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Made available in DSpace on 2022-05-01T06:02:17Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-12-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Empresa Brasileira de Pesquisa Agropecuária Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Climate change is a reality and global surface temperature is projected to rise substantially in the next 80 years. Agriculture practices will have to adapt to climate change, and also help to mitigate this effect using, among other strategies, forest conservation and management. Silvopastoral systems have been adopted in tropical climate livestock areas but their benefits on thermal comfort and reproductive performance of beef cows are not completely known. Therefore, our aims were to compare the microclimate of silvopastoral and intensive rotational unshaded grazing systems in different months and to evaluate physiological variables (Exp. 1 and 2), metabolism, and in vitro embryo production (Exp. 2) in crossbred beef females. Our hypothesis is that the silvopastoral system can improve the thermal comfort of beef heifers and cows and, consequently, also improve dry matter intake, body weight gain, and in vitro embryo production when compared to the unshaded rotational grazing system. In Exp 1, the silvopastoral system decreased body temperature and increased welfare and performance of heifers. In Exp. 2, the silvopastoral system enhanced the body weight but did not affect metabolism and the general reproductive performance, but increased the recovery rate of oocytes in primiparous cows. Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual de São Paulo “Júlio de Mesquita Filho”, Rod. Prof. Paulo Donato Castellane, s/n Embrapa Pecuária Sudeste, Rod. Washington Luiz, km 234, Fazenda Canchim, PO Box: 339 Universidade Federal Fluminense, Rua Vital Brazil, 64 Vitrogen, Av. Coronel José Nogueira Terra, 203 Texas A&M University, 400 Bizzell St University of Florida, PO Box 110910 Instituto de Zootecnia, Rod. Carlos Tonani, km 94 Faculdade de Ciências Agrárias e Veterinárias Universidade Estadual de São Paulo “Júlio de Mesquita Filho”, Rod. Prof. Paulo Donato Castellane, s/n FAPESP: Process 2015/26627-5 FAPESP: Process 2019/04528-6 CNPq: Process 307757/2019-6

Intercellular communication is an essential mechanism for development and maintenance of multicellular organisms. Extracellular vesicles (EVs) were recently described as new players in the intercellular communication. EVs are double-membrane vesicles secreted by cells and are classified according to their biosynthesis, protein markers and morphology. These extracellular vesicles contain bioactive materials such as miRNA, mRNA, protein and lipids. These characteristics permit their involvement in different biological processes. Reproductive physiology is complex and involves constant communication between cells. Different laboratories have described the presence of EVs secreted by ovarian follicular cells, oviductal cells, in vitro produced embryos and by the endometrium, suggesting that EVs are involved in the development of gametes and embryos, in animals and humans. Therefore, is important to understand physiological mechanisms and contributions of EVs in female reproduction in order to develop new tools to improve in vivo reproductive events and assisted reproductive techniques (ARTs). This review will provide the current knowledge related to EVs in female reproductive tissues and their role in ARTs.

Made available in DSpace on 2021-06-25T11:07:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-01 Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) The aims of this study were to verify the occurrence of single nucleotide polymorphisms (SNPs) in the JY-1 gene and their association with the in vivo and in vitro production of embryos in Holstein donors. The JY-1 protein is oocyte-specific and has an important regulatory role in folliculogenesis, oocyte maturation, and early embryonic development. Retrospective data of in vivo (n = 572) and in vitro (n = 1068) embryos produced between 2013 and 2015 were obtained from 144 non-lactating, Holstein cows. To identify the polymorphisms, exons 2 and 3 were studied by polymerase chain reaction (PCR) sequencing. The association of SNPs with embryo traits (number of transferable embryos produced in vivo, rate of transferable embryos in vivo, number of embryos produced in vitro, cleavage rate, and embryo rate in vitro) was determined using the GLIMMIX procedure of SAS. Eleven SNPs were detected, located in exon 2, intron 2, and exon 3. The coding genetic variants were non-synonymous, synonymous, and located in untranslated regions. Nine SNPs were associated with at least one in vitro embryo production trait, but none were associated with in vivo embryo production. JY-1 has potential markers that could be used to select donors, and its influence on in vitro embryo production was observed in the present study. Department of Pathology Theriogenology and One Health School of Agricultural and Veterinarian Science São Paulo State University (UNESP) Department of Animal Science School of Agricultural and Veterinarian Science São Paulo State University (UNESP) Clínica Veterinária Samvet São Carlos National Council for Science and Technological Development Department of Pathology Theriogenology and One Health School of Agricultural and Veterinarian Science São Paulo State University (UNESP) Department of Animal Science School of Agricultural and Veterinarian Science São Paulo State University (UNESP) CNPq: 447172/ 2014–0

Made available in DSpace on 2021-06-25T11:08:49Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-02-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) In cattle, in vitro embryo production (IVEP) is an important reproductive biotechnology responsible for the rapid expansion of the Senepol breed in our country. This breed has shown important results when used in crossbreeding and estimate IVEP in Senepol based on seminal analysis would be valuable for the semen cryopreservation industry, research institutes and breeders. Combining the evaluation of sperm subpopulations with analysis of other sperm attributes may help to improve fertility predictions in cattle. Therefore, the objectives of the present study were to: 1) identify and characterize motile sperm subpopulations in cryopreserved Senepol semen following the washing process carried out before in vitro fertilization, and 2) to determine an model for estimate IVEP based on sperm subpopulations in conjunction with other sperm quality analyzes. Samples of 38 cryopreserved semen from 28 Senepol bulls, chosen based on retrospective data from 386 IVEP routines, underwent the semen washing and were evaluated by the computer-assisted sperm analysis system. Sperm morphology was evaluated by wet preparation technique, and plasma and acrosomal membranes integrity, mitochondrial potential, oxidative status and chromatin resistance were analyzed by flow cytometry. After multivariate analysis of principal components and grouping, three sperm subpopulations were identified: SBP1 (fast and progressive motility), SBP2 (hyperactivated motility) and SBP3 (slow non-progressive motility). After categorization of IVEP in high, medium and low embryo yield, logistic regression analysis was applied to associate the results of subpopulations and other sperm quality variables with IVEP. The SBP1 and SBP2 variables affected embryo production, and an IVEP estimation model was generated for Senepol bulls based on these two subpopulations: embryo yield = 0.1563 + 0.0328 (SBP1) + 0.0173 (SBP2). SBP1 and SBP2 represents the absolute value of the percentage of subpopulations in semen. If the calculated value (by this equation) is close to 1, the embryo yield will be low; if is close to 2, will be medium; if is close to 3, will be high. In conclusion, three subpopulations were found for Senepol semen and, despite all analyzed variables, only SBP1 and SBP2 were included in the model to estimate IVEP in this breed. Universidade Estadual Paulista FCAV/UNESP Senepol 3G In Vitro Brasil Mogi Mirim Universidade de São Paulo FMVZ/USP Universidade de São Paulo FMRP/USP Centro Avançado de Pesquisa de Bovinos de Corte APTA/IZ Universidade Estadual Paulista FCAV/UNESP

Made available in DSpace on 2021-06-25T12:28:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-12-02 Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG) In vitro embryo production and embryo transfer (ET) in buffaloes has been developed for decades. However, most studies are focused on the donor or laboratory improvements, and there is a lack of reports regarding the recipients. Therefore, our aim was to investigate factors associated to pregnancy (P/ET), pregnancy loss (PL), and calving rates in buffalo recipients. The studied factors were season, recipient parity, the synchronization protocol, the CL diameter, asynchrony between the embryo and the recipient, the day of the recipient estrous cycle, the embryo (fresh vs. vitrified), the day of embryo development, and the embryo stage. These retrospective data, from a program of in vitro produced embryos, were analyzed by logistic regression, and the odds ratio was also estimated. Two factors were related to P/ET and the calving rate: (1) progesterone associated to estradiol plus eCG protocol for fixed time ET tended to affect positively P/ET on day 30 (41.9 vs. 36.1%, respectively; P = 0.07; AOR = 1.28) and P/ET on day 60 (37.8 vs. 36.1%, respectively; P = 0.09; AOR = 1.08) compared to the Ovsynch protocol; and (2) the CL diameter (>= 14.5 mm) at transfer increased P/ET on day 30 (47.4 vs. 32.5%; P < 0.01; AOR = 1.87) and on day 60 (45.3 vs. 27.7%; P < 0.01; AOR = 2.16), and also the calving rate (37.9 vs. 21.7%; P < 0.01; AOR = 2.20). PL was greater when ET was done in the nonbreeding season compared to the breeding season (PL 30-60: 12.8 vs. 0.0%, P = 0.01; AOR > 999.99; PL 60-calving: 26.8 vs. 3.6%, P = 0.03; AOR = 9.90; and PL 30-calving: 36.2 vs. 3.6%, P = 0.01; AOR = 15.30). In conclusion, the data of our study indicated that the synchronization protocol, the CL diameter, and ET during the breeding season impacted the reproductive efficiency of buffalo recipients. Univ Estadual Paulista, Dept Patol Reprod & Saude Unica, Jaboticabal, Brazil Cenatte Embrioes Soc Ltd, Pedro Leopoldo, Brazil Univ Napoli Federico II, Dipartimento Med Vet & Prod Anim, Naples, Italy Univ Illinois, Dept Anim Sci, 328 Mumford Hall, Urbana, IL 61801 USA Univ Sao Paulo, Dept Reprod Anim, Sao Paulo, Brazil Univ Fed Juiz de Fora, Dept Med Vet, Juiz De Fora, Brazil Univ Fed Minas Gerais, Dept Med Vet Prevent, Belo Horizonte, MG, Brazil Univ Estadual Paulista, Dept Patol Reprod & Saude Unica, Jaboticabal, Brazil FAPEMIG: APQ-0591-5.04/08

Cryopreservation causes damage to spermatozoa, and methods minimizing this damage are therefore needed. Although much discussed, seminal plasma removal has become an alternative to improve sperm quality and viability after freezing and has been applied to different species in attempt to obtain good results. The objective of this study was to evaluate semen quality in buffaloes submitted to two methods for seminal plasma removal (filtration and centrifugation). Semen samples were collected from seven Murrah buffalo bulls (Bubalus bubalis) once a week for 8 weeks. Each ejaculate was divided into three groups: control (presence of seminal plasma), centrifugation and filtration. Sperm kinetics was evaluated with the computer-assisted sperm analysis (CASA) system. Plasmalemma and acrosomal membrane integrity, mitochondrial membrane potential and reactive oxygen species (ROS) were measured by flow cytometry, and lipid peroxidation was evaluated by the thiobarbituric acid reactive substances (TBARS) assay. Seminal plasma removal did not improve sperm kinetics compared to the control group. Centrifugation increased the number of cells with damaged acrosomal membranes (0.77 ± 0.05) and filtration caused greater plasmalemma and acrosomal membrane damage (22.18 ± 1.07). No difference in the mitochondrial membrane potential was observed between groups. In contrast, ROS production was higher in the centrifugation group compared to the control and filtration groups, although no differences in TBARS formation were detected. In conclusion, seminal plasma removal did not improve the quality of thawed buffalo semen compared to control in terms of sperm kinetics, membrane integrity, mitochondrial membrane potential or lipid peroxidation.

Anti-Müllerian Hormone (AMH) is a protein expressed in the gonads and related to ovarian follicular development. The aim of this study was to investigate the presence of polymorphisms in the AMH gene in a Nelore herd to analyze the genetic constitution of this population and to perform association studies with early pregnancy occurrence (EPO) and age at first calving (AFC). Phenotypic data consisted of verification of early pregnancy and age at first calving of 197 unrelated heifers exposed to the mating season and aged between 15 and 17 months (precocious group) and 24 months (non-precocious group). The "precocious" group consisted of 67 heifers with age at first calving of 26.5 ± 0.59 months, and the "non-precocious" group was composed of 130 heifers with age at first calving of 36.4 ± 0.99 months. All five exons of the AMH gene were amplified by Polymerase Chain Reaction (PCR) and sequenced. A total of three SNPs were identified in this study, all of them located in exon 5 (rs527023314, rs722016629, and rs134387246), the latter one positioned in the stop codon. All three SNPs identified in exon 5 characterized synonymous mutations. Only SNP rs134387246 exhibited a significant value (P ? 0.10) for EPO and AFC. The association study of SNP rs134387246 revealed an over-dominance effect (P = 0.056), and no additive effect was observed (P = 0.67). A reduction of 2.5 months (75 days) in the age at first calving of heterozygous heifers for the SNP rs134387246 was observed. For the first time, polymorphisms of the AMH gene were described in Nelore heifers and associated with sexual precocity traits. O hormônio Anti-Mülleriano (AMH) é uma proteína expressa nas gônadas e está relacionada ao desenvolvimento folicular ovariano. O objetivo desse trabalho foi investigar a presença de polimorfismos do gene AMH em um rebanho Nelore, analisar a constituição genética da população para o gene e estudar a associação entre os polimorfismos e as características ocorrência de prenhez precoce (OPP) e idade ao primeiro parto (IPP). Os dados fenotípicos consistiram da verificação da prenhez precoce e idade ao primeiro parto de 197 novilhas, não aparentadas, expostas à estação de monta com idade entre 15 e 17 meses (grupo precoce) ou 24 meses (grupo não precoce). O grupo denominado "precoce" foi composto por 67 novilhas com idade ao primeiro parto de 26.5±0.59 meses, e o segundo denominado "não precoce" composto por 130 novilhas com idade ao primeiro parto de 36.4±0.99 meses. Os cinco éxons do gene AMH foram amplificados por meio da técnica de Reação em Cadeia da Polimerase (PCR) e ?sequenciados. Um total de três SNPs foram identificados nesse estudo, todos localizados no éxon 5 (rs527023314, rs722016629 and rs134387246), sendo o último posicionado stop codon. Os três SNPs identificados caracterizaram mutações sinônimas. Somente o SNP rs134387246 exibiu associação significante (P ? 0.10) para ambas características (OPP e IPP). O estudo de associação do SNP rs134387246 revelou um efeito de sobre dominância (P=0,056), e ausência de ação gênica aditiva (P=0,67). Foi observada redução de 2,5 meses (75 dias) na idade ao primeiro parto de novilhas heterozigotas para o SNP rs134387246. Pela primeira vez na literatura, polimorfismos do gene AMH foram descritos em novilhas da raça Nelore e associados a características indicadoras de precocidade sexual.

Made available in DSpace on 2018-12-11T16:58:07Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-01-01 The present aimed to develop a hormonal therapy based on the insertion of a progesterone (P4) insert (PI) during 10 d plus an estradiol injection (E2) at PI removal before the onset of breeding of beef heifers. In Exp. 1, the plasma P4 profile of prepubertal heifers showed that the insertion of a PI or 24 d previously used P4 insert (UPI) sustained plasma P4 above 1ng/mL for at least the first 7 d of the treatment. In Exp. 2 there was no positive effect of additional estradiol benzoate (EB) administered at the insertion of UPI on the proportion of heifers with a corpus luteum (CL/Treated) 30 d after UPI removal [UPI+EB=85.3%a (n=134); EB+UPI+EB=80.8%a (n=125)]; however, both were greater (Pb (n=129)]. In Exp. 3, a positive effect (P=0.01) of UPI treatment and both E2 supplementations [EB and estradiol cypionate (EC); P=0.10] at the UPI removal was observed on CL/Treated [Control=42.5%b (n=94); UPI=58.5%a (n=130); UPI+EB=64.0%a (n=128); UPI+EC=67.2%a (n=128)]. However, greater pregnancy per treated heifer (P/Treated) following artificial insemination (AI) upon estrus detection was achieved when EC was applied [Control=20.2%b; UPI=29.2%ab; UPI+EB=26.6%b; UPI+EC=36.7%a]. In Exp. 4, the treatment prior to the timed AI (TAI) tended to improve pregnancy per TAI [P/AI; Control 43.6% (n=298) vs. UPI+EC 51.9% (n=342); P=0.08], but increased P/Treated [26.5% vs. 43.3%; P

Made available in DSpace on 2018-11-26T17:20:51Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-01-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Zoetis Saude Animal (Campinas -SP, Brazil) The effects of addition of gonadotrophin-releasing hormone (GnRH) to a progesterone plus oestradiol-based protocol and timing of insemination in Holstein cows treated for timed AI (TAI) were evaluated. Cows (n=481) received a progesterone device and 2mg oestradiol benzoate. After 8 days, the device was removed and 25mg dinoprost was administered. Cows were allocated to one of three (Study 1; n=57) or four (Study 2; n=424) groups, accordingly to ovulation inducer alone (Study 1; oestradiol cypionate (EC), GnRH or both) or ovulation inducer (EC alone or combined with GnRH) and timing of insemination (48 or 54h after device removal; Study 2). In Study 1, the diameter of the ovulatory follicle was greater for GnRH than EC. Oestrus and ovulation rates were similar regardless of ovulatory stimuli. However, time to ovulation was delayed when GnRH only was used. In Study 2, cows treated with GnRH or not had similar pregnancy per AI (P/AI) 30 days (41.5% vs 37.3%; P=0.28) and 60 days (35.9% vs 33.0%; P=0.61) after TAI. TAI 48 and 54h after device removal resulted similar P/AI at 30 days (40.3% vs 38.5%; P=0.63) and 60 days (33.8% vs 35.1%; P=0.72). Thus, adding GnRH at TAI does not improve pregnancy rates in dairy cows receiving EC. The flexibility of time to insemination enables TAI of a large number of cows using the same protocol and splitting the time of AI. Univ Sao Paulo, Dept Reprod Anim, Fac Med Vet & Zootecnia, Ave Prof Dr Orlando Marques de Paiva 87, BR-05508270 Sao Paulo, SP, Brazil Univ Sao Paulo, Dept Med Vet, Fac Zootecnia & Engn Alimentos, Ave Duque de Caxias Norte 225,Campus USP, BR-13635900 Pirassununga, SP, Brazil MSD Saude Anim, Ave Nacoes Unidas 14171,Torre C,Crystal Tower, BR-04794000 Sao Paulo, SP, Brazil Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil Cent Pioneiros, Ave Pioneiros, BR-84145000 Carambei, PR, Brazil Univ Estadual Paulista, Dept Prod Anim, Fac Med Vet & Zootecnia, Dist Rubiao Jr S-N,Caixa Postal 560, BR-18618970 Botucatu, SP, Brazil Univ Estadual Paulista, Dept Med Vet Prevent & Reprod Anim, Fac Ciencias Agr & Vet, Via Acesso Prof Paulo Donato Castellane S-N, BR-14884900 Jaboticabal, SP, Brazil Univ Estadual Paulista, Dept Prod Anim, Fac Med Vet & Zootecnia, Dist Rubiao Jr S-N,Caixa Postal 560, BR-18618970 Botucatu, SP, Brazil FAPESP: 2009/00938-3 FAPESP: 2012/07510-1 CNPq: 486089/2013-14

The aim of the present study was to evaluate the effects of the PGF 2 α treatment given at the onset of a synchronization of ovulation protocol using a norgestomet (NORG) ear implant on ovarian follicular dynamics (Experiment 1) and pregnancy per AI ( P /AI; Experiment 2) in cyclic (CL present) Bos indicus heifers. In Experiment 1, a total of 46 heifers were presynchronized using two consecutive doses of PGF 2 α 12 days apart. At first day of the synchronization protocol the heifers received implants containing 3 mg of NORG and 2 mg of estradiol benzoate (EB). At the same time, heifers were randomly assigned to receive 150 mg of d -cloprostenol ( n = 23; PGF 2 α ) or no additional treatment ( n = 23; Control). When the ear implants were removed 8 days later, all heifers received a PGF 2 α treatment and 1 mg of EB was given 24 h later. The follicular diameter and interval to ovulation were determined by transrectal ultrasonography. No effects of PGF 2 α treatment on the diameter of the largest follicle present were observed at implant removal (PGF 2 α = 9.8 ± 0.4 vs. Control = 10.0 ± 0.3 mm; P = 0.73) or after 24 h (PGF 2 α = 11.1 ± 0.4 vs. Control = 11.0 ± 0.4 mm; P = 0.83). No differences in the time of ovulation after ear implant removal (PGF 2 α = 70.8 ± 1.2 vs. Control = 73.3 ± 0.9 h; P = 0.10) or in the ovulation rate (PGF 2 α = 87.0 vs. Control = 82.6%; P = 0.64) between treatments were observed. In Experiment 2, 280 cyclic heifers were synchronized using the same experimental design described above (PGF 2 α ; n = 143 and Control; n = 137), at random day of the estrous cycle. All heifers received 300 IU of equine chorionic gonadotropin (eCG) and 0.5 mg of estradiol cypionate (as ovulatory stimulus) when the NORG ear implants were removed. Timed artificial insemination (TAI) was performed 48 h after implant removal and the pregnancy diagnosis was conducted 30 days later. No effects on the P /AI due to PGF 2 α treatment were observed (PGF 2 α = 51.7 vs. Control = 57.7%; P = 0.29). In conclusion, PGF 2 α treatment at the onset of NORG-based protocols for the synchronization of ovulation did not alter the ovarian follicular responses or the P /AI in cyclic Bos indicus beef heifers synchronized for TAI.

Exogenous eCG for stimulation of a single dominant follicle or for superovulation are common strategies to improve reproductive efficiency by increasing pregnancy rates and embryo production, respectively. Morphofunctional changes in the CL of eCG-treated cattle include increases in CL volume and plasma progesterone concentrations. Therefore, we tested the hypothesis that eCG alters the content of luteal cells and mitochondria related to hormone production. Twelve crossbred beef cows were synchronized and then allocated into three groups (four cows per group) and received no further treatment (control) or were given eCG either before or after follicular deviation (superovulation and stimulation of the dominant follicle, respectively). Six days after ovulation, cows were slaughtered and CL collected for morphohistologic and ultrastructural analysis. Mitochondrial volume per CL was highest in superovulated followed by stimulated and then control cows (18,500 ± 2630, 12,300 ± 2640, and 7670 ± 3400 μm3; P < 0.001), and the density of spherical mitochondria and the total number of large luteal cells were increased (P < 0.05) in stimulated cows compared with the other two groups (110.32 ± 14.22, 72.26 ± 8.77, and 70.46 ± 9.58 mitochondria per μm3 and 678 ± 147, 245 ± 199, and 346 ± 38 × 106 cells, respectively. However, the largest diameters of the large luteal cells were increased in superovulated and control cows versus stimulated ones (32.32 ± 0.06, 31.59 ± 0.81, and 29.44 ± 0.77 μm; P < 0.0001). In contrast, the total number of small luteal cells was increased in superovulated cows (1456 ± 268, 492 ± 181, and 822 ± 461 × 106, P < 0.05). In conclusion, there were indications of cellular changes related to increased hormonal production (stimulatory treatment) and increased CL volume (superovulatory treatment).

The success of IVP is ultimately dependent on the number and quality of the cumulus-oocyte complexes (COC) harvested during the OPU procedure. Several factors appear to be critical to oocyte quality including follicle size, environment factors such as heatstress, genetic background, age and lactation status of donor animals, all having a remarkable influence on the results of IVP. The aim of this review is to highlight some critical areas that can help veterinary practitioners to enhance OPU efficiency and successfully implement IVP into their routine practice. Focus will be given to recent findings in the literature and underlying physiological aspects that may be interfering with the quality of oocytes addressed to IVP in cattle at younger ages (calves and prepubertal heifers), pregnant vs nonpregnant status, and possible interactions with lactation and days postpartum during OPU.

Contents Over the last several decades, a number of therapies have been developed that manipulate ovarian follicle growth to improve oocyte quality and conception rates in cattle. Various strategies have been proposed to improve the responses to reproductive biotechnologies following timed artificial insemination (TAI), superovulation (SOV) or ovum pickup (OPU) programmes. During TAI protocols, final follicular growth and size of the ovulatory follicle are key factors that may significantly influence oocyte quality, ovulation, the uterine environment and consequently pregnancy outcomes. Progesterone concentrations during SOV protocols influence follicular growth, oocyte quality and embryo quality; therefore, several adjustments to SOV protocols have been proposed depending on the animal category and breed. In addition, the success of in vitro embryo production is directly related to the number and quality of cumulus oocyte complexes harvested by OPU. Control of follicle development has a significant impact on the OPU outcome. This article discusses a number of key points related to the manipulation of ovarian follicular growth to maximize oocyte quality and improve conception rates following TAI and embryo transfer of in vivo- and in vitro-derived embryos in cattle.

The objective of this study was to evaluate the productive performance, nutrients digestion and metabolism of three different genetic groups fed with the same diet based on corn silage. 30 heifers in growth were used of three groups of cattle, the following: Nellore ( Bos taurus indicus ) (n = 10), Holstein ( Bos taurus taurus ) (n = 10), and Mediterranean buffaloes ( Bubalis bubalis ) (n = 10). The animals were fed in groups and received the same experimental diet composed of corn silage and concentrate for growing heifers. In the evaluation of animals the performance, consumption and total apparent digestibility of dry matter and nutrients with the aid of internal markers (chromic oxide) and external (iADF), rumen fermentation, excretion of purine derivatives, nitrogen balance and blood metabolites were measured. No differences were observed in animal performance. There were differences in nutrient intake and apparent digestibility of dry matter and nutrients in different groups of cattle. The concentration of ammonia nitrogen (NH3-N) and short chain fatty acids (SCFA) in the rumen were higher and lower, respectively, for the group of buffaloes in relation to other experimental groups evaluated. When considering the excretion of total purine derivatives, buffaloes showed the lowest value compared to other genetic groups evaluated; about 61.76% of the total genetic group Nellore and 57.62% of the total genetic group Holstein with an average of 33.67 mmol/day. For the buffaloes, the excretion of xanthine and hypoxanthine observed was of 5.11% of total purine derivatives. There was a better nitrogen balance (g/day) for groups of Holstein heifers and Nellore in relation to the group of buffalo heifers. There were differences in the concentrations of urea and urea nitrogen in serum and liver enzymes where the buffaloes had higher values in relation at the bovines. There is a great metabolic diversity among the experimental groups evaluated and it was more exacerbated among buffaloes and bovines, when submitted to the same diet and same management conditions.

Made available in DSpace on 2013-09-27T14:54:12Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Made available in DSpace on 2013-09-30T18:36:09Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:42:31Z No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Made available in DSpace on 2014-05-20T13:42:31Z (GMT). No. of bitstreams: 1 WOS000288627700007.pdf: 187475 bytes, checksum: f44a03d706cf45e64484e91ab7b27272 (MD5) Previous issue date: 2011-02-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) The objectives of this study were to determine the interval from ovulation to deviation and the diameter of the dominant (DF) and largest subordinate (SF) follicles at deviation in buffalo (Bubalus bubalis) heifers. Two methods of evaluation (observed vs. calculated) were used. FSH and LH profiles encompassing follicle deviation (Experiment 1), and the follicular diameter when the DF acquired ovulatory capacity (Experiment 2) were also determined. The time of deviation and the diameter of the of and the largest SF at deviation did not differ between observed and calculated methods. Overall, follicle deviation occurred 2.6 +/- 0.2 d (mean SEM) after ovulation, and the diameters of the DF and SF at deviation were 7.2 +/- 0.2 and 6.4 +/- 0.2 mm, respectively. No changes in plasma levels of FSH or LH were observed (P= 0.32 and P= 0.96, respectively). Experiment 2 was conducted in two phases according to the diameter of the of during the first wave of follicular development at the time of LH challenge (25 mg of pLH). In the first phase, follicles ranging from 5.0 to 6.0 mm (n = 7), 6.1 to 7.0 mm (n = 11), or 7.1 to 8.0 mm (n= 9) were used, and in the second phase, follicles ranging from 7.0 to 8.4 mm (n=10), 8.5 to 10.0 mm (n = 10), or 10.1 to 12.0 mm (n = 9) of diameter were used. After the pLH treatment, the of was monitored by ultrasonography every 12 h for 48 h. No ovulations occurred in heifers in the first phase. However, in the second phase, an effect of follicular diameter was observed on ovulation rate [7.0-8.4 mm (0.0%, 0/10), 8.5-10.0 mm (50.0%, 5/10), and 10.0-12.0 mm (55.6%, 5/9)]. In summary, follicle deviation occurred 2.6d after ovulation in buffalo (B. bubalis) heifers, when the diameters of the DF and SF were 7.2 and 6.4 mm, respectively. No significant changes in plasma concentrations of FSH or LH were detected. Finally, the acquisition of ovulatory capacity occurred when the of reached 8.5 mm in diameter. (C) 2010 Elsevier B.V. All rights reserved. FMVZ USP, Dept Anim Reprod, BR-05508270 São Paulo, Brazil APTA Registro, Registro, SP, Brazil Autonomous Veterinarian, Vicosa, MG, Brazil UFMA, Ctr Ciencias Agr & Ambientais, Chapadinha, MA, Brazil IB UNESP, Dept Bioestatist, Botucatu, SP, Brazil IB UNESP, Dept Farmacol, Botucatu, SP, Brazil Lab Endocrinol UNESP, Dept Apoio & Prod Saúde Anim, Aracatuba, SP, Brazil Univ Saskatchewan, Western Coll Vet Med, Saskatoon, SK S7N 0W0, Canada IB UNESP, Dept Bioestatist, Botucatu, SP, Brazil IB UNESP, Dept Farmacol, Botucatu, SP, Brazil Lab Endocrinol UNESP, Dept Apoio & Prod Saúde Anim, Aracatuba, SP, Brazil FAPESP: 03/10203-4

This review brings together information on ovarian physiology in buffaloes including folliculogenesis, ovulation, and the development and function of the corpus luteum. Features of embryonic development are also considered. The buffalo is classified as a short-day breeder but in equatorial zones can show oestrous cycles throughout the year provided that nutrition is adequate to maintain reproductive function. In sub-tropical zones and at higher latitudes, day length is often the major determinant of reproductive function including the occurrence of regular oestrous cycles, duration of oestrus, and the period to resumption of ovulation postpartum. Indeed, at higher latitudes buffaloes that give birth during the period of increasing day length may not show a resumption of ovulation until the following period of decreasing day length. This can have a major impact on the productive value of buffaloes and requires the development and utilisation of practical and effective assisted breeding technology for out-of-season breeding in buffaloes. Embryonic development in buffaloes occurs at a faster rate than in cattle and this has implications for the earlier establishment and functionality of the corpus luteum in buffaloes. It would appear that the interrelationships between the development of the early conceptus, corpus luteum function, uterine preparation, and maternal recognition of pregnancy, are more closely time-bound in buffaloes compared with cattle. The phase of embryonic attachment would seem to be a critical period for determining the reproductive outcome in buffaloes.

A total of 177 Nelore heifers were examined by ultrasonography to determine the presence or absence of a corpus luteum (CL) and received a 3 mg norgestomet ear implant plus 2 mg of estradiol benzoate i.m. On Day 8, implants were removed and 150 μg of d -cloprostenol i.m. was administered. At the time of norgestomet implant removal, heifers with or without CL at the time of initiating treatment were assigned equally and by replicate to be treated with 0 IU ( n = 87) or 400 IU ( n = 90) eCG i.m. All heifers received 1 mg of EB i.m. on Day 9 and were submitted to fixed-time artificial insemination (FTAI) 30–34 h later. The addition of eCG increased the diameter of the largest follicle (LF) at FTAI (10.6 ± 0.2 mm vs. 9.5 ± 0.2 mm; P = 0.003; mean ± SEM), the final growth rate of the LF (1.14 ± 0.1 mm/day vs. 0.64 ± 0.1 mm/day; P = 0.0009), ovulation rate [94.4% (85/90) vs. 73.6% (64/87); P = 0.0006], the diameter of the CL at Day 15 (15.5 ± 0.3 mm vs. 13.8 ± 0.3 mm; P = 0.0002), serum concentrations of progesterone 5 days after FTAI (6.6 ± 1.0 ng/ml vs. 3.6 ± 0.7 ng/ml; P = 0.0009), and pregnancy per AI [P/AI; 50.0% (45/90) vs. 36.8% (32/87); P = 0.04]. The absence of a CL at the beginning of the treatment negatively influenced the P/AI [30.2% (16/53) vs. 49.2% (61/124); P = 0.01]. Therefore, the presence of a CL (and/or onset of puberty) must be considered in setting up FTAI programs in heifers. In addition, eCG may be an important tool for the enhancement of follicular growth, ovulation, size and function of the subsequent CL, and pregnancy rates in progestin-based FTAI protocols in Bos indicus heifers.

Artificial insemination (AI) in buffalo has limited use worldwide due the difficulties in the estrus detection and in finding an adequate moment for this procedure. Therefore, an alternative to increase the number of buffalo that are inseminated is the use of protocols that allow the AI without the need of estrus detection, usually called fixed-time AI (FTAI). Our expanding knowledge of the control of follicular wave dynamics during the buffalo estrous cycle has resulted in renewed enthusiasm for the prospects of precisely control of the follicular and luteal dynamics and finely controlling the time of ovulation. Follicular wave development can be controlled by treatments with GnRH or estradiol and progestogen/progesterone in combination. Treatment of buffalo with GnRH in combination with prostaglandin F2α (PGF2α) 7 d later and a second GnRH 48 h after PGF2α (known as Ovsynch) has resulted in acceptable pregnancy rates after FTAI in cycling buffalo during the breeding season. FTAI protocols using progestin devices, estradiol and eCG have resulted in synchronous onset of a new follicular wave, synchronous ovulation and consistent pregnancy rates in anestrous buffalo during the off breeding season. The combination of these protocols permits the use of AI throughout the year, obtaining conception and calving even in anestrus buffalo during the off breeding season.

O presente estudo está apresentado em Capítulo 1 (Divergência folicular; DF) e 2 (Capacidade ovulatória), e em cada estão incluídos dois experimentos [1 - em bubalinos (Bubalus bubalis); 2 - em zebuínos (Bos indicus)]. Os objetivos do Capítulo 1 foram determinar o momento e diâmetro na DF e avaliar o perfil das concentrações plasmáticas de FSH e LH próximo à seleção folicular. No Experimento 1, a ovulação foi considerada o dia zero (D0), a partir do qual 10 novilhas Murrah foram examinadas por ultra-sonografia (US) a cada oito horas (h) até o D4 e, posteriormente, a cada 24h até o D6. A avaliação da DF foi realizada por análise visual do gráfico de cada novilha (Ginther et al., 1996) e regressão linear segmentada (Bergfelt et al. 2003). As amostras de sangue, para dosagem de FSH e LH, foram colhidas com intervalos de 8h nas primeiras 24h pós-ovulação. A partir deste momento foram feitas colheitas a cada 4h até o D4 e após, a cada 12h até o D6. No método visual, o intervalo ovulação-DF, e o diâmetro dos folículos dominante (FD) e subordinado (FS) foram de, respectivamente, 63,2±5,7h; 7,2±0,3mm e 6,4±0,3mm (média±EPM). No método matemático estes mesmos parâmetros foram de, respectivamente, 61,9±4,9h; 7,3±0,3mm e 6,3±0,3mm. Não houve diferença entre os dois métodos de avaliação da DF (P>0,05). As concentrações plasmáticas de FSH e LH não variaram ao longo do tempo (P>0,05). No Experimento 2, 12 novilhas Nelore foram submetidas à US a cada 12h até o D5 e estes mesmos intervalos foram adotados para as colheitas de sangue (dosagem de FSH e LH). No método visual, o intervalo ovulação-DF, e os diâmetros do FD e FS foram de, respectivamente, 61,0±5,8h; 6,2±0,2mm e 5,8±0,2mm. No método matemático estes mesmos parâmetros foram de, respectivamente, 57,2±6,0h; 6,2±0,3mm e 5,9±0,3mm. Não houve diferença entre os métodos (P>0,05). As concentrações plasmáticas de FSH e LH não variaram ao longo do tempo (P>0,05). No Capítulo 2, o objetivo foi avaliar o diâmetro no qual o FD adquire capacidade ovulatória após administração de 25mg de LH. Os grupos foram formados de acordo com o diâmetro do FD no momento do tratamento com LH (7,0-8,4mm; 8,5-10,0mm e >10,0mm, em ambos os experimentos). No Experimento 1, foram utilizadas 29 novilhas bubalinas Murrah x Mediterrâneo. A partir do D3 (pós-ovulação), os exames US foram realizados a cada 12h até 48h do tratamento com LH. Nos grupos 7,0-8,4; 8,5-10,0 e >10,0mm, os diâmetros foliculares no momento da aplicação do LH foram de 7,8c±0,1; 9,3b±0,2 e 11,0a±0,3mm e as taxas de ovulação de 0,0%b (0/10); 50,0%a (5/10) e 55,6%a (5/9), respectivamente. No Experimento 2, 29 novilhas Bos indicus foram monitoradas por US a cada 24h a partir da ovulação até o tratamento com LH, e então a cada 12h durante 48h. Nos grupos 7,0-8,4; 8,5-10,0 e >10,0mm, os diâmetros foliculares no momento da aplicação do LH foram de 7,6c±0,1; 9,6b±0,1 e 10,9a±0,2mm e as taxas de ovulação de 33,3%b (3/9); 80,0%a (8/10) e 90,0%a (9/10), respectivamente. Present study is presented in Chapter 1 (Follicle deviation; FD) and 2 (Ovulatory capacity). Each chapter includes two experiments [1 - in buffaloes (Bubalus bubalis); 2 - in zebu cattle (Bos indicus)]. In Chapter 1, aims were to determine moment and diameter deviation and to evaluate FSH and LH plasmatic profiles encompassing follicle selection. In Experiment 1, ovulation was considered day 0 (D0). From D0, 10 Murrah heifers were examined by ultrasound (US) each eight hours (h) until D4 and then, every 24h until D6. FD evaluation was performed by visual analysis (Ginther et al., 1996) and by segmented linear regression (Bergfelt et al., 2003). Blood samples were harvested each 8h in the first 24h post ovulation. From this moment blood collections were performed every 4h until D4 and after that, each 12h until D6. Interval ovulation-FD, dominant (DF) and subordinate follicle (SF) diameters, by visual method, were, respectively, 63.2±5.7h; 7.2±0.3mm and 6.4±0.3mm (mean±SEM). These same end points, by mathematical method, were, respectively, 61.9±4.9h; 7.3±0.3mm and 6.3±0.3mm. No differences between both methods of FD evaluation (P>0.05) were found. No changes in plasmatic FSH and LH levels along the time (P>0.05) were found. In Experiment 2, 12 Nelore heifers were submmited to US each 12h until D5, and these same intervals were used to blood harvests (FSH and LH profiles). Interval ovulation-FD, dominant (DF) and subordinate follicle (SF) diameters, by visual method, were, respectively, 61.0±5.8h; 6.2±0.2mm and 5.8±0.2mm. These same end points, by mathematical method, were, respectively, 57.2±6.0h; 6.2±0.3mm and 5.9±0.3mm. No differences between both methods of FD evaluation (P>0.05) were found. No changes in plasmatic FSH and LH levels along the time (P>0.05) were found. In Chapter 2, aim was to evaluate diameter in which DF acquires ovulatory capacity after administration of 25mg of LH. The animals were assigned in groups according to DF diameter at LH treatment (7.0-8.4mm; 8.5-10.0mm and >10.0mm, in both experiments). In Experiment 1, 29 Murrah x Mediterrâneo heifers were used. From D3 (post ovulation), US examinations were performed each 12h until 48h after LH treatment. Follicle diameters at LH chalenge in groups 7.0-8.4mm, 8.5-10.0mm and >10.0mm, were 7.8c±0.1, 9.3b±0.2 and 11.0a±0.3mm, and ovulation rates were 0.0%b (0/10), 50.0%SUP>a (5/10) and 55.6%a (5/9), respectively. In Experiment 2, 29 Bos indicus heifers were monitored by US every 24h from ovulation until LH chalenge, and then each 12h during 48h. Follicle diameters at LH treatment in groups 7.0-8.4mm, 8.5-10.0mm and >10.0mm, were 7.6c±0.1, 9.6b±0.1 and 10.9a±0.2mm, and ovulation rates were 33.3%b (3/9), 80.0%a (8/10) and 90.0%a (9/10), respectively

The aim of the present study was to evaluate the effect of bovine somatotropin (bST; 500mg) administration on lactating buffalo donors submitted to two different ovum pick-up (OPU) and in vitro embryo production schemes with a 7 or 14d intersession OPU interval. A total of 16 lactating buffalo cows were randomly assigned into one of four experimental groups according to the bST treatment (bST or No-bST) and the OPU intersession interval (7 or 14d) in a 2×2 factorial design (16 weeks of OPU sessions). The females submitted to OPU every 14d had a larger (P0.001) number of ovarian follicles suitable for puncture (15.6±0.7 vs. 12.8±0.4) and an increased (P=0.004) number of cumulus-oocyte complexes (COCs) recovered (10.0±0.5 vs. 8.5±0.3) compared to the 7d interval group. However, a 7 or 14d interval between OPU sessions had no effect (P=0.34) on the number of blastocysts produced per OPU (1.0±0.1 vs. 1.3±0.2, respectively). In addition, bST treatment increased (P0.001) the number of ovarian follicles suitable for puncture (15.3±0.5 vs. 12.1±0.4) but reduced the percentage (18.9% vs. 10.9%; P=0.009) and the number (1.4±0.2 vs. 0.8±0.1; P=0.003) of blastocysts produced per OPU session compared with the non-bST-treated buffaloes. In conclusion, the 14d interval between OPU sessions and bST treatment efficiently increased the number of ovarian follicles suitable for puncture. However, the OPU session interval had no effect on embryo production, and bST treatment reduced the in vitro blastocyst outcomes in lactating buffalo donors.

The objective of present study was to evaluate the effect of OPU interval and the effect of bST treatment on embryo production in buffalo. Sixteen females were assigned in 4 groups, in a 2x2 experimental design, with main effects of bST (0 or 500mg) and interval of OPU session (every 7 or 14 days), as follows: G-CONT7: OPU once a week; G-CONT14: OPU each 14 days; G-bST7: OPU once a week + 500mg of bST and G-bST14: OPU each 14 days + 500mg of bST. Animals of G-CONT7 and G-bST7 were submitted to 8 OPU sessions, and those of G-CONT14 and G-bST14 to 4 OPU sessions. Overall, OPU once a week (without bST) reduced the number of aspirated follicles, and the total and viable oocytes. Despite of this, OPU every 14 days + bST increased the number of degenerated oocytes, and reduced the number of blastocysts produced on days 6 and 7.

The efficiency of different doses of rBST in a MOET protocol was verified in buffaloes. The animals received an intravaginal progesterone device (DIB) plus 2.0 mg of estradiol benzoate (Ric-BE, i.m.) and 0 mg, 250 mg or 500 mg of rBST. Ovarian follicular growth was stimulated by treatment with 200 mg of FSH in decreasing amount (12/12hs). Buffaloes received injections of PGF2α on D6 and on D7. The DIB was removed on D7. On D8, 24 hours after DIB removal, ovulation was induced with GnRH. The follicular emergence and the response to FSH treatment were evaluated by ultrasound on D4 and D8. The artificial insemination (AI) was performed 12 and 24 hours after the GnRH administration. The embryonic structures were collected 5.5 days after the first AI. Data were analyzed by ANOVA. No statistical differences were found between groups. The results indicated that rBST, associated to a MOET protocol at the different dosages used, does not to improve the efficiency of the technique in buffalo.

The objectives of the present work were to determine follicle deviation assessed by ultrasonography and profile of plasmatic gonadotrophins, and ovulatory capacity in buffalo species, since these information are not available in the literature. The knowledge of follicular development and gonadotrophins profile during deviation and the follicular response to an exogenous ovulation inducer can be useful tools for follicular manipulation of estrous cycle.

In order to verify the causes of the low embryo recovery rate in superovulated buffaloes, the effect of species and of estradiol-17β (E2) treatment were evaluated on oocyte transport across the oviduct in Murrah and Nelore. The females were synchronised with progesterone plus estradiol benzoate followed by an injection of PGF2α and eCG. The ovulation was induced with GnRH and 48hs after the animals were slaughtered and the oviducts removed. The oviducts were washed with HBSS and oocytes of both species were inserted into infundibulum portion. The oviducts were put in a dish with HBSS with or without E2 and incubated for 24 h. The oviduct were then flushed with DPBS, in order to recover and count the oocytes. Data were analyzed by ANOVA. There was no effect of interaction. The total number of oocytes and the recovery rate were higher for Nelore than Murrah (P

Made available in DSpace on 2022-04-29T08:44:46Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-01-30 This study examined the usefulness of radiotelemetry for estrous detection in buffalo heifers and the impact of prostaglandin F2α (PGF2α) administration during the early or late luteal phase on estrous behavior and ovulatory follicle variables. Induction of estrus with PGF2α at a random stage of the estrous cycle was followed by the arbitrary division of heifers into groups receiving a second dose of PGF2α during either the early (n=33) or late (n=17) luteal phase (6-9 or 11-14 days after estrus, respectively) for the induction of synchronized estrus. The electronic detection of synchronized estrus by radiotelemetry was confirmed using ultrasonography every 6h until ovulation. Radiotelemetry was 90% efficient and 100% accurate for estrous detection. Intervals between the PGF2α dose and the beginning of synchronized estrus (40.7±10.9 vs. 56.7±12.8h) or ovulation (70.0±11.3 vs. 85.6±12.5h) were shorter (P0.05) results for the duration of estrus, the intervals from the beginning or end of estrus to ovulation, the number and duration of mounts per estrus, the duration of mounts, the diameter of the ovulatory follicle and the luteal profile on day 5 after estrus. In conclusion, radiotelemetry was a suitable tool for the efficient and accurate detection of estrus in buffalo heifers. Treatment with PGF2α during the early luteal phase had a shorter interval to synchronized estrus and ovulation; however, estrous behavior, ovulatory follicle dynamics and subsequent luteal activity were similar following PGF2α administration during the early or late luteal phase. © 2014 Elsevier B.V. Departamento de Reprodução Animal Faculdade de Medicina Veterinária e Zootecnia Universidade de São Paulo, São Paulo Departamento de Medicina Veterinária Preventiva e Reprodução Animal Faculdade de Ciências Agrárias e Veterinárias Jaboticabal, São Paulo Unidade de Pesquisa e Desenvolvimento de Registro-Polo Regional do D.S.A. do Vale do Ribeira/APTA Registro, São Paulo Department of Veterinary Gynaecology and Obstetrics College of Veterinary Science Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab

Equine chorionic gonadotrophin (eCG) has been widely used in superovulation and artificial insemination programmes and usually promotes an increase in corpus luteum (CL) volume and stimulates progesterone production. Therefore, to identify eCG-regulated genes in the bovine CL, the transcriptome was evaluated by microarray analysis and the expression of selected genes was validated by qPCR and western blot. Eighteen Nelore crossbred cows were divided into control (n = 5), stimulated (n = 6) and superovulated groups (n = 7). Ovulation was synchronised using a progesterone device-based protocol. Stimulated animals received 400 IU of eCG at device removal and superovulated animals received 2000 IU of eCG 4 days prior. Corpora lutea were collected 7 days after gonadotrophin-releasing hormone administration. Overall, 242 transcripts were upregulated and 111 transcripts were downregulated in stimulated cows (P ≤ 0.05) and 111 were upregulated and 113 downregulated in superovulated cows compared to the control animals (1.5-fold, P ≤ 0.05). Among the differentially expressed genes, many were involved in lipid biosynthesis and progesterone production, such as PPARG, STAR, prolactin receptors and follistatin. In conclusion, eCG modulates gene expression differently depending on the treatment, i.e. stimulatory or superovulatory. Our data contribute to the understanding of the pathways involved in increased progesterone levels observed after eCG treatment.

The present study was conducted to verify if the elevation of plasma concentrations of estradiol during superovulatory treatments affects the oocyte transport in buffalo females, as well as if the inferior quality of buffalo oocytes and/or some functional difference on the oviduct of these animals is responsible for the low embryo recovery rate in superovulated uffaloes when compared to cows subjected to the same treatment. Oviducts of 10 buffaloes and 15 of cows, treated to induce a single ovulation were used. The oviducts were placed on Petri dishes and received the following treatments: 5 buffalo oocytes with no E2 (G-BufBuf and G-BovBuf), 5 bovine oocytes with no E2 (G-BufBov and G-BovBov), 5 buffalo oocytes with E2 (G-BufE2Buf and G-BovE2Buf) and 5 bovine oocytes with E2 (G-BufE2Bov and G-BovE2Bov; factorial 2x2x2). Oocytes were incubated for 24h. Subsequently, oviducts were washed and oocytes were recovered and counted. Since no interactions were found between E2 treatment, oviducts and oocytes species, main effects were analyzed separately. Recovery rate and number of oocytes was higher on cattle compared to buffaloes (35.0+8.6% and 1.4+0.3 vs. 10.0±4.6% and 0.5±0.2, respectively; p0.05); the number of buffaloes and bovine oocytes recovered were similar (1.4±0.4 and 0.6±0.2, respectively; p>0.05). Oocytes recovery rate showed a trend (P=0.07) to be higher when buffalo oocytes were implanted when compared to bovine oocytes (35.2±9.2% vs. 12.9±5.4%). Present results suggest that oocyte transport by the oviduct of buffaloes and bovine was not dependent on oocytes species or E2 supplementation to the culture medium. O presente estudo foi realizado para verificar se a elevação das concentrações plasmáticas de estradiol durante os tratamentos superovulatórios afeta o transporte dos oócitos em fêmeas bubalinas, bem como se a qualidade inferior dos oócitos de búfalos e/ou alguma diferença funcional no oviduto destes animais é responsável pela baixa taxa de recuperação de embriões em búfalas superovuladas quando comparadas a vacas submetidas ao mesmo tratamento. Foram utilizados 10 ovidutos de búfalas e 15 de vacas, tratadas para a indução de ovulação única. Os ovidutos foram colocados em placas de Petri e receberam os seguintes tratamentos: sem E2 e inseridos com 5 oócitos de búfalas (G-BufBuf e G-BovBuf); sem E2 e com 5 oócitos de vacas (G-BufBov e G-BovBov); com E2 e com 5 oócitos de búfalas (G-BufE2Buf e G-BovE2Buf); e com E2 e com 5 oócitos de vacas (G-BufE2Bov e G-BovE2Bov; fatorial 2x2x2). Posteriormente, foram incubados por 24h e, após esse período, foram lavados para a recuperação e contagem dos oócitos. Como não foi verificado efeito de interação, foram analisados os efeitos principais. O número e a taxa de recuperação de oócitos foi maior em ovidutos de vacas que de búfalas (1,4±0,3/35,0±8,6% vs. 0,5±0,2/10,0±4,6%; P0,05). Não foi verificada diferença no número de oócitos de búfalas ou de vacas recuperados (1,4±0,4 e 0,6±0,2; P>0,05). Observou-se também que houve tendência (P=0,07) de maior taxa de recuperação de oócitos de búfalas que de vacas (35,2±9,2% vs. 12,9±5,4%). Os dados são indicativos de que o transporte de oócitos pelo oviduto de búfalas e de vacas independe da espécie do oócito e não é influenciado pelo E2.

In the current review the main fundamental biological differences in reproductive function between Bos taurus and Bos indicus cattle are discussed. Breed differences regarding puberty, estrous cycle patterns, estrous behavior, acquisition of ovulatory capacity, ovarian structures and reproductive hormones are presented. The main physiological differences that Bos indicus cattle present relative to Bos taurus cattle include: delayed age at puberty; higher circulating concentrations of hormones such as estradiol, progesterone, insulin and IGF-I, despite having smaller ovulatory follicle size and corpora lutea; greater population of small follicles and smaller size of the dominant follicle at deviation; and greater sensitivity of follicles to gonadotropins. Knowledge of the differences between Bos indicus and Bos taurus breeds help explain different management procedures and responses to hormonal treatments associated with artificial insemination, ovarian superstimulation, and in vivo and in vitro embryo production.

Currently, timed ovulation induction and fixed-time artificial insemination (FTAI) in superstimulated donors and synchronization protocols for fixed-time embryo transfer (FTET) in recipients can be performed using GnRH or estradiol plus progesterone/progestin (P4)-releasing devices and prostaglandin F2α (PGF2α). The control of follicular wave emergence and ovulation at predetermined times, without estrus detection, has facilitated donor and recipient management. However, because Bos taurus cows have subtle differences in their reproductive physiology compared with Bos indicus cattle, one cannot assume that similar responses will be achieved. The present review will focus on the importance of orchestrating donor and recipient management to assure better logistics of procedures to achieve more desirable results with embryo collection and transfer. In addition, this will provide clear evidence that the use of FTAI in superstimulated donors and FTET in embryo recipients eliminates the need to detect estrus with satisfactory results. These self-appointed programs reduce labor and animal handling, facilitating the use of embryo transfer in beef and dairy cattle.

Made available in DSpace on 2013-08-28T14:09:27Z (GMT). No. of bitstreams: 1 WOS000279402900001.pdf: 817649 bytes, checksum: 1e696de521d4d716fe83639c4a906072 (MD5) Made available in DSpace on 2013-09-30T17:54:53Z (GMT). No. of bitstreams: 2 WOS000279402900001.pdf: 817649 bytes, checksum: 1e696de521d4d716fe83639c4a906072 (MD5) WOS000279402900001.pdf.txt: 28360 bytes, checksum: 0b7872479c330605598c3c36ed62cdf4 (MD5) Previous issue date: 2010-06-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T13:15:23Z No. of bitstreams: 2 WOS000279402900001.pdf: 817649 bytes, checksum: 1e696de521d4d716fe83639c4a906072 (MD5) WOS000279402900001.pdf.txt: 28360 bytes, checksum: 0b7872479c330605598c3c36ed62cdf4 (MD5) Made available in DSpace on 2014-05-20T13:15:23Z (GMT). No. of bitstreams: 2 WOS000279402900001.pdf: 817649 bytes, checksum: 1e696de521d4d716fe83639c4a906072 (MD5) WOS000279402900001.pdf.txt: 28360 bytes, checksum: 0b7872479c330605598c3c36ed62cdf4 (MD5) Previous issue date: 2010-06-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Nuclear-mitochondrial incompatibilities may be responsible for the development failure reported in embryos and fetuses produced by interspecies somatic cell nuclear transfer (iSCNT). Herein we performed xenooplasmic transfer (XOT) by introducing 10 to 15% of buffalo ooplasm into bovine zygotes to assess its effect on the persistence of buffalo mitochondrial DNA (mtDNA). Blastocyst rates were not compromised by XOT in comparison to both in vitro fertilized embryos and embryos produced by transfer of bovine ooplasm into bovine zygotes. Moreover, offspring were born after transfer of XOT embryos to recipient cows. Buffalo mtDNA introduced in zygotes was still present at the blastocyst stage (8.3 vs. 9.3%, p = 0.11), indicating unaltered heteroplasmy during early development. Nonetheless, no vestige of buffalo mtDNA was found in offspring, indicating a drift to homoplasmy during later stages of development. In conclusion, we show that the buffalo mtDNA introduced by XOT into a bovine zygote do not compromise embryo development. on the other hand, buffalo mtDNA was not inherited by offspring indicating a possible failure in the process of interspecies mtDNA replication. USP, Dept Ciencias Basicas, FZEA, Pirassununga, SP, Brazil UNESP, FCAV, Dept Med Vet Prevent & Reprod Anim, Jaboticabal, SP, Brazil Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Pecuaria Sudeste, Sao Carlos, SP, Brazil Univ Montreal, CRRA, St Hyacinthe, PQ, Canada Vida Reprodut Consultoria, Cravinhos, SP, Brazil Univ São Paulo, Dept Anim Reprod, São Paulo, Brazil Univ Naples Federico 2, DISCIZIA, Naples, Italy UNESP, FCAV, Dept Med Vet Prevent & Reprod Anim, Jaboticabal, SP, Brazil FAPESP: 02/05054-7 FAPESP: 04/01841-0 FAPESP: 06/59074-0

The objective of this study was to evaluate the effect of bovine somatotropin (bST) on ovarian follicular population in buffalo heifers and its influence on oocyte quality, recovery rates and in vitro embryo production. We tested the hypothesis that bST treatment in buffalo females submitted to an ovum pick-up (OPU) program would improve the number of follicles recruited, oocyte quality and in vitro embryo production. A total of 10 heifers were assigned into two treatment groups: group bST (n=5; receiving 500mg of bST in regular intervals) and control group (n=5; without additional treatment). Both groups were subjected to OPU sessions twice a week (every 3 or 4 days), for a total of 10 sessions per female, although due to procedural problems, only the first five OPU sessions produced embryos. The number of follicles and the diameters were recorded at all OPU sessions. The harvested oocytes were counted and classified according to their quality as either A, B, C, D or E, with A and B considered good quality. Cleavage and blastocyst production rates were evaluated 2 and 7 days after in vitro fertilization, respectively. The bST treatment increased the total number of antral follicles (>3mm in diameter; 12.2 compared with 8.7; p0.05). The significant effects of performing repeated OPU sessions were decreasing the follicular population (p

Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-27T11:23:48Z No. of bitstreams: 0Bitstream added on 2014-05-27T14:30:16Z : No. of bitstreams: 1 2-s2.0-78149363570.pdf: 242263 bytes, checksum: a48244c57e12c118a3be14899ebe1af0 (MD5) Made available in DSpace on 2014-05-27T11:23:48Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-12-01 Present study aimed to evaluate gonadotropins profiles in 12 Nelore heifers, in order to test the hypothesis that FSH concentrations decrease and LH presents a transient increase during follicle selection. Blood samples from jugular vein were harvested twice daily starting at the time of ovulation (D0) until D5. Plasma samples were assayed for FSH and LH by double antibody radioimmunoassay method. LH and FSH assay sensitivity was 0,02ng/ml and 0,005ng/ml, respectively. The intraassay coefficient of variation was 13,6% and 18,8%, respectively. Data (mean±SEM) were normalized to follicle deviation and analyzed by ANOVA and by linear, cubic, and quadratic regressions. Comparisons between higher and lower FSH values were also performed by T-test. There was no effect of time in plasmatic FSH and LH circulating levels when variance analysis or regression analysis were performed. However, by T-test, FSH concentrationsreached the lowest plasmatic levels 36 (0,40±0,05ng/ml) and 60 hours (0,42±0,04ng/ml) after follicular deviation, comparatively to 36 hours before deviation, when the concentrations were maximal (0,63±0,08ng/ml). In conclusion, there is a FSH decrease, although a transient LH elevation has not been confirmed encompassing follicle deviation in Nelore females. Departamento de Reprodução Animal Faculdade de Medicina Veterinária e Zootecnia Universidade de São Paulo, São Paulo-SP Departamento de Apoio e Produção em Saúde Animal Laboratório de Endocrinologia Universidade Estadual Paulista, Araçatuba-SP Departamento de Apoio e Produção em Saúde Animal Laboratório de Endocrinologia Universidade Estadual Paulista, Araçatuba-SP

One important factor in the success of ovum pickup (OPU)/IVP in Bos taurus is the follicular status at OPU concerning the dominance period (Hendriksen et al. 2000 Theriogenology 53, 11-20). The hypothesis of the present study is that OPU performed after follicle deviation, when follicles show a mild level of atresia, improves competence for IVP in Nelore (NE), Holstein (HO), and buffaloes (BU). Objectives were to determine effects of OPU done at different times of synchronized follicular wave (1, 3, or 5 d after expected emergence) and of genetic group (NE, HO, and BU) on IVP. A total of 27 heifers (9 of each genetic group) were maintained in contemporary nutritional and environmental conditions during experiment, in a cross-over design, performed in 6 replicates. Recovered oocytes with at least one cumulus cell layer were matured in TCM-199 supplemented with 10% of FCS plus 50 μM of cysteamin and 0.3 mM of cystine, at 38.5°C with 5% CO2 in air for 24 h. IVF was done with 2 × 106 spermatozoa per mL of NE (for bovine oocytes) or BU semen (for BU oocytes), for 20 h at the same incubator conditions of IVM. After IVF, presumptive zygotes were denuded and cultured in SOF under the same previous atmosphere conditions. Medium was changed 3 d after IVF when cleavage rate (CR) was assessed. Blastocyst (BR) and hatching rates (HR) were evaluated 7 and 9 days after IVF, respectively. About 50% of hatched blastocysts were fixed until nuclei counting. Data were analyzed by ANOVA using the Proc Mixed model. No effects of interaction or time of synchronization were observed in any of the variables. Concerning genetic group, NE had better results than HO and BU (mean ± SEM / heifer / replicate), respectively, for visualized follicles (41.0a ± 2.1, 22.1b ± 1.3, 18.8b ± 0.9), total oocytes (37.1a ± 2.5, 15.4b ± 1.2, 14.8b ± 1.0), oocytes at IVM (30.8a ± 2.4, 10.7b ± 1.0, 7.9b ± 0.7), oocytes at IVC (18.7a ± 0.8, 8.0b ± 0.5, 7.5b ± 0.4), cleaved embryos (15.4a ± 0.7, 4.6b ± 0.4, 4.4b ± 0.3),CR(81.8a, 59.1b, 62.3b), blastocysts on Day 7 (5.1a ± 0.6, 1.0b ± 0.2, 0.6b ± 0.1), BR (25.8a, 13.6b, 9.1b), and hatched blastocysts on Day 9 (2.6a ± 0.4, 0.3b ± 0.1, 0.3b ± 0.1). Recovery rate and HR were greater for NE (89.4 and 50.6%, respectively) than for HO (73.3 and 23.2%), but neither differed from BU (82.8 and 31.9%). Also, the percentage of viable was greater for NE (83.0) than for HO (66.9) and BU (53.1). No effects were observed for nuclei counting (NE = 176.6 ± 5.3, HO = 168.9 ± 13.7 and BU = 206.1 ± 23.1). Results demonstrate that Nelore had a better efficiency for IVP than Holstein and buffaloes. OPU performed at different times of synchronized follicular wave did not influence IVP, conversely to the initial hypothesis of this study. FAPESP (06/59550-6, 07/04782-2), Tortuga Cia Zootecnica®, Santa Adele and São Caetano Farms, LMMD, PCAPS, HOVET (Dr. Ubiraem Schalch), VRA, VNP (Prof. Dr. Francisco de Palma Rennó).

In Holstein cows, the diameter of the dominant follicle (DF) at the time of follicle deviation is 8.5 mm and the subordinate follicle (SF) is 7.2 mm (Ginther et al. 1996 Biol. Reprod. 55, 1187–1194). However, follicular responsiveness to an ovulatory treatment occurs only with 10.0-mm-diameter follicles (Sartori et al. 2001 Biol. Reprod. 65, 1403–1409). The current study tested the hypothesis that, in Bos indicus (Nelore and crossbred Nelore � Gir) females, the follicular diameters at the time of deviation and ovulation responsiveness are smaller than those in Holstein cows. The experiment was performed in two phases. In the first phase, 12 Nelore heifers were previously synchronized with a protocol using progestagen and estradiol benzoate. After implant removal, all heifers were evaluated by transrectal ultrasonography (Aloka SSD-500, Tokyo, Japan) every 12 h until Day 5 of the estrous cycle (Day 0 = Day of the ovulation) to assess the time of ovulation, the time of follicle deviation, and the follicular diameter at the deviation. In the second phase, 29 Bos indicus heifers (Nelore and crossbred Nelore � Gir) were previously synchronized with the same protocol as cited above. After the ovulations (Day 0), the follicles were evaluated by transrectal ultrasonography every 24 h, until they reached the diameter of 7.0–8.4 mm (n = 9); 8.5–10.0 mm (n = 10); and >10.0 mm (n = 10). In order to assess the ovulatory capacity, all animals were treated with 25 mg of LH (Lutropin-V�; Bioniche Animal Health, Inc., Belleville, Ontario, Canada) at these follicle diameter ranges. After the LH treatment, all animals were monitored by ultrasonography every 12 h for 48 h. ANOVA, Bartlett, and chi-square tests were used in the statistical analyses. In the first phase, the diameters of the DF and SF at the time of follicular deviation (61.9 � 4.9 h after ovulation) were 6.2 � 0.2 and 5.8 � 0.2 mm, respectively. In the second phase, the the average follicular diameters at the time of LH administration in the groups 7.0–8.4 mm, 8.5–10.0 mm, and >10.0 mm were 7.6a � 0.1 mm, 9.6b � 0.1 mm, and 10.9c � 0.2 mm; and their ovulation rates were 33.3%a (3/9), 80.0%b (8/10), and 90.0%b (9/10), respectively (P < 0.05). The interval from LH treatment to ovulation was 38.0 � 4.0 h, 31.5 � 2.7 h, and 30.0 � 2.0 h, respectively (P > 0.05). In conclusion, in Bos indicus heifers, follicle deviation occurred with smaller diameters than previously reported in Bos taurus breeds. In addition, Bos indicus heifers are able to ovulate in response to 25 mg of LH with smaller diameters compared to those of Bos taurus breeds. Moreover, in Bos indicus heifers, ovulatory capacity is acquired by follicles as small as 7.0–8.4 mm, but this responsiveness significantly increases after follicles reach 8.5–10.0 mm. This work was supported by FAPESP (Proc:03/10203-4); Bioniche Animal Health, Inc., Belleville, Ontario, Canada; and Tecnopec, S�o Paulo, Brazil.

Estradiol given during progestin-treatment promotes emergence of a new follicular wave in cattle. The interval from estradiol treatment to follicular wave emergence is dependent on the estradiol preparation, the dose, and possibly the animal category (cows or heifers). Therefore, this experiment aimed to compare the effects of either estradiol benzoate (EB) or estradiol valerate (EV) on the synchrony of ovarian follicular wave emergence in Bos indicus (Nelore) heifers and cows treated with a Norgestomet implant (Crestar�; Intervet, Boxmeer, The Netherlands; factorial 3 � 2 design). Lactating cows (n = 30; 70-90 days postpartum) and cycling heifers (n = 29) were homogeneously assigned, according to the presence of corpora lutea, to six experimental groups (G2EB, G2.5EV, and G5EV for each animal category). At random stages of the estrous cycle (Day 0), the females received an auricular implant and were assigned to receive 2 mg of EB (Farmavet, S�o Paulo, Brazil; 10 cows and 10 heifers; G2EB), 2.5 mg of EV plus 1.5 mg of Norgestomet (10 cows and 10 heifers; G2.5EV; half of a recommended dose), or 5 mg of EV plus 3 mg of Norgestomet (10 cows and 10 heifers; G5EV, full recommended dose) i.m. Transrectal ultrasonography was performed once daily from Day 0 to confirm the follicular dominance (new growing dominant follicle >8 mm in diameter). The day of wave emergence was retrospectively defined as the day when the dominant follicle was first detected at a diameter of 3 mm. Two-way analysis of variance was used to compare the effects of treatments (G2EB, G2.5EV, and G5EV), category (cows and heifers), and the treatments-by-category interaction on the interval from beginning of the treatment to wave emergence. The LSD was used to locate differences among groups and the Bartlett's test revealed heterogeneity of variance. Statistical analysis was conducted by SAS for Windows (SAS Institute, Inc., Cary, NC, USA). There was a significant effect of treatment and category (P < 0.001), but there were no treatments-by-category interactions (P > 0.05). A new follicular wave was induced in all animals after treatment with EB or EV. The heifers given G2EB (2.50 � 0.19a; 2 to 3x days) had a reduced (P < 0.01) and least variable interval (P < 0.05) than those given G2.5EV (4.20 � 0.27b; 3 to 6xy days) or G5EV (6.10 � 0.57c; 3 to 8y days). The cows given G5EV (4.00 � 0.52b; 2 to 6y days) had the longest (P = 0.05) and more variable interval (P < 0.001) than those given G2EB (2.50 � 0.22a; 2 to 4x days), but were similar to those given G2.5EV (3.10 � 0.41ab; 2 to 6xy days). The effect of EB was similar in heifers and cows (P > 0.05); however, the EV or 2.5EV promoted a longer emergence wave interval in the heifers than in the cows (P < 0.05). Analyzing the main effects, cows and heifers given G2EB (2.50 � 0.15a; 2 to 4x days) had a smaller and less variable interval than those given G2.5EV (3.65 � 0.27b; 2 to 6y) or G5EV (5.05 � 0.44c; 2 to 8z). In conclusion, the EV treatment, in both doses (5 or 2.5 mg), resulted in a delayed and more variable interval to emergence of a new follicular wave than 2 mg of EB in Bos indicus cows and heifers treated with a Norgestomet auricular implant. This work was supported by Intervet, Brazil.

O artigo não apresenta resumo.

The aim of the present study was to determine the optimal phase of the follicular wave to perform ovum pickup (OPU) for in vitro embryo production (IVEP) in various genetic groups. For this purpose, 27 heifers-nine Bos taurus (Holstein), nine Bos indicus (Nelore), and nine Bubalus bubalis (Mediterranean)-were maintained under the same nutritional, management, and environmental conditions. Heifers within each genetic group were submitted to six consecutive OPU trials with 14-day intersession intervals, at three different phases of the pharmacologically synchronized follicular wave (Day 1, 3, or 5 after follicular wave emergence), in a 3 × 3 crossover design. When OPU was performed at different phases of the pharmacologically synchronized follicular wave (Day 1, 3, or 5), no differences were found in the percent of oocytes recovered (70.5 ± 3.1%, 75.0 ± 3.1%, 76.0 ± 3.2%, respectively; P = 0.41) or blastocyst production rates (19.4 ± 2.9%, 16.6 ± 2.9%, 15.9 ± 2.6%, respectively; P = 0.36). Comparing genetic groups, B indicus showed a higher blastocyst rate (28.3(a) ± 2.8%; P 0.01) than B taurus and B bubalis (14.1(b) ± 2.9% and 10.2(b) ± 2.0%, respectively). However, only B indicus heifers showed a variation in the number of visualized follicles and the total and viable oocytes along consecutive OPU sessions. In conclusion, different phases of the pharmacologically synchronized ovarian follicular wave did not affect OPU-IVEP in B indicus, B taurus, and B bubalis heifers. Additionally, B indicus heifers showed greater OPU-IVEP efficiency than did the other genetic groups, under the same management conditions.

The emergent concepts on ovary environment, reproductive physiology and the development of pharmacology are constantly supporting the advance of assisted reproduction. Within the last years, the biotechnics related to the synchronization of follicular development and the manipulation of bovine estrus cycle have progressed rapidly and consistently. The combined use of timed-artificial insemination (TAI), superovulation (SOV), ovum pick up (OPU), in vitro embryo production (IVEP) and timed-embryo transfer (TET) has a great potential to improve reproductive outcomes and disseminate selected genetics, diminishing the interval of generations and improving herds genetic gain. However, several factors can potentially affect the efficiency of these procedures. The knowledge of the particularities of the genetic groups, follicular growth manipulation, follicular population predictors, and metabolic and environmental aspects that interfere with ovarian environment and, consequently, oocyte quantity and quality is crucial to optimize the reproductive programs. This review aims to elucidate some factors that affect the ovarian environment and must be well known in order to improve the efficiency of reproduction in cattle.