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Falha na sexagem por inibição do desenvolvimento de embriões bovinos produzidos in vitro com anticorpos anti H-Y; Failure of sexing by developmental arrest of bovine embryos in vitro produced with H-Y antisera

RESENDE, M.V.; MOREIRA-FILHO, C.A.; LEAL, C.L.V.; RAMALHO, M.F.P.D.; ALMEIDA, A.O.; VANTINI, R.; HOSSEPIAN DE LIMA, V.F.M.
Fonte: Universidade Federal de Minas Gerais, Escola de Veterinária Publicador: Universidade Federal de Minas Gerais, Escola de Veterinária
Tipo: Artigo de Revista Científica
POR
Relevância na Pesquisa
37.34%
Embriões bovinos produzidos in vitro, em estádio de mórula, foram cultivados em meio contendo anticorpos anti H-Y de alto título proveniente de ratos por 24h e, após este tempo, classificados em dois grupos: 1) embriões inibidos em estádio de mórula (classificados como machos) e 2) embriões que se desenvolveram e formaram a blastocele (classificados como fêmeas). O sexo de 311 embriões, distribuídos em três grupos de concentração dos anticorpos, 3%, 5% ou 7%, foi identificado pela reação em cadeia da polimerase. Não houve desvio da proporção entre machos e fêmeas (P>0,05) nos grupos em que se utilizaram os anticorpos anti H-Y, quando comparadas ao grupo-controle, sem adição de anticorpos anti H-Y. Diferentemente dos resultados obtidos utilizando-se embriões bovinos produzidos in vivo, a sexagem com anticorpos anti H-Y de alto título em embriões produzidos in vitro não propiciou sucesso.; In vitro produced bovine embryos at morula stage were cultured in medium containing high titer of rat H-Y antisera for 24h. The embryos were classified in two groups: 1) embryos arrested at morula stage (classified as males); and 2) embryos that developed and formed a blastocoele (classified as female). The sex of 311 embryos...

MHM assay: molecular sexing based on the sex-specific methylation pattern of the MHM region in chickens

CAETANO, Lisandra Cristina; RAMOS, Ester Silveira
Fonte: SPRINGER Publicador: SPRINGER
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
37.73%
Bird sex determination using molecular methods has proved to be a valuable tool in different studies. Although it is possible to sex most birds by coupling the CHD assay with others available methods, no sex-determining gene like SRY in mammalians has been identified in birds. The male hypermethylated (MHM) region on the Z chromosome has been found to be hypermethylated in males and hypomethylated in females in birds of the order Galliformes. We analyzed the DNA from feathers of 50 adult chickens to verify the methylation pattern of the MHM region by PCR and the restriction enzyme HpaII (a method named MHM assay). The results, visualized in agarose gel, were compared with PCR amplification of the CHD-Z and CHD-W genes (polyacrylamide gel) and with the birds` phenotype. All males (25) showed hypermethylation of the MHM region, and all females (25) showed hypomethylation. The sexing by MHM assay was in according with phenotype and CHD sexing. To our knowledge, this is the first study that uses the MHM region for sexing birds. Although the real role of the MHM region in the sex determination is still unclear, this could be a universal marker for sexing birds and may be involved in sex determination by its influence on transcriptional processes. The MHM assay could be a good alternative for CHD assay in developmental studies.

Sexing of murine and bovine embryos by developmental high-titer arrest induced by H-Y antisera

Ramalho, MFPDT; Garcia, J. M.; Esper, C. R.; Vantini, R.; Alves, BCA; Almeida, I. L.; de Lima, VFMH; Moreira, C. A.
Fonte: Elsevier B.V. Publicador: Elsevier B.V.
Tipo: Artigo de Revista Científica Formato: 1569-1576
ENG
Relevância na Pesquisa
37.64%
Murine and bovine embryos at the late morula stage were cultured in medium containing high-titer rat H-Y antisera. After 12 h of incubation, embryos blocked at the late morulae stage were classified as males and those at the blastocyst stage were classified as females. Sexing of murine embryos by PCR and cytogenetics revealed that 83% of the embryos classified as males and 82% of those classified as females had their sex correctly predicted (P < 0.05). Bovine embryos were transferred to recipient females. Pregnancy rates were 71.4% (10/14) for embryos classified as males and 68.8% (11/16) for embryos classified as females. The sex was correctly predicted for 80% (8/10) of the embryos classified as males and for 81.8% (9/11) of those classified as females (overall accuracy, 80.9%, P < 0.05). Therefore, the induction of developmental arrest by high-titer male-specific antisera was an efficient strategy for non-invasive embryo sexing. The procedure was straightforward and has considerable commercial potential for sexing bovine embryos. (c) 2004 Elsevier B.V. All rights reserved.

Development of a real-time PCR method for rapid sexing of human preimplantation embryos

Martinhago, C. D.; Vagnini, L. D.; Petersen, C. G.; Mauri, A. L.; Baruffi, R. L.; de Oliveira, R. M.; Franco, J. G.
Fonte: Reproductive Healthcare Ltd Publicador: Reproductive Healthcare Ltd
Tipo: Artigo de Revista Científica Formato: 75-82
ENG
Relevância na Pesquisa
37.34%
Genes on the X chromosome are known to be responsible for more than 200 hereditary diseases. After IVF, the simple selection of embryo sex before uterine transfer can prevent the occurrence of affected offspring among couples at risk for these genetic disorders. The aim of this investigation was to develop a rapid method of preimplantation genetic diagnosis (PGD) using real-time polymerase chain reaction (PCR) for the sexing of human embryos, and to compare it to the fluorescence in-situ hybridization technique, considered to be the gold standard. After biopsies were obtained from 40 surplus non-viable embryos for transfer, a total of 98 blastomeres were analysed. It was possible to analyse 24 embryos (60%) by both techniques, generating a total of 70 blastomeres (35 per technique), white 28 blastomeres from 16 embryos (40%) were analysed only by real-time PCR. A rapid and safe method was developed in the present study for the sexual diagnosis of a single human cell (blastomere and buccal cell) using the emerging technology of real-time PCR. (C) 2009, Reproductive Healthcare Ltd. Published by Elsevier Ltd. All rights reserved.

Bovine carcass sexing by PCR method

Curi, Rogério Abdallah; Mota, Lígia Souza Lima Silveira da; Amarante, Mônica Regina Vendrame; Lopes, Catalina Romeiro
Fonte: Universidade de São Paulo (USP), Faculdade de Medicina Veterinária e Zootecnia (FMVZ) Publicador: Universidade de São Paulo (USP), Faculdade de Medicina Veterinária e Zootecnia (FMVZ)
Tipo: Artigo de Revista Científica Formato: 147-148
ENG
Relevância na Pesquisa
37.34%
Objetivou-se, nesta pesquisa, o desenvolvimento de uma metodologia que permitisse a sexagem de carne bovina pronta para comercialização. Para tanto, utilizou-se primers seqüência macho-específica e posterior análise do produto amplificado. O método proposto mostrou-se eficiente para verificar o sexo, bem como sua utilização prática, a fim de evitar fraudes na comercialização de carne bovina.; The objective of the present study was to develop a methodology that would permit sexing bovine meat ready for commercialization. A male-specific primer sequence was used, followed by analysis of the amplified product. The method proved to be efficient for sex verification and is of practical utility in the prevention of fraud in beef sale.

Molecular sexing of birds: a comparative review of PCR-based methods.

Morinha, Francisco; Cabral, João Alexandre; Bastos, Estela
Fonte: Universidade de Trás-os-Montes e Alto Douro Publicador: Universidade de Trás-os-Montes e Alto Douro
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
27.64%
Accurate identification of sex in birds is important for the management and conservation of avian wildlife in several ways, namely in the development of population, behavioral and ecological studies, as well as in the improvement of ex situ captive breeding programs. In general, nestlings, juveniles and adult birds of a wide number of sexually monomorphic species cannot be sexed based on phenotypic traits. The development of molecular methodologies for avian sexing overcame these difficulties, allowing a reliable gender differentiation for these species. The polymerase chain reaction (PCR)-based methods have been widely applied in molecular sexing of birds, using a large diversity of sex-linked markers. During the last 15 yrs, there was a continuous improvement in the PCR-based protocols for bird sexing, increasing the accuracy, speed and high-throughput applicability of these techniques. The recent advances in real-time PCR platforms and whole genome analysis methods provided new resources for the detection and analysis of novel specific markers and protocols. This review presents a comparative guide of classical and recent advances in PCR-based methods for avian molecular sexing, highlighting its strengths and limitations. Future research opportunities in this field are also addressed. © 2012 Elsevier Inc. All rights reserved.

Bovine carcass sexing by PCR method

Curi,Rogério Abdallah; Mota,Lígia Souza Lima Silveira da; Amarante,Mônica Regina Vendrame; Lopes,Catalina Romeiro
Fonte: Faculdade de Medicina Veterinária e Zootecnia / Universidade de São Paulo Publicador: Faculdade de Medicina Veterinária e Zootecnia / Universidade de São Paulo
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/01/2002 EN
Relevância na Pesquisa
37.34%
The objective of the present study was to develop a methodology that would permit sexing bovine meat ready for commercialization. A male-specific primer sequence was used, followed by analysis of the amplified product. The method proved to be efficient for sex verification and is of practical utility in the prevention of fraud in beef sale.

Use of the TSPY gene for sexing cattle

Lemos,Daniela Cristina; Rios,Álvaro Fabrício Lopes; Caetano,Lisandra Cristina; Lôbo,Raysildo Barbosa; Vila,Reginaldo Aparecido; Martelli,Lúcia; Takeuchi,Paula Lumy; Ramos,Ester Silveira
Fonte: Sociedade Brasileira de Genética Publicador: Sociedade Brasileira de Genética
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/03/2005 EN
Relevância na Pesquisa
37.34%
The Y-encoded, testis-specific protein (TSPY) is a Y-specific gene. The copy numbers of TSPY range from 20 to 60 in men and up to 200 in bulls. In this study, we examined the possibility of using the TSPY gene to sex cattle. DNA from blood samples of 100 Nelore cattle (50 males and 50 females) from the Nelore Cattle Breeding Program (PMGRN) was screened for TSPY by PCR using TSPY-specific primers. The assay was highly specific since all male samples were TSPY-positive and all female samples were negative. Positive results were also obtained at low DNA concentrations (less than 1 rhog/muL). These results showed that TSPY was a good male-specific marker, the usefulness of which was enhanced by the high copy number of the gene. This is the first report to demonstrate the applicability of TSPY for sexing cattle.

Embryo Sexing and Sex Chromosomal Chimerism Analysis by Loop-Mediated Isothermal Amplification in Cattle and Water Buffaloes

HIRAYAMA, Hiroki; KAGEYAMA, Soichi; MORIYASU, Satoru; SAWAI, Ken; MINAMIHASHI, Akira
Fonte: The Society for Reproduction and Development Publicador: The Society for Reproduction and Development
Tipo: Artigo de Revista Científica
EN
Relevância na Pesquisa
27.73%
In domestic animals of the family Bovidae, sex preselection of offspring has been demanded for convenience of milk/beef production and animal breeding. Development of the nonsurgical embryo transfer technique and sexing methods of preimplantation embryos made it possible. Sexing based on detection of Y chromosome-specific DNA sequences is considered the most reliable method to date. PCR enables amplification of a target sequence from a small number of blastomeres. However, it requires technical skill and is time consuming. Furthermore, PCR has the risk of false positives because of DNA contamination during handling of the PCR products in duplicate PCR procedures and/or electrophoresis. Therefore, for embryo sexing to become widely used in the cattle embryo transfer industry, a simple, rapid and precise sexing method needs to be developed. Loop-mediated isothermal amplification (LAMP) is a novel DNA amplification method, and the reaction is carried out under isothermal conditions (range, 60 to 65 C) using DNA polymerase with strand displacement activity. When the target DNA is amplified by LAMP, a white precipitate derived from magnesium pyrophosphate (a by-product of the LAMP reaction) is observed. It is noteworthy that LAMP does not need special reagents or electrophoresis to detect the amplified DNA. This review describes the development and application of an embryo sexing method using LAMP in cattle and water buffaloes.

Successful Nonsurgical Transfer of Bovine Elongating Conceptuses and Its Application to Sexing

KIMURA, Koji; MATSUYAMA, Shuichi
Fonte: The Society for Reproduction and Development Publicador: The Society for Reproduction and Development
Tipo: Artigo de Revista Científica
EN
Relevância na Pesquisa
27.52%
The objectives of the present study were to establish a nonsurgical transfer method for elongating bovine conceptuses and to combine this method with biopsy and sexing. Bovine conceptuses were recovered from donor cows on days 13–14 of the estrus cycle. In experiment 1, day 13 conceptuses were transferred to recipient cows using a standard day 7 embryo transfer (ET) method. The pregnancy rate of day 13 conceptus transfer (CT) is comparable to that of day 7 ET. In experiment 2, day 14 conceptuses were transferred using modified methods (balloon catheters or ET guns with modified sheaths). Using the standard ET method, no pregnancies were obtained; however, when balloon catheters or ET guns with modified sheaths were used, the pregnancy rates after CT were 48.0% and 44.8%, respectively. In experiment 3, day 14 conceptuses were biopsied without a micromanipulator, sexed using the loop-mediated isothermal amplification method and transferred to recipient cows. The pregnancy rate of biopsied conceptuses was 46.2% and did not differ significantly from that of unbiopsied conceptuses. Moreover, all pregnant cows transferred conceptuses following biopsy and sexing delivered calves with the expected sexes. These results suggested that the nonsurgical bovine CT method was comparable to day 7 ET and that this technique enables biopsy and sexing without expensive equipment such as a micromanipulator or specialized skills.

Investigating the effectiveness of manual sexing, behavioural sexing and sexing by growth rate for Oreochromis niloticus (Nile Tilapia).

Kochel, Anya
Fonte: Quens University Publicador: Quens University
Tipo: Tese de Doutorado
EN
Relevância na Pesquisa
37.99%
Male tilapia are more valued in aquaculture because they grow faster than female tilapia. Stunting of tilapia populations usually result from male and female tilapia being raised in the same tank. Monosex male cultures are regularly used in tilapia aquaculture to increase production yield. However, a reliable sexing method is required to maximize production yields. This study looked into three different methods of sexing tilapia: manual sexing, behavioural sexing and sexing by growth rate. Tilapia were manually sexed by observing the genital papilla for the presence of an oviduct opening in females and the lack of an opening in males. For the behavioural trials tilapia were divided into two glass aquaria, one with gravel and one without. They were observed for aggressive behaviours such as charging, biting, chasing, quivering, nest excavation, next circling, and leading. The lengths and weights of juvenile tilapia were also observed over the course of three months to determine if there were differences between male and female growth rates. It was determined that overall manual sexing was the most reliable and a significant (p-value = 2.3x10-13) predictor of tilapia sex. Furthermore, tilapia are able to be significantly (p-value = 4.8x10-2...

Sexual size dimorphism and assortative mating in the short- tailed shearwater puffinus tenuirostris

Einoder, L.; Page, B.; Goldsworthy, S.
Fonte: African Seabird Group Publicador: African Seabird Group
Tipo: Artigo de Revista Científica
Publicado em //2008 EN
Relevância na Pesquisa
27.52%
To improve methods for sexing live birds in field studies, we assessed sexual size dimorphism in the Short-tailed Shearwater Puffinus tenuirostris and produced a sex-discriminating function. Despite a degree of overlap in body size, males were significantly larger than females. A stepwise discriminant function analysis of five morphometric characters indicated that bill depth and head length were the most dimorphic characters, and the resultant sex model correctly discriminated 92.0% of known males (23 of 25), and 92.3% of known females (24 of 26). The model was validated by applying it to an additional group of birds whose sex was assumed, based on their pairing with known-sex individuals. Of the assumed females, 93% were correctly classified (n = 15), as were 96% of males (n = 15). Application of the sex model to another breeding colony reduced its performance to 70%–82% accuracy because of the existence of significant geographic variation in body size in this species. For individuals in which certainty was low (i.e. when small males are confused with large females), sexing could be improved by measuring the body size of the breeding partner. This improvement was a result of significant positive assortative mating with respect to bill depth and a body size index. This sex model provides a quick and easy means of sexing in instances in which molecular methods and other techniques are not feasible.; Luke D. Einoder...

Pregnancy diagnosis, fetal quantification and gender estimation by ultra-sonography in ewes; Diagnóstico de gestação, quantificação e sexagem fetal por ultra-sonografia em ovelhas

DIAS, Lilian Mara Kirsch; SOUZA, José Camisão de; ASSIS, Roberta de Moura; RAYMUNDO, Camila de Moraes
Fonte: Editora da Universidade Federal de Lavras Publicador: Editora da Universidade Federal de Lavras
Tipo: Artigo de Revista Científica
ENG
Relevância na Pesquisa
27.52%
The objective of this experiment was to evaluate the accuracy of gestation, fetal sexing and quantification diagnoses in ewes. Pregnancy and fetal quantification were diagnosed in 105 ewes at 35 days of pregnancy. For the fetal gender diagnosis sexing diagnose 55 ewes between 49 and 59 days of pregnancy were used. All exams were recorded on DVD for posterior analysis. After birth, lamb sex was recorded to determine fetal sexing precision. Data were analyzed by chisquare (χ2) or Fisher's test, with a significance of 0.05. One hundred percent of pregnancy ultrasound diagnoses were correct. As for the fetal quantification diagnoses, there was an error of 12%. It was possible to diagnose the fetal sex in 87% of the 69 examined fetuses, and 90% of these were estimated correctly. The real-time ultrasound diagnoses were not different from the recorded DVD image diagnoses. Therefore, pregnancy diagnosis accuracy may reach 100%, differing from fetal gender estimation and quantification, which are dependent upon other variables such as fetal gender and examiner experience.; O objetivo deste experimento foi avaliar a acurácia do diagnóstico de gestação, quantificação e sexagem fetal em ovelhas. Foram realizados o diagnóstico de gestação e a quantificação fetal em 105 ovelhas aos 35 dias de gestação. Para o diagnóstico da sexagem fetal foram utilizadas 55 ovelhas com período de gestação entre 49 e 59 dias. As imagens de todos os exames foram gravadas em DVD para permitir posterior análise. Após o nascimento dos cordeiros...

A transgenic embryonic sexing system for the Australian sheep blow fly Lucilia cuprina

Yan, Ying; Scott, Maxwell J.
Fonte: Nature Publishing Group Publicador: Nature Publishing Group
Tipo: Artigo de Revista Científica
Publicado em 05/11/2015 EN
Relevância na Pesquisa
27.52%
Genetic approaches, including the sterile insect technique (SIT), have previously been considered for control of the Australian sheep blow fly Lucilia cuprina, a major pest of sheep. In an SIT program, females consume 50% of the diet but are ineffective as control agents and compete with females in the field for mating with sterile males, thereby decreasing the efficiency of the program. Consequently, transgenic sexing strains of L. cuprina were developed that produce 100% males when raised on diet that lacks tetracycline. However, as females die mostly at the pupal stage, rearing costs would not be significantly reduced. Here we report the development of transgenic embryonic sexing strains of L. cuprina. In these strains, the Lsbnk cellularization gene promoter drives high levels of expression of the tetracycline transactivator (tTA) in the early embryo. In the absence of tetracycline, tTA activates expression of the Lshid proapoptotic gene, leading to death of the embryo. Sex-specific RNA splicing of Lshid transcripts ensures that only female embryos die. Embryonic sexing strains were also made by combining the Lsbnk-tTA and tetO-Lshid components into a single gene construct, which will facilitate transfer of the technology to other major calliphorid livestock pests.

Pregnancy diagnosis, fetal quantification and gender estimation by ultra-sonography in ewes

Dias,Lilian Mara Kirsch; Souza,José Camisão de; Assis,Roberta de Moura; Raymundo,Camila de Moraes
Fonte: Editora da Universidade Federal de Lavras Publicador: Editora da Universidade Federal de Lavras
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/06/2009 EN
Relevância na Pesquisa
27.52%
The objective of this experiment was to evaluate the accuracy of gestation, fetal sexing and quantification diagnoses in ewes. Pregnancy and fetal quantification were diagnosed in 105 ewes at 35 days of pregnancy. For the fetal gender diagnosis sexing diagnose 55 ewes between 49 and 59 days of pregnancy were used. All exams were recorded on DVD for posterior analysis. After birth, lamb sex was recorded to determine fetal sexing precision. Data were analyzed by chisquare (χ2) or Fisher's test, with a significance of 0.05. One hundred percent of pregnancy ultrasound diagnoses were correct. As for the fetal quantification diagnoses, there was an error of 12%. It was possible to diagnose the fetal sex in 87% of the 69 examined fetuses, and 90% of these were estimated correctly. The real-time ultrasound diagnoses were not different from the recorded DVD image diagnoses. Therefore, pregnancy diagnosis accuracy may reach 100%, differing from fetal gender estimation and quantification, which are dependent upon other variables such as fetal gender and examiner experience.

Sexagem de carcaça bovina pelo método de PCR; Bovine carcass sexing by PCR method

Curi, Rogério Abdallah; Mota, Lígia Souza Lima Silveira da; Amarante, Mônica Regina Vendrame; Lopes, Catalina Romeiro
Fonte: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia Publicador: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; ; ; ; ; Formato: application/pdf
Publicado em 01/01/2002 ENG
Relevância na Pesquisa
37.34%
Objetivou-se, nesta pesquisa, o desenvolvimento de uma metodologia que permitisse a sexagem de carne bovina pronta para comercialização. Para tanto, utilizou-se primers seqüência macho-específica e posterior análise do produto amplificado. O método proposto mostrou-se eficiente para verificar o sexo, bem como sua utilização prática, a fim de evitar fraudes na comercialização de carne bovina.; The objective of the present study was to develop a methodology that would permit sexing bovine meat ready for commercialization. A male-specific primer sequence was used, followed by analysis of the amplified product. The method proved to be efficient for sex verification and is of practical utility in the prevention of fraud in beef sale.

Controle ultra-sonográfico de gestações, de mortalidades embrionárias e fetais e do sexo de fetos bovinos zebuínos; Ultrasonic control of early pregnancies, embryonic and fetal mortalities and fetal sex in zebu cattle

Barros, Breno José Pelozo de; Visintin, José Antonio
Fonte: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia Publicador: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; Formato: application/pdf
Publicado em 01/01/2001 POR
Relevância na Pesquisa
27.73%
Este trabalho avaliou a eficácia do ultra-som no diagnóstico precoce de prenhez e nas avaliações das mortalidades embrionárias e fetais e dos sexos de fetos em dois grupos de vacas zebuínas. Os animais que não retornaram ao estro aos 21 dias da Inseminação Artificial (G1) ou aos 14 dias da Transferência de Embriões (G2B) foram examinados aos 25 dias de gestação para o diagnóstico precoce de prenhez, aos 45 dias para avaliação das perdas embrionárias entre 26 e 45 dias e aos 60 dias para avaliações das perdas fetais entre 45 e 60 dias e dos sexos de fetos. O Grupo G2A foi examinado por palpação retal aos 45 dias para diagnóstico de prenhez e aos 60 dias para avaliações das perdas fetais e dos sexos de fetos. As taxas de prenhez foram, respectivamente, 94,6%, 88,1% e 83,4% aos 25, 45 e 60 dias de gestação. As taxas de perdas embrionárias e fetais e de abortos foram, respectivamente, 4,6%, 4,7% e 1,2%. As taxas de acertos do diagnóstico precoce de gestação e dos sexos de fetos foram 88,5% e 90,7%, respectivamente. A ultra-sonografia mostrou-se eficaz no diagnóstico precoce de prenhez aos 25 dias, nas avaliações das perdas embrionárias aos 45 dias e fetais aos 60 dias e no diagnóstico do sexo de fetos a partir dos 60 dias de gestação. Os exames ultra-sonográficos não causaram perdas gestacionais nos animais Bos indicus ou Bos indicus X Bos taurus.; This study analysed the efficiency of early pregnancy diagnosis and embryo and fetal mortality and sexing evaluation by ultrasound in two groups of zebu cows submitted to artificial insemination (G1) or embryo transfer (G2 = G2A and G2B). The animals that not returned to estrus at 21th day after artificial insemination (G1) and 14th day after embryo transfer (G2B) were examinated at 25th day of gestation for early pregnancy diagnosis...

Sexagem de embriões bovinos fecundados in vitro pela técnica de PCR multiplex; Sexing of in vitro fertilized bovine embryos by multiplex PCR

Luz, Marcelo Rezende; Watanabe, Yeda Fumie; Ferro, Jesus Aparecido; Ferro, Maria Inês T.; Mauro, Sônia Marli Singaretti de; Hossepian de Lima, Vera Fernanda Martins; Franceschini, Paulo Henrique
Fonte: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia Publicador: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; Formato: application/pdf
Publicado em 01/12/2000 POR
Relevância na Pesquisa
37.34%
Neste trabalho, a técnica de PCR ("polymerase chain reaction") foi utilizada para a sexagem de 92 embriões bovinos fertilizados in vitro. Os embriões originaram-se de fertilização in vitro de oócitos aspirados de ovários de fêmeas bovinas, provenientes de abatedouros comerciais. Os oócitos foram maturados, fertilizados e cultivados até o estádio de blastocisto. Os embriões foram lavados em solução de PBS, transferidos para tubos de polipropileno contendo água ultrapura, e imediatamente congelados a -196ºC. Os embriões foram descongelados sobre isopor contendo gelo picado e tratados com proteinase K. Para a reação de PCR, utilizaram-se alíquotas de 34 µl de cada tudo, onde foram acrescidos dois pares de primers, seqüência BC1.2 e seqüência satélite 1.715, desoxinucleotídeos, MgCl2, tampão PCR 10X, TaqDNA polimerase e água, em um volume final de 50 µl. As amostras foram amplificadas e a eletroforese realizada em gel de poliacrilamida a 8%. Os géis foram corados com solução de brometo de etídio e analisados em transiluminador de luz ultravioleta. Um índice de 93,47% de amplificação foi atingido, com 41 embriões (47,67%) machos e 45 (52,32%) embriões fêmeas. O uso de gel de poliacrilamida a 8% foi eficaz na separação de fragmentos de DNA muito próximos.; In the present study the polymerase chain reaction (PCR) was used for sexing ninety-two in vitro fertilized bovine embryos. The embryos were obtained after in vitro fertilization of oocytes from slaughterhouses. The oocytes were matured...

Identificação do sexo e variabilidade genética em uma população de Astronotus ocellatus (Agassiz, 1831) por marcadores ISSR

Paiva, Isadora Marques
Fonte: Universidade Federal de Lavras; Programa de Pós-Graduação em Ciências Veterinárias; UFLA; brasil; Departamento de Medicina Veterinária Publicador: Universidade Federal de Lavras; Programa de Pós-Graduação em Ciências Veterinárias; UFLA; brasil; Departamento de Medicina Veterinária
Tipo: Dissertação
Publicado em 17/12/2015 POR
Relevância na Pesquisa
27.64%
Studies conducted in order to investigate alternative methods for sexing fish species, such as Astronotus ocellatus, using reduced sexual dimorphism, are extremely important to facilitate reproduction management techniques. The objective of this study was to determine the feasibility of manual sexing, widely used by fish farmers and hobbyists, and the identification of specific molecular markers for a particular sex using ISSR markers. The latter technique also generates genetic diversity and similarity data, and is important for conservation studies. Manual sexing was performed by macroscopic analysis of the urogenital papilla. Fin and gonad samples of 30 A. ocellatus (±83,32g e ±15, 96 cm) were collected for DNA extraction and histology, respectively. For DNA extraction, we adopted the NaCl protocol (SAMBROOK, 1989). Quality samples were used for amplification, using universal ISSR primers with subsequent separation of the generated fragments by electrophoresis, and assessment of similarity and genetic diversity levels. The manual sexing did not appear as a viable technique to distinguish the sexes for this species, given the occurrence of 37% of errors during selection. Likewise...

Iris colour as an indicator of age feature in female Brazilian tanagers (Passeriformes: Emberizidae) confirmed by a molecular sexing technique

Monnerat Nogueira,Denise; S. Alves,Maria Alice
Fonte: Revista de Biología Tropical Publicador: Revista de Biología Tropical
Tipo: Artigo de Revista Científica Formato: text/html
Publicado em 01/12/2008 EN
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The Brazilian tanager, Ramphocelus bresilius is an endemic species from Brazil that is sexually dimorphic in adult plumage. Young males are similar to adult and young females until their second year. Adults and young females are not distinguishable in plumage. We tested whether iris colour can be used to separate adult females from immature females. We used for the first time the molecular sexing technique based on CHD-genes to confirm the sex of the individuals classified as "female plumage with red iris", and to identify the sex of individuals classified as "female plumage and brown iris". The adult males were used as a positive control. DNA samples from 190 individuals were analysed. The sizes of the PCR products were identified as 350 base pairs (bp) for CHD-Z and 388 bp for CHD-W. We confirmed that adult females have a red iris and the young females a brown iris. We could also separate young males and females which present the same iris colour and plumage. Although there are indications that the iris colour can be used by birds to identify the adults in co-operative breeding species such as the Brazilian tanager, more behavioural data are required to understand the role of iris coloration in this species. Rev. Biol. Trop. 56 (4): 1629-1633. Epub 2008 December 12.