A preliminary approach to embryonic development of Austrolebias wolterstorffi, an endangered neotropical annual fish species / Desenvolvimento embrionário de Austrolebias wolterstorffi, uma espécie de peixe anual neotropical criticamente ameaçada de extinção

Annual fish live exclusively in temporary wetlands and are among the shortest-lived vertebrates in the world. These fish persist in these habitats due to drought-resistant eggs, that which, through diapauses are able to detect stimuli from the environment to start the development processes. They are also able to direct their embryonic development in different trajectories with different development times. Our objective in this paper was to describe the different stages of embryonic development of Austrolebias wolterstorffi, a critically endangered annual fish. A total of 27 stages of embryonic development were identified for the two observed developmental pathways (direct and diapause 2). Of these 27 developmental stages observed, 24 were identical between the two types of trajectories and three different. A total of 90% of the embryos that completed their development proceeded normally, without pauses. One embryo had a different development from the others, entering diapause 2, with a pause in development for 48 h. Although the embryonic development of A. wolterstorffi is similar to that of other Neotropical annual fish species, the diapause 2 occurs when the embryo has a large body size and a well-developed brain, indicating that the lack of embryonic information of the genus Austrolebias may hide characteristics still undescribed or even different survival strategies than what has been observed for other annual fish.

Os peixes anuais vivem exclusivamente em áreas úmidas temporárias e estão entre os vertebrados de vida mais curta do mundo. Esses peixes persistem nesses habitats devido a ovos resistentes à seca, que, por meio de diapausas, são capazes de detectar estímulos do ambiente para iniciar os processos de desenvolvimento embrionário. Eles também são capazes de direcionar seu desenvolvimento embrionário em diferentes trajetórias com diferentes tempos de desenvolvimento. O objetivo deste trabalho foi descrever os diferentes estágios do desenvolvimento embrionário de Austrolebias wolterstorffi, um peixe anual criticamente ameaçado. Um total de 27 estágios de desenvolvimento embrionário foram identificados para as duas trajetórias de desenvolvimento observadas (direto e diapausa 2). Desses 27 estágios de desenvolvimento observados, 24 eram idênticos entre os dois tipos de trajetórias e três diferentes. Um total de 90% dos embriões que completaram seu desenvolvimento seguiu normalmente, sem pausas. Um embrião teve um desenvolvimento diferente dos demais, entrando na diapausa 2, com pausa no desenvolvimento de 48h. Embora o desenvolvimento embrionário de A. wolterstorffi seja semelhante ao de outras espécies de peixes anuais neotropicais, a diapausa 2 ocorre quando o embrião tem um corpo grande e um cérebro bem desenvolvido, indicando que a falta de informação embrionária do gênero Austrolebias pode escondem características ainda não descritas ou mesmo estratégias de sobrevivência diferentes das observadas para outros peixes anuais.

Lab partners: oocytes, embryos and company. A personal view on aspects of oocyte maturation and the development of monozygotic twins

The present review addresses the oocyte and the preimplantation embryo, and is intended to highlight the underlying principle of the "nature versus/and nurture" question. Given the diversity in mammalian oocyte maturation, this review will not be comprehensive but instead will focus on the porcine oocyte. Historically, oogenesis was seen as the development of a passive cell nursed and determined by its somatic compartment. Currently, the advanced analysis of the cross-talk between the maternal environment and the oocyte shows a more balanced relationship: Granulosa cells nurse the oocyte, whereas the latter secretes diffusible factors that regulate proliferation and differentiation of the granulosa cells. Signal molecules of the granulosa cells either prevent the precocious initiation of meiotic maturation or enable oocyte maturation following hormonal stimulation. A similar question emerges in research on monozygotic twins or multiples: In Greek and medieval times, twins were not seen as the result of the common course of nature but were classified as faults. This seems still valid today for the rare and until now mainly unknown genesis of facultative monozygotic twins in mammals. Monozygotic twins are unique subjects for studies of the conceptus-maternal dialogue, the intra-pair similarity and dissimilarity, and the elucidation of the interplay between nature and nurture. In the course of in vivo collections of preimplantation sheep embryos and experiments on embryo splitting and other microsurgical interventions we recorded observations on double blastocysts within a single zona pellucida, double inner cell masses in zona-enclosed blastocysts and double germinal discs in elongating embryos. On the basis of these observations we add some pieces to the puzzle of the post-zygotic genesis of monozygotic twins and on maternal influences on the developing conceptus.(AU)

Desafios na andrologia de espécies selvagens, com ênfase em felídeos / Challenges in wild species andrology, with an emphasis on felids

Na prática, compreender a anatomia reprodutiva, a fisiologia e o comportamento do macho, da espécie selvagem com a qual se está trabalhando, é essencial para a avaliação andrológica e para a reprodução bem sucedida. Neste artigo damos especial ênfase à andrologia de felídeos selvagens. O profissional pode ser chamado para solucionar problemas quando a espécie apresenta dificuldade de reprodução em cativeiro e/ou encontra-se ameaçada de extinção. Baixas taxas de fertilidade e de prenhez podem estar associadas ao macho, com influências do estresse pelo cativeiro, nutrição inadequada e erros de manejo reprodutivo. E também, as instituições que mantêm essas espécies sob cuidado humano poderão ser auxiliadas no manejo reprodutivo com o exame andrológico periódico, que pode ser acompanhado de criopreservação de sêmen para formação ou manutenção de um banco de reserva genômica. Como componente de um programa holístico de conservação, incluindo os estudos de ecologia de campo, esta estratégia fortalece ainda mais o vínculo entre as populações ex situ e in situ.(AU)

In practice, understanding the male reproductive anatomy, physiology and the behavior, of the wild species that you are working, is essential for andrological assessment and successful reproduction. In this article we give special emphasis to the andrology of wild felids. The professional can be called to solve problems when the species has difficulty reproducing in captivity and/or is threatened with extinction. Low fertility and pregnancy rates may be associated with the male, with influences from stress in captivity, inadequate nutrition and reproductive management errors. Also, the institutions that keep these species under human care can be assisted in reproductive management with periodic andrological examination, which can be accompanied by semen cryopreservation for the formation or maintenance of a genome resource bank. As a component of a holistic conservation program, including field ecology studies, this strategy further strengthens the link between ex situ and in situ populations.(AU)

Controles gestacionales y salud materno-neonatal en gatas / Gestational checking and maternal-neonatal health in cats

El primer punto a tener en cuenta es determinar si el estado de salud de la gata permite incluirla en un plan reproductivo y de ser así cuál es su probabilidad de distocia. Es así que se planifican 3 consultas: la primera durante el celo previo al servicio, la segunda 20 días después del día de los servicios momento en el que se realiza el diagnóstico de gestación y se evalúa el desarrollo de los conceptos gestados si la gata quedó preñada, y la tercera al día 50 de gestación en la cual se realiza un examen de salud y el control del número de fetos gestados mediante Rx. La evaluación del estado de salud de la madre y de los conceptos gestados, así como el desarrollo de los mismos permitirá estimar la fecha probable de parto, y programar la asistencia al parto o una cesárea con mínimos riesgos según sea el caso.(AU)

Processos que afetam a taxa de gestação pós-inovulação do embrião equino / Processes that affect pregnancy rate after embryo transfer

A transferência de embriões equinos é uma biotecnologia que traz inúmeros benefícios reprodutivos para um haras. Entretanto, os índices reprodutivos obtidos numa estação de monta podem ser afetados por diversos fatores que interferem tanto na recuperação embrionária que normalmente são relacionados à doadora de embriões, quanto na taxa de gestação na receptora de embriões. O objetivo do presente artigo foi revisar os fatores relacionados à receptora de embriões em programas de transferência de embriões equinos.(AU)

Equine embryo transfer is a biotechnology that brings reproductive benefits to a stud farm. However, the reproductive rates obtained in a breeding season can be affected by several factors that interfere both in the embryonic recovery that are normally related to the embryo donor, and in the pregnancy rate in the embryo recipient. The aim of this article was to review the factors related to the recipient of embryos in equine embryo transfer programs.(AU)

Differentially methylated regions identified in bovine embryos are not observed in adulthood

The establishment of epigenetic marks during the reprogramming window is susceptible to environmental influences, and stimuli during this critical stage can cause altered DNA methylation in offspring. In a previous study, we found that low levels of sulphur and cobalt (low S/Co) in the diet offered to oocyte donors altered the DNA methylome of bovine embryos. However, due to the extensive epigenetic reprogramming that occurs during embryogenesis, we hypothesized that the different methylation regions (DMRs) identified in the blastocysts may not maintain in adulthood. Here, we aimed to characterize DMRs previously identified in embryos, in the blood and sperm of adult progenies of two groups of heifers (low S/Co and control). We used six bulls and characterized the DNA methylation levels of KDM2A, KDM5A, KMT2D, and DOT1L genes. Our results showed that all DMRs analysed in both groups and tissues were hypermethylated unlike that noticed in the embryonic methylome profiles. These results suggest that embryo DMRs were reprogrammed during the final stages of de novo methylation during embryogenesis or later in development. Therefore, due to the highly dynamic epigenetic state during early embryonic development, we suggest that is essential to validate the DMRs found in embryos in adult individuals.(AU)

Efeitos da melatonina exógena na viabilidade embrionária e no ambiente uterino de ovelhas / Effects of exogenous melatonin on embryo viability and uterine environment of ewes

O objetivo deste artigo é revisar o efeito da melatonina na viabilidade embrionária e ambiente uterino em ovinos, tanto in vivo quanto in vitro, considerando as vias pelas quais esse hormônio pode ter influência no embrião ou o mecanismo de reconhecimento materno e estabelecimento de gravidez em ovelhas.(AU)

The aim of this article is to review the effect of melatonin on embryo viability and uterine environment in sheep, both in vivo and in vitro, considering the pathways by which this hormone could have influences at the embryo or the mechanism of maternal recognition and establishment of pregnancy in sheep.(AU)

Maternal contributions to pregnancy success: from gamete quality to uterine environment

The establishment and maintenance of a pregnancy that goes to term is sine qua non for the long-term sustainability of dairy and beef cattle operations. The oocyte plays a critical role in providing the factors necessary for preimplantation embryonic development. Furthermore, the female, or maternal, environment where oocytes and embryos develop is crucial for the establishment and maintenance of a pregnancy to term. During folliculogenesis, the oocyte must sequentially acquire meiotic and developmental competence, which are the results of a series of molecular events preparing the highly specialized gamete to return to totipotency after fertilization. Given that folliculogenesis is a lengthy process in the cow, the occurrence of disease, metabolic imbalances, heat stress, or other adverse events can make it challenging to maintain oocyte quality. Following fertilization, the newly formed embryo must execute a tightly planned program that includes global DNA remodeling, activation of the embryonic genome, and cell fate decisions to form a blastocyst within a few days and cell divisions. The increasing use of assisted reproductive technologies creates an additional layer of complexity to ensure the highest oocyte and embryo quality given that in vitro systems do not faithfully recreate the physiological maternal environment. In this review, we discuss cellular and molecular factors and events known to be crucial for proper oocyte development and maturation, as well as adverse events that may negatively affect the oocyte; and the importance of the uterine environment, including signaling proteins in the maternal-embryonic interactions that ensure proper embryo development. We also discuss the impact of assisted reproductive technologies in oocyte and embryo quality and developmental potential, and considerations when looking into the prospects for developing systems that allow for in vitro gametogenesis as a tool for assisted reproduction in cattle.(AU)

Efeitos imediatos, tardios e transgeracionais do estresse térmico em gametas / Immediate, late and transgenerational effects of heat stress on gametes

As perdas de produtividade e fertilidade animal associadas ao estresse térmico durante os meses mais quentes do ano é um dos maiores desafios do setor pecuário. Na indústria de leite as perdas econômicas causadas pelo estresse térmico foram estimadas em mais de 1,5 bilhões de dólares por ano. No que tange a reprodução, já foi demonstrado que o estresse térmico exerce múltiplos efeitos deletérios, causando disfunções endócrinas e alterando a sequência orquestrada de eventos importantes para a gametogênese e para o desenvolvimento embrionário inicial. Estudos recentes têm esclarecido o padrão temporal no qual os danos são estabelecidos e carreados dependendo da intensidade do estresse. Enquanto os efeitos imediatos do estresse térmico nos gametas já são bem caracterizados, existem evidências de que alguns danos podem ser carreados de forma tardia e possivelmente entre gerações. Além disso, dados emergentes indicam que o estresse térmico compromete a reprogramação da metilação do DNA que ocorre durante a gametogênese e a programação do desenvolvimento in utero. Dessa forma, esse artigo visa explorar os efeitos imediatos, tardios e transgeracionais do estresse térmico nos gametas.(AU)

The drop on animal productivity and fertility associated with heat stress during the hot months of the year is one of the biggest challenges for the livestock sector. For the dairy industry the economic losses caused by heat stress have been estimated over 1.5 billion dollars per year. It has already been demonstrated that heat stress exerts multiple deleterious effects on reproductive function, causing endocrine dysfunctions as well as changes in the sequence of events required for gametogenesis and early embryonic development. Recent studies have shed a light in the temporal pattern in which heat-induced damage is established and carried forward depending on the intensity of stress. While the immediate effects of heat stress on gametes are well characterized, there is evidence that some damage can be carried over for longer periods and even across generations. Furthermore, emerging data indicate that heat stress compromises DNA methylation reprogramming that occurs during gametogenesis and developmental programming in utero. Thus, this paper aims to explore the immediate, late and transgenerational effects of heat stress on gametes.(AU)

Effect of temperature on embryonic development and first exogenous feeding of goldfish Carassius auratus (Linnaeus, 1758) / Efeito da temperatura no desenvolvimento embrionário e na primeira alimentação exógena do goldfish Carassius auratus (Linnaeus, 1758)

Goldfish or Kinguio is a widely marketed species worldwide due to the ornamental market. There is some lack of acknowledgment of the production of the species under specific climatic conditions. To evaluate the effect of temperature on embryonic development and the first exogenous feeding of goldfish, an experiment was proposed. Fifteen incubators, organized in five treatments (18, 22, 26, 30, and 34 °C) with three replications each, were used to keep the fertilized goldfish eggs until the first exogenous feeding of the larvae. The main development events were observed to understand the possible effects of these temperatures on embryos and larvae of the species. Temperature influences embryo development and the time of first exogenous feeding of goldfish. The temperature of 34 °C was lethal to the species causing 100% of anomalies in the embryos and larvae. The experiment data allow us to conclude that the species presents a maximum thermal limit during embryogenesis, and these data are important to the aquaculture industry and to understand the effect of climate changes on goldfish. The data obtained in this experiment will assist in the management of invasive species and production of the species (aquaculture).

O peixe dourado ou Kinguio é uma espécie amplamente comercializada em todo o mundo devido ao mercado ornamental. Existe alguma falta de conhecimento da produção da espécie em condições climáticas específicas. Para avaliar o efeito da temperatura no desenvolvimento embrionário e na primeira alimentação exógena do Kinguio, um experimento foi proposto. Quinze incubadoras, organizadas em cinco tratamentos (18, 22, 26, 30 e 34 °C) com três repetições cada, foram utilizadas para manter os ovos fertilizados de Kinguio até a primeira alimentação exógena das larvas. Os principais eventos do desenvolvimento foram observados para entender os possíveis efeitos dessas temperaturas em embriões e larvas da espécie. A temperatura influencia o desenvolvimento do embrião e o tempo da primeira alimentação exógena do Kinguio. A temperatura de 34°C foi letal para a espécie causando 100% de anomalias nos embriões e larvas. Os dados do experimento permitem concluir que a espécie apresenta um limite térmico máximo durante a embriogênese, sendo esses dados importantes para a indústria da aquicultura e para entender o efeito das mudanças climáticas no Kinguio. Os dados obtidos neste experimento auxiliarão no manejo de espécies invasoras e na produção da espécie (aquicultura).

The interaction between the environment and embryo development in assisted reproduction

It can be assumed that the natural processes of selection and developmental condition in the animal provide the best prerequisites for embryogenesis resulting in pregnancy and subsequent birth of a healthy neonate. In contrast, circumventing the natural selection mechanisms and all developmental conditions in a healthy animal harbors the risk of counteracting, preventing or reducing the formation of embryos or substantially restricting their genesis. Considering these facts, it seems to be obvious that assisted reproductive techniques focusing on early embryonic stages serve an expanded and unselected germ cell pool of oocytes and sperm cells, and include the culture of embryos outside their natural habitat during and after fertilization for manipulation and diagnostic purposes, and for storage. A significant influence on the early embryonic development is seen in the extracorporeal culture of bovine embryos (in vitro) or stress on the animal organism (in vivo). The in vitro production per se and metabolic as well as endocrine changes in the natural environment of embryos represent adequate models and serve for a better understanding. The purpose of this review is to give a brief presentation of recent techniques aimed at focusing more on the complex processes in the Fallopian tube to contrast in vivo and in vitro prerequisites and abnormalities in early embryonic development and serve to identify potential new ways to make the use of ARTs more feasible.(AU)

Pontos críticos na produção de embriões equinos com enfoque na doadora / Critical points in the production of equine embryos with a donor approach

Inúmeras são as vantagens da utilização da técnica de transferência de embriões (TE) em equinos. No entanto, alguns fatores podem afetar a taxa de recuperação embrionária: dia da colheita do embrião; condição uterina e idade da doadora; manejos reprodutivo, sanitário e nutricional; garanhão/qualidade do sêmen; habilidade do técnico; condições climáticas, número de ovulações e adaptação da doadora. O objetivo deste artigo é revisar os fatores que podem exercer influência sobre a taxa de recuperação embrionária tendo impacto direto nos índices reprodutivos de um programa de TE.(AU)

There are countless advantages of using the embryo transfer (ET) technique in horses. However, some factors may affect the embryonic recovery rate: day of embryo collection; uterine condition and donor age; reproductive, health and nutritional management; stallion/semen quality; skill of the technician; climatic conditions, number of ovulations and adaptation of the donor. The objective of this article is to review those factors that can influence the embryonic recovery rate, having a direct impact on the reproductive rates of an ET program.(AU)

Impacto da qualidade seminal e dos fatores relativos ao macho na produção in vitro de embriões bovinos / Impact of semen quality and other male-related factors on in vitro embryo production in cattle

A importância da qualidade do sêmen no processo comercial de produção in vitro de embriões (PIVE) é bem conhecida, ainda que não devidamente relatada na literatura. Existe não apenas uma significativa diferença entre touros nas taxas de clivagem e de blastocistos, mas também nas taxas de prenhezes subsequentes. Adicionalmente, há evidências de interação entre touro e tecnologia de processamento do sêmen (particularmente na separação de espermatozoides por sexo), entre touro e protocolo de preparação do sêmen para fertilização in vitro, e ainda entre touro e doadora. Controlar estes efeitos em uma rotina comercial tem sido um desafio crescente para os laboratórios, particularmente com a alta oferta de novos touros decorrente da recente adoção da seleção genômica. O presente trabalho aborda algumas destas questões, com base na experiência da Bio Biotecnologia da Reprodução Animal nesta área.(AU)

The importance of semen quality in a commercial in vitro embryo production (IVEP) routine is well-known, although underreported in the literature. There is not only a significant difference among sires on cleavage and blastocyst rates, but also on subsequent pregnancy rates. Moreover, there are evidences of interaction between sire and sperm processing technology (particularly in the case of sex-sorted semen), between sire and the protocol for sperm preparation for in vitro fertilization, and between sire and donor. Controlling such effects in a commercial routine has been a growing challenge for the laboratories, especially due to the high turnover of sires caused by the recent adoption of genomic selection in most breeds. The current study discusses some of these aspects, from the perspective of the experience of Bio Biotecnologia da Reprodução Animal in this field.(AU)

In vivo embryo development in bitches inseminated laparoscopically after ovulation time estimated based on a single progesterone determination

Logistic and economical limitations are often the causes of dog owners not accurately monitoring the estrous cycle and the optimal insemination time. The aim of this study was to evaluate in vivo early embryonic development in bitches, after the analysis of sequential vaginal cytologies associated to single progesterone measurement and single laparoscopic insemination with high quality semen (fresh and with high spermatozoa concentration) or low-quality semen (frozen/thawed and with low spermatozoa concentration) at 48 h post- ovulation time predicted on a single progesterone measurement. Ten bitches were inseminated with 250 x 106 fresh spermatozoa (80% motility), and ten with 80 x 106 frozen/thawed spermatozoa (60% motility) in the cranial part of each uterine horn. Seven days later, ovariohysterectomy was performed and the oviducts and uterine horns and body were flushed to recover embryos and unfertilized oocytes. In 80% of the bitches inseminated with fresh and 50% of bitches inseminated with frozen/thawed semen, embryos at 2 to 8 cells stage were recovered mostly from the, oviducts. This study indicates that pregnancies can be obtained with a single laparoscopic intrauterine insemination after single serum progesterone measurement, although with a low number of embryos. This result should be taken into account in case economic or logistic restrictions that affect the possibility of owners to plan an accurate monitoring of the optimal breeding time using fresh and frozen semen.(AU)

The waveless model: clinical and research applications of follicular wave suppression in cattle / Modelo waveless: aplicações clínicas e na pesquisa da supressão das ondas foliculares em bovinos

The follicular growth waves are directly linked to the fluctuations in plasma gonadotrophins, which are controlled by the hypothalamic GnRH release pattern. Therefore, if the actions of the GnRH are inhibited or blocked, the final stages of the antral follicle growth are suppressed, resulting in an induced anestrus (a.k.a. waveless model). In the human medicine, GnRH agonists or antagonists are broadly used in the control of ovarian disfunctions, as well as in the preparation of women for assisted reproductive cycles. In cattle, a similar effect can be obtained by active immunization against GnRH. This was shown to be a viable strategy, for example, for the control of chronic cases of cystic ovarian disease in oocyte donors. However, on shall take into account the substantial individual variation on the immune response and, consequently, the lack of control of the duration of the anestrus induced. The waveless model is also very useful as a research model, once it controls the potential interference of the endogenous FSH and LH, improving the sensitivity of essays with exogenous hormones and consequently reducing the required number of replicas within studies.(AU)

O padrão de crescimento folicular em ondas está diretamente associado às flutuações nas concentrações plasmáticas de gonadotrofinas, controladas por sua vez pelo padrão de liberação de GnRH hipotalâmico. Desta forma, a inibição ou bloqueio da ação do GnRH suprime as etapas finais do crescimento folicular, resultando em anestro induzido (também chamado modelo waveless). Na medicina humana, agonistas ou antagonistas de GnRH são utilizados tanto no controle de disfunções ovarianas quanto na preparação de pacientes para procedimentos de reprodução assistida. Em bovinos, este efeito pode ser obtido pela imunização ativa contra GnRH, e mostrou-se estratégia viável, por exemplo, no controle de casos crônicos de doença ovariana cística em doadoras de oócitos. Contudo, é importante considerar a grande variação individual na resposta à imunização e consequente impossibilidade de controlar a duração do anestro induzido. O modelo waveless também é de grande utilidade na pesquisa, uma vez que elimina a potencial interferência do FSH e LH endógenos, aumentando a sensibilidade nos ensaios com hormônios exógenos e consequentemente reduzindo o número de réplicas necessárias nos estudos.(AU)

Biologia reprodutiva das abelhas africanizadas (Apis melífera) e biotecnologias aplicadas à sua conservação / Reproductive biology of Africanized bees (Apis mellifera) and biotechnologies applied to their conservation

A biodiversidade das abelhas africanizadas (Apis melífera) encontra-se sob grande ameaça e, neste cenário as investigações acerca da biologia reprodutiva, alguns aspectos ainda incipientes, bem como métodos aprimorados para a inseminação instrumental e criopreservação de gametas podem ser ferramentas preciosas para a conservação in situ e ex situ de subespécies e ecótipos. O entendimento e adoção de ferramentas como estas mencionadas, podem auxiliar na seleção de características de interesse para os criadores, assim como, nos esforços para a conservação de populações de abelhas ameaçadas.(AU)

The biodiversity of Africanized bees (Apis mellifera) is under great threat and, in this scenario, investigations about reproductive biology, some aspects are still incipient, as well as improved methods for instrumental insemination and cryopreservation of gametes can be precious tools for ex situ and in conservation. situ of subspecies and ecotypes can be contemplated. The understanding and adoption of tools such as those mentioned can help in the selection of characteristics of interest to breeders, as well as those committed to the conservation of endangered bee populations.(AU)

Embryo biopsies for genomic selection in tropical dairy cattle

Genomic selection has transformed the livestock industry, enabling early-life selection of animals. Biopsy sampling of pre-implantation embryos has been described since 1968. However, it was only after 2010, with the advancement of molecular biology techniques such as whole genomic amplification and SNP Chips, that next-generation sequencing became commercially available for bovine embryos. It is now possible to make decisions about which embryos to transfer not only based on recipients' availability or embryo morphology but also on genomic estimates. This technology can be implemented for a wide spectrum of applications in livestock. In this review, we discuss the use of embryo biopsy for genomic selection and share our experience with Gir and Girolando Brazilian breeding programs, as well as future goals for implementing it in Brazilian bovine in vitro embryo production practices.(AU)

Mecanismos epigenéticos nos espermatozoides e o impacto sobre a fertilidade masculina / Epigenetic mechanisms on sperm cells and the impact on male fertility

Por ser uma célula altamente especializada, o espermatozoide apresenta diferentes mecanismos epigenéticos, sendo os principais as metilações do DNA, o código de histonas, os ncRNAs (RNAs não codificadores), e a alta condensação da cromatina pela presença das protaminas. Estes mecanismos interagem entre si, contribuindo para a formação do epigenoma espermático, que modela a carga molecular espermática, que, por sua vez, pode impactar sobre as características do desenvolvimento embrionário e da progênie. Dessa forma, atualmente é consenso que o papel do espermatozoide ultrapassa a entrega de DNA de qualidade para o oócito no momento da fecundação. Pesquisas recentes de diversos grupos, incluindo o nosso, mostram que além da contribuição com DNA de qualidade, o espermatozoide entrega moléculas ao oócito no momento da fecundação que influenciam o desenvolvimento do embrião. Recentemente, essas moléculas de origem espermática (Em inglês: sperm-borne) também são associadas com alterações metabólicas e cognitivas da progênie. Embora ainda pouco se entenda como esses mecanismos podem persistir mesmo com o ciclo de reprogramação celular que ocorre logo após a fecundação, é evidente que estes podem impactar as características da progênie. Nesta revisão abordaremos sobre a modulação do epigenoma espermático e seus efeitos no desenvolvimento embrionário.(AU)

Since it is a highly specialized cell, the spermatozoa display different epigenetic mechanisms; the main ones are DNA methylation, histone code, ncRNAs (non-coding RNAs), and high chromatin condensation by the presence of protamines. These mechanisms act in synergy contributing to forming the sperm epigenome, which modulates the spermatic molecular cargo, and, may impact embryo and offspring development features. Thus, it is currently a consensus that the role of spermatozoa goes beyond delivering quality DNA to the oocyte at fertilization. Relevant findings from several research groups, including ours, have shown that sperm delivers several molecules to the oocyte at fertilization, beyond the contribution to DNA, which influences the development of the embryo. Recently, these sperm-borne molecules have also been associated with metabolic and cognitive changes in the offspring. Although the mechanism by which these changes can persist even after embryo reprogramming is not completely understood, evidence shows that sperm cell molecular content impacts embryo and offspring development. This review will mainly focus on the modulation of the sperm epigenome and its effects on embryo development.(AU)

Paternal effects on fetal programming

Paternal programming is the concept that the environmental signals from the sire's experiences leading up to mating can alter semen and ultimately affect the phenotype of resulting offspring. Potential mechanisms carrying the paternal effects to offspring can be associated with epigenetic signatures (DNA methylation, histone modification and non-coding RNAs), oxidative stress, cytokines, and the seminal microbiome. Several opportunities exist for sperm/semen to be influenced during development; these opportunities are within the testicle, the epididymis, or accessory sex glands. Epigenetic signatures of sperm can be impacted during the pre-natal and pre-pubertal periods, during sexual maturity and with advancing sire age. Sperm are susceptible to alterations as dictated by their developmental stage at the time of the perturbation, and sperm and seminal plasma likely have both dependent and independent effects on offspring. Research using rodent models has revealed that many factors including over/under nutrition, dietary fat, protein, and ingredient composition (e.g., macro- or micronutrients), stress, exercise, and exposure to drugs, alcohol, and endocrine disruptors all elicit paternal programming responses that are evident in offspring phenotype. Research using livestock species has also revealed that sire age, fertility level, plane of nutrition, and heat stress can induce alterations in the epigenetic, oxidative stress, cytokine, and microbiome profiles of sperm and/or seminal plasma. In addition, recent findings in pigs, sheep, and cattle have indicated programming effects in blastocysts post-fertilization with some continuing into post-natal life of the offspring. Our research group is focused on understanding the effects of common management scenarios of plane of nutrition and growth rates in bulls and rams on mechanisms resulting in paternal programming and subsequent offspring outcomes. Understanding the implication of paternal programming is imperative as short-term feeding and management decisions have the potential to impact productivity and profitability of our herds for generations to come.(AU)

Beneficial Impact of Hypercapnic Conditions During Early Incubation on Broiler Hatchability. Embryo Mortality and Postnatal Performance

We assessed the extent to which CO2 levels altered different hatching and chick parameters. In Experiments 1 and 2, a total of 16,184 eggs from Cobb 500 breeders were incubated in single stage incubators under three different conditions: (a) standard ventilated incubator (CON, Exp.1 and 2); (b) increasing CO2 levels during the first 10 days of incubation until 0.7% (V7000, Exp. 1) and (c) until 0.8% (V8000, Exp. 2). High levels of CO2 improved hatchability, possibly due to lower embryo mortality from ED18 to ED21. Internal and external pipping in experiment V8000 started later than in CON; nevertheless, the hatch still occurred before in V8000 as a result of the shorter durations of external pipping and hatch. In Experiment 3, a total of 12,138 eggs from Cobb 500 were incubated in single stage incubators under three different conditions: (a) standard ventilated incubator (CON); (b) increasing CO2 levels until 1.0% with ventilation (V10000); and (c) increasing CO2 levels until 1.0% without ventilation (NV10000). Hypercapnic conditions led to better hatchability and lower embryo mortality from ED18 to ED21. Internal pipping started earlier in NV10000, but only V10000 differed from CON in terms of the average time for hatch. Hypercapnic groups also showed shorter durations of external pipping and hatch when compared to CON. Post-hatch analysis revealed no differences among incubation conditions in terms of body weight gain, feed conversion ratio, mortality by sudden death syndrome, and production factor. Nevertheless, V10000 showed a lower mortality by ascites and a better viability when compared to CON, while NV10000 presented a higher mortality by other causes. Altogether, our findings indicate that in addition to not being detrimental to embryo survival, high CO2 levels reduce embryonic mortality at 18-21 days of incubation and increase hatchability.(AU)