In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos / Desempenho de vacas doadoras zebuínas (Bos indicus) e taurinas (Bos taurus) na produção de embrião in vitro

In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P < 0.0001; n = 681 vs. 509 for grade 2 COC, P = 0.010, for zebu and taurine donors, respectively). There were differences in embryo production percentages obtained from OM (0.44% from zebu and 6.42% from taurine, P = 0.017), BL (14.18% from zebu and 3.74% from taurine, P < 0.0001), and BX (81.43% from zebu and 75.13% from taurine, P < 0.0001). No significant difference was observed for embryo production from BI and pregnancy rate (P > 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.(AU)

O objetivo foi avaliar a produção in vitro de embriões bovinos a partir de doadoras zebuína e taurina. Os complexos cumulus oócitos (COCs) foram obtidos de 167 doadoras Bos taurus e 161 Bos indicus, por meio de Ovum pick-up. Os COCs foram classificados quanto à qualidade morfológica, maturados em incubadora por 22 a 24h em meio de maturação, e encaminhados à fertilização entre 18 e 22h. Os zigotos foram transferidos para meio de cultivo, onde permaneceram durante sete dias. Os embriões foram classificados em mórula (MO), blastocisto inicial (BI), blastocisto (BL) e blastocisto expandido (BX), envasados em palhetas contendo meio de cultivo, e inovulados em receptoras sincronizadas. Após 30 e 45 dias da inovulação, realizou-se o diagnóstico de gestação. Os animais Bos indicus apresentaram maior produção oocitária (n = 2556) em comparação aos Bos taurus (n = 1903) (P = 0,008). Os COCs das doadoras zebu apresentaram melhor qualidade morfológica do que das doadoras taurina (n = 689 vs. 444 para COCs grau I, P < 0,0001; n = 681 vs. 509 para COCs grau II, P = 0,010, para doadoras zebu e taurina, respectivamente). Quanto à produção de embriões, houve diferença nas porcentagens obtidas de MO (0,44% zebuínas e 6,42% taurinas, P = 0,017), BL (14,18% zebuínas e 3,74% taurinas, P < 0,0001) e BX (81,43% zebuínas e 75,13% taurinas, P < 0,0001). Não foi observado diferença para BI e taxa de prenhez (P > 0,05). Os animais Bos indicus apresentaram maior recuperação oocitária e maior número de oócitos viáveis em comparação às vacas Bos taurus, como também, maior produção de embriões viáveis.(AU)

In vitro performance of Zebu (Bos indicus) and Taurus (Bos taurus) donor cow embryos / Desempenho de vacas doadoras zebuínas (Bos indicus) e taurinas (Bos taurus) na produção de embrião in vitro

In this study, the in vitro production of bovine embryos from zebu and taurine donors was compared. Cumulus-oocyte complexes (COCs) were obtained from 167 Bos taurus and 161 Bos indicus donors by ovum pick-up. COCs were classified based on their morphological quality, matured in incubators for 22 to 24 h in maturation medium, and then fertilized for 18 to 22 h. The zygotes were transferred to the culture medium for seven days. The embryos were classified as morula (OM), initial blastocyst (BI), blastocyst (BL), and expanded blastocyst (BX), before being transferred to synchronized recipient cows. Pregnancy was diagnosed 30-45 days post-transfer. The Bos indicus donors had a higher oocyte yield (n = 2556) than Bos taurus donors (n = 1903) (P = 0.008). The COCs from zebu donors had a better morphological quality than those from taurine donors (n = 689 vs. 444 for grade 1 COC, P 0.05). The Bos indicus cows showed greater oocyte recovery, number of viable oocytes, and production of viable embryos than the Bos taurus cows.

O objetivo foi avaliar a produção in vitro de embriões bovinos a partir de doadoras zebuína e taurina. Os complexos cumulus oócitos (COC’s) foram obtidos de 167 doadoras Bos taurus e 161 Bos indicus, por meio de Ovum pick-up. Os COC’s foram classificados quanto à qualidade morfológica, maturados em incubadora por 22 a 24h em meio de maturação, e encaminhados à fertilização entre 18 e 22h. Os zigotos foram transferidos para meio de cultivo, onde permaneceram durante sete dias. Os embriões foram classificados em mórula (MO), blastocisto inicial (BI), blastocisto (BL) e blastocisto expandido (BX), envasados em palhetas contendo meio de cultivo, e inovulados em receptoras sincronizadas. Após 30 e 45 dias da inovulação, realizou-se o diagnóstico de gestação. Os animais Bos indicus apresentaram maior produção oocitária (n = 2556) em comparação aos Bos taurus (n = 1903) (P = 0,008). Os COCs das doadoras zebu apresentaram melhor qualidade morfológica do que das doadoras taurina (n = 689 vs. 444 para COCs grau I, P 0,05). Os animais Bos indicus apresentaram maior recuperação oocitária e maior número de oócitos viáveis em comparação às vacas Bos taurus, como também, maior produção de embriões viáveis.

Transferência Nuclear de Células Somáticas interespecífica (TNCSi) na conservação de cervídeos em risco de extinção / Interspecific Somatic Cell Nuclear Transfer (TNCSi) in the conservation of endangered species of deer

A Transferência Nuclear de Células Somáticas interespecífica (TNCSi) envolve a transferência de um núcleo ou célula de uma espécie para o citoplasma de um oócito enucleado de outra espécie. Após ativação, os complexos carioplasto-citoplasto reconstruídos podem ser cultivados in vitro até o blastocisto, o estádio final de desenvolvimento pré-implantação. Esta pode ser uma estratégia interessante na tentativa de conservação de espécies em risco de extinção. Esta revisão tem por objetivo apresentar alguns detalhes da técnica e relatar a experiência de nosso grupo usando como modelo o veado-catingueiro.(AU)

Interspecies Somatic Cell Nuclear Transfer (iSCNT) involves the transfer of a nucleus or cell from one species into the cytoplasm of an enucleated oocyte from another. Once activated, reconstructed caryoplast-cytoplast complexes can be cultured in vitro to blastocyst, the final stage of preimplantation development. This may be an interesting strategy in attempting to conserve endangered species. The objective of this review is to present some details of the technique and to report the experience of our group using as model the brown brocket deer.(AU)

Transferência Nuclear de Células Somáticas interespecífica (TNCSi) na conservação de cervídeos em risco de extinção / Interspecific Somatic Cell Nuclear Transfer (TNCSi) in the conservation of endangered species of deer

A Transferência Nuclear de Células Somáticas interespecífica (TNCSi) envolve a transferência de um núcleo ou célula de uma espécie para o citoplasma de um oócito enucleado de outra espécie. Após ativação, os complexos carioplasto-citoplasto reconstruídos podem ser cultivados in vitro até o blastocisto, o estádio final de desenvolvimento pré-implantação. Esta pode ser uma estratégia interessante na tentativa de conservação de espécies em risco de extinção. Esta revisão tem por objetivo apresentar alguns detalhes da técnica e relatar a experiência de nosso grupo usando como modelo o veado-catingueiro.

Interspecies Somatic Cell Nuclear Transfer (iSCNT) involves the transfer of a nucleus or cell from one species into the cytoplasm of an enucleated oocyte from another. Once activated, reconstructed caryoplast-cytoplast complexes can be cultured in vitro to blastocyst, the final stage of preimplantation development. This may be an interesting strategy in attempting to conserve endangered species. The objective of this review is to present some details of the technique and to report the experience of our group using as model the brown brocket deer.

How can we improve embryo production and pregnancy outcomes of Holstein embryos produced in vitro? (12 years of practical results at a California dairy farm)

Genomic evaluations have revolutionized dairy cattle breeding, and the demand for embryos produced from very young heifers with high genetic merit has increased over time. The combination of low oocyte recovery, young age of donors, and milk production status can make the in vitro embryo production (IVP) of Holstein cattle incredibly challenging. Several factors need to be coordinated to obtain a live calf from an IVP embryo, but the quality of the oocyte at the start of the process is one of the key factors. Aspects related to oocyte quality, laboratory quality control, embryo quality and recipient selection are addressed here, based on the measures that the RuAnn Genetics Laboratory (Riverdale, California, USA) adopted in the last 12 years, with the goal of improving production of live, healthy calves from Holstein embryos. Follicular wave synchronization and stimulation with follicular stimulating hormone (FSH) is necessary to improve oocyte quality and consequently embryo production. Laboratory quality control and the use of high-quality supplies are essential to reduce variability in production and facilitate identification of other factors that might interfere with embryo production. High pregnancy rates can be achieved with good quality embryos selected at optimal time and stage of development, transferred by an experienced embryo transfer (ET) technician, to well managed recipients 7 or 8 days after estrus. Attention to detail at every step of the process is crucial to success.

How can we improve embryo production and pregnancy outcomes of Holstein embryos produced in vitro? (12 years of practical results at a California dairy farm)

Genomic evaluations have revolutionized dairy cattle breeding, and the demand for embryos produced from very young heifers with high genetic merit has increased over time. The combination of low oocyte recovery, young age of donors, and milk production status can make the in vitro embryo production (IVP) of Holstein cattle incredibly challenging. Several factors need to be coordinated to obtain a live calf from an IVP embryo, but the quality of the oocyte at the start of the process is one of the key factors. Aspects related to oocyte quality, laboratory quality control, embryo quality and recipient selection are addressed here, based on the measures that the RuAnn Genetics Laboratory (Riverdale, California, USA) adopted in the last 12 years, with the goal of improving production of live, healthy calves from Holstein embryos. Follicular wave synchronization and stimulation with follicular stimulating hormone (FSH) is necessary to improve oocyte quality and consequently embryo production. Laboratory quality control and the use of high-quality supplies are essential to reduce variability in production and facilitate identification of other factors that might interfere with embryo production. High pregnancy rates can be achieved with good quality embryos selected at optimal time and stage of development, transferred by an experienced embryo transfer (ET) technician, to well managed recipients 7 or 8 days after estrus. Attention to detail at every step of the process is crucial to success.(AU)

Strategies to increment in vivo and in vitro embryo production and transfer in cattle

Knowledge of follicular wave dynamics obtained through the use of real-time ultrasonography and the development of the means by which follicular wave dynamics can be controlled have provided practical approaches for the in vivo and in vitro production and transfer of embryos in cattle. The elective control of follicular wave emergence and ovulation has had a great impact on the application of on-farm embryo transfer, especially when large groups of donors need to be superstimulated at the same time. Although estradiol and progestins have been used for many years, practitioners in countries where estradiol cannot be used have turned to alternative treatments, such as mechanical follicle ablation or the administration of GnRH for the synchronization of follicle wave emergence. In vitro embryo production also benefits from the synchronization of follicle wave emergence prior to Cumulus Oocyte Complexes (COCs) recovery. As Bos indicus cattle have high antral follicle population, large numbers of oocytes can be obtained by ovum pick-up (OPU) without superstimulation. However, synchronization of follicular wave emergence and superstimulation is necessary to obtain high numbers of COCs by OPU and blastocysts following in vitro fertilization in Bos taurus donors. Finally, embryos can now be transferred in commercial beef or dairy herds using efficacious synchronization and resynchronization protocols that are easily implemented by farm personnel. These technologies can also be used to resolve reproductive problems such as the reduced fertility observed during summer heat stress and/or in repeat-breeder cows in commercial dairy herds.

Strategies to increment in vivo and in vitro embryo production and transfer in cattle

Knowledge of follicular wave dynamics obtained through the use of real-time ultrasonography and the development of the means by which follicular wave dynamics can be controlled have provided practical approaches for the in vivo and in vitro production and transfer of embryos in cattle. The elective control of follicular wave emergence and ovulation has had a great impact on the application of on-farm embryo transfer, especially when large groups of donors need to be superstimulated at the same time. Although estradiol and progestins have been used for many years, practitioners in countries where estradiol cannot be used have turned to alternative treatments, such as mechanical follicle ablation or the administration of GnRH for the synchronization of follicle wave emergence. In vitro embryo production also benefits from the synchronization of follicle wave emergence prior to Cumulus Oocyte Complexes (COCs) recovery. As Bos indicus cattle have high antral follicle population, large numbers of oocytes can be obtained by ovum pick-up (OPU) without superstimulation. However, synchronization of follicular wave emergence and superstimulation is necessary to obtain high numbers of COCs by OPU and blastocysts following in vitro fertilization in Bos taurus donors. Finally, embryos can now be transferred in commercial beef or dairy herds using efficacious synchronization and resynchronization protocols that are easily implemented by farm personnel. These technologies can also be used to resolve reproductive problems such as the reduced fertility observed during summer heat stress and/or in repeat-breeder cows in commercial dairy herds.(AU)

Embryo competence and cryosurvival: Molecular and cellular features

Global cattle genetic market is experiencing a change of strategy, large genetic companies, traditionally recognized in the artificial insemination field, have also begun to operate in the embryo market. Consequently, the demand for in vitro produced (IVP) embryos has grown. However, the overall efficiency of the biotechnology process remains low. Additionally, the lack of homogeneity of post-cryopreservation survival results of IVP embryos still impairing a massive dissemination of this biotechnology in the field. A great challenge for in vitro production labs is to increase the amount of embryos produced with exceptional quality after each round of in vitro fertilization. Herein, we discuss the molecular and cellular features associated with the competence and cryosurvival of IVP embryos. First, morphofunctional, cellular and molecular competence of the embryos were addressed and a relationship between embryo developmental ability and quality were established with cryosurvival and pregnancy success. Additionally, determinant factors of embryo competence and cryosurvival were discussed including the following effects: genotype, oocyte quality and follicular microenvironment, in vitro production conditions, and lipids and other determining molecules. Finally, embryo cryopreservation aspects were addressed and an embryofocused approach to improve cryosurvival was presented.

Embryo competence and cryosurvival: Molecular and cellular features

Global cattle genetic market is experiencing a change of strategy, large genetic companies, traditionally recognized in the artificial insemination field, have also begun to operate in the embryo market. Consequently, the demand for in vitro produced (IVP) embryos has grown. However, the overall efficiency of the biotechnology process remains low. Additionally, the lack of homogeneity of post-cryopreservation survival results of IVP embryos still impairing a massive dissemination of this biotechnology in the field. A great challenge for in vitro production labs is to increase the amount of embryos produced with exceptional quality after each round of in vitro fertilization. Herein, we discuss the molecular and cellular features associated with the competence and cryosurvival of IVP embryos. First, morphofunctional, cellular and molecular competence of the embryos were addressed and a relationship between embryo developmental ability and quality were established with cryosurvival and pregnancy success. Additionally, determinant factors of embryo competence and cryosurvival were discussed including the following effects: genotype, oocyte quality and follicular microenvironment, in vitro production conditions, and lipids and other determining molecules. Finally, embryo cryopreservation aspects were addressed and an embryofocused approach to improve cryosurvival was presented.(AU)

Goat incubator: Can bovine oocytes be matured in the uterine horn of a goat? / Cabra incubadora: os oócitos bovinos podem ser maturados no corno uterino de uma cabra?

We used a goat as a live incubator, along with associated nonsurgical embryo transfer techniques, to perform ex situ (in vivo) maturation of bovine oocytes. Immature bovine cumulus-oocyte complexes (COCs) aspirated from 3-8 mm follicles from slaughterhouse ovaries were randomly split into two groups for in vitro (IVM; n = 38) and ex situ maturation (ESM; n = 40). IVM was performed for a period of 24 h at 38.5 ºC and with 5% CO2 in the air of maximum humidity. For ESM, a presynchronized nulliparous goat (12 months old) received 40 immature COCs in the uterine horn apiece, via the transcervical route. After 24 h the structures were retrieved through uterine flushing. Analyses of nuclear maturation and lipid quantification were performed on oocytes from both groups. Fluorescent intensity was compared using the Student’s t-test. Forty-seven percent of the structures were recovered after uterine flushing (19/40). The nuclear maturation rate was 94.5% (18/19) and 81.6% (31/38) for the ESM and IVM groups, respectively. In vitro-matured COCs contained more lipid droplets, expressed as a higher amount (p < 0.05) of emitted fluorescent light than ex situ-matured COCs (858 ± 73 vs. 550 ± 64 arbitrary fluorescence units, respectively). This is the first report to associate nonsurgical embryo transfer techniques and a goat as a live incubator for the maturation of bovine oocytes. We

Utilizamos uma cabra como incubadora viva, juntamente com técnicas de transferência de embriões não cirúrgicas associadas, para realizar a maturação ex situ (in vivo) de oócitos bovinos. Os complexos cumulus-oócitos bovinos (COCs), aspirados de folículos de 3-8 mm de ovários de abatedouro foram aleatoriamente divididos em dois grupos para maturação in vitro (IVM; n = 38) e ex situ (ESM; n = 40). A MIV foi realizada por um período de 24 h no meio TCM-199, a 38,5 ºC, e com 5% de CO2 em umidade máxima. Para o ESM, uma cabra nulípara pré-sincronizada (12 meses de idade) recebeu 40 COCs imaturos no ápice do corno uterino, por via transcervical. Após 24 h as estruturas foram recuperadas através de lavagem uterina. Análises de maturação nuclear e quantificação de lipídios foram realizadas em oócitos de ambos os grupos. A intensidade de fluorescente foi comparada usando o teste t de Student. Quarenta e sete por cento das estruturas foram recuperadas após lavagem uterina (19/40). A taxa de maturação nuclear foi de 94,5% (18/19) e 81,6% (31/38) para os grupos ESM e IVM, respectivamente. Os COCs maturados in vitro continham mais gotículas lipídicas, expressos como uma quantidade maior (p < 0,05) da luz fluorescente emitida do que os COCs ex situ (858 ± 73 vs 550 ± 64 unidades de fluorescência arbitrárias, respectivamente). Este é o primeiro relató

Approaches to reduce lipids: a review of its impacts on in vitro embryo production / Abordagens para reduzir lipídeos: uma revisão sobre seus impactos na produção de embriões in vitro

In vitro embryo production (IVP) has limitations for better outcomes in blastocyst production, cryopreservation efficiency and pregnancy rates. Among the factors impairing IVP, high lipid content in both oocytes and embryos, and consequent reactive oxygen species (ROS) production, have a negative impact in embryo development and further biotechnologies. The present review aims to address techniques and strategies that collaborate to improve embryo development rates through reduction of lipid content either in the oocyte or in the embryo.(AU)

A produção in vitro de embriões (PIVE) possui limitações para melhoras na produção de blastocistos, eficiência da criopreservação e taxas de prenhez. Dentre os fatores limitantes à PIVE, o alto conteúdo lipídico tanto em oócitos quanto em embriões e a consequente produção de espécies reativas a oxigênio (ROS), possuem impacto negativo no desenvolvimento embrionário e subsequentes biotecnologias. A presente revisão visa tratar sobre tecnologias e estratégias capazes de colaborar com a melhora nas taxas de desenvolvimento embrionário através da redução do conteúdo lipídico tanto em oócitos quanto em embriões.(AU)

Approaches to reduce lipids: a review of its impacts on in vitro embryo production / Abordagens para reduzir lipídeos: uma revisão sobre seus impactos na produção de embriões in vitro

In vitro embryo production (IVP) has limitations for better outcomes in blastocyst production, cryopreservation efficiency and pregnancy rates. Among the factors impairing IVP, high lipid content in both oocytes and embryos, and consequent reactive oxygen species (ROS) production, have a negative impact in embryo development and further biotechnologies. The present review aims to address techniques and strategies that collaborate to improve embryo development rates through reduction of lipid content either in the oocyte or in the embryo.

A produção in vitro de embriões (PIVE) possui limitações para melhoras na produção de blastocistos, eficiência da criopreservação e taxas de prenhez. Dentre os fatores limitantes à PIVE, o alto conteúdo lipídico tanto em oócitos quanto em embriões e a consequente produção de espécies reativas a oxigênio (ROS), possuem impacto negativo no desenvolvimento embrionário e subsequentes biotecnologias. A presente revisão visa tratar sobre tecnologias e estratégias capazes de colaborar com a melhora nas taxas de desenvolvimento embrionário através da redução do conteúdo lipídico tanto em oócitos quanto em embriões.

Developments of reproductive management and biotechnology in the pig

This review aims to describe changes in production environment, management tools and technology to alleviate problems seen with the present hyperprolific sow model. Successful parturition in the pig includes the possibility to express adequate maternal behaviour, rapid expulsion of piglets, complete expulsion of placenta, elimination of uterine contamination and debris, neonatal activity and colostrum intake. We focus on management of large litters, including maternal behaviour, ease of parturition, colostrum production, piglet quality parameters and intermittent suckling. There are also some interesting developments in technology to assess colostrum and immune state of the piglet. These developments may be utilized to improve the success rate of reproductive management around farrowing, lactation and after weaning. We also discuss new insights in how to examine the health of the mammary gland, uterus and ovaries of hyperprolific sows. Finally, we assess the latest developments on breeding and technology of hyperprolific sows, including artificial insemination (AI), real-time ultrasound of the genital tract and embryo transfer (ET). We conclude that 1) for the sow to produce sufficient colostrum, both the behavioural and physiological needs of the sow need to be met before and after parturition. Furthermore, 2) new ultrasound and biopsy technology can be effectively applied for accurate diagnosis of inflammatory processes of the udder and uterus and timing of AI regarding ovulation to improve insemination efficiency. Finally, 3) developments in cryopreservation of germ cells and embryos appear promising but lack of valid oocyte collection techniques and nonsurgical ET techniques are a bottleneck to commercial ET. These latest developments in management of parturition and reproductive technology are necessary to cope with the increasing challenges associated with very large litter sizes.

Developments of reproductive management and biotechnology in the pig

This review aims to describe changes in production environment, management tools and technology to alleviate problems seen with the present hyperprolific sow model. Successful parturition in the pig includes the possibility to express adequate maternal behaviour, rapid expulsion of piglets, complete expulsion of placenta, elimination of uterine contamination and debris, neonatal activity and colostrum intake. We focus on management of large litters, including maternal behaviour, ease of parturition, colostrum production, piglet quality parameters and intermittent suckling. There are also some interesting developments in technology to assess colostrum and immune state of the piglet. These developments may be utilized to improve the success rate of reproductive management around farrowing, lactation and after weaning. We also discuss new insights in how to examine the health of the mammary gland, uterus and ovaries of hyperprolific sows. Finally, we assess the latest developments on breeding and technology of hyperprolific sows, including artificial insemination (AI), real-time ultrasound of the genital tract and embryo transfer (ET). We conclude that 1) for the sow to produce sufficient colostrum, both the behavioural and physiological needs of the sow need to be met before and after parturition. Furthermore, 2) new ultrasound and biopsy technology can be effectively applied for accurate diagnosis of inflammatory processes of the udder and uterus and timing of AI regarding ovulation to improve insemination efficiency. Finally, 3) developments in cryopreservation of germ cells and embryos appear promising but lack of valid oocyte collection techniques and nonsurgical ET techniques are a bottleneck to commercial ET. These latest developments in management of parturition and reproductive technology are necessary to cope with the increasing challenges associated with very large litter sizes.(AU)

Reproductive physiology of the heat-stressed dairy cow: implications for fertility and assisted reproduction

Heat stress causes a large decline in pregnancy success per insemination during warm times of the year. Improvements in fertility are possible by exploiting knowledge about how heat stress affects the reproductive process. The oocyte can be damaged by heat stress at the earliest stages of folliculogenesis and remains sensitive to heat stress in the peri-ovulatory period. Changes in oocyte quality due to heat stress are the result of altered patterns of folliculogenesis and, possibly, direct effects of elevated body temperature on the oocyte. While adverse effects of elevated temperature on the oocyte have been observed in vitro, local cooling of the ovary and protective effects of follicular fluid may limit these actions in vivo. Heat stress can also compromise fertilization rate. The first seven days of embryonic development are very susceptible to disruption by heat stress. During these seven days, the embryo undergoes a rapid change in sensitivity to heat stress from being very sensitive (2- to 4-cell stage) to largely resistant (by the morulae stage). Direct actions of elevated temperature on the embryo are likely to be an important mechanism for reduction in embryonic survival caused by heat stress. An effective way to avoid effects of heat stress on the oocyte, fertilization, and early embryo is to bypass the effects through embryo transfer because embryos are typically transferred into females after acquisition of thermal resistance. There may be some opportunity to mitigate effects of heat stress by feeding antioxidants or regulating the endocrine environment of the cow but neither approach has been reduced to practice. The best long-term solution to the problem of heat stress may be to increase genetic resistance of cows to heat stress. Thermotolerance genes exist within dairy breeds and additional genes can be introgressed from other breeds by traditional means or gene editing.

Reproductive physiology of the heat-stressed dairy cow: implications for fertility and assisted reproduction

Heat stress causes a large decline in pregnancy success per insemination during warm times of the year. Improvements in fertility are possible by exploiting knowledge about how heat stress affects the reproductive process. The oocyte can be damaged by heat stress at the earliest stages of folliculogenesis and remains sensitive to heat stress in the peri-ovulatory period. Changes in oocyte quality due to heat stress are the result of altered patterns of folliculogenesis and, possibly, direct effects of elevated body temperature on the oocyte. While adverse effects of elevated temperature on the oocyte have been observed in vitro, local cooling of the ovary and protective effects of follicular fluid may limit these actions in vivo. Heat stress can also compromise fertilization rate. The first seven days of embryonic development are very susceptible to disruption by heat stress. During these seven days, the embryo undergoes a rapid change in sensitivity to heat stress from being very sensitive (2- to 4-cell stage) to largely resistant (by the morulae stage). Direct actions of elevated temperature on the embryo are likely to be an important mechanism for reduction in embryonic survival caused by heat stress. An effective way to avoid effects of heat stress on the oocyte, fertilization, and early embryo is to bypass the effects through embryo transfer because embryos are typically transferred into females after acquisition of thermal resistance. There may be some opportunity to mitigate effects of heat stress by feeding antioxidants or regulating the endocrine environment of the cow but neither approach has been reduced to practice. The best long-term solution to the problem of heat stress may be to increase genetic resistance of cows to heat stress. Thermotolerance genes exist within dairy breeds and additional genes can be introgressed from other breeds by traditional means or gene editing.(AU)

In vitro embryo production in buffaloes: from the laboratory to the farm

Transvaginal follicular aspiration technique together with in vitro embryo production are the biotechnological alternatives currently available to support genetic improvement breeding programs in buffalo species. However, aspects related to animal management, lack of knowledge of the metabolic needs and biochemical peculiarities of gametes and embryos, as well as the reproductive physiology characteristics have hampered progress in the results. Despite the low availability of good quality oocytes collected after OPU in donors as a physiological characteristic of buffalo species, high rates of oocyte maturation, modest embryo cleavage, blastocyst production and pregnancy rates after transvaginal embryo transfer in recipients could be obtained in buffalo in vitro embryo production programs. The results of implementing an in vitro embryo production program in buffaloes in the northern region of Pará state, Brazil, and results published by other groups demonstrate the feasibility of implementing this biotechnology in the routine of breeding programs. Nevertheless, in order to achieve better and consistent results, it is necessary to deepen the knowledge on the peculiarities of reproductive biology in this specie. Selection of donor animals based on ovarian size and ovarian follicular reserve and on the rate of blastocyst production is presented as an effective alternative to increase the efficiency of the in vitro embryo production technique applied to the buffalo species.

In vitro embryo production in buffaloes: from the laboratory to the farm

Transvaginal follicular aspiration technique together with in vitro embryo production are the biotechnological alternatives currently available to support genetic improvement breeding programs in buffalo species. However, aspects related to animal management, lack of knowledge of the metabolic needs and biochemical peculiarities of gametes and embryos, as well as the reproductive physiology characteristics have hampered progress in the results. Despite the low availability of good quality oocytes collected after OPU in donors as a physiological characteristic of buffalo species, high rates of oocyte maturation, modest embryo cleavage, blastocyst production and pregnancy rates after transvaginal embryo transfer in recipients could be obtained in buffalo in vitro embryo production programs. The results of implementing an in vitro embryo production program in buffaloes in the northern region of Pará state, Brazil, and results published by other groups demonstrate the feasibility of implementing this biotechnology in the routine of breeding programs. Nevertheless, in order to achieve better and consistent results, it is necessary to deepen the knowledge on the peculiarities of reproductive biology in this specie. Selection of donor animals based on ovarian size and ovarian follicular reserve and on the rate of blastocyst production is presented as an effective alternative to increase the efficiency of the in vitro embryo production technique applied to the buffalo species.(AU)

Inflammation: friend or foe of bovine reproduction?

Inflammation is not only the first line of defense of the organism but is also required in many reproductive processes such as ovulation, corpus luteum development, luteolysis, uterine clearance after insemination and post partum. Nevertheless, if excessive or persistent, inflammation can switch from a positive mechanism to a deleterious process, impairing oocyte quality and embryo development. Not only uterine but also non genital inflammatory sites can depreciate reproductive performances, with a carry over effect of 2 to 4 months. Since the metabolic challenges of the peripartum transition period make difficult for the cow to control inflammation, dairy cows are frequently in a pro-inflammatory stage, suggesting that inflammation, rather than infection, is a limiting factor of fertility in modern dairy cows. Within the first week after calving, cows have to mount an intense inflammatory response to the bacterial invasion of the uterine cavity with the challenge of being able to switch it off in no more than 5-6 weeks. The absence of neutrophils on endometrial smear is associated with the highest success rate at insemination. Since a fine tuning – rather than an absence - of inflammation is required along the reproductive cycle, anti-inflammatory drugs do not allow any improvement of pregnancy rate, except in the specific case of embryo transfer. Appropriate management of the transition period (especially nutritional) and in a long term perspective, genetic selection contribute to improve the aptitude of cows to controls the intensity of inflammatory process.