TY - JOUR
T1 - Methodological guidelines for minimally invasive tail-clipping
T2 - a case study on Rana huanrenensis tadpoles
AU - Othman, Siti N.
AU - Chuang, Ming Feng
AU - Kang, Hakyung
AU - Bae, Yoonhyuk
AU - Kim, Ajoung
AU - Jang, Yikweon
AU - Borzee, Amael
N1 - Funding Information:
ments on this manuscript. We are grateful to the local authorities of Nowon-gu for the permission to conduct this study on the tadpoles. This study was supported by a research grant from the Korean Environmental Industry and Technology Institute through ‘Development of behavioural ecological methods for population regulation of invasive species’ funded by Korea Ministry of Environment (KEITI 2017002270003), and a research grant from Rural Development Administration (PJ01507102) to YJ. The funders had no role in the design and conduct of the study.
Publisher Copyright:
© 2020 TCRP Foundation. All Rights Reserved.
PY - 2020
Y1 - 2020
N2 - Tail clipping of amphibian tadpoles is one of the minimally invasive, non-lethal procedures, (apart from skin swabbing) used to collect tissues without euthanising the target individual. It is commonly used for species identification, especially when the continuity between the tadpoles and adult stages is not known. However, there is a lack of published standard and safe protocol for tail clipping of anuran tadpoles. To determine the efficiency of the protocol defined herein, we tail clipped 3.0 mm of four Rana huanrenensis tadpoles (Fei, Ye & Huang, 1990), two at each of the Gosner stages 34 and 41. We observed the tails resorbing from tail length = 20.625 ± 0.64 mm on day 0 post-clipping to 5.75 ± 3.49 mm on day 6 post-clipping. During this period, metamorphosis progressed for individuals tail-clipped at Gosner stage 34 (total length: 33.75 ± 2.35 mm; day 0 post-clipping) to Gosner stage 43 (total length: 28.5 ± 3.47 mm; day 6 post-clipping); and individuals tail-clipped at Gosner stage 41 (total length: 35.75 ± 0.35 mm; day 0 post-clipping) to Gosner stage 46 (total length: 15.00 ± 0.00 mm; day 6 post-clipping). We did not record any fatality during the experiment. DNA extracted from 3.0 mm of tail tip tissue yielded gDNA concentrations between 10 and 32 ng/µl. a sufficient amount for barcoding and fingerprinting. We conclude that this protocol is adequate for R. huanrenensis and Ranidae in general, and it is safe for tadpoles at Gosner stage 34 and above.
AB - Tail clipping of amphibian tadpoles is one of the minimally invasive, non-lethal procedures, (apart from skin swabbing) used to collect tissues without euthanising the target individual. It is commonly used for species identification, especially when the continuity between the tadpoles and adult stages is not known. However, there is a lack of published standard and safe protocol for tail clipping of anuran tadpoles. To determine the efficiency of the protocol defined herein, we tail clipped 3.0 mm of four Rana huanrenensis tadpoles (Fei, Ye & Huang, 1990), two at each of the Gosner stages 34 and 41. We observed the tails resorbing from tail length = 20.625 ± 0.64 mm on day 0 post-clipping to 5.75 ± 3.49 mm on day 6 post-clipping. During this period, metamorphosis progressed for individuals tail-clipped at Gosner stage 34 (total length: 33.75 ± 2.35 mm; day 0 post-clipping) to Gosner stage 43 (total length: 28.5 ± 3.47 mm; day 6 post-clipping); and individuals tail-clipped at Gosner stage 41 (total length: 35.75 ± 0.35 mm; day 0 post-clipping) to Gosner stage 46 (total length: 15.00 ± 0.00 mm; day 6 post-clipping). We did not record any fatality during the experiment. DNA extracted from 3.0 mm of tail tip tissue yielded gDNA concentrations between 10 and 32 ng/µl. a sufficient amount for barcoding and fingerprinting. We conclude that this protocol is adequate for R. huanrenensis and Ranidae in general, and it is safe for tadpoles at Gosner stage 34 and above.
KW - tail clip. Huanren Frog. larvae. DNA collection. minimally invasive
UR - http://www.scopus.com/inward/record.url?scp=85100645277&partnerID=8YFLogxK
M3 - Article
AN - SCOPUS:85100645277
SN - 2278-7666
VL - 9
SP - 188
EP - 195
JO - Asian Journal of Conservation Biology
JF - Asian Journal of Conservation Biology
IS - 2
ER -