Inland Reef / Reefs.org Salt Analysis Project

[Habib (Salifert)]

............
The fact that manufacturer's calims were used for analysis is relatively unimportnat to the validity of the study, although analysis would go a long way to confirming a significant correlation between composition and larvotoxicity. Personally, the assays seem more important and may be cheaper than complicated analysis of each salt. If the salts are ultimately toxic, I don;t want them no matter what they contain. They would just be "out". Then, of the ones that appear to be relatively safe, those could be used as subjects of the more intesive and expensive analysis.
..........
.......

I agree with Eric.
If toxicity is the issue then a salt analysed for copper showing say 5 ppb could be more toxic than a salt having a much higher copper concentration of say 40 ppb.

Or is there somewhere proof that a certain metals concentration higher than a certain value is ALWAYS toxic? Don't forget that techniques such as ICP and ICP-MS cannot determine the metal's speciation.

The bioassay as done by Ron Shimek was able to discrimate between saltmixes and even between two tankwaters. This is IMO fantastic.

The biosensors told us which water they liked and which not.

They might have not liked it because of just a single metal, a combination of metals, a plasticizer (thanks Jerel aka Bomber), an anticaking agent, ammonia, highly oxidixing substances in the salt, ........

The bioassay can provide us IMO very useful and important information.

Bioassay done using water of tanks using just NSW and a good ASW might tell us if there is a difference after a certain period of time.

FWIW, and I think it is worth a lot, tanks using NSW acquire a metal concentration comparable to tanks using IO or other salt mixes. That is at least true for the tanks Ron has analysed and the tanks analysed by us.

An ICP-MS analyses would tell us that all the tanks are (approximately) the same while we know that they are not. We need something which can discrimate between those tanks which according to one technique (e.g. ICP-MS) are the same.

The bioassay technique done by Ron Shimek has IMO the potential to discrimate between such tanks but also between virgin salt mixes.

 

[jamesw]

Oy, I go to sleep for a few hours and look what I come back to! I'm glad to see you folks participating over here Eric, Randy, and Habib. Keep the good feedback coming!

I noticed that there is one particular poster that is being a "seagull" on this thread - flying in and *****ing on everything but not making any constructive contribution. Since this is reefs.org we are not going to delete his posts - so just try to ignore him (would we be extended the same courtesy? Nope, we'd be censored!). Sorry about that gang.

Cheers
James

 

[baseman]

I think that doing a dry analysis is just as important as doing a bioassay. Dry analysis will not determine 'bad' and 'good' salt mixes, but it will provide a valuable datapoint for a correlation between the composition and the bioassay results.

 

[Bill2]

I will be going to French Polynesia from March 30th through April 13th on a biz trip. Gotta love it I will be happy to pick up some NSW samples. I might be able to get some samples from some of the outlying islands. I'm pretty sure I can get Tahiti, Moorea, Bora Bora, maybe Huahine. If someone can give me instructions on what is needed so it will be a valid sample that would be great.

 

[MiNdErAsR]

I noticed that there is one particular poster that is being a "seagull" on this thread - flying in and *****ing on everything but not making any constructive contribution. Since this is reefs.org we are not going to delete his posts - so just try to ignore him (would we be extended the same courtesy? Nope, we'd be censored!). Sorry about that gang.

I'm sorry but I disagree. I've seen nothing but good suggestions and valid points made here. This childish game you're playing does nothing but attempt to widen the gap between boards while others are trying to work together for the common good. We all (supposedly) have the same goals, the spreading of knowledge in order to help the hobby. Let's try to remain focused here, shall we?

IMO

 

[kazzoo]

I've been away from the board for a while, and noticed this thread. Being an environmental toxicologist by education (PhD), an ecological risk assessor, experienced in toxicity testing and reviewing toxicity tests, I found this thread intriguing. I went back and read Dr. Shimek's article in reefkeeping. There are some points I'd like to discuss.

He stated in the introduction that organisms typically detoxify metals by binding them, and that there will ultimately be accumulation of those metals. This is partially true. Metals may be detoxified in ways including what he mentioned, but that does not automatically mean bioaccumulation of the metal complex. Many detoxification mechanisms enhance excretion. The degree of bioaccumulation depends on the formed complex and its lipophilicity, or affinity for fatty or fat-like substances: higher lipophilicity, higher bioaccumulation potential, and visa versa. These lipophilic compounds will accumulate in non-polar environments such as animal lipids and organic rich soils and sediments. If it suspected that metal bioaccumulation is taking place, check for metal bioaccumulation in the organisms by analyzing their tissues. However, information of selected metal bioaccumulation potential may very well be in the appropriate literature.

Significant differences among treatments were observed. The conclusion was that some of the ASWs were toxic. I would be cautious with that conclusion. Most people think of toxicity due to the presence of some bioactive compound. What if there was an absence of a or some bioessential compounds that inhibited larval survival? Are we certain that everything necessary for larval survival is in those salts? Somebody correct me if I'm wrong, but I assume the Marinemix bioassay was specifically forumulated for larval survival while the other salts were not. Consider the possibility that something is missing from those salts that is necessary for larval survival as opposed to something being present that is toxic. I would do a thorough analysis of the salts to determine all components, not just the metals.

Risk assessment assesses the risks of an action or substance to something of personal or societal value. Risk is a function of exposure and toxicity. We always compare exposure levels, whether measured or predicted, to actual toxicity data indicating some sort of exposure response relationship, i.e. the LC50 (in aquatic risk assessments), LOAEC, and so on to determine if there is a risk. In this test, Dr. Shimek performed a modified urchin embryo/larval toxicity test on various commercial salts. Given the results of the toxicity tests, it would be most accurate to say that some commercial sea salts do not favor urchin embryo larval development. The reason for the poor performance should be investigated.

These salts have a number of components. The relative amounts of these components compared to NSW indicate elevated levels for many of the components, even in the mixes that were not "toxic" to the larvae. I don't think we can safely say that the poor larval performance is due to elevated metal concentrations, unless we have some data indicating what the metal toxicity levels are for marine invertebrates, or ideally for larval urchins. If we had those toxicity data, then we'd compare the exposure levels to the known metal toxicity and derive a risk or hazard quotient. The value of that quotient may or may not indicate a risk depending on whether or not it exceeds a level of concern (LOC). The LOC can vary depending on the regulatory law. In any event, it is hypothetical to single out any components, other than the salt itself, as a risk. Even then, be careful extrapolating that risk to other organisms.

Somebody had mentioned that there needs to be solid hypotheses before this project commences. I agree. We need to find out why the larvae did not perform well in some salts as opposed to others. Elevated metal content is one hypothesis. We have the exposure data, what we need is the toxicity data for the risk analysis. You don't want to spend the money on toxicity tests for each metal found in the salt, $$$$$. The literature and federal databases are the sources for that information. Another hypothesis is that there is some component missing from the salts that adversely affect larval survival. A complete analysis of various salts in that study would begin to answer that question. The answer would be further served by knowing the critical elements for larval development.

There is also concern for metal accumulation within a tank. Analyze the water and compare it to freshly prepared ASW.

Don't spend money on analyzing all those salts until you are fairly certain you know the cause(s) of the poor larval performance. I hope this discussion helps.

Tim

 

[JohnRogan]

wow...well you people have officially lost the interest of the "common man"

At the beginning this sounded like a great idea, but it has become far too complex for most people. If you are trring to raise money from the "average" person, it is best to keep it so an "average" person can understand it. Unless, of course, you don't care about funding the project.

 

[Anonymous]

Speaking on behalf of Reefs.org....

There's a few things that need to be made clear. First, this is NOT the James Wiseman Salt Study.

Second, this is NOT the John Link Salt Study.

Third, it should also go without saying that it also is not the Ron Shimek Salt Study, the Eric Borneman Salt Study or the Randy Holmes-Farley Salt Study.

This is the Inland Reef salt study, and I find it in exceptionally poor taste that those who fronted, supported, reviewed and published Dr Shimek's salt article are attempting to take control over it (John Link said, "I think Eric and Randy would be much better choices to lead this effort." whereas Eric said, "I also strongly suggest that Randy, Habib and Craig should ultimately lead this project").

Tom and Matt of Inland have been extremely gracious throughout this, eagerly accepting suggestions, advice and input. For this they have been rewarded by having their competency not only questioned, but actually dismissed. "Good intentions aside", Eric indicates, this project should be taken away from them.

Tom and Matt have also been accused of a lack of integrity by JohnL, who characterized them as "unfair" and "self-serving" (Posted: 14 Mar 2003 03:50). This, in response to Tom and Matt stating, "We are the ones who proposed doing this test and will carry it out." Even in the face of this blatant provocation, Tom and Matt have remained graceful and receptive to further input from John.

May I make a suggestion? No, better yet, allow me to let Dr Ron Shimek make a suggestion:

"Fine, do some tests and write up your results like this if you wish."
excerpt from this thread

and

"If you or anyone wishes to try to disprove my hypothesis, go for it."
excerpt from this thread

Ron has very clearly taken the attitude on several occasions in the past that when it comes to his experiments, he will do them as he chooses and if someone doesn't like it, tough, it's not their experiment. Tom and Matt haven't taken that line. Instead, they've actively sought input from others -- yet the basic respect awarded to Ron, that his project is his project, no one elses, is curiously withheld from Tom and Matt by the same people who so strenuously defended Ron's right to that position.

It certainly appears from JohnL's posts in this thread that he wishes RC and his paid staff (Eric, Ron, Randy) to assume a leadership role in this project. However, it's not their project. It's not Reefs.org's project, although we support it as much as we can and have lent our resources to providing Tom and Matt as much help as we can. It's Inland Reef's project, no one elses.

Allow me to publicly and directly ask JohnL the following questions:

1. Will you support Tom and Matt's study, under their vision and leadership?
2. You have been accepting donations from members, both ours and yours, pledged towards Inland's study. As I'm sure some of those who have donated are wondering, will you still be making the donation (and matching it from your own funds) in their name to the Inland project? If not, will you be refunding donations to those who used you to donate through?
3. A few days ago one of our ops, Shane Graber (Liquid), posted similiar questions on Reef Central. His post was deleted in a matter of hours. Can you explain that behaviour, when we've let you and your paid employees (Eric, for example) and your volunteers (Jim Fox) take shots at us in this very thread?

John, I know it's usually your policy to ignore any hard questions that are asked of you publicly. However, based on the sheer number of your own posts within this thread, it will be impossible for you to pretend to have not seen these questions. Will you not extend the courtesy to us of actually providing straight answers to these questions?

The beneficiaries of the study are hobbyists that frequent boards such as these. There is no reason to let site rivalry get in the way of science. In that case, not only the hobbyist loses, but also the very organisms we
intend to keep healthy in our tanks.

I feel quite certain that Tom & Matt will continue to take all the feedback, criticism and advice that's offered to them, and I encourage that to continue. What Reefs.org does not encourage is the attitude that Tom & Matt somehow do not measure up to the task of taking on this project -- after all, they have an excellent history of and reputation for professional product testing and review.

As RC Moderator Jim Fox said in this thread -- albeit immediately after making a personal attack of his own -- "We all (supposedly) have the same goals, the spreading of knowledge in order to help the hobby. Let's try to remain focused here, shall we? "

 

[dragon0121]

JohnRogan,
And, ultimately, that is the point of the experts. What we are asking to accomplish, is really beyond the scope of the "average" man, and I put myself squarely in the "average" man camp! :lol: If we want results that are truely scientific, not anecdotal, then we should heed the advice of the experts. Of course, if you read the boards, there are a lot of bad feelings between "experts" and very little recognition of work done. But, maybe with enough people pushing, and enough interest the whole thing will continue to move forward.

Personally,
I don't care what is in the salts. Find me salts that do well, and tell me why. If the salts don't do well, why even analyse them. I feel urchin embryos was an effective method of finding a good salt, but, others seem to feel this is the ravings of a madman based on their comments. So be it, find something that "good" enough for the masses. Berghia have also been tried in at least one salt, and performed as dismally as urchin embryos. Also, if a month after mixing new salt water, we have significantly changed the composition of the water via feeding and additives, such that they are indistinguishable from the "bad" salts. Who cares what salt you start with, we have other problems to address first.

 

[Len]

John,
Understandably, this thread may have become a bit too technical for the average hobbyist, but rest assured, the aim of the study has not changed. I'm sure I speak for everyone in saying it is no one's intention to dissuade the average hobbyists from helping fund this project. The bottom line is Inland's salt study serves to better every reefkeeper for generations to come, and I hope we all base our decision to donate based solely on this fact.

Personally, I think it's quite healthy for these technical issues be discussed publically and uncensored; it will only serve to broaden everyone's horizon regarding the scope of Inland's study, even if the concepts being discussed may be too complex for you or I.

FWIW, I'm unclear what all this fuss is about. I think baseman said it best:

I think that doing a dry analysis is just as important as doing a bioassay. Dry analysis will not determine 'bad' and 'good' salt mixes, but it will provide a valuable datapoint for a correlation between the composition and the bioassay results.

Inland's study will provide an important reference (one currently lacking) for any sucessive studies to come. The values derived from this study, if nothing else, are datapoints for open interpretation and future reference.

 

[Anonymous]

cjdevito




kudos!

very well said

 

[kazzoo]

John,

Think about why you were considering sending your money. If it is only to find out ASW composition, then great. That is an easy enough proposition.

What I said is be careful about what you can infer from Dr. Shimek's study. The way I read it, metals were the implied culprit for the poor larval performance. While some metals, under the right conditions, are toxic, I pointed out that there may be other possible culprits for the results, with the bottom line conclusion being that some of the ASWs were not the best for urchin larvae development. The question is why. I'm not convinced yet that the metals are the culprit.

I'm not against a study of the various commercial ASWs. If upon further data evaluation, the culprit can be determined, then further studies might be warranted because we all as reef aquarists would like to be sure that we provide the best conditions for our tank inhabitants.

Perhaps it is best to simply do the assays on the different ASWs, not concerning ourselves with the why question, but only with the bottom line, "will the larvae do well in the artificial water?" That is a simple question to answer. That is what Dr. Shimek did. Leave it up to the ASW manufacturer to determine why. I think a drop in product sales could spur the manufacturers to do such studies to answer why. But again, be careful. Does poor urchin larval survival mean poor performance for other organisms? This would require more research comparing the "good" ASWs against the "not so good" ASWs for organism (other than urchin larvae) performance. There has been alot of anecdotal evidence posted on this site over the years by hobbyists of organism performance as well as the salt they used. But there are too many variables to draw any sort of solid conclusion. Controlled studies would help much here.

Tim

 

[Baalz]

As RC Moderator Jim Fox said in this thread -- albeit immediately after making a personal attack of his own

Pot kettle

 

[Anonymous]

Don't the RC mods and members of Team RC have better things to do then come here and engage in personal attacks? John, is this encouraged by you, or are they doing it on their own?

 

[Baalz]

My reply had nothing to do with Johnl.. it was directed at your scathing attacks of everything thats not reefs.org..

You have become the master of personal attacks in this thread.

 

[liquid]

Baalz, if you have nothing constructive to add to this thread, I respectfully request that you refrain from posting.

Shane

 

[Anonymous]

I have? I quote others comments and somehow I'm guilty of their actions? I have called no one childish, self-serving, unfair. Nor have I called anyone unprofessional, bitter, tempermental or arrogant.

I have said nothing negative about any board or organization, merely defended the right of two individuals to engage in a study that they thought up, organized and began.

John, is this the purpose for Team RC? To provide sniping attacks when you'd rather not get your hands dirty personally? If not, I have to wonder at your selection process for members. Is this really the face you want RC to have?

 

[randy holmes-farley]

3. A few days ago one of our ops, Shane Graber (Liquid), posted similiar questions on Reef Central. His post was deleted in a matter of hours. Can you explain that behaviour, when we've let you and your paid employees (Eric, for example) and your volunteers (Jim Fox) take shots at us in this very thread?

This is ridiculous. Everyone grow up and focus on the study, not the petty politics.

I, as the most frequent contributor to RDO's magazine and a moderator of my author's forum at RDO, and author of more than 2000 posts at RDO (who, as Advanced Aquarist, pays me more than RC does), acted at RC in my capacity as a moderator there to take some posts out of a thread.

The posts taken out were exclusively about the handling of money by RC management, not about the design or functionality of the tests in question. So the questions the he and James posted were not similar to anything here. No one here from RC is arguing about handling of money, but rather discussing the scientific aspects of the test.

No, I have no interest in any direct involvement in these tests as they been laid out so far. I gave some advice on how I'd do it, since Tom O'Toole asked me to comment. Any further direct or indirect comments about why I've posted here, from people who know nothing about my motives, are also ridiculous.

 

[MattM]

Maybe a little background on our thought process will help to clear the air here.

I read Dr. Shimek's study with interest. My initial thought was that it was quite a bold statement to say that the most popular salt mix in the world, and the second most popular, are both poisonous to marine life.

Reading further and thinking about it more, I found that the most amazing thing to me was that Dr. Shimek used analyses supplied by the manufacturers, even that from their advertising literature, for the two salts that faired the best in his study. Let's recall the last time that we discovered the value of a manufacturer's analysis of their own product -- Combi-San. It's contents, based on ICPMS analysis, were not even close to the "guaranteed analysis" printed on the bottle. Dr. Shimek should be very familiar with this, since he did the initial analysis of it! We at Inland Reef found the huge discrepancy between the analyses from a respected researcher and a respected manufacturer astounding and we were not sure who to believe. So we took it on ourselves to have the product analyzed and we confirmed Dr. Shimek's results. This led to an almost immediate change in the product labelling, and eventually to a whole new supplement product from that manufacturer.

So I immediately wanted to get an analysis done for the two salts that did well. If they are truly different in a quantifiable way from other salts, they should be the ones we recommend to our customers.

The first problem is one of comparison. If we analyze these 2, then we should do the other 2 at the same time, with the same procedure, from the same lab so we can compare them in a valid manner.

The second issue is consistancy. What if the samples Dr. Shimek used did not represent the normal bag-to-bag salt from these manufacturers? How much does each salt vary from bag-to-bag? To get this data I thought that three tests, from three different lots should get us some feel for the consistancy from each manufacturer.

Third, we need some sort of control. Natural sea water has been analyzed extensively by many researchers over the years. If we have the same lab perform a test on a local seawater sample, we can at least be sure they are "in the ballpark" in the testing. We would not be trying to establish definitively what is in NSW, that has already been done. We just want some assuance that that sample comes out near what it should, which would increase our confidence in the validity of the ASW samples. FYI - there is no particular reason to test 3 different NSW samples -- we were testing three of everything else, so I just "went with it!"

So, 4 salts plus NSW times three samples each would be 15 tests. The lab we normally use charges $350 per ICPMS test for these quantities, so that's $5,250. We decided right away, that we are not prepared to spend that amount ourselves right now.

Then the thought occured to me that if we try to raise the money, maybe this could be a one time opportunity to get every popular salt tested. Additionally, quantity discounts might apply. Well, it turns out that we can get 17 salts plus NSW tested for a total of just over $10,000. More than four times the number of salts for about twice the price.

And that brings us to where we are today.

We don't intend that this is the be-all end-all of salt mix analysis. More testing as has been suggested above, via further bio-assay, or other methods will probably be required to fully determine the reason for Dr. Shimek's results. But I continue to believe that getting the full elemental analysis for all these salts is an essential first step, and that future studies, of whatever nature, will find these results valuable.

 

[Anonymous]

Randy:
The posts taken out were exclusively about the handling of money by RC management, not about the design or functionality of the tests in question. So the questions the he and James posted were not similar to anything here. No one here from RC is arguing about handling of money, but rather discussing the scientific aspects of the test.

Thank you for setting the record straight. I asked a legitimate question, this is a legitimate answer. I can accept that you removed the posts in question because they were not relevent to the discussion there. I do feel the questions are relevent, here if nowhere else, if for no other reason then some who have donated have raised them. I can understand you wishing to keep them out of a thread on scientific methodology, however.

In the interests of giving those posts a place where the legitimate questions they raised can be addressed, I will include Shane's deleted post here. I don't have James', or I'd include it as well....

">>How about updating RC's PayPal link to direct all monies collected from RC
>>to go directly to Inland Reef's PayPal account instead of your own? It
>would
>>make things much easier for Inland Reef to keep an accurate tally on how
>>much money has been donated to the project. I know that there are many
>>people that would like to see where everyone's at in regards to the total.
>>
>>For those of you interested, you can PayPal Inland Reef using the
following
>>address: [email protected].
>>
>>Shane
>>Admin, reefs.org
>>=============
>>03/16/2003 11:43 PM
>>
>>Also, this would help maximize contributor's donations as donated money is
>>already losing 3% due to PayPal's fees to a business account. If the money
>>goes first to RC via PayPal and then to Inland reef via PayPal, it will be
>>charged 3% the first time around, and then 3% again. We all need to be
good
>>stewards with everyone's money.
>>
>>FWIW,
>>
>>Shane"


No, I have no interest in any direct involvement in these tests as they been laid out so far

I'm sorry, I had thought that all those throwing your name out as the logical person to take over and lead had at least discussed this with you. This was an assumption on my part. I apologize for it. I know Tom and Matt are greatful for your input and I wouldn't wish any comments of mine to dissuade you from continuing to offer it.