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The objective of this study was to evaluate the effects of different concentrations of antifreeze protein (AFP) extracted from the larva of the beetle, Tenebrio molitor (TmAFP), on vitrification of in vitro-produced bovine embryos. In vitro-produced blastocysts were divided into three experimental groups and vitrified using a cryotop. TmAFP was added to the equilibrium solution (ES) and vitrification solution (VS) at a concentration of 0 ng/mL (control), 500 ng/mL (500TmAFP), or 1000 ng/mL (1000TmAFP). Vitrification was carried out by first placing the blastocysts in ES for 2 minutes (7.5% ethylene glycol [EG] and 7.5% dimethyl sulfoxide [DMSO]). The blastocysts were then transferred to VS (15% EG and 15% DMSO) and promptly deposited on a cryotop stem and submerged in liquid nitrogen. Warming was carried out in three steps with decreasing sucrose concentrations. After warming, the blast cells were cultured for 24 hours for subsequent survival analysis and ultrastructural evaluation. There was a significant difference in the survival rate and expansion in the 500TmAFP group compared with the other groups. The ultrastructural analysis revealed intracellular lesions in all vitrified embryos; however, the embryos of the 500TmAFP and 1000TmAFP groups showed fewer cytoplasmic lesions compared with the control group. Taken together, addition of TmAFP can mitigate cellular changes that involve organelles and cellular components essential for proper functioning and improve the viability of warmed and vitrified in vitro-produced bovine embryos.

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The aim of this study was to evaluate the effect of the replacement of soybean meal by increasing levels of sugarcane yeast and urea on the energy, enzymatic, protein, mineral, and hormonal profiles of buffalo heifers. Twenty Murrah heifers with an average body weight of 168 ± 2.0 kg and 1 year old were used in a completely randomized design. The treatments consisted of the replacement of soybean meal by increasing levels of sugarcane yeast and urea (0, 33, 67, and 100% dry matter). The animals were weighed at the beginning and end of the experimental period, and blood was collected on the final day. The replacement of soybean meal with sugarcane yeast influenced the performance and metabolic dynamics, with total weight gain (P = 0.005) and average daily gain (P = 0.015), presenting, for both variables, a quadratic effect. Considering the biochemical profile, there was an influence on the serum concentration of triglycerides (P = 0.055), the serum activities of aspartate aminotransferase, gamma-glutamyl transferase, and alkaline phosphatase (P < 0.05), in addition to the serum concentration of phosphorus (P = 0.007) and potassium (P = 0.053), which showed positive linear effects. The hormonal profile was not influenced by diets (P > 0.05). The elements copper, iron, and zinc were able to be quantified at all levels of sugarcane yeast and urea in the diets offered. The use of sugarcane yeast with urea promotes adequate metabolic response of buffalo heifers and can replace up to 100% of soybean meal in the concentrate.

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The study aimed to verify the influence of the FecGE mutation in superovulated ewes and to evaluate the probability of logistic models to determine the response capacity of these ewes to superovulatory treatment. Santa Inês ewes (n = 29) were genotyped for the FecGE mutation and separated for their genotype group in carriers of the mutant E allele (FecGE/E, FecG+/E) and non-carrier (FecG+/+) alleles. The ewes underwent hormonal treatment for superovulation. Aside from the genotypes, variables included in the statistical model were reproductive status (empty, early lactation, or late lactation), age (> or < 6 years), and number of births (nulliparous, primiparous, multiparous). The carriers of the mutation could be discriminated from the non-carriers based on the number of corpora lutea, rate of frozen embryos, and fecundity. Recovery rate was significantly higher (P < 0.05) in FecGE/E (94.31%) compared to FecG+/E (63.15%) and FecG+/+ (61.90%) (P < 0.05), whereas fecundity rate of FecG+/+ ewes (50.76%) was significantly higher than FecG+/E (18.96%) and FecGE/E (32.53%) (P < 0.05). We determined in this study that the response to superovulation and embryo production can be discriminated between FecGE/E and FecG+/E ewes in relation to the FecG+/+ genotype. Logistic models that included reproductive status and mutation, or reproductive status and age, or reproductive status and number of births were effective in predicting the response to superovulatory treatment.

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We investigated whether the recombinant leptin (1, 10, 100 ng/mL) influences the meiotic maturation of goat oocytes, whether the MAPK and JAK2/STAT3 pathways mediate the effects of leptin during in-vitro maturation, and whether leptin differently affects the abundance of mRNAs relevant to leptin signal transduction and apoptosis in oocytes and cumulus cells. The addition of leptin to the maturation medium positively affected the number of oocytes that completed nuclear maturation. Nuclear oocyte maturation stimulated by leptin was significantly impaired when we added the specific inhibitors of MAPK (U0126) and JAK2/STAT3 (AG490) to the maturation medium. The addition of leptin (10 ng/mL) during maturation did not affect the expression of AMPKα1, PPARα, Caspase 3, and BCL2 genes in oocytes or cumulus cells. The PPARγ and BAX mRNA abundances were significantly reduced in cumulus cells in the leptin group compared to the control group. Our results demonstrate that supplementation of the in-vitro maturation medium with leptin significantly improves nuclear maturation and reveal the important role of the MAPK and JAK2/STAT3 signaling pathways in establishing the leptin-mediated nuclear maturation of goat oocytes. Moreover, leptin treatment affects PPARγ and BAX gene expression in cumulus cells.

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This study was conducted to characterize the morphology and morphometry of follicles containing multiple oocytes (MOFs) and determine the association with the FecGE mutation in Santa Inês ewes. Based on the genotypes, 65 ewes were characterized as being homozygous wild-type (n = 25; FecG+/+), heterozygous mutant (n = 27, FecG+/E), and homozygous mutant (n = 13, FecGE/E). The variables evaluated were follicle developmental stage, number of oocytes per follicle, morphology, and morphometry of MOFs. The FecGE mutation did not affect the frequency of MOFs (P > 0.05) (3.0 % in FecG+/+; 3.3 % in FecG+/E; and 3.5 % in FecGE/E). The greater viability (P < 0.05) of MOFs was identified in transitory stage of the FecGE/E (95.0 %) and FecG+/E (90.9 %) when compared to the FecG+/+ genotype (73.3 %). Furthermore, the morphology of transitory follicles with two oocytes was the variable and when evaluated was the most reliable determinant for predicting which ewes had an FecGE mutation. In conclusion, the FecGE mutation did not affect the frequency of MOFs. The ewes with FecGE mutation had a greater frequency of morphologically normal MOFs in the transitory stage. Furthermore, the ewes with the FecGE mutation had a greater likelihood of having MOFs containing two morphologically normal oocytes.

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Housekeeping genes (HKG) are paramount for accurate gene expression analysis during preimplantation development. Markedly, quantitative reverse transcription polymerase chain reaction (RT-qPCR) in ovine embryos currently lacks HKGs. Therefore, we tested 11 HKGs for RT-qPCR normalization during ovine parthenogenetic preimplantation development. Seven HKGs reached the qPCR efficiency threshold (97.20-105.96%), with correlation coefficients ranging from -0.922 to -0.998 and slopes from -3.22 to -3.59. GeNorm ranked glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and TATA-binding protein (TBP) as the best HKG pair, while H3 histone, family 3A (H3F3A) was the third HKG. Relative gene expression was measured for zinc finger protein X-linked (ZFX) and developmental pluripotency-associated 3 (DPPA3) transcripts during ovine parthenogenetic preimplantation development. ZFX did not show any transcript abundance fluctuation among oocytes, cleavage-stage embryos, and morulae. DPPA3 transcript abundance was also similar among all developmental stages, therefore suggesting that it may not display a maternal gene expression profile. In silico analysis of ovine DPPA3 mRNA and protein showed high conservation to bovine orthologues. However, DPPA3 orthologues differed in regulatory motifs. In conclusion, GAPDH, TBP and H3F3A are stable HKGs in ovine parthenogenetic embryos and allow accurate RT-qPCR-based gene expression analysis.

RESUMO

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106), G2 (2*106) and G3 (4*106) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chi-square (P<0.05) and correlated with CASA and fluorescence data using Person Correlation (P<0.05). The IVF efficiency, cleavage and total blastocyst rates did not show any significant difference (P>0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P<0.05) without difference (P>0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P<0.05), without significant difference (P>0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0.05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106), G2 (2*106) e G3 (4*106) espermatozoides/mL. Às 18h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o Computer-Assisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P<0,05). A eficiência da FIV, taxas de clivagem e blastocisto total não mostraram diferença significativa (P>0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P<0,05), sem diferença (P>0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P<0,05), sem apresentar diferença significativa (P>0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.

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RESUMO

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106 ), G2 (2*106 ) and G3 (4*106 ) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chisquare (P0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0,05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106 ), G2 (2*106 ) e G3 (4*106 ) espermatozoides/mL. Às 18 h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o ComputerAssisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.

Assuntos

RESUMO

In the present study we aimed to test the best insemination dose for in vitro embryo production (IVEP) and to correlate sperm traits in bovine. In vitro matured oocytes were inseminated with three different sperm concentrations of the same bull: G1 (1*106 ), G2 (2*106 ) and G3 (4*106 ) sperm/mL. At 18 h post-insemination (hpi), presumptive zygotes [G1 (n=114), G2 (n=139) and G3 (n=136)] were stained to evaluate the pronuclei numbers, or continued to in vitro culture [G1 (n=102), G2 (n=111) and G3 (n=106)]. Sperm kinetics were analyzed using Computer-Assisted Semen Analysis (CASA). Sperm plasma membrane, acrosome integrity and mitochondrial activity were analyzed using fluorescent probes. In vitro fertilization (IVF) and IVEP data were compared using chisquare (P0.05) among the groups. In G3, the polyspermy rate was the highest (7.4%; P0.05) between G1 (0%) and G2 (0%). In G1, the early blastocyst rate was the highest (7.8%; P0.05) between G2 (1.8%) and G3 (0.9%). The IVF efficiency and total blastocyst rates were positively correlated with curvilinear velocity (VCL) (r≃+1; P<0,05). We concluded that the reduction of insemination dose may negatively affect embryo development and VCL may be used as a parameter to improve the IVEP outcomes.(AU)

O objetivo deste estudo foi testar a melhor dose inseminante para a produção de embriões in vitro (IVEP) e sua correlação com as características espermáticas na espécie bovina. Oócitos maturados in vitro foram inseminados com três concentrações diferentes de espermatozoides de único touro: G1 (1*106 ), G2 (2*106 ) e G3 (4*106 ) espermatozoides/mL. Às 18 h pós-inseminação (hpi), os presumíveis zigotos [G1 (114), G2 (139) e G3 (136)] foram corados para avaliar o número de pronúcleos, ou continuaram para o cultivo in vitro [G1 (102), G2 (111) e G3 (106)]. Os parâmetros da cinética espermática foram analisados usando o ComputerAssisted Semen Analysis (CASA). A integridade de membrana plasmática espermática, acrossomal e a atividade mitocondrial foram analisadas usando sondas fluorescentes. Os dados da fertilização in vitro (FIV) e IVEP foram comparadas com qui-quadrado (P=0,05) e correlacionados com dados de CASA e Fluorescência usando Correlação de Pearson (r=±1; P0,05) entre os grupos. Em G3, a taxa de polispermia foi a maior (7,4%; P0,05) entre G1 (0%) e G2 (0%). Em G1, a taxa de blastocisto inicial foi a maior (7,8%; P0,05) com G2 (1,8%) e G3 (0,9%). A eficiência de FIV e a taxa de blastocisto total foram positivamente correlacionadas com velocidade curvilinear (VCL) (P<0,05). Concluímos que a dose inseminante reduzida pode negativamente afetar o desenvolvimento embrionário e VCL pode ser usada como parâmetro para melhorar os resultados da PEIV.(AU)

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RESUMO

Based on ovarian and follicular variables, there was determination of ewes with different FecGE genotypes. Based on the FecGE genotype, 65 Santa Inês ewes were assigned to three experimental groups: homozygous wild-type (n = 25; FecG+/+), mutant heterozygous (n = 27; FecG+/E) and mutant homozygous (n = 13; FecGE/E). The ewe's ovaries were weighed and measured, then the follicles (oocyte, nucleus and nucleolus) were histologically evaluated for morphometry and morphology. Morphologically normal follicles, in the primordial and transitional stages, explained 70.18% of the variability morphological characteristics between mutant and wild-type ewes. Conducting the morphometric evaluation resulted in a more precise determination of the genotype groups when there was assessment of the primordial and secondary follicular developmental stages. The diameter of the oocyte and the oocyte nucleus of the primordial follicles explained 36.76% of the variability in follicular morphology between ewes with the mutation and those with the wildtype group. Similarly, the core diameter of oocytes in secondary follicles explained 10.63% of the variability in follicular morphology among FecGE/E, FecG+/E and FecG+/+ ewes. Thus, morphologically normal follicles in the primordial and transitional stages of development are the variables that allow for a more precise differentiation of Santa Inês ewes with the FecGE mutation. These variables may be evaluated to make more efficient the adoption of biotechniques that when conducted there is utilisation of follicles in the initial developmental stages as a physiological basis for classifying whether specific follicles are useful when conducting the techniques.

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The aim of this work was to determine the effect of homeopathic supplementation on both ovarian dynamics and conception rate in Nellore cows subjected to fixed-time artificial insemination (FTAI). Cows (n = 150) were randomly distributed to the control (CG) and the homeopathy group (HG). The HG cows were supplemented with Pró-cio in the mineral salt for 60 days and both experimental groups were further subjected to FTAI. Cows were evaluated for ovarian dynamics (n = 16), progesterone (P4) concentration (n = 16), and conception rates (n = 150). Ovarian dynamics determined by ultrasonography and showed similar findings for CG and HG, respectively. Thus follicular diameter (8.7 ± 1.0 mm vs. 10.0 ± 0.8 mm), mean pre-ovulatory follicle volume (0.46 ± 0.15 mL vs. 0.61 ± 0.12 mL), and mean follicular growth (3.65 ± 1.41 mm vs. 4.60 ± 1.21 mm) did not differ between groups. Moreover, corpus luteum diameter was similar between groups (CG: 16.28 ± 0.7 mm vs. HG: 15.6 ± 0.8 mm; P > 0.05), although P4 levels did differ (CG: 2.55 ± 0.85 ng mL-1 vs. HG: 6.52 ± 1.19 ng mL-1; P < 0.05). The conception rate after FTAI was not affected by homeopathic supplementation (CG: 74.67 %, and did HG: 77.33 %; P > 0.05). In conclusion, the homeopathic supplementation Pró-cio increases P4 concentrations but does improve the reproductive efficiency of Nellore cows subject to FTAI.

O objetivo foi determinar o efeito da suplementação homeopática na dinâmica ovariana e taxa de concepção em vacas Nelore cows submetidas à inseminação artificial em tempo fixo (IATF). As vacas (n = 150) foram distribuídas aleatoriamente nos grupos controle (GC) e grupo homeopático (GH). As vacas do GH foram suplementadas com Pró-cio® no sal mineral mineral por 60 dias. Ambos os grupos foram submetidos à IATF. As vacas foram avaliadas quanto à dinâmica ovariana (n = 16), concentração de progesterona (P4; n = 16) e taxa de concepção (n = 150). A dinâmica ovarina foi determinada por ultra-sonografia e mostrou resultados semelhantes para o GC e o GH, respectivamente. Portanto, para diâmetro folicular (8,7 ± 1,0 mm vs. 10,0 ± 0,8 mm), volume médio do folículo pré-ovulatório (0,46 ± 0,15 mL vs. 0,61 ± 0,12 mL) e crescimento folicular médio (3,65 ± 1,41 mm vs. 4,60 ± 1,21 mm) não diferiram entre os grupos. Além disso, o diâmetro do corpo lúteo foi semelhante entre os grupos (CG: 16,28 ± 0,7 mm vs. HG: 15,6 ± 0,8 mm; P > 0.05), apesar dos níveis de P4 diferirem (CG: 2,55 ± 0,85 ng mL-1 vs. GH: 6,52 ± 1,19 ng mL-1; P < 0.05). A taxa de concepção após a IATF não foi afetada pela suplementação homeopática (GC:74.67 % vs. GH: 77.33 %; P > 0.05). Em conclusão, a suplementação homeopática com Pró-cio aumenta a concentração de P4 mas não melhora a eficiência reprodutiva de vacas Nelore cows submetidas à IATF.

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Recent studies have shown that adiponectin, an adipokine predominantly produced by adipose tissue, regulates several reproductive processes. However, the mechanisms of action of adiponectin on the maturation of goat oocytes remain to be determined. The aim of this study was to investigate whether (a) adiponectin influences the meiotic maturation of goat oocytes; (b) MAPK MEK 1/2 mediates the effects of adiponectin; and 3) adiponectin differentially affects mRNA relative abundance of genes relevant for adiponectin signal transduction in goat oocytes. The addition of adiponectin (5 µg/ml) during the maturation of goat oocytes resulted in a higher percentage of successful nuclear maturation compared to those of the group without adiponectin (p < 0.05). Adiponectin-stimulated nuclear oocyte maturation was significantly impaired by a mitogen-activated protein kinase MEK 1/2 inhibitor, U0126 (p < 0.05). There was no evidence of any adiponectin-induced difference in the relative transcript abundances of AdipoR1, AdipoR2, AMPKα1, AMPKα2, PPARα and PPARγ genes. In conclusion, these results indicate that adiponectin has a positive effect on the meiotic maturation of goat oocytes through the MAPK MEK 1/2 pathway. Furthermore, the adiponectin does not affect the relative abundance of genes relevant for adiponectin signal transduction in goat oocytes.

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O objetivo deste estudo foi avaliar a utilização da ultrassonografia e do dispositivo intravaginal Metricheck® na seleção de vacas leiteiras para um programa de inseminação artificial em tempo fixo (IATF). Foram avaliadas vacas Holandesas (n = 138) lactantes com média de 54 dias em lactação (DEL), ECC médio de 2,5 e mantidas em sistema semi-intensivo de criação. Após o período voluntário de espera (PVE) de 45 dias, foram distribuidas em dois grupos (G1 e G2). No G1 (n = 72) foram submetidas ao exame ginecológico porpalpação transretal auxiliado por ultrassonografia, sendo selecionadas apenas aquelas que não apresentavam secreção intrauterina hiperecoica. As vacas do G2 (n = 66) também foram submetidas ao exame ginecológico por palpação transretal auxiliado pelo Metricheck® para avaliação do muco vaginal, sendo atribuídos escores de 0 a 3 (0 = translucido, 1 = contendo poucas manchas de pus branco, 2 = contendo até 50% de pus branco e 3 =contendo mais 50% de pus branco, creme ou sanguinolento). Todos os animais foram submetidos a IATF e ao diagnóstico de prenhez 35 e 60 dias após a inseminação. Os dados foram submetidos ao teste Z a 5%. Na avaliação ginecológica 11,1% das vacas do G1 e 7,6% no G2 apresentaram endometrite e foram consideradasinaptas para reprodução (P > 0,05). Em relação à taxa de concepção também não houve diferença estatística entre os grupos (P > 0,05), sendo 46,9% para o G1 e 57,4% para o G2. Em relação ao muco vaginal avaliado no G2, as vacas apresentaram 78,8%,13,6%, 4,5% e 3,0%, respectivamente para os escores 0, 1, 2 e 3. Conclui-se, que ambos os métodos de diagnóstico podem ser utilizados com eficiência na avaliação de vacas leiteiras para programas reprodutivos. Bem como, vacas apresentando muco vaginal com escore 1 ao exame com Metricheck®não sofrem prejuízos para concepção, quando submetidas a protocolo de IATF.

The objective of this study was to assess the use of ultrasound and Metricheck® intravaginal device toselect dairy cows for a timed artificial program (TAI).There were evaluated lactating Holstein cows (n=138) with an average of 54 days in milk (DIM) and average BCS 2.5maintained in semi-intensive breeding system. After a voluntary waiting period (VWP) of 45 days, cows were distributed into two groups (G1 and G2). In G1 (n= 72) cows were submitted to gynecological examination by transrectal palpation aided by ultrasonography,being selected only those which did not present hyperechoic intrauterine secretion. G2 cows (n = 66) were also submitted to gynecological examination by transrectal palpation aided by the Metricheck® for evaluation of vaginal mucus, being assigned scores of 0 to 3 (0 = translucent, 1 = containing flecks of white pus, 2 =containing less than 50% white pus and 3 = containing more than 50% white, yellow or sanguineous pus). All animals underwent TAI and pregnancy diagnoses were performed on days 35 and 60 after insemination. Thedata were subjected to Z test at 5% significance. In the gynecological evaluation, 11.1% of G1 cows and 7.6% in G2 presented endometritis and were considered unfit for reproduction (P > 0.05). Regarding the conception rate, there was also no statistical difference between groups, being 46.9% for G1 and 57.4% for G2 (P > 0.05). Regarding vaginal mucus evaluation in G2, cows presented 78.8, 13.6 and 3.0% for scores 0, 1, 2 and 3,respectively. It was concluded that both diagnostic methods can be used efficiently in the evaluation of dairy cows for reproductive programs. As well, cows presenting vaginal mucus with score 1 on the Metricheck® test does not impair conception when submitted to TAI protocol.

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O objetivo deste estudo foi avaliar a utilização da ultrassonografia e do dispositivo intravaginal Metricheck® na seleção de vacas leiteiras para um programa de inseminação artificial em tempo fixo (IATF). Foram avaliadas vacas Holandesas (n = 138) lactantes com média de 54 dias em lactação (DEL), ECC médio de 2,5 e mantidas em sistema semi-intensivo de criação. Após o período voluntário de espera (PVE) de 45 dias, foram distribuidas em dois grupos (G1 e G2). No G1 (n = 72) foram submetidas ao exame ginecológico porpalpação transretal auxiliado por ultrassonografia, sendo selecionadas apenas aquelas que não apresentavam secreção intrauterina hiperecoica. As vacas do G2 (n = 66) também foram submetidas ao exame ginecológico por palpação transretal auxiliado pelo Metricheck® para avaliação do muco vaginal, sendo atribuídos escores de 0 a 3 (0 = translucido, 1 = contendo poucas manchas de pus branco, 2 = contendo até 50% de pus branco e 3 =contendo mais 50% de pus branco, creme ou sanguinolento). Todos os animais foram submetidos a IATF e ao diagnóstico de prenhez 35 e 60 dias após a inseminação. Os dados foram submetidos ao teste Z a 5%. Na avaliação ginecológica 11,1% das vacas do G1 e 7,6% no G2 apresentaram endometrite e foram consideradasinaptas para reprodução (P > 0,05). Em relação à taxa de concepção também não houve diferença estatística entre os grupos (P > 0,05), sendo 46,9% para o G1 e 57,4% para o G2. Em relação ao muco vaginal avaliado no G2, as vacas apresentaram 78,8%,13,6%, 4,5% e 3,0%, respectivamente para os escores 0, 1, 2 e 3. Conclui-se, que ambos os métodos de diagnóstico podem ser utilizados com eficiência na avaliação de vacas leiteiras para programas reprodutivos. Bem como, vacas apresentando muco vaginal com escore 1 ao exame com Metricheck®não sofrem prejuízos para concepção, quando submetidas a protocolo de IATF.(AU)

The objective of this study was to assess the use of ultrasound and Metricheck® intravaginal device toselect dairy cows for a timed artificial program (TAI).There were evaluated lactating Holstein cows (n=138) with an average of 54 days in milk (DIM) and average BCS 2.5maintained in semi-intensive breeding system. After a voluntary waiting period (VWP) of 45 days, cows were distributed into two groups (G1 and G2). In G1 (n= 72) cows were submitted to gynecological examination by transrectal palpation aided by ultrasonography,being selected only those which did not present hyperechoic intrauterine secretion. G2 cows (n = 66) were also submitted to gynecological examination by transrectal palpation aided by the Metricheck® for evaluation of vaginal mucus, being assigned scores of 0 to 3 (0 = translucent, 1 = containing flecks of white pus, 2 =containing less than 50% white pus and 3 = containing more than 50% white, yellow or sanguineous pus). All animals underwent TAI and pregnancy diagnoses were performed on days 35 and 60 after insemination. Thedata were subjected to Z test at 5% significance. In the gynecological evaluation, 11.1% of G1 cows and 7.6% in G2 presented endometritis and were considered unfit for reproduction (P > 0.05). Regarding the conception rate, there was also no statistical difference between groups, being 46.9% for G1 and 57.4% for G2 (P > 0.05). Regarding vaginal mucus evaluation in G2, cows presented 78.8, 13.6 and 3.0% for scores 0, 1, 2 and 3,respectively. It was concluded that both diagnostic methods can be used efficiently in the evaluation of dairy cows for reproductive programs. As well, cows presenting vaginal mucus with score 1 on the Metricheck® test does not impair conception when submitted to TAI protocol.(AU)

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tThis study aimed to compare the accuracy of two diagnostic methods, rectal palpation and transrectalultrasound of the ovaries, and assess the reliability of the diagnosis by palpation method. A total of sixteen (16)dairy farm cows underwent ultrasonographic examinations performed soon after rectal palpation. The accuracyof palpation may seem low, but when we analyze the data, we observed that the main structures diagnosed byultrasound were almost always diagnosed by palpation. Ultrasonography, however, has its strong point in theidentification of complex structures such as hemorrhagic body and follicular cysts. We conclude that ultrasoundas well as rectal palpation can be used as a diagnostic tool, although ultrasound provides good visualization ofovarian structures, allowing their measurement.

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This study evaluated the efficacy of a long term progesterone protocols associated with PGF2a andGnRH by measurement of follicular diameter (FD) in D14, D22 and D30; time between the end of treatment andthe onset of estrus and conception rate in groups of Girolando heifers and Girolando cows (N1 and N2, V1 andV2). On D30, the diameter of FD for the groups V1, V2, N1 and N2 was 13.3 ± 0,11mm, 13.8 ± 0.14 mm, 0.10mm and 13.7 ± 12.7 ± 0.49mm respectively. The time between the end of the treatment and the onset of estrus forthe groups V1, V2, and N1, N2 was 115,2hs, 103,2hs, 127,2hs, 129,6hs, respectively. Based on the results it canbe concluded that the protocols were efficient for obtaining a dominant follicle in the PGF2a application dayfavoring a good conception rate.

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tThis study aimed to compare the accuracy of two diagnostic methods, rectal palpation and transrectalultrasound of the ovaries, and assess the reliability of the diagnosis by palpation method. A total of sixteen (16)dairy farm cows underwent ultrasonographic examinations performed soon after rectal palpation. The accuracyof palpation may seem low, but when we analyze the data, we observed that the main structures diagnosed byultrasound were almost always diagnosed by palpation. Ultrasonography, however, has its strong point in theidentification of complex structures such as hemorrhagic body and follicular cysts. We conclude that ultrasoundas well as rectal palpation can be used as a diagnostic tool, although ultrasound provides good visualization ofovarian structures, allowing their measurement.(AU)

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This study evaluated the efficacy of a long term progesterone protocols associated with PGF2a andGnRH by measurement of follicular diameter (FD) in D14, D22 and D30; time between the end of treatment andthe onset of estrus and conception rate in groups of Girolando heifers and Girolando cows (N1 and N2, V1 andV2). On D30, the diameter of FD for the groups V1, V2, N1 and N2 was 13.3 ± 0,11mm, 13.8 ± 0.14 mm, 0.10mm and 13.7 ± 12.7 ± 0.49mm respectively. The time between the end of the treatment and the onset of estrus forthe groups V1, V2, and N1, N2 was 115,2hs, 103,2hs, 127,2hs, 129,6hs, respectively. Based on the results it canbe concluded that the protocols were efficient for obtaining a dominant follicle in the PGF2a application dayfavoring a good conception rate.(AU)

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We evaluated the "in vitro" blastocyst rate production using bovine sexed semen. Semen from three bulls was used to verify the individual"s semen variation, cleavage rates and embryo production. In this study, we employed reproductive biotechnologies, computer analysis of post-thawed semen and fluorescent probes for sperm cells integrity analysis (plasma membrane, acrosome membrane and mitochondrial potential). A total of 959 oocysts went through in vitro maturation steps for in vitro fertilization and cultivation, being 473 with sexed semen and 486 with conventional semen. The cleavage rate was observed in blastocysts on D2 and D7. Data were analyzed by SPSS 16.0 software using analysis of variance (ANOVA), Student"s t-test was used to detect differences between groups, and chi-square analysis for in vitro production results (P 0.05 ). The results differed between conventional (31.06%) and sexed semen (21.10%) in the obtainment of blastocyst. When the blastocyst production was individually compared in sexed semen samples (27.69%, 17.93% and 25.56%, bulls 1, 2 and 3, respectively) we verified T2 T1 and T1 = T3 and T2 = T3. [...](AU)

Avaliou-se a taxa de produção de blastocisto in vitro utilizando-se o sêmen bovino sexado. Foram utilizados três reprodutores para verificar a variação individual do sêmen, taxas de clivagem e produção embrionária. O trabalho utilizou-se de biotécnicas reprodutivas, análise computadorizada do sêmen pós-descongelação e sondas fluorescentes para análises de integridade da célula espermática (membrana plasmática, membrana acrossomal e potencial mitocondrial). Um total de 959 oócitos passou por etapas de maturação in vitro, fertilização in vitro (sexado, n= 473; convencional, n = 486) e cultivo in vitro. A taxa de clivagem foi observada no D2 e a de blastocistos no D7. Os dados foram analisados pelo programa SPSS 16.0 empregando-se a análise de variância (ANOVA), sendo o teste t-Student usado para detectar diferenças entre os grupos e o Qui-quadrado  para análise dos resultados da produção in vitro (P 0,05). Os resultados diferiram entre o sêmen convencional (31,06%) e sexado (21,10%) para produção de blastocisto. Quando comparada a produção de blastocisto individualmente nas amostras de sêmen sexado (27,69%; 17,93% e 25,56%, touros 1, 2 e 3, respectivamente), percebeu-se que T2 T1 e T1=T3 e T2=T3. Quanto às análises de cinética espermática, as amostras de sêmen sexado mostraram diferençasentre os touros nas variáveis velocidade curvilínea, velocidade linear e velocidade do trajeto em que o T1(117,7±1,6 µm/s; 60,0±0,3 µm/s; 73,6±0,4 µm/s, respectivamente) quando comparado aostouros T2 (80,2±2,3 µm/s; 47,0±2,0 µm/s; 57,7±0,9 µm/s, respectivamente) e T3 (86,4±5,7 µm/s; 46,2±2,7 µm/s; 53,8±2,8 µm/s, respectivamente) obteve valores mais elevados. As análises daintegridade da célula espermática não diferiram entre as amostras de sêmen convencional, já no sêmen sexado a integridade de membrana foi a variável que diferiu estatisticamente entre os touros, em que o T1 (38 ±2,7) diferiu do T3(53,8± 1,8) (P=0,009), mas não divergiu do T2 (44,1±4,4).[...](AU)

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