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.

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)

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)

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)

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)

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)

Taxa de recuperação embrionária em éguas Mangalarga Marchador inseminadas com sêmen refrigerado / Embryonic recovery rate in Mangalarga Marchador mares inseminated with refrigerated semen

O acompanhamento reprodutivo da espécie equina tem sido cada vez mais aplicado na aplicade de diversos países, resultando em melhores índices reprodutivos para a espécie e em animais de linhagens selecionadas. Nos programas de transferência de embrião, a taxa de recuperação embrionária consiste em um parâmetro essencial a fim de verificar o sucesso da técnica. Por conseguinte, este trabalho propõe-se a avaliar as taxas de recuperação embrionária de éguas doadoras submetidas à inseminação artificial com sêmen refrigerado de maneira a complementar os dados relacionados ao assunto. Conforme os dados coletados durante a estação de monta de agosto de 2018 a fevereiro de 2019, obteve-se 129 lavados uterinos provenientes de 20 éguas e 83 embriões recuperados. Desse modo, a taxa de recuperação embrionária obtida foi de 64,34%. Diante dos dados apresentados neste estudo, conclui-se que a aplicabilidade da técnica de recuperação embrionária visando o uso do sêmen refrigerado na inseminação artificial de éguas se mostra promissora, pois permite o transporte de material genético de alto valor entre propriedades sem interferir drasticamente nos índices dos programas de reprodução.

The reproductive monitoring of the equine species has been increasingly applied in several countries, resulting in better reproductive rates for the species and in animals from selected strains. In embryo transfer programs, the embryo recovery rate is an essential parameter to verify the technique success. Therefore, the present work proposes to evaluate the embryo recovery rates of donor mares submitted to artificial insemination with refrigerated semen to complement the data related to the subject. According to the data collected during the breeding season for the period from August 2018 to February 2019, 129 uterine washes were obtained from 20 mares and 83 recovered embryos. Thus, the embryonic recovery rate obtained was 64.34%. Given the data presented in this study, it is concluded that the applicability of the embryonic recovery technique aiming at the use of refrigerated semen in artificial insemination of mares is promising, as it allows the transport of high-value genetic material between properties without interfering dramatically in breeding program indexes.

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)

Estudos para o incremento da produção de embriões in vivo e in vitro em vacas de corte taurinas e sintéticas / Studies to improve in vivo and in vitro embryo production in beef cows from taurine and synthetic breeds

O Brasil destaca-se por ser um dos líderes mundiais na produção in vitro de embriões, o que é decorrente do rebanho nacional ser predominantemente composto por raças zebuínas, que possuem maior número de folículos antrais aspiráveis. Por outro lado, vacas taurinas apresentam menor população folicular antral, o que limita o número de ovócitos obtidos por seção de aspiração folicular (ovum pick-up; OPU). Portanto, na região Sul do Brasil há demanda para produção tanto in vivo como in vitro de embriões de doadoras de raças taurinas e sintéticas. Este texto discute estudos recentes que buscam estratégias para aperfeiçoar as etapas envolvidas na produção de embriões bovinos, bem como para aumentar o aproveitamento de receptoras utilizadas nos programas de transferência de embriões (TE), com foco em vacas taurinas e sintéticas.(AU)

Brazil stands among the leaders on in vitro embryo production, as the Brazilian herd is predominantly composed by Zebu breeds, which have a greater number of antral follicles available to ovum pick-up (OPU). On the other hand, taurine cows have a lower antral follicle population, which limits the number of oocytes obtained by each OPU section. Therefore, in the Brazilian Southern region there is a demand for both in vivo and in vitro production of embryos from donors of taurine and synthetic breeds. This text discusses recent studies that seek strategies to improve the steps involved in the production of bovine embryos, as well as to increase the use of recipients in embryo transfer (ET) programs, focusing on taurine and synthetic cows.(AU)

Prostaglandin F2α treatment concurrent with artificial insemination does not affect bovine embryo production / Administração de prostaglandina F2 alfa no momento da inseminação artificial não afeta a produção de embriões bovinos

Treatment with prostaglandin F2α (PGF) induces ovulation and increases conception rates in cows, while improving embryo production in buffalos. However, its effect on superovulated cows is unknown. This study verified whether single PGF administration concurrent with artificial insemination (AI) improves fertilization and embryo production rates in superovulated cows. In each replicate, embryo donor cows were equally allocated to two groups: the untreated control and PGF groups. The latter of which received 482 µg of cloprostenol concurrent with the first AI. Each cow (n = 35) was subjected to two superovulations (SOV) in a crossover design (total = 70 embryo collections). In the control and PGF groups, respectively, the observed responses were [median (95% CI)]: 12 (10-18) and 15 (12-18) total structures, 9 (7-11) and 7 (6-10) viable embryos, 1 (0-1) and 1 (1-3) degenerated embryos, and 1 (0-3) and 2 (0-5) oocytes (P > 0.05). In conclusion, single PGF treatment concurrent with the first AI did not affect embryo production in superovulated cows.

A prostaglandina F2α (PGF) pode induzir a ovulação e melhorar tanto a concepção em vacas, como a produção de embriões em búfalas, mas o efeito em vacas superovuladas é desconhecido. Esse estudo teve como objetivo verificar se a administração de uma dose de PGF na inseminação artificial (IA) após a superovulação (SOV) melhora as taxas de fecundação e produção embrionária em vacas. Em cada replicação, vacas doadoras de embriões foram equilibradamente alocadas em dois grupos: controle, não tratado, ou PGF, que recebeu 482 µg de cloprostenol no momento da primeira IA. Cada doadora (n = 35) foi submetida a duas SOV em um delineamento crossover (total = 70 coletas de embriões). Nos grupos controle e PGF, respectivamente, foram observados [medianas (IC 95%)]: 12 (10-18) e 15 (12-18) estruturas totais; 9 (7-11) e 7 (6-10) embriões viáveis; 1 (0-1) e 1 (1-3) embriões degenerados; e 1 (0-3) e 2 (0-5) ovócitos (P > 0,05). Conclui-se que uma única administração de PGF no momento da primeira IA não afeta a produção embrionária de vacas superovuladas.

Assessing Differences in the Quality Properties and Ultrastructure of Eggshell as Affected by Chicken Strain and Flock Age During Incubation Period

Three hundred hatching eggs from two layer breeder flocks (Hy-Line Brown and hy-Line W-36; 150 each) were obtained at three different periods of laying cycle (early, middle and late stages). The hatching eggs were randomly taken to assess the impact of chicken strain, age, and hatching time on some eggshell measurements and structures. Results indicate that egg weight, shell percentage and shell thickness were significantly affected by strain, breeder age, and incubation time. Increasing rate of calcium absorption and utilization by the embryos during the embryonic stage clearly reflects the decrease occurred in eggshell strength. The differences of eggshell breaking strength during incubation time periods were 0.41 and 0.55 kg/cm2, the differences due to the incubation time may be due to the higher demand for calcium utilization in the late incubation period (18 days), observed on the10th day of incubation. Brown eggs recorded significant higher calcium percentage in eggshell compared to the white ones. The brown breeder hens recorded significant higher total pores per egg compared to the white egg. Concerning eggshell ultrastructure, total thickness of the brown eggshell was significantly higher than those of the white eggshell by about 3.7%. All parameters, except for total thickness, were significantly affected by the interaction between strain and breeder age. Finally, we conclude that some traits including egg weight, shell percentage, shell thickness, calcium percentages in eggshell, and total pores per egg were significantly affected by strain, breeder age, and incubation time. Eggshell ultrastructure of brown egg was better compared to the white ones.(AU)

Reproductive and economic evaluation of sexual stimulation during the anestrous period in a commercial farm with neonatal lamb losses / Avaliação reprodutiva e econômica da estimulação sexual durante o período antrópico em uma fazenda comercial com perdas neonatais de cordeiros

This study was performed during the anestrous, involving 140 Akkaraman Kangal ewes whose lambs had died in the neonatal stage due to pneumonia and enteritis. Intravaginal sponge containing progesterone was placed to the animals (Group 1, n = 70) on day 0 and removed after 7 days, following which 263 µg PGF2α and 500 IU eCG were administered to the sheep. Ram introduction was performed for 7 days (days 8-14), starting from the day after the removal of the intravaginal sponge (day 8). The animals in Group 2 (n = 70) were not exposed to any treatment. Ram introduction was performed simultaneously in both the groups. To determine the reproductive response, reproductive parameters such as estrous, pregnancy, multiple pregnancy, and embryonic mortality rates, number of births, number of offspring, and fertility, as well as their economic implications, were compared between groups. Each reproductive parameter exhibited a statistical difference between groups. An economically positive trend was observed in the study group compared with the control group. It was concluded that in case of lamb losses in commercial farms that derive profit from lambing, pregnancy of ewes can be achieved via sexual stimulation without waiting for the next breeding season.

Este estudo foi realizado durante o anestrous, envolvendo 140 ovelhas Akkaraman Kangal cujos cordeiros haviam morrido no estágio neonatal devido a pneumonia e enterite. A esponja intragaginal contendo progesterona foi colocada aos animais (Grupo 1, n = 70) no dia 0 e removida após 7 dias, após os quais 263 µg PGF2α e 500 UI eCG foram administrados às ovelhas. A introdução do carneiro foi realizada por 7 dias (dias 8-14), a partir do dia seguinte à remoção da esponja intravaginal (dia 8). Os animais do Grupo 2 (n = 70) não foram expostos a nenhum tratamento. A introdução do carneiro foi realizada simultaneamente em ambos os grupos. Para determinar a resposta reprodutiva, foram comparados entre os grupos parâmetros reprodutivos, tais como estrogênio, gravidez, gravidez múltipla e taxas de mortalidade embrionária, número de nascimentos, número de descendentes e fertilidade, bem como suas implicações econômicas. Cada parâmetro reprodutivo exibia uma diferença estatística entre os grupos. Uma tendência economicamente positiva foi observada no grupo de estudo em comparação com o grupo de controle. Concluiu-se que no caso de perdas de cordeiros em fazendas comerciais que obtêm lucros com a parição, a gravidez de ovelhas pode ser obtida através de estimulação sexual sem esperar pela próxima estação de reprodução.

Relationships of morphological and phototextural attributes of presumptive ovine zygotes and early embryos to their developmental competence in vitro: a preliminary assessment using timelapse imaging

The assessment of morphology and digital image opacity may provide valuable information on the present embryo quality. Time-lapse imaging has been employed in research to establish a means of monitoring the dynamic nature of preimplantation embryo development. The aim of present study was to use time-lapse imaging for assessing various prospective morphometric and phototextural markers of the developmental potential of in vitro-derived ovine embryos. Oocytes were obtained by scarification of ovaries from nine Polish Longwool ewes. After in vitro maturation (IVM) and fertilization (IVF) of oocytes with fresh ram semen, the development of embryos to the blastocyst stage was monitored and evaluated using Primo Vision time-lapse imaging technology. Commercially available Image-Pro® Plus software was used to measure zona pellucida thickness, embryo diameter, total area of the perivitelline space, cellular grey-scale pixel intensity and cellular pixel heterogeneity. Statistical assessment of all attributes was done at various time points during embryo development (i.e., presumptive zygote stage: t(0); first cleavage detected at t(2) or t(3); and second cleavage detected at t(4) or t(6)). Out of thirty-seven zygotes analyzed in this study, five did not divide, 26 arrested before and six developed to the blastocyst stage. Our present results indicate that most parameters analyzed did not differ among embryos varying in their developmental fate except for the perivitelline space area that was greater (P<0.05) for non-dividing zygotes than future blastocysts at the presumptive zygote stage (4040±1850 vs. 857±262 µm2, respectively; means±SEM). Consequently, the measurement of perivitelline space at t(0) can potentially be used to prognosticate developmental potential of in vitro-produced ovine embryos albeit further confirmational studies are needed.(AU)

Effect of the presence and location of corpus luteum on competence of bovine cumulus-oocyte complexes

This study aimed to determine the effect of presence of the corpus luteum (CL) and its influence on cumulus­oocyte complexes (COCs) obtained from the ipsilateral or contralateral ovary in bovine on the recovery and capacity of the oocytes to sustain mono-spermic fertilization, undergo preimplantation development, and develop to the blastocyst stage. Ovaries were collected at a local slaughterhouse and kept in pairs corresponding to the same animal. In the first experiment the variables evaluated were compared between cows with (CCL+) and without (CCL- ) CL, and for the second experiment, comparisons were made between ovaries with an ipsilateral (CL+), contralateral (CL−), and no (NCL). The recovery rate of COCs was higher in ovaries from CCL− cows, and a higher proportion of grade 1 COCs were recovered from this group. A higher proportion of metaphase I oocytes at 7 h of maturation, and a higher rate of cleavage were observed in the CCL+ group; however, a higher proportion of embryos were obtained from the CCL− group. Besides, COCs from the CL+ group had a lower proportion of grades 1 and 2 morphological qualities, lower rate of metaphase II oocytes at 22 h of maturation, and lower rate of formation of two pronuclei, whereas a higher proportion of unfertilized oocytes after in vitro fertilization. On the other hand, the COCs from the CL− group displayed a lower proportion of oocytes with more than two pronuclei, higher cleavage rate, and higher final blastocyst production were obtained when compared to CL+. Thus, the effects of CL on the competence of bovine COCs are different depending on the anatomical proximity of their location in the animal, negatively affecting the quality of COCs located in the same ovary, but not having negative effects on the competence of COCs in the ovaries contralateral to their location.(AU)

Larval development of Characidium orientale (Actinopterygii: Crenuchidae) a small Neotropical fish

ABSTRACT The Characidium orientale Buckup & Reis, 1997 larvae development is described using specimens collected in the natural environment of Antas River, in the Taquari-Antas river basin, Rio Grande do Sul, Brazil. Were considerate ontogenetic changes in morphology, pigmentation, fin morphology, and meristic characters. Thirty-six larvae (33 in the preflexion stage, two in flexion, and one in the postflexion stage), with a standard length between 4.24 and 11.26 mm were analyzed. The larvae of C. orientale are altricial and present fusiform body, subterminal mouth, long intestine with anal opening posterior to the vertical line over the median region of the body. Two chromatophores are evident in the interorbital region, as well pigments concentrated on the side of the body, forming a longitudinal stripe, and between the rays of the caudal fin, which intensify during the development. Also, a conspicuous remnant of the embryonic membrane (finfold) in front of the dorsal fin appears to be characteristic of species of the genus during larval development. The total number of myomeres varies from 32 to 35 (18-23 pre; 11-16 postanal). The main changes in morphometry occur during the preflexion stage and suggest physiological and behavioral changes.

RESUMO O desenvolvimento das larvas de Characidium orientale Buckup & Reis, 1997 é descrito a partir de espécimes coletados no rio Antas, na bacia do rio Taquari-Antas, Rio Grande do Sul, Brasil. Foram consideradas mudanças ontogenéticas na morfologia, pigmentação, morfologia das nadadeiras e caracteres merísticos. Foram analisadas 36 larvas (33 na fase de pré-flexão, duas em flexão e uma em pós-flexão), com comprimento padrão entre 4,24 e 11,26 mm. As larvas de C. orientale são altriciais e apresentam corpo fusiforme, boca subterminal, intestino longo com abertura anal posterior à linha vertical sobre a região mediana do corpo. Dois cromatóforos são evidentes na região interorbital, bem como pigmentos concentrados na lateral do corpo, formando uma faixa longitudinal e entre os raios da nadadeira caudal, que se intensificam durante o desenvolvimento. Além disso, um remanescente conspícuo da membrana embrionária na frente da nadadeira dorsal parece ser característico de espécies do gênero durante o desenvolvimento larval. O número total de miômeros varia de 32 a 35 (18-23 pré; 11-16 pós-anal). As principais mudanças na morfometria ocorrem durante o estágio de pré-flexão e sugerem mudanças fisiológicas e comportamentais.

Larval development of Characidium orientale (Actinopterygii: Crenuchidae) a small neotropical fish / Desenvolvimento larval de Characidium orientale (Actinopterygii: Crenuchidae)

The Characidium orientale Buckup & Reis, 1997 larvae development is described using specimens collected in the natural environment of Antas River, in the Taquari-Antas river basin, Rio Grande do Sul, Brazil. Were considerate ontogenetic changes in morphology, pigmentation, fin morphology, and meristic characters. Thirty-six larvae (33 in the preflexion stage, two in flexion, and one in the postflexion stage), with a standard length between 4.24 and 11.26 mm were analyzed. The larvae of C. orientale are altricial and present fusiform body, subterminal mouth, long intestine with anal opening posterior to the vertical line over the median region of the body. Two chromatophores are evident in the interorbital region, as well pigments concentrated on the side of the body, forming a longitudinal stripe, and between the rays of the caudal fin, which intensify during the development. Also, a conspicuous remnant of the embryonic membrane (finfold) in front of the dorsal fin appears to be characteristic of species of the genus during larval development. The total number of myomeres varies from 32 to 35 (18-23 pre; 11-16 postanal). The main changes in morphometry occur during the preflexion stage and suggest physiological and behavioral changes.

O desenvolvimento das larvas de Characidium orientale Buckup & Reis, 1997 é descrito a partir de espécimes coletados no rio Antas, na bacia do rio Taquari-Antas, Rio Grande do Sul, Brasil. Foram consideradas mudanças ontogenéticas na morfologia, pigmentação, morfologia das nadadeiras e caracteres merísticos. Foram analisadas 36 larvas (33 na fase de pré-flexão, duas em flexão e uma em pós-flexão), com comprimento padrão entre 4,24 e 11,26 mm. As larvas de C. orientale são altriciais e apresentam corpo fusiforme, boca subterminal, intestino longo com abertura anal posterior à linha vertical sobre a região mediana do corpo. Dois cromatóforos são evidentes na região interorbital, bem como pigmentos concentrados na lateral do corpo, formando uma faixa longitudinal e entre os raios da nadadeira caudal, que se intensificam durante o desenvolvimento. Além disso, um remanescente conspícuo da membrana embrionária na frente da nadadeira dorsal parece ser característico de espécies do gênero durante o desenvolvimento larval. O número total de miômeros varia de 32 a 35 (18-23 pré; 11-16 pós-anal). As principais mudanças na morfometria ocorrem durante o estágio de pré-flexão e sugerem mudanças fisiológicas e comportamentais.

Occurrence of Hymenolepis diminuta: a potential helminth of zoonotic importance in murid rodents

Abstract The study reveals the prevalence of a potential rodent-borne zoonotic helminth species, Hymenolepis diminuta in commensal rodents caught from irrigated and rain-fed areas of Swat, Pakistan. Three hundred and fifty rodents (269 rats and 81 mice) trapped during vegetative, flowering/fruiting and mature/harvesting stages of crops were studied from 2011-2013. Hymenolepisdiminuta eggs were identified on the basis of their shape, size, colour and markings on the surface of the egg shell and three pairs of embryonic hook-lets. Overall prevalence of H. diminuta was 3.14% (n=11/350). The highest prevalence 3.49% (n=5/143) of H. diminuta was noted at harvesting stages of the crops whereas the lowest 2.59% (n=2/77) during vegetative stage. Infection was higher in males 3.25% (n=7/215) than females 2.96% (n=4/135). Adult rodents were highly infected while no sub-adult was found infected. Infection was higher in mice 3.70% (3/81) than rats 2.69% (8/269) while no significance (p=1.0000:0.1250 to 32.00 CI). Rats and mice appears to show the most suitable reservoirs by hosting H. diminuta a zoonotic helminth. The presence of these rodents in all possible habitats can act as a main channel of transferring parasites through various habitats and can pose a hazard to humans in the area.

Resumo O estudo revela a prevalência de uma espécie potencial de helmintos zoonóticos transmitidos por roedores, Hymenolepis diminuta, em roedores comensais capturados em áreas irrigadas e alimentadas pela chuva de Swat, Paquistão. Trezentos e cinquenta roedores (269 ratos e 81 camundongos) presos durante os estágios vegetativo, floração/frutificação e maturidade/colheita das safras foram estudados de 2011-2013. Ovos de Hymenolepisdiminuta foram identificados com base em sua forma, tamanho, cor e marcações na superfície da casca do ovo e três pares de anzóis embrionários. A prevalência geral de H. diminuta foi de 3,14% (n = 11/350). A maior prevalência 3,49% (n = 5/143) de H. diminuta foi observada na fase de colheita das lavouras, enquanto a menor 2,59% (n = 2/77) durante a fase vegetativa. A infecção foi maior nos homens 3,25% (n = 7/215) do que nas mulheres 2,96% (n = 4/135). Roedores adultos foram altamente infectados, enquanto nenhum subadulto foi encontrado infectado. A infecção foi maior em camundongos 3,70% (3/81) do que em ratos 2,69% (8/269), embora sem significância (p = 1,0000: 0,1250 a 32,00 CI). Ratos e camundongos parecem mostrar os reservatórios mais adequados hospedando H. diminuta um helmintos zoonótico. A presença desses roedores em todos os habitats possíveis pode atuar como um canal principal de transferência de parasitas através de vários habitats e pode representar um perigo para os humanos na área.

Biomorphometry of Automeris liberia Cramer (Lepidoptera: Saturniidae) in oil palm in the Amazon / Biomorfometria de Automeris liberia Cramer (Lepidoptera: Saturniidae) em palma de óleo, Amazônia Oriental

Automeris liberia Cramer are found in South America, with documented sightings in Ecuador, Peru, some regions of Mexico and, more recently, the Brazilian Amazon. These moths cause damage to several plant species. We designed an experiment to study the biology and morphometry of A. liberia in the laboratory and provide information for the management of the pest in oil palm cultivation in the Brazilian Amazon. Caterpillars for rearing were collected from commercial areas of oil palm cultivation, where they cause defoliation of the plant. They were fed a natural diet of oil palm leaves of the Tenera variety and observed from second generation onwards. Sex, behavior, posture, mass, body length, wingspan, body diameter, sex ratio, average growth ratio, and antenna were quantified. The embryonic stage lasted 14 days, followed by seven larval instar stages over 36 days. The pupal stage lasted 21 days. Adults had a longevity of 4.5 and 6 days and a total biological cycle of 78.5 and 80 days, for males and females, respectively. The growth ratio was 1.49. The antennae showed sexual dimorphism, with bipectinate morphology in males and filiform in females. In the adult stage, morphological variables were evaluated (body length, thoracic diameter, wingspan, forewing length, forewing height, hindwing length, hindwing height, antenna length, number of antennomeres, and body mass), and they showed significant differences between males and females (Newman-Keuls test, P < 0.05).(AU)

A espécie Automeris liberia Cramer tem ocorrência na América do Sul, com registro no Equador, Peru, algumas regiões no México e, mais recentemente, na Amazônia brasileira, causando danos a várias espécies de plantas. Objetivou-se estudar pioneiramente a biologia e a morfometria de A. liberia em laboratório, visando subsidiar informações para o manejo da praga na cultura da palma de óleo, na Amazônia brasileira. A criação, foi oriunda de lagartas coletadas em áreas comerciais de palma de óleo, onde as quais estavam causando desfolhamentos. Foram alimentadas com dieta natural a base de folhas de palma de óleo da var. Tenera. As observações foram iniciadas a partir da segunda geração. Foram avaliados sexagem, comportamento, posturas, massa, comprimento, envergadura, diâmetro do corpo, razão sexual, razão da média de crescimento e antena. O estágio embrionário durou 14 dias e o larval apresentou sete instares em 36 dias. O estágio pupal foi de 21 dias. Os adultos apresentaram longevidade de 4,5 e 6 dias e ciclo biológico total de 78,5 e 80 dias, para machos e fêmeas, respectivamente. A razão de crescimento foi de 1,49. As antenas apresentaram dimorfismo sexual, sendo do macho tipo bipectinada e da fêmea filiforme. Na fase adulta foram avaliadas variáveis morfológicas (comprimento do corpo, diâmetro do tórax, envergadura, comprimento da asa anterior, altura da asa anterior, comprimento da asa posterior, altura da asa posterior, comprimento da antena, número de antenômeros e massa corpórea) entre machos e fêmeas, as quais apresentaram diferença estatística pelo teste Newman-Keuls (P < 0,05).(AU)

Uso de éguas receptoras acíclicas em programas de transferência de embrião / Anovulatory recipient mares in embryo transfer programs

O maior entrave para o êxito de um programa de transferência de embrião (TE) está relacionado às receptoras, tanto a qualidade de características reprodutivas, quanto a sincronização entre doadoras e receptoras. Éguas doadoras, geralmente, apresentam melhor escore corporal e disponibilidade alimentar nos períodos de seca, fazendo com que iniciem o seu ciclo estral precocemente enquanto éguas receptoras se encontram ainda em anestro ou período transicional, o que dificulta a TE. Vários estudos foram realizados com o objetivo de utilizar receptoras acíclicas como receptoras de embrião. Assim, podem ser utilizadas nos programas de TE quando submetidas a protocolos hormonais que mimetizam o ciclo estral natural. Uma diversidade de protocolos é encontrada na literatura, usando diferentes doses, posologias e intervalos do uso de estradiol associado a progestágenos. Portanto, objetivou-se analisar resultados de pesquisas com protocolos hormonais que poderiam ajudar a solucionar a escassez de receptoras cíclicas e maximizar os resultados da biotécnica de transferência de embriões.(AU)

There are obstacles to the success of equine embryo transfer programs, concomitantly there has been an increase in the popularity of the technique. One of the most urgent issues to overcome is related to the recipient mare availability and the donor-recipient synchrony. Usually, the donor mares have a higher body condition score and are fed a higher quality diet than recipient mares during the non-breeding season. Therefore, the donor mares start cycling earlier than recipient mares, which makes embryo transfer challenging. Several studies have been carried on finding hormonal protocols that mimic the natural oestrous cycle to allow the use of noncycling recipient mares for embryo transfer programs. Many protocols can be found in the literature, using different doses and intervals. Among the main difference is the use of progestogens, which can be used alone or in combination with oestradiol. The aim of this review was to analyse results that could help solve the lack of recipient mares and improve the results of the embryo transfer technique.(AU)

Use of melatonin as an inhibitor of apoptotic process for cryopreservation of zebrafish (Danio rerio) embryos / Uso da melatonina como inibidor do processo apoptotico para a criopreservação de embriões de zebrafish (Danio rerio)

This study investigated the use of melatonin to arrest the effects of apoptosis in vitrified zebrafish (D. rerio) embryos. Dechorionated embryos at 22-24 somite-stage were divided (n = 60/treatment) into a non-vitrified (Control Group, 0 M melatonin) and vitrified treatments with 0 M (T1), 1 µM (T2) and 1 mM of melatonin (T3). For vitrified treatments, a solution methanol/propylene glycol based was used and the embryos stored in -196 °C for a week. After thaw, survival rate, scanning electron microscopy, expression of anti (bcl-2) and pro-apoptotic (bax/caspase-3) genes, reactive oxygen species (ROS) formation and DNA fragmentation analyses were performed. No live embryos were obtained from vitrified treatments, observing a rapid degeneration immediately after thawing, with the vitelline layer rupture and leakage of its content, followed by breakdown of epithelial cells and melanisation of the tissue. Regarding the apoptotic process, T3 had the highest relative gene expression, for the three genes (P 0.05). The inclusion of 1 µM of melatonin in the vitrifying solution, countered the effects of apoptotic process in post-thaw embryos, suggesting its utility in cryopreserving fish embryos.

Este estudo investigou o uso da melatonina para conter os efeitos da apoptose em embriões vitrificados de zebrafish (D. rerio). Embriões descorionados no estágio de 22-24 somitos foram divididos (n = 60 / tratamento) em tratamento não vitrificado (Grupo Controle, melatonina 0 M) e tratamentos vitrificados com 0 M (T1), 1 µM (T2) e 1 mM de melatonina (T3). Para os tratamentos vitrificados, utilizou-se uma solução à base de metanol/propilenoglicol e os embriões foram armazenados em -196 °C por uma semana. Após o descongelamento, foram realizadas análises de taxa de sobrevivência, microscopia eletrônica de varredura, expressão dos genes anti (bcl-2) e pró-apoptóticos (bax/caspase-3), formação de espécies reativas de oxigênio (EROS) e análises de fragmentação de DNA. Não foram obtidos embriões vivos a partir dos tratamentos vitrificados, observando uma rápida degeneração imediatamente após o descongelamento, com ruptura da camada vitelina e vazamento de seu conteúdo, seguida de quebra das células epiteliais e melanização do tecido. Em relação ao processo apoptótico. T3 apresentou expressão gênica relativa alta para os três genes (P 0,05). A inclusão de 1 µM de melatonina na solução de vitrificação, contrariou os efeitos do processo apoptótico em embriões pós-descongelamento, sugerindo sua utilidade na criopreservação de embriões de peixes.