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Dna Barcoding Aquarium Cyprinids
January 21, 2012
1:44 am
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racoll
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Open access (free download) article in PLoS ONE: Barcoding and Border Biosecurity: Identifying Cyprinid Fishes in the Aquarium Trade. Seriouslyfish.com gets a mention!

If anyone is interested, there is a large amount of supplementary information associated with this paper, including a huge table of morphological data on distinguishing aquarium cyprinids, and Web-based interactive trees where taxon names are clickable, and the links take you to database pages with photos of the fishes (unfortunately this interactivity is not available in Internet Explorer, but you can still see the trees).

There were some interesting findings here, as common misidentifications were observed, and a whole host of familiar aquarium species may comprise multiple species. Some of these were already known, and this was confirmed with DNA.

Danio choprae, D. dangila, D. kyathit, Devario devario, Epalzeorhynchos kalopterus, Microdevario kubotai, Microrasbora rubescens, Puntius assimilis, P. denisonii, P. fasciatus, P. gelius, P. lateristriga, P. stoliczkanus, Rasbora dorsiocellata (now cheeya), R. einthovenii, R. heteromorpha, R. maculata, R. pauciperforata and Sundadanio axelrodi (now multiple spp.).

One was particularly interesting, being the harlequin, Rasbora/Trigonostigma heteromorpha. I am not sure if anyone has noticed, but there appears to be two types. One a large, deep bodied fish with a plain anal fin (R. heteromorpha), and another more delicate fish with a orange blotch on the anal fin (R. cf. heteromorpha). There were other morphological differences too.

Puntius gelius was also very interesting, with at least two types found in the trade (being easily distinguished by presence or absence of black spots at base of pelvic fins), and differing so much in DNA (both mitochondrial and nuclear), that they don't even appear particularly closely related.

The images are all Creative Commons licensed, so can be copied and altered freely providing the source is acknowledged. Would be great to see some of these on SF! The image library is somewhat hidden though, but can be found eventually. Go to BOLD, then "published projects" on the left of the top menu bar, then search (ctrl+f) for "Ornamental Cyprinidae", then click the "Image library" link at the bottom of the left hand menu. There are two pages of images, but the link to the second page is only at the top.

January 21, 2012
10:41 am
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Matt
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At last!!!! Congratulations Rupert and thanks for posting and the mention!

We'd love to use the images and I'll sort it out asap.

Is the other form of P. gelius the larger one? Think we talked about the possibility of it being P. canius?

Still haven't been able to bring myself to change the Trigonostigma profiles to Rasbora. /tongue.gif" style="vertical-align:middle" emoid=":p" border="0" alt="tongue.gif" />

Cake or death?
January 22, 2012
2:10 am
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racoll
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Thanks Matt!

QUOTE ("Matt")
Is the other form of P. gelius the larger one? Think we talked about the possibility of it being P. canius?

I found three types of "P. gelius". Two from the trade, and another kindly donated to me by Heok Hee. I don't know where he got that one from, but I didn't find any others like it in the trade.

This is it Heok Hee's. The specimens matched published P. gelius characters, but it is a darker and larger fish than the "regular" P. gelius. The difference in DNA is consistent with it being a separate species from the regular one.
Image

This is the "regular" P. gelius from the trade, and the one that is called P. canius in your P. gelius profile. This one is most consistent with the description of P. gelius.
Image

Now this one has completely different DNA to both those above, and does not appear to be closely related to either (I didn't do a phylogeny, so this remains to be tested more thoroughly). It is common in the trade, and is the one labelled P. gelius in your profile. It is not P. gelius, and is distinct in numerous morphological characters: most notably, a lack of spots on pelvics, and the spot on the anal fin extending well onto the body. I believe this is undescribed, although P. ornatus Vishwanath & Laisram is a possibility. I referred to it as P. aff. gelius.
Image

Now, which of these is P. canius? That species is shown to have distinct spots on the pelvics, as illustrated by Hamilton in McClelland (1839). That rules out the last one. Puntius canius is described as a smaller fish (than P. gelius) with a reddish hue; I did not record this colouration in any my specimens, and it may be a seasonal breeding effect. Bordoloi and Baishya (2006) report this colouration from specimens of "P. ornatus" from Assam, and the specimens they picture appear similar, but I am not sure these are P. ornatus as described. The description of that species being not being terribly informative, however. There are certainly more forms of this complex out there, and it will require a more study before we can get to the bottom of it (especially regarding the real identity of the elusive P. canius)

QUOTE
Still haven't been able to bring myself to change the Trigonostigma profiles to Rasbora.

That would be a personal choice I think. I was asked about this on PC. Seems taxonomists are ignoring the work of Tang et al. (2010), so it might be better to stick with Liao's genera? I don't personally have any favouritism to either lab group, or with molecules vs. morphology being superior. It is my philosophy that seeing as there's conflict between the data, then it is best to be conservative until this issue is resolved by a study directly focussing on that. Treating them all as Rasbora seems to me like the safest option for now. I am probably in a minority with that kind of opinion, however.

January 23, 2012
10:02 am
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Matt
Málaga, Spain
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Thanks for the detailed explanation Rupert - all very interesting stuff and it appears I've some editing to do and at least one additional profile to add! Do you have locality details for the three Puntius specimens pictured?

I actually agree with you with regards to adopting a conservative approach towards Rasbora in the absence of a full revision, but post-publication of Tang et al. most people I asked preferred to retain Trigonostigma, Boraras, Brevibora, etc. so that's what we did.

Cake or death?
January 24, 2012
12:09 am
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racoll
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QUOTE
Do you have locality details for the three Puntius specimens pictured?

Sadly not. They were bought from retailers. Maybe some of the Indian members can help out here?

QUOTE
it appears I've some editing to do and at least one additional profile to add!

You could also consider adding a few more. For example:

I found Puntius hexazona was well differentiated from P. pentazona in terms of DNA, and can readily be distinguished by the spot at the end of the dorsal fin. Funnily, they didn't seem that closely related, not grouping together in either mtDNA or nDNA trees (again, however, an explicit phylogenetic analysis was not carried out).

The striped and spotted Danio kyathit were very different in terms of DNA, and this was consistent with them being different species. There were other morphological differences too. The name D. kyathit applies to the spotted one. I referred to the striped one as D. aff. kyathit.

The fishes I called D. aff. choprae and D. aff. dangila were also different in terms of DNA and morphological characters. They are likely different species too.

There are more, and I can update when these come up in discussion.

January 24, 2012
8:47 am
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Matt
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QUOTE (racoll @ Jan 24 2012, 12:52 AM) < {POST_SNAPBACK}>
The striped and spotted Danio kyathit were very different in terms of DNA, and this was consistent with them being different species. There were other morphological differences too. The name D. kyathit applies to the spotted one. I referred to the striped one as D. aff. kyathit.

Ok will make this change.

QUOTE (racoll @ Jan 24 2012, 12:52 AM) < {POST_SNAPBACK}>
The fishes I called D. aff. choprae and D. aff. dangila were also different in terms of DNA and morphological characters. They are likely different species too.

Is aff. choprae the 'northern glowlight danio' that was in the trade a few years back?

QUOTE (racoll @ Jan 24 2012, 12:52 AM) < {POST_SNAPBACK}>
There are more, and I can update when these come up in discussion.

Yes please!

Cake or death?
January 25, 2012
12:04 am
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racoll
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QUOTE (Matt @ Jan 24 2012, 09:30 PM) < {POST_SNAPBACK}>
Is aff. choprae the 'northern glowlight danio' that was in the trade a few years back?

Yes, the larger grey one.
Image

There was also, surprisingly, a heck of a lot of molecular diversity in "regular" D. choprae. Unfortunately neither my specimens or my images were particularly good for this species, so I could not tell if any of these splits were backed up by morphological differences. Seems strange given their relatively restricted distribution.

January 25, 2012
7:09 pm
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Matt
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That's the one. /smile.gif" style="vertical-align:middle" emoid=":)" border="0" alt="smile.gif" /> Were the 'regular' D. choprae you used from the aquarium trade?

Cake or death?
January 25, 2012
10:37 pm
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racoll
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QUOTE
Were the 'regular' D. choprae you used from the aquarium trade?

Correct, almost every fish in the study was sourced from the aquarium trade.

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