Sexing as a tool for reinsertion of Amazona aestiva parrots to nature: use of less invasive technique
DOI:
https://doi.org/10.33448/rsd-v10i12.20601Keywords:
CHD-Z/W; PCR; Molecular sexing; Primer 2550F/2718R.Abstract
The sexing birds is considered an important tool for behavioral studies and programs for the reintroduction of animals into the wild. Several techniques are used for this purpose, such as laparoscopy, magnetic resonance and molecular sexing. The first are considered more invasive and stressful for the animal, and the last is considered the most accurate. According to it, the aim of this study was to compare the effectiveness of using three sets of primers in the molecular sexing process of true parrots (Amazona aestiva). Blood samples from 10 animals were collected at a Wildlife Screening Center (CETAS) in Bahia, Brazil. The DNA was extracted and the molecular markers amplified by Polymerase Chain Reaction (PCR) using primer pairs P2/P8, 1237L/1272H and 2250F/2718R. The amplified material was visualized with electrophoresis performed at 2% agarose and 12.5% polyacrylamide gels. Among the primer sets used, the 2250F/2718R pair showed the best results for the sexing process, including visualization of the amplified products on an agarose gel. Agarose gel electrophoresis is considered to be faster and cheaper. The results revealed a sample composition of 5 males (0.5) and 5 females (0.5).
References
Ágh, N., Kovács, Sz., Nemesházi, E., & Szabó, K. (2018). Univerzális, ivarhatározáshoz használt CHD1 markerek alkalmazhatósága különböző madárrendekben. Magyar állatorvosok lapja, 140(1), 47-59.
Allgayer, M. D. C., & Cziulik, M. (2007). Reprodução de psitacídeos em cativeiro. Revista Brasileira de Reprodução Animal, 31(3), 344-350.
Baker, A. J., Piersma, T., & Greenslade, A. D. (1999). Molecular vs. phenotypic sexing in Red Knots. The Condor, 101(4), 887-893.
Bosnjak, J., Stevanov-Pavlovic, M., Vucicevic, M., Stevanovic, J., Simeunovic, P., Resanovic, R., & Stanimirovic, Z. (2013). Feasibility of non-invasive molecular method for sexing of parrots. Pakistan Journal Zoology, 45(3), 715-720.
Brasil. (2015). Instrução Normativa ICMBio nº 23, de 31 de dezembro de 2014. Define as diretrizes e os procedimentos para a destinação de animais silvestres apreendidos, resgatados por autoridade competente ou entregues voluntariamente pela população, bem como para o funcionamento dos Centros de Triagem de Animais Silvestres do IBAMA - CETAS. Diário Oficial da União, seção 1, Brasília, DF, 115-118, 02 jan. 2015.
Casey, A. E., Jones, K. L., Sandercock, B. K., & Wisely, S. M. (2009). Heteroduplex molecules cause sexing errors in a standard molecular protocol for avian sexing. Molecular Ecology Resources, 9(1), 61-65.
Dalfovo, M. S., Lana, R. A., & Silveira, A. (2008). Métodos quantitativos e qualitativos: um resgate teórico. Revista Interdisciplinar Científica Aplicada, 2(4), 01-13.
Dash, S. K., Kumarasamy, P., Malik, H. N., & Thiagarajan, V. (2013). Phylogenetic analysis of ratite bird emu (Dromaius novaehollandiae) using sex specific sequences. Indian Journal of Animal Research, 47(3), 247-250.
Dawson, D. A., Darby, S., Hunter, F. M., Krupa, A. P., Jones, I. L., & Burke, T. (2001). A critique of avian CHD‐based molecular sexing protocols illustrated by a Z‐chromosome polymorphism detected in auklets. Molecular Ecology Notes, 1(3), 201-204.
Del Puerto, F., Pésole, D., Molina, S., Vera, K., Arias, M., Sosa, J., Ortiz, M. L., Fernández, J., & Garay, A. (2017). Identificación molecular del sexo en 9 especies de aves del Centro de Investigación en Animales Silvestres de la hidroeléctrica de ITAIPU, lado paraguayo. Memorias del Instituto de Investigaciones en Ciencias de la Salud, 15(3), 89-92.
Desjardins, P., & Conklin, D. (2010). NanoDrop microvolume quantitation of nucleic acids. Journal of visualized Experiments, 45, e2565.
Dos Remedios, N., Lee, P. L. M., Szekely, T., Dawson, D. A., & Kupper C. (2010). Molecular sex‐typing in shorebirds: a review of an essential method for research in evolution, ecology and conservation. Wader study group bulletin, 117(2), 109‐118.
Ellegren, H. (1996). First gene on the avian W chromosome (CHD) provides a tag for universal sexing of non-ratite birds. Proceedings of the Royal Society of London Series B, 263(1377), 1635-1641.
Franco-Gutiérrez, L. J., Álvarez-Cardona, J., & Soto-Calderón, I. D. (2017). Sex identification of neotropical macaws (Ara spp.) from invasive and non-invasive samples. Ornitología Colombiana, (16), eNB03, 01-07.
Freitas, A. C. P., Oviedo-Pastrana, M. E., Vilela, D. A. R., Pereira, P. L. L., Loureiro, L. O. C., Haddad, J. P. A., Martins, N. R. S., & Soares, D. F. M. (2015). Diagnóstico de animais ilegais recebidos no centro de triagem de animais silvestres de Belo Horizonte, Estado de Minas Gerais, no ano de 2011. Ciência Rural, 45(1), 163-170.
Fridolfsson, A-K., & Ellegren, H. (1999). A simple and universal method for molecular sexing of non-ratite birds. Journal of avian biology, 30(1), 116-121.
Griffiths, R., Daan, S., & Dijkstra, C. (1996). Sex identification in birds using two CHD genes. Proceedings of the Royal Society of London. Series B: Biological Sciences, 263, 1251-1256.
Griffiths, R., Double, M. C., Orr, K., & Dawson, R. J. G. (1998). A DNA test to sex most birds. Molecular ecology, 7(8), 1071-1075.
Gonçalves, B. P. (2013). Sexagem molecular em aves: contribuições para a conservação biológica e divulgação científica. Master’s thesis, Universidade Estadual Paulista, Botucatu, SP, Brasil. http://hdl.handle.net/11449/108613
Ibama. (2016). O que são CETAS? http://www.ibama.gov.br/fauna-silvestre/cetas/o-que-sao-os-cetas
Jensen, T., Pernasetti, F. M., & Durrant, B. (2003). Conditions for rapid sex determination in 47 avian species by PCR of genomic DNA from blood, shell-membrane blood vessels, and feathers. Zoo Biology, 22(6), 561-571.
Khan, N. W., St. John, J., & Quinn, T. W. (1998). Chromosome-specific intron size differences in the avian CHD gene provide an efficient method for sex identification in birds. The Auk, 115(4), 1074-1078.
Maheshkumar, G., Saravanan, R., Mani, K., & Murali, N. (2017). Molecular marker based sex identification in ratite and non-ratite type of birds: A comparative study. Indian Veterinary Journal, 94(5), 11-13.
Malaitad, T., Poeaim, S., & Eiamampai, K. (2015). Sex identification in barn swallows (Hirundo rustica Linnaeus) by molecular technique. Journal of Agricultural Technology, 11(8), 2411-2418.
Miyaki, C. Y., Griffiths, R., Orr, K., Nahum, L. A., Pereira, S. L., & Wanjtal, A. (1998). Sex identification of parrots, toucans, and curassows by PCR: perspectives for wild and captive population studies. Zoo Biology, 17(5), 415-423.
Monge, O., Dumas, D., & Baus, I. (2018). Environmental DNA from avian residual saliva in fruits and its potential uses in population genetics. Conservation Genetics Resources, 12, 131-139.
Morinha, F., Cabral, J. A., & Bastos, E. (2012). Molecular sexing of birds: A comparative review of polymerase chain reaction (PCR) -based methods. Theriogenology, 78(4), 703-714.
Ong, A. H. K., & Vellayan, S. (2008). An evaluation of CHD-Specific primer sets for sex typing of birds from feathers. Zoo Biology, 27(1), 62-69.
Pereira, D. C., Mirabal, B., França, A. M. M., Antonio, E. S., Fraga, R. E., & Tomazi, L. (2021). Sexagem molecular na formação de pares de arara-canindé (Ara ararauna) em programas de reintrodução. Research, Society and Development, 10(10), 570101019330.
Pereira, A. S., Shitsuka, D. M., Parreira, F. J., & Shitsuka R. (2018). Metodologia da pesquisa científica (1a ed.). [free e-book]. Santa Maria: Núcleo de Tecnologia Educacional da Universidade Federal de Santa Maria. https://repositorio.ufsm.br/handle/1/15824
Piacentini, V. Q., Aleixo, A., Agne, C. E., Maurício, G. N., Pacheco, J. F., Bravo, G. A., Brito, G. R. R., Naka, L. N., Olmos, F., Posso, S., Silveira, L. F., Betini, G.S., Carrano, E., Franz, I., Lees, A. C., Lima, L. M., Pioli, D., Schunck, F., Amaral, F. R., Bencke, G. A., Cohn-Haft, M., Figueiredo, L. F. A., Straube, F. C., & Cesari, E. (2015). Annotated checklist of the birds of Brazil by the Brazilian Ornithological Records Committee. Revista Brasileira de Ornitologia, 23(2), 91-298.
Renctas – Rede Nacional de Combate ao Tráfico de Animais Silvestres. Brasília. (2016). I Relatório Sobre Gestão e Uso Sustentável da Fauna Silvestre. http://www.renctas.org.br/i-relatorio-sobre-gestao-e-uso-sustentavel-da-fauna-silvestre/
Ribeiro, L. B., & Silva, M. G. (2007). O comércio ilegal põe em risco a diversidade das aves no Brasil. Ciencia e Cultura, 59(4), 4-5.
Sambrook, J., Fritsch, E. F., & Maniatis, T. (1989). Molecular cloning: a laboratory manual (Vol. 3, 2a ed.). New York: Cold Spring Harbor Laboratory Press.
Shizuka, D., & Lyon, B. E. (2008). Improving the reliability of molecular sexing of birds using a W-specific marker. Molecular Ecology Resources, 8(6), 1249-1253.
Silva, R. M. N., Figueiredo, P. O., & Santana, M. I. (2015). Development and distribution of the brachial plexus in Blue-fronted Parrot (Amazona aestiva, Linnaeus, 1758). Ciência Animal Brasileira, 16(3): 464-473.
Vieira, J. N. (2009). Sexagem molecular em aves via PCR: avaliação de três técnicas de extração de DNA. Master’s thesis, Universidade Federal de Minas Gerais, Belo Horizonte, MG, Brasil. http://www.dominiopublico.gov.br/pesquisa/DetalheObraForm.do?select_action=&co_obra=135898
Vieira, J. N., Coelho, E. G. A., Teixeira, C. S., & Oliveira, D. A. A. (2012). PCR as a technique for molecular sexing in birds. Revista brasileira de Reprodução Animal, 36(3), 199-201.
Vucicevic, M., Stevanov‐Pavlovic, M., Stevanovic, J., Bosnjak, J., Gajic, B., Aleksic, N., & Stanimirovic, Z. (2012). Sex determination in 58 bird species and evaluation of CHD gene as a universal molecular marker in bird sexing. Zoo biology, 32(3), 269-276.
Wang, L., Chen, C., Lee, H., Li, S., Lir, J., Chin, S., Pu, C., & Wang, C. (2007). Sexing a wider range of avian species based on two CH3D1 introns with a unified reaction condition. Zoo Biology, 26(5), 425-431.
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2021 Carine Novato Ramos; Tácita Borges Barros; Edma Santos de Antonio; Bernardo Mirabal; Márcio Borba da Silva; Ricardo Evangelista Fraga; Laize Tomazi
This work is licensed under a Creative Commons Attribution 4.0 International License.
Authors who publish with this journal agree to the following terms:
1) Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
2) Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
3) Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work.
