Comparing Organo-MS results across five different reef tanks

[jeremie]

1) Very similar results despite diff dosing regime.

The concentration of many labile organics, such as amino acids, does seem to be a result of various biological processes, such as excretion, decomposition, or consumption. How each of these factors contributes to the pattern observed here is certainly still mostly a black box.

2) Very concerned about the antibiotics that not vanish. No new info though, cipro is not decomposed at all in nature and reason why it's forbidden in Europe.

As for cipro, most of what I added does seem to leave the water column, either by binding to particulate organics, being skimmed out (which probably also happens when it binds to other organics), or perhaps through some degradation.

I´m sorry to say but I do not think they are able to do such analyses in samples not analysed directly after sampling. NH3/NH4 changes to much during storage - in both ways. 4 years ago there was an urgent need of analysing NH3/NH4 in one tank at my previous workplace and I got that answer from @Christoph and I understand the reasons.

Sincerely Lasse

Oh, that's unfortunate. He only told me that he could give me the data if I wanted. I wasn't aware of the storage issue.

 

[Lasse]

Oh, that's unfortunate. He only told me that he could give me the data if I wanted. I wasn't aware of the storage issue.

Its the transport time - from sampling to the analyse that´s critical - but they may have a solution for it today. This was 4 years ago.

Sincerely Lasse

 

[Dan_P]

Thanks for the analysis. I would also add that all of my samples were collected about two hours after the lights were turned off.

Another possible factor that might have quite a large impact on the diel variation in amino acid composition would be excretion from photosynthetic organisms.

Cellular and extracellular production of carbohydrates and amino acids by the marine diatom Skeletonema costatum: Diel variations and effects of N depletion
This study found that the extracellular amino acid composition of a species of diatom varied both throughout the light cycle and depending on the growth phase it was in.

“Composition of the extracellular free amino acids changed significantly from exponential to stationary growth phase (Table 4). The proportions of acidic and small amino acids (Asp, Glu, Ala, Gly and Ser) decreased, while large hydrophobic amino acids (Ile, Leu, Phe and Val) increased in accordance with intracellular changes. But in contrast with the intracellular pool, extracellular Gln was below detection level during exponential growth, and then slightly increased during the stationary growth phase.”

"The composition of the cellular free amino acids also showed striking diel variation during exponential growth. Glutamine emerged as the principal amino acid during the photophase, effectively increasing the Gln:Glu ratio (Fig. 5). This ratio probably reflects the main route of NH4+ incorporation, the glutamine synthetase-glutamate synthase pathway. Thus, the high rate of N assimilation leads to an increasing Gln:Glu ratio in the photophase, and vice versa in the scotophase."

The mention of the GS-GOGAT pathway also got me thinking about whether the Gln/Glu ratio could be used as an indicator of nitrogen limitation. Most inorganic nitrogen assimilation in organisms in our tanks involves this pathway, in which glutamate is used to assimilate ammonium and form glutamine.

⁠
I plotted the relationship between the log10-transformed Gln/Glu ratio and PO4 concentration, and it did show a significant positive relationship (p = 0.0105).

I have also requested ammonium measurements for my samples from Oceamo, but it might take a while since they are currently on vacation.

Thanks for the reference and your ideas. Both support the type information gathered and ideas proposed by Chat GPT as being reasonably accurate. The AI has been useful but I remain skeptical about what it produces.

I also observed the positive correlation with PO4, but did not come up with any brilliant ideas about what it meant. I do want to point out that the paper reports extracellular Gln is detected during stationary phase growth. This make sense that such an important amino acid for ammonia uptake is found outside the cell during stationary growth. I also observed a change in amino acids during vinegar dosing aquarium bacteria that were in stationary phase growth. I need to revisit that data to see if the extracellular amino acids match those observed in the paper you referenced.

The ammonia analysis of your aquaria will be of interest. I am currently observIng two experimental aquaria being dosed with the same amount of ammonium chloride, one getting a single dose, the other a continuous dose throughout the day. Both are dosed with a single dose of the same amount of PO4.

 

[JonasRoman]

OK, but your absolute statement that it is "not decomposed in nature" is too strong, IMO.

https://strathprints.strath.ac.uk/16363/

In contrast, a follow-up mesocosm-scale field study using low POC water showed that photodegradation could also dominate cipro fate. In conclusion, both adsorption and photodegradation strongly influence cipro fate in aquatic systems, although the dominant mechanism appears to depend upon the ambient POC level.

OK, but your absolute statement that it is "not decomposed in nature" is too strong, IMO.

https://strathprints.strath.ac.uk/16363/

In contrast, a follow-up mesocosm-scale field study using low POC water showed that photodegradation could also dominate cipro fate. In conclusion, both adsorption and photodegradation strongly influence cipro fate in aquatic systems, although the dominant mechanism appears to depend upon the ambient POC level.

The importance is that the degradation of cipro in nature is unsignificant and therefore shouldn't be used by amateurs in a hobby field. We even in human care uses this with restrictions.(I work as a doctor as professional) You message could be misunderstood in opposite directions where we under no circumstances should defend the use of cipro.

 

[Randy Holmes-Farley Verified Community Expert]

The importance is that the degradation of cipro in nature is unsignificant and therefore shouldn't be used by amateurs in a hobby field. We even in human care uses this with restrictions.(I work as a doctor as professional) You message could be misunderstood in opposite directions where we under no circumstances should defend the use of cipro.

OK, I understand what you are saying. I'm not certain I fully agree that hobby use of cipro is a substantial issue (especially for folks on septic systems), but the point is well taken. Overuse of antibiotics by farmers and people is, IMO, a far larger issue, as is the lack of a suitable business model to develop new antibiotics.

Do you have an alternative antibiotic that you would suggest for, say, treatment of a new magnifica anemone that is suffering after transport?

 

[JonasRoman]

OK, I understand what you are saying. I'm not certain I fully agree that hobby use of cipro is a substantial issue (especially for folks on septic systems), but the point is well taken. Overuse of antibiotics by farmers and people is, IMO, a far larger issue, as is the lack of a suitable business model to develop new antibiotics.

Do you have an alternative antibiotic that you would suggest for, say, treatment of a new magnifica anemone that is suffering after transport?

Definitely agree with you that our hobbys potential contribution to resistance development of bacteria are very small, probably negligible at the moment. Let's keep it that way
No unfortunately not, I am not the right person answering that. I guess a veterinary may help us for that question, or if someone know in detail which microbial pattern this disease is caused by, we can figure out.

 

[Lasse]

OK, I understand what you are saying. I'm not certain I fully agree that hobby use of cipro is a substantial issue (especially for folks on septic systems), but the point is well taken. Overuse of antibiotics by farmers and people is, IMO, a far larger issue, as is the lack of a suitable business model to develop new antibiotics.

Do you have an alternative antibiotic that you would suggest for, say, treatment of a new magnifica anemone that is suffering after transport?

Definitely agree with you that our hobbys potential contribution to resistance development of bacteria are very small, probably negligible at the moment. Let's keep it that way
No unfortunately not, I am not the right person answering that. I guess a veterinary may help us for that question, or if someone know in detail which microbial pattern this disease is caused by, we can figure out.

First - the type of arguments in bold - its the typical answer when every one meets demands to change a behaviour - I do not think that my use of that or that will contribute to the actual problem - however If every one other than me can have it or use it - that´s a problem

However there is one more dimension to this problem - and that´s the individual aquarium and the risk that something bad would develop inside this bacteria farm that we name an aquarium. Use of broad spectra antibiotics in a closed bacteria driven system as an aquarium can very fast cause that various bacteria (and other microorganisms with short generation cycles) can quickly develop resistance and if the misfortune comes, it is a zoonotic organism that acquires these properties. Zoonotic means - fatal for other organism including humans.

It is also the case that if microorganisms acquire resistance to various substances early in the distribution chain - then this resistant bacterial culture will spread further to wholesalers, shops and individual aquariums. During the years that I have been active in the hobby, I have seen tendencies towards this - not only when it comes to antibiotics but also an increased resistance to other substances.

Sincerely Lasse

 

[Dan_P]

, I have seen tendencies towards this - not only when it comes to antibiotics but also an increased resistance to other substances.

Lasse,

I am interested in learning about examples of the increased resistance to other substances.

Dan

 

[Lasse]

Lasse,

I am interested in learning about examples of the increased resistance to other substances.

Dan

Can´t quantify this - its more of a feeling - it does not need to be true but I think the recommended dos of Cu has rise a little with time. Also when we use formaldehyde against gill worms (egg laying) in a couple of eel farms - we had to rise the dosage by a time scale. Its also known that Norwegian Salmon farmers today have to use higher dosage of H2O2 against "Laxeluse" compared when the method was first introduced This treatment happens in cages in open sea - not indoors!

Sincerely Lasse

 

[Randy Holmes-Farley Verified Community Expert]

First - the type of arguments in bold - its the typical answer when every one meets demands to change a behaviour - I do not think that my use of that or that will contribute to the actual problem - however If every one other than me can have it or use it - that´s a problem

However there is one more dimension to this problem - and that´s the individual aquarium and the risk that something bad would develop inside this bacteria farm that we name an aquarium. Use of broad spectra antibiotics in a closed bacteria driven system as an aquarium can very fast cause that various bacteria (and other microorganisms with short generation cycles) can quickly develop resistance and if the misfortune comes, it is a zoonotic organism that acquires these properties. Zoonotic means - fatal for other organism including humans.

It is also the case that if microorganisms acquire resistance to various substances early in the distribution chain - then this resistant bacterial culture will spread further to wholesalers, shops and individual aquariums. During the years that I have been active in the hobby, I have seen tendencies towards this - not only when it comes to antibiotics but also an increased resistance to other substances.

Sincerely Lasse

Yes, thanks.

For a number of years I worked on an antimicrobial drug program for exactly this reason: development of resistance to current antibiotics. I agree that resistance is a big issue.

 

[JonasRoman]

First - the type of arguments in bold - its the typical answer when every one meets demands to change a behaviour - I do not think that my use of that or that will contribute to the actual problem - however If every one other than me can have it or use it - that´s a problem

However there is one more dimension to this problem - and that´s the individual aquarium and the risk that something bad would develop inside this bacteria farm that we name an aquarium. Use of broad spectra antibiotics in a closed bacteria driven system as an aquarium can very fast cause that various bacteria (and other microorganisms with short generation cycles) can quickly develop resistance and if the misfortune comes, it is a zoonotic organism that acquires these properties. Zoonotic means - fatal for other organism including humans.

It is also the case that if microorganisms acquire resistance to various substances early in the distribution chain - then this resistant bacterial culture will spread further to wholesalers, shops and individual aquariums. During the years that I have been active in the hobby, I have seen tendencies towards this - not only when it comes to antibiotics but also an increased resistance to other substances.

Sincerely Lasse

My statement that you choose to bold is not a "typical answer" as its meaning here is emphasis exactly the same concern as you have, risk of resistance development.

Randy is right though that the hobby contribution to this is at this moment very small though.

Concerning cipro, we use this as a standard antibiotics in abdominal infections. Just an example to clarify that aquarium use of this is not measurable compared to human medicin. Randy is right there aswell.
But again, we shall ofcourse anyway do our own contributions to this and avoid when possible.

I do not think any of us disagree.

Jonas

 

[JonasRoman]

Lasse,

I am interested in learning about examples of the increased resistance to other substances.

Dan

Me too. Despite I work with this I have never heard about that that Lasse claims. Maybe my microbiology friends or colleagues working with infectious disease knows, but I doubt we have seen such crossresistance developments.

 

[Lasse]

Me too. Despite I work with this I have never heard about that that Lasse claims. Maybe my microbiology friends or colleagues working with infectious disease knows, but I doubt we have seen such crossresistance developments.

I´ll think you totally missunderstand me - I did not mean that I have a feeling that acquired resistance against antibiotics make the organism resistant to other compounds - what I try to say that I have a feeling that other compounds than antibiotics can trigger resistance towards the actual compound among short life cycle organisms like bacteria and other microorganisms - as my examples in post #29.

About crossresistance development - interesting question because development of group-resistance in short life-cycle organisms is either favour of individuals that already have an genetical resistance or favour mutations in the gen-pool. And we do not know which gen-pools the surviving individuals have in addition to resistance against a special compound. There are examples of genetical factors for one thing that can ad a type of resistance against something else - Individuals with only one copy of the gene for Sickle cell anemia and malaria is one schoolbook example

Sincerely Lasse

 

[JonasRoman]

I´ll think you totally missunderstand me - I did not mean that I have a feeling that acquired resistance against antibiotics make the organism resistant to other compounds - what I try to say that I have a feeling that other compounds than antibiotics can trigger resistance towards the actual compound among short life cycle organisms like bacteria and other microorganisms - as my examples in post #29.

About crossresistance development - interesting question because development of group-resistance in short life-cycle organisms is either favour of individuals that already have an genetical resistance or favour mutations in the gen-pool. And we do not know which gen-pools the surviving individuals have in addition to resistance against a special compound. There are examples of genetical factors for one thing that can ad a type of resistance against something else - Individuals with only one copy of the gene for Sickle cell anemia and malaria is one schoolbook example

Sincerely Lasse

yes you are correct concerning sickle cell etc. We have more examples of genetic varations that can give favour for the individual. But thats far away from the subject in this thread.

Thanks for clarification. Developing higher tolerance against other compounds I do not think is nothing new.

 

[Dan_P]

Can´t quantify this - its more of a feeling - it does not need to be true but I think the recommended dos of Cu has rise a little with time. Also when we use formaldehyde against gill worms (egg laying) in a couple of eel farms - we had to rise the dosage by a time scale. Its also known that Norwegian Salmon farmers today have to use higher dosage of H2O2 against "Laxeluse" compared when the method was first introduced This treatment happens in cages in open sea - not indoors!

Sincerely Lasse

Thanks Lasse. These are interesting examples to consider.

 

[ReneReef]

I'm not certain I fully agree that hobby use of cipro is a substantial issue (especially for folks on septic systems), but the point is well taken. Overuse of antibiotics by farmers and people is, IMO, a far larger issue, as is the lack of a suitable business model to develop new antibiotics.

I trust you know your argument here is a fallacy.

 

[Randy Holmes-Farley Verified Community Expert]

I trust you know your argument here is a fallacy.

Maybe. IMO, one has to consider cost vs benefit for a lot of things. I think it is entirely possible there is no or maybe very low environmental cost from the levels of Cipro released by hobbyists.

In tank use and release of those bacteria seems more likely to be an issue, IMO.