Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Tipo de documento: | Dissertação |
Idioma: | eng |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da USP |
Texto Completo: | https://www.teses.usp.br/teses/disponiveis/11/11139/tde-14022022-131254/ |
Resumo: | Studies were conducted to validate a kit for measuring P4 in cattle and to evaluate the circulating progesterone (P4) profile and ovarian dynamics after treatment with long-acting injectable progesterone (P4i) in postpartum cows. In the first study, tests were conducted to validate a commercial kit based on chemiluminescent technology, Immulite 1000 Progesterone (LKPW1), to measure P4 in cattle. For this purpose, standard curves were generated from plasma samples of steers enriched with known concentrations of P4, and the precision, accuracy, anticoagulant effect, and residual effect were evaluated. The method proved to be accurate for samples with a P4 concentration between 0.3 and 10.0 ng/mL and precise for samples between 0.5 and 20.0 ng/mL. In addition, the results provided by the equipment are reliable to the concentrations in the samples, whether using serum or plasma, and it is not necessary to analyze standard curves for each assay. In the second study, Nelore primiparous (n = 28) in postpartum anestrus were randomly assigned to receive 1 mL of oily vehicle (Con; n = 14) or 1 mL of oily vehicle with 150 mg of P4i (P4i; n = 14) on D0. Thereafter, for 21 d, blood samples and ovarian ultrasonography were performed in all animals, in addition to observation of expression of estrus between D8 and D15. In general, no cow expressed estrus, remaining anovulatory until D20. On D1, cows in the P4i group had a peak of circulating P4 (3.0 ng/mL), remaining above 0.5 ng/mL for 7 d, while in the Con group, circulating P4 remained below 0.2 ng/mL throughout the study. Cows in the P4i group had a larger diameter of the dominant follicle during the study (12.8 ± 0.5 vs. 10.6 ± 0.6 mm), a longer interval between waves (9.4 ± 1.0 vs. 6.5 ± 0.7 d) and more days with follicles with ovulatory capacity in the ovaries compared to the Con group (12.5 ± 0.5 vs. 8.3 ± 1.3 d). Thus, despite being inefficient in inducing cyclicity in postpartum cows, treatment with P4i promoted changes in ovarian dynamics and circulating profile of P4 that could serve as a basis for further studies and use strategies. Furthermore, the tested kit showed high potential for use in the quantification of circulating P4 in cattle. However, further tests and analyses of a larger number of samples are needed to confirm its robustness. |
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Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesteroneValidação de um sistema de imunoensaio para dosagem de progesterona e respostas fisiológicas de vacas no pós-parto tratadas com progesterona injetável de longa açãoBos indicusBos indicusAnestroAnestrusChemiluminescenceImmuliteImmuliteProgesteronaProgesteroneQuimioluminescênciaStudies were conducted to validate a kit for measuring P4 in cattle and to evaluate the circulating progesterone (P4) profile and ovarian dynamics after treatment with long-acting injectable progesterone (P4i) in postpartum cows. In the first study, tests were conducted to validate a commercial kit based on chemiluminescent technology, Immulite 1000 Progesterone (LKPW1), to measure P4 in cattle. For this purpose, standard curves were generated from plasma samples of steers enriched with known concentrations of P4, and the precision, accuracy, anticoagulant effect, and residual effect were evaluated. The method proved to be accurate for samples with a P4 concentration between 0.3 and 10.0 ng/mL and precise for samples between 0.5 and 20.0 ng/mL. In addition, the results provided by the equipment are reliable to the concentrations in the samples, whether using serum or plasma, and it is not necessary to analyze standard curves for each assay. In the second study, Nelore primiparous (n = 28) in postpartum anestrus were randomly assigned to receive 1 mL of oily vehicle (Con; n = 14) or 1 mL of oily vehicle with 150 mg of P4i (P4i; n = 14) on D0. Thereafter, for 21 d, blood samples and ovarian ultrasonography were performed in all animals, in addition to observation of expression of estrus between D8 and D15. In general, no cow expressed estrus, remaining anovulatory until D20. On D1, cows in the P4i group had a peak of circulating P4 (3.0 ng/mL), remaining above 0.5 ng/mL for 7 d, while in the Con group, circulating P4 remained below 0.2 ng/mL throughout the study. Cows in the P4i group had a larger diameter of the dominant follicle during the study (12.8 ± 0.5 vs. 10.6 ± 0.6 mm), a longer interval between waves (9.4 ± 1.0 vs. 6.5 ± 0.7 d) and more days with follicles with ovulatory capacity in the ovaries compared to the Con group (12.5 ± 0.5 vs. 8.3 ± 1.3 d). Thus, despite being inefficient in inducing cyclicity in postpartum cows, treatment with P4i promoted changes in ovarian dynamics and circulating profile of P4 that could serve as a basis for further studies and use strategies. Furthermore, the tested kit showed high potential for use in the quantification of circulating P4 in cattle. However, further tests and analyses of a larger number of samples are needed to confirm its robustness.Estudos foram desenvolvidos com o objetivo de validar um kit para dosagem de P4 em bovinos e avaliar o perfil de progesterona (P4) circulante e dinâmica ovariana após tratamento com progesterona injetável de longa ação (P4i) em vacas pós-parto. No primeiro estudo, foram realizados testes para validação de um kit comercial baseado em tecnologia quimioluminescente, Immulite 1000 Progesterone (LKPW1), para dosagem de P4 em bovinos. Para tal, curvas padrão foram produzidas a partir de amostras de plasma de garrotes castrados enriquecidas com concentrações conhecidas de P4, e a precisão, exatidão, efeito de anticoagulante e efeito residual foram avaliados. O método se mostrou exato para amostras com concentração de P4 entre 0.3 e 10.0 ng/mL e preciso para amostras entre 0,5 e 20,0 ng/mL. Além disso, os resultados fornecidos pelo kit são fidedignos às concentrações nas amostras, sejam elas soro ou plasma, não sendo necessária a análise de curvas padrão a cada ensaio. No segundo estudo, primíparas Nelore (n = 28) em anestro pós-parto foram aleatoriamente designadas a receber 1 mL de veículo oleoso (Con; n = 14) ou 1 mL de veículo com 150 mg de P4i (P4i; n = 14) no D0. A partir de então, durante 21 d, foram realizadas colheitas de sangue e ultrassonografia ovariana em todos os animais, além de observação da expressão de cio entre D8 e D15. De modo geral, nenhuma vaca expressou cio, permanecendo anovulatórias até o D20. No D1, vacas do grupo P4i tiveram um pico de P4 circulante (3,0 ng/mL), permanecendo acima de 0,5 ng/mL por 7 d, enquanto no grupo Con a P4 circulante se manteve abaixo de 0,2 ng/mL durante todo o estudo. Vacas do grupo P4i apresentaram maior diâmetro do folículo dominante durante o estudo (12,8 ± 0,5 vs. 10,6 ± 0,6 mm), maior intervalo entre ondas (9,4 ± 1,0 vs. 6,5 ± 0,7 d) e mais dias com folículos com capacidade ovulatória nos ovários em comparação ao grupo Con (12,5 ± 0,5 vs. 8,3 ± 1,3 d). Sendo assim, apesar de ineficiente na indução de ciclicidade em vacas pós-parto, o tratamento com P4i promoveu alterações na dinâmica ovariana e perfil circulante de P4 que podem servir de base para novos estudos e estratégias de uso. Além disso, o kit testado apresentou alto potencial para uso na quantificação de P4 circulante em bovinos. Contudo, ainda são necessários outros testes e análises de maior número de amostras para confirmar sua robustez.Biblioteca Digitais de Teses e Dissertações da USPSartori Filho, RobertoSilva, Taynara Jaqueline Barreiro da2021-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisapplication/pdfhttps://www.teses.usp.br/teses/disponiveis/11/11139/tde-14022022-131254/reponame:Biblioteca Digital de Teses e Dissertações da USPinstname:Universidade de São Paulo (USP)instacron:USPLiberar o conteúdo para acesso público.info:eu-repo/semantics/openAccesseng2022-02-16T18:29:07Zoai:teses.usp.br:tde-14022022-131254Biblioteca Digital de Teses e Dissertaçõeshttp://www.teses.usp.br/PUBhttp://www.teses.usp.br/cgi-bin/mtd2br.plvirginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.bropendoar:27212022-02-16T18:29:07Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP)false |
dc.title.none.fl_str_mv |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone Validação de um sistema de imunoensaio para dosagem de progesterona e respostas fisiológicas de vacas no pós-parto tratadas com progesterona injetável de longa ação |
title |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
spellingShingle |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone Silva, Taynara Jaqueline Barreiro da Bos indicus Bos indicus Anestro Anestrus Chemiluminescence Immulite Immulite Progesterona Progesterone Quimioluminescência |
title_short |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
title_full |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
title_fullStr |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
title_full_unstemmed |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
title_sort |
Validation of an immunoassay system for progesterone dosage and physiological responses of postpartum cows treated with long-acting injectable progesterone |
author |
Silva, Taynara Jaqueline Barreiro da |
author_facet |
Silva, Taynara Jaqueline Barreiro da |
author_role |
author |
dc.contributor.none.fl_str_mv |
Sartori Filho, Roberto |
dc.contributor.author.fl_str_mv |
Silva, Taynara Jaqueline Barreiro da |
dc.subject.por.fl_str_mv |
Bos indicus Bos indicus Anestro Anestrus Chemiluminescence Immulite Immulite Progesterona Progesterone Quimioluminescência |
topic |
Bos indicus Bos indicus Anestro Anestrus Chemiluminescence Immulite Immulite Progesterona Progesterone Quimioluminescência |
description |
Studies were conducted to validate a kit for measuring P4 in cattle and to evaluate the circulating progesterone (P4) profile and ovarian dynamics after treatment with long-acting injectable progesterone (P4i) in postpartum cows. In the first study, tests were conducted to validate a commercial kit based on chemiluminescent technology, Immulite 1000 Progesterone (LKPW1), to measure P4 in cattle. For this purpose, standard curves were generated from plasma samples of steers enriched with known concentrations of P4, and the precision, accuracy, anticoagulant effect, and residual effect were evaluated. The method proved to be accurate for samples with a P4 concentration between 0.3 and 10.0 ng/mL and precise for samples between 0.5 and 20.0 ng/mL. In addition, the results provided by the equipment are reliable to the concentrations in the samples, whether using serum or plasma, and it is not necessary to analyze standard curves for each assay. In the second study, Nelore primiparous (n = 28) in postpartum anestrus were randomly assigned to receive 1 mL of oily vehicle (Con; n = 14) or 1 mL of oily vehicle with 150 mg of P4i (P4i; n = 14) on D0. Thereafter, for 21 d, blood samples and ovarian ultrasonography were performed in all animals, in addition to observation of expression of estrus between D8 and D15. In general, no cow expressed estrus, remaining anovulatory until D20. On D1, cows in the P4i group had a peak of circulating P4 (3.0 ng/mL), remaining above 0.5 ng/mL for 7 d, while in the Con group, circulating P4 remained below 0.2 ng/mL throughout the study. Cows in the P4i group had a larger diameter of the dominant follicle during the study (12.8 ± 0.5 vs. 10.6 ± 0.6 mm), a longer interval between waves (9.4 ± 1.0 vs. 6.5 ± 0.7 d) and more days with follicles with ovulatory capacity in the ovaries compared to the Con group (12.5 ± 0.5 vs. 8.3 ± 1.3 d). Thus, despite being inefficient in inducing cyclicity in postpartum cows, treatment with P4i promoted changes in ovarian dynamics and circulating profile of P4 that could serve as a basis for further studies and use strategies. Furthermore, the tested kit showed high potential for use in the quantification of circulating P4 in cattle. However, further tests and analyses of a larger number of samples are needed to confirm its robustness. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12-01 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://www.teses.usp.br/teses/disponiveis/11/11139/tde-14022022-131254/ |
url |
https://www.teses.usp.br/teses/disponiveis/11/11139/tde-14022022-131254/ |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
|
dc.rights.driver.fl_str_mv |
Liberar o conteúdo para acesso público. info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
Liberar o conteúdo para acesso público. |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.coverage.none.fl_str_mv |
|
dc.publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
publisher.none.fl_str_mv |
Biblioteca Digitais de Teses e Dissertações da USP |
dc.source.none.fl_str_mv |
reponame:Biblioteca Digital de Teses e Dissertações da USP instname:Universidade de São Paulo (USP) instacron:USP |
instname_str |
Universidade de São Paulo (USP) |
instacron_str |
USP |
institution |
USP |
reponame_str |
Biblioteca Digital de Teses e Dissertações da USP |
collection |
Biblioteca Digital de Teses e Dissertações da USP |
repository.name.fl_str_mv |
Biblioteca Digital de Teses e Dissertações da USP - Universidade de São Paulo (USP) |
repository.mail.fl_str_mv |
virginia@if.usp.br|| atendimento@aguia.usp.br||virginia@if.usp.br |
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1815256957436559360 |