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Leishmania spp. are important agents of human and animal leishmaniases that have an important impact on public health. In this study, we aimed to detect the circulation of Leishmania spp. in cattle from a visceral leishmaniasis non-endemic area of the state of São Paulo, Brazil. DNA was extracted from blood samples from 100 heifers in the municipality of Pirassununga and was amplified using primers specific for the first internal transcriber spacer (ITS1), to assess the presence of trypanosomatids. The assays revealed that one sample presented bands of between 300 and 350 base pairs. In GenBank, this sample matched 100% with Leishmania infantum (314 base pairs). The results suggest that cattle can be infected by Leishmania infantum in Brazil.

Two experiments were conducted to investigate the effects of equine chorionic gonadotropin (eCG) at progestin removal and gonadotropin-releasing hormone (GnRH) at timed artificial insemination (TAI) on ovarian follicular dynamics (Experiment 1) and pregnancy rates (Experiment 2) in suckled Nelore (Bos indicus) cows. Both experiments were 2 × 2 factorials (eCG or No eCG, and GnRH or No GnRH), with identical treatments. In Experiment 1, 50 anestrous cows, 134.5 ± 2.3 d postpartum, received a 3 mg norgestomet ear implant sc, plus 3 mg norgestomet and 5 mg estradiol valerate im on Day 0. The implant was removed on Day 9, with TAI 54 h later. Cows received 400 IU eCG or no further treatment on Day 9 and GnRH (100 μg gonadorelin) or no further treatment at TAI. Treatment with eCG increased the growth rate of the largest follicle from Days 9 to 11 (means ± SEM, 1.53 ± 0.1 vs. 0.48 ± 0.1 mm/d; P < 0.0001), its diameter on Day 11 (11.4 ± 0.6 vs. 9.3 ± 0.7 mm; P = 0.03), as well as ovulation rate (80.8% vs. 50.0%, P = 0.02), whereas GnRH improved the synchrony of ovulation (72.0 ± 1.1 vs. 71.1 ± 2.0 h). In Experiment 2 (n = 599 cows, 40 to 120 d postpartum), pregnancy rates differed (P = 0.004) among groups (27.6%, 40.1%, 47.7%, and 55.7% for Control, GnRH, eCG, and eCG + GnRH groups). Both eCG and GnRH improved pregnancy rates (51.7% vs. 33.8%, P = 0.002; and 48.0% vs 37.6%, P = 0.02, respectively), although their effects were not additive (no significant interaction). In conclusion, eCG at norgestomet implant removal increased the growth rate of the largest follicle (LF) from implant removal to TAI, the diameter of the LF at TAI, and rates of ovulation and pregnancy rates. Furthermore, GnRH at TAI improved the synchrony of ovulations and pregnancy rates in postpartum Nelore cows treated with a norgestomet-based TAI protocol.

We evaluated the efficiency of replacing GnRH with LH in the ovulation synchronization protocol in buffaloes. Buffaloes received GnRH on Day 0, (Buserelin; Conceptal, 20 microg), PGF2alpha (Luprostiol; Prosolvin, 15 mg) on Day 7 and GnRH (Buserelin; Conceptal, 10 microg; Group 1) or porcine LH (LH; Lutropin-V, 12.5 mg; Group 2) on Day 9. In Experiment 1, we studied the follicular dynamics of 30 buffaloes (Group 1, n = 15 and Group 2, n = 15). We performed ultrasonography every 12 h from Days 0 to 2, then on Day 7 and then every 6 h from the time of GnRH or LH treatment (Day 9) until the time of ovulation. All females not ovulating by 48 h after the second GnRH or LH injection were considered as nonresponders. In Experiment 2, we evaluated 305 buffaloes (Group 1, n = 154; Group 2, n = 151), using the same two treatments studied in Experiment 1. We also recorded and evaluated aspects like parity, lactational status, the presence of mucus, and uterine tone at the time of artificial insemination (Al). In Experiment 1, ovulation rate after the first GnRH was 86.6% (26/30). Ovulation rates were 93.3% (14/15; Group 1) after the second dose of GnRH and 93.3% (14/15) after LH (Group 2). Ovulation occurred 36.4+/-10.4 h after the first GnRH. The interval for treatment to ovulation was 26.5+/-9.6 h for buffaloes treated with GnRH (Group 1) and 24.4+/-7.9 h for buffaloes treated with LH (Group 2); the time of ovulation did not differ statistically between the two groups (GnRH versus LH; P0.05). In Experiment 2, conception rates of the animals AI in the field were 56.5% (Group 1) and 64.2% (Group 2), respectively (P = 0.08). The response to the treatment with LH was not different to the treatment with GnRH; however, multiparous buffaloes had higher conception rates than the primiparous buffaloes in both groups (P0.05). Buffaloes with mucus at the time of AI in Group 2 had higher conception rates than the buffaloes that had mucus in Group 1 (P0.05). Uterine tone and lactational status did not influence conception rates (P0.05). In summary, the results showed that both treatments resulted in synchronization of ovulation and acceptable conception rates. Therefore, the exogenous injection of LH can substitute the GnRH injections in the Ovsynch program in buffaloes.

The negative effect of some diseases, such as bovine viral diarrhea (BVD), infectious bovine rhinotracheitis (IBR), and Leptospirosis, on bovine reproduction rates are well known. The uses of vaccines are considered to be an important tool available in order to control reproductive losses but their efficiency is still controversial. The aim of this study was to evaluate the effects of vaccination against BVD, IBR, and Leptospirosis to improve pregnancy rate in beef cattle submitted to fixed-timed AI (FTAI). Nelore cows (n = 1172) from 4 beef cattle farms in Brazil were randomly distributed in two experimental groups: Group 1 (treated, n = 584) received the first dose of the inactivated vaccines (Bioleptogen and Bioabortogen H, Biogénesis Bagó, Garín, Argentina) at the beginning of the FTAI protocol and the second dose on the pregnancy diagnostic 40 days later; Group 2 (control group, n = 588) received 0.9% saline solution. Serum samples from 3–5% of animals in each farm herd were collected to determinate IBR, BVD, and Leptospirosis challenges, by using ELISA protocol for BVD and IBR (Synbiotics BVD p80 ab monoblocking test and Synbiotics BoHV-1 gB monoblocking test, respectively; Synbiotics Corp., Kansas City, MO, USA); and microaglutination test for Leptospirosis. All animals were submitted to the FTAI protocol: D0 = intravaginal P4 device (Cronipress, Biogenesis Bagó) and application of 2 mg oestradiol benzoate (Bioestrogen, Biogénesis Bagó); Day 8 = intravaginal device removal + 0.5 mg oestradiol cipionate (E.C.P, Zoetis Inc., Florham Park, NJ, USA) and 25 mg D-cloprostenol sodium (Croniben, Biogénesis Bagó) and FTAI after 48 h. Body condition scores (BCS) were measured on Day 0 and pregnancy diagnostic were performed on Day 40. Data were analysed by logistic regression using PROC LOGISTIC procedure of the SAS software (SAS 9.3, SAS Institute, Inc., Cary, NC, USA) as well as the significant differences between the factors was analysed to nonparametric statistical frequency (chi-square test; PROC FREQ). Pregnancy rates on day 40 were greater (P

O objetivo desse estudo foi avaliar eficiência do protocolo de sincronização da ovulação (GnRH/PGF2alfa/GnRH) para inseminação artificial em tempo fixo em bubalinos. No Experimento 1, trinta e três búfalas com 60 dias pós-parto, foram separadas em dois grupos para avaliação da dinâmica folicular (Grupo 1, n = 16, 10mg de GnRH/15mg de PGF2alfa/10mg de GnRH; Grupo 2, n = 17, 20mg de GnRH/PGF2alfa/10mg de GnRH). Exames ultra-sonográficos foram realizados de 12 em 12 horas para verificar a resposta ovariana à primeira aplicação de GnRH. Outras avaliações foram realizadas nos dias da aplicação da PGF2alfa e do segundo GnRH. Em seguida, foram realizados exames de 6 em 6 horas até o momento da ovulação. Foram colhidas amostras de sangue nos dias das aplicações para avaliar as concentrações de progesterona no momento das administrações hormonais. No Experimento 2, foi verificada a eficiência da sincronização da ovulação para inseminação artificial em tempo fixo (16 horas após o segunda dose de GnRH, n=1053), durante as estações reprodutiva favorável e desfavorável, nos anos de 1998 e 1999. Os animais receberam 20mg de GnRH/PGF2alfa/10mg de GnRH. Ainda, foi analisada a influência da condição corporal, ordem de partos, período pós-parto ao início do tratamento, ano e a estação reprodutiva (favorável ou desfavorável), sobre a taxa de concepção. No Experimento 1, verificou-se que 33,0 ± 8,3h após a primeira aplicação de GnRH, 60,60% dos animais ovularam (Grupo 1 = 70,5 vs. Grupo 2 = 50,00%; P>;0,05). A dose de GnRH (10mg vs. 20mg) e a concentração de progesterona no início do tratamento não interferiu na ovulação (P>;0,05). Os animais que ovularam (n = 20) após a primeira aplicação de GnRH apresentaram maior diâmetro folicular (P;0,05). No Experimento 2, observou-se taxa de concepção de 45,40% (n = 1053). Animais sincronizados durante a estação reprodutiva favorável apresentaram melhores taxas de concepção quando comparados ao período desfavorável (48,80% vs. 6,90%; P < 0,05). A taxa de concepção foi influenciada pela condição corporal dos animais e pela ordem de partos durante a estação reprodutiva favorável (P;0,05). Em conclusão, os Experimentos indicaram que a utilização da sincronização da ovulação para inseminação artificial em tempo fixo, durante a estação reprodutiva favorável, apresenta resultados satisfatórios em bubalinos, podendo ainda ser melhorados quando utilizadas búfalas multíparas com bom estado corporal no início do tratamento. The objective of this study was to evaluate the efficiency of synchronization ovulation protocol (GnRH/PGF2alpha/GnRH) for fixed timed insemination in buffalo. In Experiment 1, thirty-three female buffaloes with 60 days postpartum, were divided in two groups for evaluation of the follicular dynamics (Group 1, n = 16, 10mg of GnRH/15mg of PGF2alpha/10mg of GnRH; Group 2, n = 17, 20mg of GnRH/15mg of PGF2alpha/10mg of GnRH). Ultrasonography exams were performed every 12 hours to verify the ovarian response to the first GnRH injection. Another evaluations were performed on PGF2a injection and second GnRH injection. After last hormonal injection, ultrasonography exams were performed every 6 hours to verify the moment of the ovulation. Blood samples were collected on Days 0, 7 and 9 to evaluate the progesterone concentrations at the moment of the hormonal injections. In Experiment 2, the efficiency of the synchronization of ovulation was verified for fixed timed insemination (16 hours after second GnRH injection, n=1053), during the favorable and unfavorable reproductive season, in the years of 1998 and 1999. Animals received 20mg of GnRH/15mg of PGF2alpha/10mg of GnRH. Besides, also recorded and evaluated aspects like body condition score, parity, postpartum period, year and the reproductive season (favorable or unfavorable season), on the conception rate. In Experiment 1, it was verified that 33.0 ± 8.3h after the first GnRH injection, 60.60% of the animals ovulated (Group 1 = 70.5 vs. Group 2 = 50.00%; P>;0.05). The dose of GnRH (10mg vs. 20mg) and the progesterone concentration did not interfere in ovulation in the beginning of the treatment (P>;0.05). Animals that ovulated (n = 20) after the first GnRH injection presented larger follicular diameter (P ;0.05). In Experiment 2, it was observed conception rate of 45.40% (n = 1053). Animals synchronized during the favorable reproductive season presented better conception rate than the unfavorable season (48.80% vs. 6.90%; P ;0.05). In summary, the Experiments indicated that the use of synchronization of the ovulation for fixed timed insemination, during the favorable reproductive season, presents satisfactory results in buffalo and could still be better when used multiparous buffaloes with good body conditon score in the beginning of the treatment.

O objetivo deste estudo foi avaliar a eficiência do controle da ovulação através da aplicação de PMSG, GnRH, Benzoato de Estradiol e PGF2a junto ao protocolo comercial Crestar®, para viabilizar a IA em tempo fixo. Foram utilizadas 348 vacas multíparas, cruzadas Nelore (Bos taurus indicus) X Charolês (Bos taurus taurus), divididas em dois grupos: 179 vacas paridas com bezerros de 90 a 120 dias de idade e outras 169 vacas solteiras. Estes animais foram submetidos a cinco tratamentos de controle farmacológico do ciclo estral e ovulação para a inseminação artificial em tempo fixo. Dentro dos cinco tratamentos todas as vacas receberam o protocolo Crestar â como agente sincronizador do crescimento folicular. Este protocolo consiste na colocação implante subcutâneo com 3mg de norgestomet e na injeção 3mg de norgestomet + 5mg de valerato de estradiol (i.m. - simultânea a colocação do implante). Após a remoção do implante (9 dias), as vacas foram submetidas aos cinco tratamentos de controle da ovulação: T1 - (n=70): uma injeção de solução fisiológica 48h após a retirada do implante (D 11); T2 - (n=68): 0,75mg de benzoato de estradiol 24h após a retirada do implante (D 10); T3 - (n=70): aplicação de 7,5mg de PGF2a, no dia da retirada (D 9) e 0,75mg de benzoato de estradiol 24h após a retirada do implante (D 10); T4 - (n=70): 500 UI de PMSG na retirada do implante (D 9) e T5 - (n=70): 500mg de GnRH 48h após a retirada dos implante (D 11). Todos os animais foram inseminados 54-56h após a retirada do implante. As taxas de prenhez foram analisadas estatisticamente por regressão logística. Não houve diferença entre os tratamentos (p>0,05) onde: 35,7, 31,4, 22,0, 37,0 e 42,8% para T1, T2, T3, T4 e T5 respectivamente. Entretanto a taxa de prenhez das vacas solteiras foi maior (39,6% - P0.05) 35.7, 31.4, 22.0, 37.0 and 42.8% for T1, T2, T3, T4 and T5, respectively. However, there was difference (p

Os resultados das taxas de fertilidade e dos tamanhos das leitegadas foram analisados na granja de suínos da Faculdade de Medicina Veterinária e Zootecnia da Universidade de São Paulo no Campus de Pirassununga, durante o período de 4 anos (1992 a 1995), com o objetivo de comparar a inseminação artificial (IA) e a cobertura natural (CN) em diferentes épocas do ano, procurando estudar as influências sazonais sobre os índices reprodutivos. Foram avaliadas 799 coberturas em fêmeas das raças Landrace (L), Large White (LW) e mestiças, sendo 539 de IA e 260 de CN. Reprodutores (L e LW) de comprovada fertilidade foram utilizados tanto para CN quanto para IA. As doses de sêmen apresentavam concentração mínima de 3 bilhões de espermatozóides em volume de 100 ml. As IA foram realizadas às 12 e às 24 horas após reflexo positivo de tolerância ao macho, enquanto as montas foram realizadas no momento e às 24 horas após o diagnóstico do cio. Os índices de fertilidade foram de 72,9% e 75,8% e o número de leitões nascidos de 12,4 e 12,1, respectivamente, para inseminação artificial e monta natural, não mostrando diferença significativa. A estação climática influenciou a taxa de parição (71,2%, 81,4%, 76,9% e 66,4%, para verão, outono, inverno e primavera, respectivamente; p < 0,05), mas não mostrou efeito sobre o tamanho da leitegada. Não houve influência do tipo de cobertura (IA ou CN) e da interação entre estação climática e tipo de cobertura sobre as taxas de parições e os tamanhos das leitegadas. As médias de temperatura máxima e mínima se correlacionaram negativamente com a taxa de parição. The data of parturition rate and litter size from the herd of the Veterinary School of São Paulo University were analysed during a four years period (1992 to 1995) in order to compare the data from artificial insemination (IA) and natural mating (CN) in different seasons, trying to study the seasonal influences and define which of these methods would be better in order to reach adequate reproductive rates. Data from 799 breeding (539 IA and 260 CN) in Landrace (L), Large White (LW) females and the cross-breed from both were analysed. Boars (L and LW) with known fertility were used either for IA or for CN. The semen doses used had at least 3 billions spermatozoa in 100 ml. The inseminations were performed 12 and 24 hours after the positive back pressure test in response to the boar, and natural mating were performed at the moment of positive back pressure test and 24 hours later. The fertility rates were 72.9% and 75.8% and the litter size was 12.4 and 12.1, for artificial insemination and natural mating, respectively, which were not different. The season influenced the parturition rate (71.2%, 81.4%, 76.9% and 66.4%, for summer, autumn, winter and spring, respectively; p < 0.05) but not the litter size.

O uso de suplementacao com gordura no pre e pos-parto mostra-se uma alternativa para incrementar a densidade energetica da dieta. Entretanto, a suplementacao com gordura resulta em efeitos inconsistentes e variados sobre a funcao reprodutiva. Esclarecer como esta suplementacao pode influenciar a reproducao e um processo delicado. A complexidade do sistema reprodutivo assim como das fontes de suplementacao com gordura sao influenciadas pelas condicoes de manejo e da qualidade da forrageira. SUMMARY Fat supplementation, before and after calving, is a good alternative to increase energy density of the diet. However, feeding supplemental fat has resulted in varied and inconsistent effects on the reproductive function of cattle. Furthermore, understanding how fat supplementation can influence the reproductive function has been a difficult process. Reproductive system and the fat supplementation sources are influenced by management conditions and forage quality.

The growth, selection, regression and ovulation of ovarian follicles was ultrasonically monitored in 30 Murrah buffalo throughout a spontaneous estrous cycle during the breeding season (autumn). Examinations revealed that follicular growth during the estrous cycle occurs in waves; the buffalo showed 1-wave (3.3%, n = 1), 2-wave (63.3%, n = 19) or 3-wave (33.3%, n = 10) follicular growth. The first wave began at 1.00, 1.16 +/-0.50 and 1.10 +/- 0.32 d in buffalo with 1, 2 and 3 waves, respectively (ovulation = Day 0). The second wave appeared at 10.83 +/- 1.09 and 9.30 +/- 1.25 d (P0.01) for the 2 and 3 wave cycle animals, respectively. The third wave started at 16.80 +/- 1.22 d. Structural persistence of the first dominant follicle was longer in the 2- than 3-wave cycles (20.67 +/- 1.18 vs 17.90 +/- 3.47 d ; P0.05). The duration of the growth and static phases of the first dominant follicle differed between the 2 and 3 wave cycles (P0.05), whereas there were no differences in linear growth rates (cm/d). Two and three wave cycles differed (P0.05) with respect to the maximum diameter of both the first dominant follicle (1.51 +/- 0.24 vs 1.33 +/- 0.18 cm) and the ovulatory follicles (1.55 +/- 0.16 vs 1.34 +/- 0.13 cm). No relationship was found between dominant follicle development and the presence of either a CL or a previous dominant follicle in either ovary. Two and three wave cycles also differed with respect to the mean length of intervals between ovulation (22.27 +/- 0.89 vs 24.50 +/- 1.88 d; P0.01) and the mean length of luteal phases (10.40 +/- 2.11 vs 12.66 +/- 2.91 d; P0.05). These results demonstrate that buffalo have estrous cycles with 1, 2 or 3 follicular waves; that 2-wave cycles are the most common; and that the number of waves in a cycle is associated with the luteal phase and with estrous cycle length.

Intravaginal progesterone (P4) releasing devices are widely used in hormonal protocols for fixed-time artificial insemination (FTAI). To reduce production cost in Brazilian farms, after first use these devices are currently reused once or twice, providing good reproductive efficiency in cattle. The aim of this study was to compare P4 plasma concentrations among time in ovariectomized cows receiving 3 different commercially available devices: Sincrogest® (SIN, 1 g of P4), Cronipres® (CRO, 1 g of P4 and 3 rings of 0,1 g of P4 for the third use) and Primer® (PRI, 1 g of P4). For each type, new (1st use, n = 2), once-used (2nd use, n = 2) and twice-used (3rd use, n = 2) devices were tested. Blood samples were collected at times 0 (before device insertion), 2, 4, 8, 10, 12, 24, 48, 72, 96, 120, 144, 168, and 192 h (Day 0 to Day 8). P4 plasma concentrations were measured by radioimmunoassay and the averages for each experimental group were submitted to analysis of variance adding repeated-measures in time factor. Comparisons were made among 3 types of device in each of 3 uses. P4 concentrations (ng mL–1) and standard deviation in the periods in which there was statistical difference (P

Foram avaliados os períodos pós-parto de vacas primíparas mestiças Gir x Caracu e Nelore x Caracu mantidas em regime extensivo de pastagem. Para a determinação dos níveis hormonais de progesterona, amostras de leite foram coletadas semanalmente até que os animais apresentassem o 1º estro perceptível, quando foram inseminadas artificialmente. Decorridos 21 dias da inseminação, amostras foram coletadas e dosadas para fazer o diagnóstico precoce de gestação. Os resultados mostraram início de ciclicidade ovariana com 9,89 semanas após o parto para vacas Gir x Caracu, com valores médios de progesterona de 0,28 nmol/l no dia do estro e 7,62 nmol/1 aos 21 dias após a inseminação. Para vacas Nelore x Caracu, o estro se deu com 12,08 semanas após o parto e com valores médios de progesterona de 0,46 nmol/l e 9.23 nmol/1 para o dia do estro e no 21º dia, respectivamente. Dez gestações de doze diagnosticadas no 21º dia foram confirmadas por palpação retal, tendo ocorrido um caso de mortalidade embrionária precoce. Outras três vacas foram inseminadas durante a fase luteínica. A eficiência na observação de estros foi de 52.94%, pois de 34 identificados pelos níveis hormonais, 18 foram percebidos visualmente. A variação entre os cruzamentos não alterou significativamente o nível de progesterona no dia do estro, tampouco no 21º dia após inseminação e também quanto ao período necessário ao reinício da atividade ovariana pós - parto. Progesterone concentrations were measured by radioimmunoassay in fat-free milk samples obtained weekly from 26 primiparous, suckling cows, raised in range conditions. Samples were collected in post partum period up to observation of first signs of estrus. Profiles of progesterone concentrations for early pregnancy diagnosis were also examined on milk samples taken 21 days after artificial insemination. Results showed ovarian ciclicity starting 9.89 weeks post partum in crossed Gir x Caracu cows, with 0.28 nmol/1 progesterone on the day of estrus anti 7.62 nmol/1 21 days post Al. In crossed Nelore x Caracu cows, these results were 12.08 weeks, 0.46 nmol/1 and 9.23 nmol/1, respectively. Ten pregnancies out of 12 diagnosed were confirmed by rectal palpation. There was one case of embryo mortality and three AI were practiced during luteal phase. Estrus observation efficiency was 52.94%, since out of 34 heats identified through hormonal determinations, 18 were detected visually. Variations between crossings did not significantly alter progesterone profiles on day of estrus, nor on 21th day after AI or as for ovarian activity post partum.

Duzentos e vinte e uma vacas (78 paridas com bezerros entre 40 e 90 dias de idade e as demais secas vazias) Nelore (Bos taurus indicus) e mestiças Charolês (Bos taurus taurus) foram utilizadas para avaliar a reutilização de implantes auriculares de silicone impregnados com progestágenos para controle farmacológico do ciclo estral e ovulação sobre a taxa de prenhez. Os tratamentos experimentais consistiram de três protocolos para sincronização do cio e inseminação artificial em tempo fixo (IATF). No tratamento 1 (T1; n = 73) as vacas receberam um implante auricular de Crestar® (3 mg de norgestomet); no tratamento 2 (T2; n = 75) as vacas receberam um implante auricular de Crestar® já utilizado previamente; e no tratamento 3 (T3; n = 73) as vacas receberam dois implantes auriculares de Crestar® também utilizados previamente. Além dos implantes, as vacas receberam aplicação intramuscular (i.m) 2 mL de progesterona (25 mg/mL) + 2 mL de benzoato de estradiol i.m (1 mg/mL) no momento da colocação dos implantes. Os implantes foram removidos após oito dias junto a uma aplicação de 2,0 mL de Preloban® i.m (150 µg de D-cloprostenol). Após 24 h da remoção do implante, aplicou-se 1 mL de Estrogin® i.m (1 mg de benzoato de estradiol). A IATF foi realizada 54-56 h após a retirada dos implantes. Não houve diferença na taxa de prenhez das vacas, sendo 39,72%, 34,21% e 36,98% para os tratamentos T1, T2 e T3, respectivamente. A reutilização de implantes com progestágeno (Crestar®) não alterou a taxa de prenhez de vacas aptas à reprodução.

The potential to use a GnRH agonist bioimplant and injection of exogenous LH to control the time of ovulation in a multiple ovulation and embryo transfer (MOET) protocol was examined in buffalo. Mixed-parity buffalo (Bubalus bubalis; 4-15-year-old; 529 13 kg LW) were randomly assigned to one of five groups (n = 6): Group 1, conventional MOET protocol; Group 2, conventional MOET with 12 It delay in injection of PGF(2alpha); Group 3, implanted with GnRH agonist to block the pre-ovulatory surge release of LH; Group 4, implanted with GnRH agonist and injected with exogenous LH (Lutropin(R), 25 mg) 24 h after 4 days of superstimulation with FSH; Group 5, implanted with GnRH agonist and injected with LH 36 h after superstimulation with FSH. Ovarian follicular growth in all buffaloes was stimulated by treatment with FSH (Folltropin-V(R), 200 mg) administered over 4 days, and was monitored by ovarian ultrasonography. At the time of estrus, the number of follicles greater than or equal to8 mm. was greater (P < 0.05) for buffaloes in Group 2 (12.8) than for buffaloes in Groups 1 (8.5), 3 (7.3), 4 (6.1) and 5 (6.8), which did not differ. All buffaloes were mated by AI after spontaneous (Groups 1-3) or induced (Groups 4 and 5) ovulation. The respective number of buffalo that ovulated, number of corpora lutea, ovulation rate (%), and embryos + oocytes recovered were: Group 1 (2, 1.8 +/- 1.6, 18.0 +/- 13.6, 0.2 +/- 0.2); Group 2 (4, 6.1 +/- 2.9, 40.5 +/- 17.5, 3.7 +/- 2.1); Group 3 (0, 0, 0, 0); Group 4 (6, 4.3 +/- 1.2, 69.3 +/- 14.2, 2.0 +/- 0.9); and Group 5 (1, 2.5 +/- 2.5, 15.5 +/- 15.5, 2.1 +/- 2.1). All buffaloes in Group 4 ovulated after injection of LH and had a relatively high ovulation rate (69%) and embryo recovery (46%). It has been shown that the GnRH agonist-LH protocol can be used to improve the efficiency of MOET in buffalo.