Use of cooled buffalo semen as a strategy to increase conception rates in fixed-time artificial insemination programs during unfavorable reproductive periods / [Uso de sêmen resfriado de búfalo como estratégia para aumentar as taxas de concepção em programas de inseminação artificial em tempo fixo, durante períodos reprodutivos desfavoráveis]

The objective of this study was to compare the reproductive efficiency of dairy buffaloes undergoing fixed-time artificial insemination (FTAI) protocols based on progesterone/estrogen (P4/E2) and eCG during unfavorable breeding season using cooled (CS) and frozen semen (FS). A total of 446 buffaloes (> 40 days postpartum) were randomly distributed into four blocks (years): B1-2014 (n = 143), B2-2015 (n = 34), B3-2016 (n = 90), and B4-2017 (n = 179). Each block was subdivided into two (AI with CS and FS using the same ejaculate of each bull). Thus, the block subdivision was as follows: B1 (CS = 71 and FS = 72); B2 (CS = 18 and FS = 16); B3 (CS = 47 and FS = 43); and B4 (CS = 90 and FS = 89). The ejaculates of eight Murrah bulls collected using an artificial vagina were divided into two aliquots: one aliquot was diluted in Botu-Bov® commercial extender and cooled (BB-CS), and the other was diluted in the same extender and frozen (BB-FS). BB-CS aliquots were cooled at 5 °C/24 h using a refrigerator. BB-FS group aliquots were also cooled, and after equilibrating at 5 °C for 4 h, were placed in a 21-L Styrofoam box, 5 cm above the surface of liquid nitrogen. In the afternoon (A) on D0 (2:00 p.m.) the animals received EB 2.0 mg IM (Estrogin®) and an ear implant (CRESTAR® 3.0 mg P4). At D9 (A), the implant was removed, and the animals received eCG 400 IU IM (Folligon® 5000) + Cloprostenol PGF2α 0.530 mg IM (Sincrocio®). At D10 (A), the animals received EB 1.0 mg IM (Estrogin®), and at D12 (8:00 a.m.), AI was performed. At D42, pregnancy was diagnosed via ultrasonography. Total CRs were 48.2% CS and 34.6% FS for years 2014 to 2017, with a significant difference of 13.7% (P<0.05). In conclusion, cooled semen resulted in higher CR than frozen semen in dairy buffaloes under the P4/E2 and eCG FTAI during the unfavorable reproductive season.(AU)

O objetivo deste estudo foi comparar a eficiência reprodutiva de búfalas leiteiras submetidas a protocolos de inseminação artificial em tempo fixo (IATF) à base de progesterona/estrogênio (P4/E2) e eCG, durante a estação reprodutiva desfavorável, usando-se sêmen resfriado (SR) e congelado (SC) Um total de 446 búfalas (> 40 dias após o parto) foi distribuído aleatoriamente em quatro blocos (anos): B1-2014 (n = 143), B2-2015 (n = 34), B3-2016 (n = 90) e B4-2017 (n = 179). Cada bloco foi subdividido em dois (IA com SR e SC utilizando-se a mesma ejaculação de cada touro). Assim, a subdivisão do bloco foi a seguinte: B1 (SR = 71 e SC = 72); B2 (SR = 18 e SC = 16); B3 (SR = 47 e SC = 43); e B4 (SR = 90 e SC = 89). Os ejaculados de oito touros Murrah coletados com vagina artificial foram divididos em duas alíquotas: uma alíquota diluída em diluente comercial Botu-Bov® e resfriada (BB-SR), e a outra diluída no mesmo diluente e congelada (BB-SC). As alíquotas de BB-SR foram resfriados a 5°C/24h usando-se um refrigerador. As alíquotas do grupo BB-SC também foram resfriadas e, após equilíbrio a 5°C por 4h, foram colocadas em uma caixa de isopor de 21L, 5 cm acima da superfície do nitrogênio líquido. À tarde (A), no D0 (14h), os animais receberam BE 2,0 mg IM (Estrogin®) e um implante auricular (Crestar® 3,0 mg P4). No D9 (A), o implante foi retirado e os animais receberam eCG 400 UI IM (Folligon® 5000) + cloprostenol PGF2α 0,530 mg IM (Sincrocio®). No D10 (A), os animais receberam BE 1,0mg IM (Estrogin®), e, no D12 (8h da manhã), foram realizadas as IAs. No D42, a gestação foi diagnosticada por ultrassonografia. As taxas de concepção (TC) totais foram 48,2% SR e 34,6% SC para os anos de 2014 a 2017, com uma diferença significativa de 13,7% (P<0,05). Em conclusão, o sêmen resfriado resultou em maior TC do que o sêmen congelado em bubalinos leiteiros sob P4/E2 e eCG FTAI durante a estação reprodutiva desfavorável.(AU)

Use of estradiol cypionate to induce ovulation in buffalo raised in floodplain areas of Itacoatiara, Amazonas / [Uso do cipionato de estradiol na indução da ovulação de búfalas criadas em várzea da região de Itacoatiara, Amazonas]

This study aimed to determine the efficiency of estradiol cypionate (EC) as an ovulation inducer in a Timed Artificial Insemination protocol. 69 buffalo cows received an intravaginal progesterone device and 2mg of estradiol benzoate (EB) at D0. On D9, the intravaginal device was removed and 0.53mg of prostaglandin (PGF2α) and 400UI of equine chorionic gonadotrophin (eCG) were applied. The cows were distributed into two groups: the first group received 1mg of EC (ECG) in D9, and the second group received 1mg of EB (EBG) in D10. Inseminations occurred on D11. Ovarian activity and pregnancy diagnosis were analyzed by ultrasonography. There was no difference (P>0.05) in follicular diameter (9.6 ± 0.89mm vs. 10.7 ± 1.12mm; P=0.06), in ovulation rate (90.9% vs. 90.9%; P=1) and pregnancy rate (58.8% vs. 62.9%; P=0.79), however, buffalo cows from the ECG treatment have less time between P4 removal and ovulation when compared to EBG buffalos (37.4h vs. 52.8h; P=0.001), respectively. Thus, it was concluded that the implantation of TAI in the floodplain of Amazonas is feasible and the use of EC results in successful rates, similar to EB.(AU)

O objetivo deste trabalho foi determinar a eficiência do cipionato de estradiol (CE) como indutor de ovulação em um protocolo de inseminação artificial de tempo fixo. Para isso, 69 búfalas receberam no D0 um dispositivo intravaginal de progesterona e 2mg de benzoato de estradiol (BE). No D9, o dispositivo intravaginal foi removido e foram aplicados 0,53mg de prostaglandina (PGF2α) e 400UI de gonadotrofina coriônica equina (eCG), para, então, os animais serem divididos em dois grupos: um deles (GCE) recebeu 1mg de CE no D9, e o outro (GBE) recebeu 1mg de BE 24h após. As inseminações ocorreram no D11. A atividade ovariana e o diagnóstico de prenhez foram avaliados por ultrassonografia. Não houve diferença (P>0,05) no diâmetro folicular (9,6 ± 0,89mm vs. 10,7 ± 1,12mm; P=0,06), na taxa de ovulação (90,9% vs. 90,9%; P=1) e na taxa de prenhez (58,8% vs. 62,9%; P=0,79), no entanto búfalas do tratamento GCE apresentaram menor tempo entre a remoção da P4 e a ovulação, quando comparadas com as búfalas do GBE (37,4h vs. 52,8h; P=0,001), respectivamente. A implantação da IATF nas várzeas do Amazonas é viável e a utilização do CE resulta em taxas de sucesso similares ao BE.(AU)

Influence of seasonality on the physiological and seminal parameters of buffaloes in the western region of Pará1 / Influência da sazonalidade nos parâmetros fisiológicos e seminais de búfalos da região oeste do Pará

Awareness of the physiological changes that occur when animals are subjected to climatic changes that are considered stressful is essential to maintain animal welfare and to be able to exploit their reproductive potential efficiently and rationally. The present study was carried out to evaluate climatic variables' influence on physiological parameters, and Murrah buffalo ejaculates reared in a humid tropical climate in the Amazon. The immediate analyzes pertinent to the physical and morphological characteristics of the ejaculates were carried out and corresponded in the rainy season (RS) volume of 3.4±2.0mL; the mass activity of 4.4±0.5; motility of 80.4±5.6%; vigor of 4.4±0.4; concentration of 657,300±237,865.1 x 106sptz/mL; major defects of 9.0±2.6%; minor defects of 11.2±3.9%; total defects 20.2±5.3% and sperm plasma membrane integrity (SPMI) 84.8±5.6%, whereas in the non-rainy season (nRS), the results were 4.0±2.1mL; the mass activity of 3.0±1.0; motility of 56.2±13.4%; vigor of 3.0±1.0; concentration of 586,000±291,925.9 x 106sptz/mL; major defects of 20.8±9.9%; minor defects of 27.5±6.3%; total defects 48.3±9.3% and SPMI of 57.9±12.4%. Furthermore, a statistical difference (P<0.05) was observed for the parameters mass activity, motility, vigor, major defects, minor defects, total defects, and sperm plasma membrane integrity between both periods. The data on heart frequency, superficial temperature (head, back, groin, and scrotal pouch) showed a statistical difference between both periods (P<0.05). To conclude is necessary specific management in the non-rainy season that thermal stress is not a determining factor in reducing the reproductive quality of buffaloes; it is necessary to use means to improve animal welfare; one alternative is to use baths regularly for these animals or provide constant access to areas of rivers or lakes, as well as shading, preventing the buffaloes from being directly exposed to the unfavorable thermal environment.(AU)

O conhecimento das alterações fisiológicas que ocorrem quando os animais são submetidos a alterações climatológicas consideradas estressantes é fundamental para manter o bem-estar animal, e poder explorar o seu potencial reprodutivo de forma eficiente e racional. O presente estudo foi realizado com o objetivo de avaliar a influência de variáveis climáticas sobre parâmetros fisiológicos e de ejaculados de búfalos, da raça Murrah, criados em clima tropical úmido da Amazônia. As análises imediatas pertinentes às características físicas e morfológicas dos ejaculados foram realizadas e corresponderam no período chuvoso (PCh) o volume de 3,4±2,0mL, turbilhonamento de 4,4±0,5; motilidade de 80,4±5,6%; vigor de 4,4±0,4; concentração de 657.300±237.865,1 x 106sptz/mL; defeitos maiores de 9,0±2,6%; defeitos menores de 11,2±3,9%; defeitos totais de 20,2±5,3% e integridade da membrana plasmática (IMP) de 84,8±5,6%, enquanto que no período não chuvoso (PnCh), os resultados foram de 4,0±2,1mL; turbilhonamento de 3,0±1,0; motilidade de 56,2±13,4%; vigor de 3,0±1,0; concentração de 586.000±291.925,9 x 106sptz/mL; defeitos maiores de 20,8±9,9%; defeitos menores de 27,5±6,3%; defeitos totais de 48,3±9,3% e IMP de 57,9±12,4%. Observou-se diferença estatística (P<0,05) para os parâmetros movimento de massa, motilidade, vigor, defeitos maiores, defeitos menores, defeitos totais e integridade da membrana plasmática entre os dois períodos. Dados de frequência cardíaca, temperatura superficial (cabeça, dorso, virilha e bolsa escrotal) diferiram estatisticamente entre os períodos (P<0,05). Conclui-se que se faz necessário usar de um manejo específico no período não chuvoso para que o estresse térmico não seja um fator determinante na redução da qualidade reprodutiva dos búfalos, para isto se faz necessário utilizar de meios para melhorar o bem-estar animal, sendo uma das alternativas fazer uso de banhos regularmente para estes animais, ou disponibilizar acesso constante destes a áreas de rios ou lagos, assim como sombreamentos, evitando que os búfalos fiquem expostos diretamente ao ambiente térmico desfavorável.(AU)

Avaliação termográfica do desenvolvimento mamário de búfalas e sua regulação endócrina em distintos estágios fisiológicos / Thermographic evaluation of the mammary development of buffaloes and their endocrine regulation in different physiological stages

O conhecimento do desenvolvimento mamário de um rebanho leiteiro é fundamental, pois relaciona-se à eficiência produtiva. Objetivou-se avaliar a associação dos parâmetros térmicos mamários com concentrações hormonais de búfalas em distintos estágios fisiológicos. Foram utilizadas 24 búfalas mestiças Murrah, em quatro grupos (n= 6): grupo 1 (bezerras), grupo 2 (novilhas), grupo 3 (gestantes) e grupo 4 (lactantes). A cada 28 dias, durante 4 meses, realizaram-se exames de termografia digital por infravermelho para verificar temperatura superficial dos corpos mamários craniais (CMCr) e caudais (CMC), das cisternas craniais (CGMCr) e caudais (CGMC) e tetas craniais (TGMCr) e caudais (TGMC). Foi aferida temperatura retal (TR) e colhido sangue para mensuração das concentrações plasmáticas do fator semelhante a insulina tipo-I, insulina (INS), hormônio do crescimento (GH), progesterona (P4) e estradiol. Grupos 1 e 2 apresentaram correlação de TR com CGMCr. No grupo 3, TR correlacionou-se com TGMCr, TGMC e concentrações plasmáticas de P4. No grupo 4, houve correlação de TR com CGMC, TGMCr e concentrações plasmáticas de INS e GH, e de TGMC com concentrações plasmáticas de GH. Nos quatro grupos, CGMCr correlacionou-se com CMCr e TGMCr, semelhantemente ao observado na porção caudal. Variações térmicas mamárias refletiram alterações fisiológicas aguardadas nos períodos avaliados.(AU)

The knowledge of the mammary development of a dairy herd is key, since it is related to its productive efficiency. The objective of this study was to evaluate the association of mammary thermal parameters with hormonal concentrations of buffaloes at different physiological stages. Twenty-four Murrah crossbred buffaloes were used in four groups (n= 6): group 1 (calves), group 2 (heifers), group 3 (pregnant animals) and group 4 (lactating animals). Every 28 days, for 4 months, infrared digital thermography was performed to check the surface temperature of cranial (CrCM) and caudal (CCM) corpus mammae, cranial (CrC) and caudal (CC) cisterns and cranial (CrT) and caudal teats (CT). Rectal temperature (RT) was measured and blood was collected to measure plasma concentrations of insulin-like factor I, insulin (INS), growth hormone (GH), progesterone (P4) and estradiol. Groups 1 and 2 presented correlation of RT with CrC. In group 3, RT correlated with CrT, CT and plasma P4 concentrations. In group 4, there was correlation of RT with CC, CrT and plasma concentrations of INS and GH, and CT with plasma GH concentrations. In all four groups, CrC correlated with CrCM and CrT, similar to that observed in the caudal portion. The thermal changes in the mammary glands reflected the expected physiological changes in the evaluated periods.(AU)

Teores de cobre, zinco e ferro no fígado de búfalos (Bubalus bubalis) com paratuberculose / Copper, zinc and iron content in the liver of buffaloes (Bubalus bubalis) with paratuberculosis

Com o objetivo de realizar um estudo dos teores de cobre (Cu), zinco (Zn) e ferro (Fe) em búfalas com paratuberculose (PTB) foram utilizadas 13 búfalas, das raças Murrah, Mediterrâneo e seus mestiços acima de três anos de idade, pertencentes a duas propriedades localizadas nos municípios de São Luiz e São Mateus, no Estado do Maranhão. Os animais foram selecionados de acordo com a presença de sinais clínicos sugestivos de paratuberculose, caracterizados por estado nutricional regular a ruim, diarreia crônica líquida a semi-líquida, desidratação, edema submandibular, anestro prolongado, mastites e verminose gastrintestinal. Foi realizada biópsia retal em todos os animais, para detecção de Mycobacterium avium subsp. paratuberculosis (Map) por meio da qPCR, e exames histopatológicos (HE e Ziehl-Neelsen)...

In order to study copper, zinc and iron concentration in the liver of buffaloes with paratuberculosis (PTB), 13 buffalo cows above three years of age of the Murrah and Mediterranean races or their crosses were used. They originated from two farms in the municipalities of São Luís and São Mateus, state of Maranhão, Brazil. The animals were selected according to clinical signs of paratuberculosis, as diarrhea, dehydration and submandibular edema. Rectal biopsies for detection of Mycobacterium avium subsp. paratuberculosis (Map) through PCR in real time (qPCR) and Ziehl-Neelsen (ZN) stain were performed on all animals...

Carcass quality of buffalo (Bubalus bubalis) finished in silvopastoral system in the Eastern Amazon, Brazil / Qualidade da carcaça de búfalos (Bubalus bubalis) terminados em sistema silvipastoril na Amazônia Oriental, Brasil

The influence of the diet on the quality of buffalo (Bubalus bubalis) carcasses of crossbred Murrah and Mediterranean, concentrate fed on traditional (corn/soybean) or on agro-industrial residues (palm oil and coconut cakes) was studied. The animals were sacrificed in a slaughterhouse, and physical and physical-chemical analyses in the carcasses were carried out after 24 hours, and instrumental color and texture analyses of the Longissimus dorsi were carried out 48 hours after cooling. The animals supplemented with palm oil cake had better carcass conformation and fat trim, and differences (P<0.05) in the percentage of fat in the carcass fat thickness and coverage were observed.

Avaliou-se a influência da dieta sobre a qualidade de carcaça de búfalos mestiços das raças Murrah e Mediterrâneo, alimentados com concentrado tradicional - milho/soja - ou resíduos agroindustriais - tortas de coco e dendê. Os animais foram sacrificados em abatedouro comercial, e realizadas análises físicas e físico-químicas na carcaça, após 24 horas, e de cor e textura instrumentais no músculo Longissimus dorsi, após 48 horas de resfriamento. Os animais suplementados com a torta de dendê adquiriram melhor conformação de carcaça e acabamento de gordura, e observaram-se diferenças (P<0,05) no percentual de gordura na carcaça e na espessura de gordura de cobertura.

Influence of nitric oxide on in vitro growth, survival, steroidogenesis, and apoptosis of follicle stimulating hormone stimulated buffalo (Bubalus bubalis) preantral follicles

Effect of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on in vitro survival, growth, steroidogenesis, and apoptosis of buffalo preantral follicles (PFs) was investigated. PFs (200~250 microm) were isolated by micro-dissection and cultured in 0 (control), 10(-3), 10(-5), 10(-7), and 10(-9) M SNP. To examine the reversible effect of SNP, PFs were cultured with 10(-5) M SNP + 1 mM Nomega-nitro-L-arginine methyl ester (L-NAME) or 1.0 microg hemoglobin (Hb). The results showed that greater concentrations of SNP (10(-3), 10(-5), 10(-7) M) inhibited (p < 0.05) FSH-induced survival, growth, antrum formation, estradiol production, and oocyte apoptosis in a dose-dependent manner. However, a lower dose of SNP (10(-9) M) significantly stimulated (p < 0.05) the survival, growth, antrum formation, follicular oocyte maturation, and stimulated progesterone secretion compared to the control. A combination of SNP + L-NAME promoted the inhibitor effect of SNP while a SNP + Hb combination reversed this effect. Nitrate and nitrite concentrations in the culture medium increased (p < 0.05) in a dose-dependent manner according to SNP concentration in the culture medium. At higher concentrations, SNP had a cytotoxic effect leading to follicular oocyte apoptosis whereas lower concentrations have stimulatory effects. In conclusion, NO exerts a dual effect on its development of buffalo PFs depending on the concentration in the culture medium.

Conforto ambiental de bezerros bubalinos (Bubalus bubalis Linnaeus, 1758) em sistemas silvipastoris na Amazônia Oriental / Environmental comfort of buffalo calves (Bubalus bubalis Linnaeus, 1758) on silvopastoral systems in the Eastern Amazon

Foi analisado o efeito de dois sistemas silvipastoris nos índices de conforto ambiental e alterações nos parâmetros fisiológicos de bezerros bubalinos criados na Embrapa Amazônia Oriental, Belém-PA (clima Afi), no Período 1 (abril a setembro/2007) e Período 2 (outubro/2007 a março/2008). Foram inseridos onze bezerros no Sistema Silvipastoril 1 (SSP1), que apresentava sombreamento útil nas pastagens de 18 a 21%, e oito no Sistema Silvipastoril 2 (SSP2), sem sombreamento, com lago para banho. Foram mensuradas as variáveis fisiológicas: temperatura retal (TR), frequência respiratória (FR), temperatura da pele (TP), e calculados o Índice de Temperatura e Umidade (ITU) e Índice de Conforto de Benezra (ICB), nos dois períodos experimentais, comparados pelo Teste Tukey (P < 0,05). O ITU apresentou diferença estatística entre horários (P < 0,05) e período do ano (P < 0,05), e oscilou de 73,5 ± 1,3 até 82,2 ± 0,8. A TR apresentou diferença estatística entre horários e períodos do ano (P < 0,05), com amplitude de 38,3 ± 0,26 a 39,3 ± 0,38 °C. A FR apresentou diferença significativa entre horários (P < 0,05), com amplitude de 32,2 ± 9,2 a 56,5 ± 19,0 mov min-1, consideradas acima dos níveis normais, enquanto a TP foi diferente estatisticamente entre períodos e horários (P < 0,05) e variou de 23,6 ± 8,3 a 31,7 ± 5,4 °C. Nos Períodos 1 e 2 e nos dois SSP's, os ICBs estiveram acima do valor ideal, variando de 2,46 ± 0,33 a 3,31 ± 0,62 (SSP1) e 2,42 ± 0,30 a 3,45 ± 0,66 (SSP2).

This aim of the study was evaluating effects of two different silvopastoral systems on environmental comfort and changes in physiological parameters of buffalo calves bred in the Eastern Amazon. The experiment was conducted at Embrapa Amazônia Oriental, Belém-PA, (Afi climate type) in two periods of the year: Period 1 (April to Sep/2007) and Period 2 (Oct/2007 to March/2008). Eleven calves were included in the Silvopastoral System 1 (SSP1), with a useful shaded pasture area of 18 to 21%; and eight calves were included in the Silvopastoral System 2 (SSP2), without shading, but with a lake for swimming. Physiological status of animals bred in the two SSP's were measured (rectal temperature-RT; respiratory frequency-RF and skin temperature-ST). The Temperature and Humidity Index (THI) and Benezra´s Animal Comfort Index (ACI) were calculated, in two phases of the year, and were compared by the Tukey test (P < 0.05). THI ranged from 73.5 ± 1.3 to 82.2 ± 0.8. RT was 38.3 ± 0.26 to 39.3 ± 0.38 °C, but within the normal range for buffalo. RF ranged from 32.2 ± 9.2 to 56.5 ± 19.0 mov min-1, above the levels considered normal, and ST ranged from 23.6 ± 8.3 and 31.7 ± 5.4 °C. In both periods of the year and the two SSP's, ACIs were considered higher than ideal, ranging from 2.46 ± 0.33 to 3.31 ± 0.62 (SSP1) and 2.42 ± 0.30 to 3.45 ± 0.66 (SSP2).

Development of the body condition score system in Murrah buffaloes: validation through ultrasonic assessment of body fat reserves

The body condition score (BCS) system is a subjective scoring method of evaluating the energy reserves of dairy animals to provide better understanding of biological relationships between body fat, milk production and reproduction. This method helps in adopting the optimum management practices to derive maximum production and maintain optimum health of the livestock. In this study, a new BCS system was developed for Murrah buffaloes. The skeletal check points were identified by studying the anatomical features and amount of fat reserves in slaughtered animals. The scores were assigned from 1 to 5 based on the amount of fat reserves in slaughtered animals. A score of 1 represents least and 5 represents most amount of fat. The skeletal check points identified were ordered based on the amount of carcass fat reserves and scores assigned to prepare a preliminary BCS chart on a 1 to 5 scale at 0.25 increments. The BCS chart was further modified by eliminating the skeletal check points at which the fat reserves were less evident on palpation in most of the buffaloes and a new BCS chart on a 1 to 5 scale at 0.5 increments examining eight skeletal check points was developed. The new BCS system developed was tested for precision in 10 buffaloes for each point of the 1-5 scale by ultrasonographic measurements of body fat reserves. Ultrasonographic measurements showed that as the BCS increased, the amount of fat reserves also increased (p < 0.01), indicating that the BCS adequately reflected the amount of actual fat reserves. BCS was significantly correlated (r = 0.860) with the carcass fat reserves as well as the ultrasonographic fat reserves (r = 0.854).

Thickness of cumulus cell layer is a significant factor in meiotic competence of buffalo oocytes

This study evaluated the meiotic competence of buffalo oocytes with different layers of cumulus cells. A total of 588 oocytes were collected from 775 ovaries averaging 0.78 oocytes per ovary. Oocytes with homogenous cytoplasm (n = 441) were selected for in vitro maturation (IVM) and divided into four groups based on their cumulus morphology: a) oocytes with > or == 3 layers of cumulus cells, b) 1-2 layers of cumulus cells and oocytes with partial remnants or no cumulus cells to be cocultured c) with or d) without cumulus cells. Oocytes in all four groups were matured in 100 microL drop of TCM-199 supplemented with 10microgram/mL follicle stimulating hormone (FSH), 10microgram/mL luteinizing hormone (LH), 1.5microgram/mL estradiol, 75microgram/mL streptomycin, 100 IU/mL penicillin, 10 mM Hepes and 10% FBS at 39degrees C and 5% CO2 for 24 hours. After IVM, cumulus cells were removed from oocytes using 3 mg/mL hyaluronidase, fixed in 3% glutaraldehyde, stained with DAPI and evaluated for meiotic competence. The oocytes with > or ==3 layers of cumulus cells showed higher maturation rates (p <0.05: 64.5%) than oocytes with partial or no cumulus cells (8.6%) and oocytes co-cultured with cumulus cells (34.5%) but did not differ from oocytes having 1-2 layers of cumulus cells (51.4%). The degeneration rates were higher (p < 0.05) for oocytes with partial or no cumulus cells (51%) than rest of the groups (range: 13.8% to 17.4%). These results suggest that buffalo oocytes with intact layers of cumulus cells show better IVM rates than oocytes without cumulus cells and the co-culture of poor quality oocytes with cumulus cells improves their meiotic competence.

Effect of naloxone on GnRH-induced LH and FSH release in buffalo, Bubalus bubalis.

The effect of naloxone on GnRH-induced LH and FSH release was measured in buffaloes in luteal phase of estrous cycle. Animals were administered intravenously, naloxone/saline (50 mg/injection) every 15 min for 3 hr followed by GnRH (100 micrograms). Peripheral plasma LH and FSH concentrations were measured in blood samples collected at 15 min intervals from 1 hr prior to beginning of naloxone/saline treatment up to 3 hr post GnRH administration and every 30 min for the subsequent 3.5 hr. Between the animals of Group I administered naloxone and those of Group II given saline, GnRH-induced peak LH and FSH concentrations, the total LH and FSH released in response to GnRH, and the time to peak LH and FSH concentrations were not significantly different. The results of the present study suggest the absence of a direct effect of naloxone on pituitary responsiveness to GnRH.

Ascorbic acid in buffalo ovary in relation to oestrous cycle.

Concentration of ascorbic acid was determined in different parts of buffalo ovary at four different stages of oestrous cycle viz. early luteal, mid luteal, late luteal and follicular. The stages were decided from the physical and morphological examinations of corpora lutea. The ovary was dissected in three components viz. corpus luteum, follicular fluid and ovarian stromal tissue for ascorbic acid assay. Corpus luteum showed significant change in concentration of ascorbic acid with the advancement of oestrous cycle, value being highest in late- luteal stage. Follicular fluid and ovarian stromal tissue did not show significant changes in ascorbic acid at any stage of the oestrous cycle. Small follicles, irrespective of the stage of oestrous cycle had, however, significantly higher ascorbic acid content than large follicles.

Radiographical study on the time of complete post-natal union of the ossification loci of the bones of the elbow joint in Egyptian buffalo Bubalus

The bones of the elbow joint of the Egyptian buffalo were examined radiographically to determine their ossification loci. This work recorded that there are five separate centers [three of them are located on the distal epiphysis of the humerus, the medial epicondyle, lateral epicondyle and the humeral condyles, while the other two centers are located on the radius and ulna at the proximal epiphysis of the ulna and proximal epiphysis of the radius]. The ossification loci of humeral condyles were fused at the age of 6 months and the proximal epiphysis of the radius were completely fused at the age of 8 months, while that of the medial and lateral epicondyles of the humerus were fused at the age of 1.5 year. The last ossification locus has been fused was the proximal epiphysis of the ulna at the age of six years old

Testosterone concentration in blood serum of buffalo bulls during 24 hours in winter and summer seasons

Concentraçöes de testosterona foram determinadas por radioimunoensaio em 30 amostras de soro sanguíneo obtidas cinco vezes durante 24 horas, de 6 búfalos adultos Jaffarabadi x Mediterrâneo. As amostras foram obtidas durante um dia do inverno e um dia do veräo de 1991. As concentraçöes de testosterona variaram de 0,10 a 1,36 ng/ml no inverno e de 0,10 a 2,454 ng/ml no veräo. Valores máximos foram obtidos às 6,00 horas (0,52 ng/ml) no veräo e 0,82 ng/ml no inverno, ocorrendo entäo duas quedas abruptas, a primeira às 12 horas (0,37 ng/ml) no veräo e 0,21 ng/ml no inverno e a segunda 24 horas (0,17 ng/ml) no veräo e 0,49 ng/ml no inverno. No veräo, níveis mais altos de testosterona sérica foram observados durante o dia

Effect of temperature of thawing and diluent on the post--thaw physiological changes of buffalo frozen semen.

The effect of thawing was studied in buffalo semen diluted in three diluents (Tris egg-yolk, Egg-yolk citrate and Citric Acid whey) at three temperatures (5 degrees C, 35 degrees C and 75 degrees C) on motility, eosin staining, morphological and acrosomal changes, hyaluronidase, glutamic oxalacetic transaminase and glutamic pyruvic transaminase activities. The motility and lack of staining of sperm by eosin were maximum on thawing at 35 degrees C and in tris egg-yolk diluent followed by egg-yolk citrate and citric acid whey. Hyaluronidase, glutamic oxalacetic transaminase and glutamic pyruvic transaminase increased significantly in the extra-cellular fluid on thawing of semen diluted with all the three diluents. The buffalo semen diluted in tris egg-yolk and thawed at 35 degrees C for 30 seconds gave the best results.

Effect of vitamin E on lipid peroxidation in buffalo Bubalus bubalis L.

Vitamin E significantly (P less than 0.01), inhibited lipid peroxidation as indicated by malonaldehyde (MDA) production and improved significantly (P less than 0.01) motility and percent live spermatazoa of B. bubalis semen. Bulls with higher MDA formation had lower sperm motility and percent live count. Variance due to bulls for all the three parameters were significant (P less than 0.05).