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Master's Dissertation
DOI
https://doi.org/10.11606/D.10.2020.tde-12052020-144315
Document
Author
Full name
Flavia Morag Elliff
E-mail
Institute/School/College
Knowledge Area
Date of Defense
Published
São Paulo, 2020
Supervisor
Committee
Baruselli, Pietro Sampaio (President)
Batista, Emiliana de Oliveira Santana
Gimenes, Lindsay Unno
Title in Portuguese
Estratégias para melhorar a produção in vitro de embriões de doadoras jovens das raças Holandês (Bos taurus) e Gir (Bos indicus)
Keywords in Portuguese
Bezerras
BST
Embrião
FSH
Oócito
Abstract in Portuguese
O uso de animais jovens como doadores de oócitos contribui para acelerar o ganho genético através da redução do intervalo de gerações. Entretanto, doadoras jovens possuem baixa eficiência na produção in vitro de embriões (PIVE). Com isso, o presente estudo objetivou avaliar estratégias para melhorar a PIVE dessa categoria de doadoras. No Experimento 1, 59 bezerras Holandesas foram divididas em: Controle (CTL; n=15), FSH (140mg de FSH, n=15), BST (500mg de rBST, n=14) e BST_FSH (associação de rBST+FSH, n=15) e 15 novilhas púberes (GNP) como controle positivo. No Experimento 2, 90 doadoras Gir foram utilizadas (30 bezerras, 30 novilhas púberes e 30 vacas). Cada categoria foi subdividida em: Controle (sem tratamento, n=15 por categoria) e FSH (tratadas com FSH, n=15 por categoria). Os embriões produzidos foram transferidos e foi avaliada a taxa de prenhez por transferência de embriões (P/TE). Ainda, foi realizada dosagem de IGF1 e de glicose de todos os animais. Os resultados foram analisados utilizando o software SAS. No Experimento 1, o tratamento com FSH elevou a quantidade de folículos médios (Com FSH: 7,2±1,4; Sem FSH: 1,0±0,2; P<0,0001) e grandes (Com FSH: 2,4±0,6; Sem FSH: 0,7±0,1; P=0,0042). O tratamento com FSH apresentou tendência (P=0,08) em aumentar a taxa de oócitos viáveis (Com FSH: 62,1% vs. Sem FSH: 54,3%). Foi observada interação BST*FSH (P=0,0007) para taxa de clivagem (GCTL: 40,5%b ; GFSH: 45,2%ab; GBST: 48,4%a ; GBST_FSH: 40,9%ab). Animais tratados com rBST apresentaram tendência (P=0,06) em aumentar o número de blastocistos (Com rBST: 0,86±0,24; Sem rBST: 0,39±0,14) e taxa de blastocisto (Com rBST: 7,3%; Sem rBST: 3,7%; P=0,09). O tratamento com rBST elevou a concentração plasmática de IGF1 (Com rBST: 311,1±24,9; Sem rBST: 231,3±15,5 ng/mL; Pbst=0,0031). No Experimento 2 não foi verificada interação Trat*Cat para nenhuma variável. Entretanto, verificou-se efeito de categoria para: população folicular (P<0,0001), oócitos totais (P=0,0002) e oócitos clivados (P=0,0004), sendo que as novilhas foram superiores. Verificou-se que as bezerras apresentaram maior concentração de glicose (P=0,0157) que as novilhas e vacas. Ainda, as bezerras e as novilhas apresentaram maiores concentrações de IGF1 (P=0,0001) que vacas. O tratamento com FSH elevou a população folicular (Com FSH: 25,9±0,76; Sem FSH: 21,9±0,70, P=0,0002) e o número de folículos médios (P>0,0001) e grandes (P>0,0001). Animais tratados com FSH apresentaram menor taxa de recuperação oocitária (Com FSH: 51,8%; Sem FSH: 65,2%; P=0,0136). A taxa de clivagem foi influenciada pelo tratamento (Com FSH: 59,5%; Sem FSH: 50,7%; P=0,0163). Houve tendência de maior P/TE aos 30 (Bezerras: 40,0%b; Novilhas: 44,5%b; Vacas: 57,6%a; P=0,0623) e aos 60 dias (Bezerras: 35,3%b; Novilhas: 38,1%b; Vacas: 50,4%a; P=0,0950) para os embriões de vacas. O tratamento com FSH influenciou negativamente as concentrações de IGF1 (Com FSH: 238,7±25,2; Sem FSH: 296,8±25,5 ng/mL; P=0,0312), não influenciando os níveis de glicose (P=0,8593). Desta forma, conclui-se que o tratamento com rBST em bezerras holandesas e o tratamento com FSH em bezerras, novilhas e vacas Gir aumentaram a eficiência da PIVE.
Title in English
Strategies to improve the in vitro embryo production of young Holstein (Bos taurus) and Gyr (Bos indicus) donors
Keywords in English
BST
Calves
Embryo
FSH
Oocyte
Abstract in English
Use of young animals as oocyte donors contributes to genetic gain enhancement through reduction of the generation interval. However, young donors have low efficiency on the in vitro embryo production (IVEP). Thus, the objective of the present study was to evaluate strategies to improve the IVEP of these animals. In Experiment 1, 59 prepubertal Holstein heifers were used, divided into 4 groups: Control (CTL; n=15), FSH (140 mg of FSH; n=15), BST (500 mg of rbST, n=14) and BST_FSH (association of both treatments; n=15) and 15 pubertal heifers (PH) as positive control. In Experiment 2, 90 Gyr donors were used (30 calves, 30 pubertal heifers and 30 cows). Each category was subdivided into 2 groups: Control (no treatment, n=15 per category) and FSH (treated with FSH, n=15 per category). The IVEP parameters were evaluated, the embryos produced were transferred and the pregnancy rate per embryo transfer (P/ET) was evaluated. Also, IGF1 and glucose were assayed for all animals. Data were analyzed using SAS software. In Experiment 1, treatment with FSH increased the number of medium follicles (With FSH: 7.23±1.43; Without FSH: 1.03±0.22; P<0.0001) and large follicles (With FSH: 2.39±0.56; Without FSH: 0.66±0.12; P=0.0042). Treatment with FSH showed a tendency (P=0.08) to increase the rate of viable oocytes (With FSH: 62%; Without FSH: 54%). There was a BST*FSH interaction (P=0.0007) for cleavage rate (CTL: 40.5%b; FSH: 45.2%ab; BST: 48.4%a; BST_FSH: 40.9%ab). Animals treated with rBST showed a tendency (P=0.06) to have increased number of blastocysts (With rBST: 0.86±0.24; Without rBST: 0.39±0.14) and blastocyst rate (With rBST: 7.3%; Without rBST: 3.7%; P=0.09). Treatment with rBST increased the levels of IGF1 (With rBST: 311.06±24.9; Without rBST: 231.28±15.54; Pbst=0.0031). In Experiment 2, there was no Treat*Cat interaction for any of the variables. There was a category effect for the following variables: follicle population (P>0.0001), total oocytes (P=0.0002) and cleaved oocytes (P=0.0004), such that pubertal heifers had higher values for all these variables. In addition, calves and heifers presented higher concentration of IGF1 (P=0.0001) than cows. Treatment with FSH increased the follicle population (With FSH: 25.9±0.76; Without FSH: 21.9±0.70, P=0.0002) and the number of medium (P>0.0001) and large (P>0.0001) follicles. Animals treated with FSH had lower recovery rate (With FSH: 51.8%; Without FSH: 65.2%; P=0.0136). The cleavage rate was influenced (P=0.0163) by the treatment (With FSH: 59.5% vs. Without FSH: 50.7%). There was a tendency for increased pregnancy rate at 30 days (Calves: 40.0%b; Heifers: 44.5%b; Cows: 57.6%a; P=0.0623) and 60 days (Calves: 35.3%b; Heifers: 38.1%b; Cows: 50.4%a; P=0.0950) for embryos produced by cow donors. Treatment with FSH had a negative influence on the IGF1 concentration (With FSH: 238.7±25.2; Without FSH: 296.8±25.5; P=0.0312), with no effect on the glucose levels (P=0.8593). Thus, it is concluded that treatment with rBST for Holstein calves and treatment with FSH for Gyr calves, heifers and cows were effective for increasing the IVEP efficiency.
 
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Publishing Date
2020-08-13
 
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