animals, and if the tests show that the levels are above those levels, and animals do die then it is reasonable to suggest that the metal is responsible.

Is this a TIE?

Yes, in part. There are several different types of tests, some involve direct toxicity to selected animals, others involve bacteriological testing.

Also, the TIEs have not been in use long enough or in enough cases to draw many conclusions from them. In most of the areas where they have been developed and used, the metals concentrations are relatively low compared to the organics, for example. In a lot of the areas where I worked, the situation was reversed.

We typically found free metal concentrations in oxygenated pore water to be low - probably due to organic complexing, or to binding with iron. These waters were toxic, but not as much as was expected. We attributed the toxicity to the organo-metal complexes, rather than the metals themselves. Directly above the sediments, the metals concentrations in the water were far lower, and the toxicities were correspondingly lower. The animal density in the upper (1-3 cm) layers of these sediments was often really quite high, but the animals present were largely those that could ventilate their burrows with the overlying waters.

As one went deeper into the sediments (4-6 cm), and the sediments became anaerobic, the metals concentrations in the waters rose dramatically, and the pore water became extremely toxic. However, as few organisms could tolerate this metallic tea, there was little water movement in these sediments and the metals were contained. If the sediments were disturbed, signficant, but transient mortalities were seen (sediment faunal kills, mass mortality of epifaunal animals).

I think you can see where I am going with this.

I think a comparable situation exists in our tanks, particularly after several years. The sediments build up a signficant load of chemicals that - if they become soluble - will be very toxic. Disturbances of the sediment will release toxins. Additionally, the salt water, as mixed, has higher concentations of copper and some other metals than we found in smelter slag. If the aquaria have highly "polished" water with low organic levels, this water will kill some organisms. If the water has the organics to bind the metals, these may form particulates which are eaten and in the acid phases of digestion they may become toxic. Additionally, the metals or metal/organic complexes in the water are very likely toxic to many animals. A lot of the mortality in aquaria attributed to incorrect acclimation is, I think due to incorrect acclimation alright, but to acclimation not to salinity or temperature but rather to heavy metals.