Meiotic inhibition of bovine oocytes in medium supplemented with a serum replacer and hormones: effects on meiosis progression and developmental capacity

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Heat or cold chronic stress affects organ weights and Hsp70 levels in chicken embryos

This study was carried out with the objective of evaluating the effect of heat (38.8 degreesC) or cold (35.8 degreesC) stress on chicken embryo development and tissues Hsp70 levels, after the 13th day of incubation. Embryo weight (percent egg weight), organ weight (percent embryo weight) and Hsp70 levels (ng Hsp70 mug(-1) total protein) in different tissues (liver, breast muscle, heart, lungs, brain and kidney) were studied at the end of incubation. Cold stress induced a lower embryo weight and lower kidney and lungs weights, whereas heart and liver were lighter in heat-stressed embryos. An interaction between temperature and age was obtained only for Hsp70 levels in kidney and heart. Cold-stressed embryos showed higher Hsp70 levels in the brain, lungs and liver; a decrease in brain and breast muscle Hsp70 levels was seen from the 19th to 20th days in control embryos. Hsp70 levels increased with age in kidneys of control embryos and in heart of heat- and cold-stressed embryos. In conclusion, this study showed that chicken embryo organ weights are affected by incubation temperature, and that Hsp70 expression is tissue dependent (higher levels being seen in the brain) being cold-stress more effective in increasing Hsp70 levels in most studied tissues.

Desenvolvimento do Trato Gastrointestinal de Embriões Oriundos de Matrizes Pesadas de 30 e 60 Semanas de Idade

O objetivo deste trabalho foi investigar o efeito da idade das matrizes pesadas sobre o desenvolvimento do trato gastrointestinal (TGI) dos embriões no terço final do período de incubação, bem como a utilização das reservas do saco vitelino nas 24 h pós-eclosão, em pintos alimentados ou em jejum. Foram utilizados ovos férteis da linhagem Cobb 500, oriundos de matrizes pesadas com 30 e 60 semanas de idade. O desenvolvimento do TGI (proventrículo+moela, segmentos do intestino delgado e saco vitelino) foi estudado entre o 17º e 21º dias de incubação (Experimento 1). Nas 24 h pós-eclosão foi pesquisado o efeito da presença ou não de alimento no lúmen intestinal sobre a utilização das reservas do saco vitelino (Experimento 2). Os achados deste trabalho mostraram que, ao contrário do embrião, o desenvolvimento do intestino delgado e o peso do saco vitelino não sofreram influência da idade das matrizes. Na fase pós-eclosão, na ausência de alimento, o desenvolvimento do intestino delgado foi maior nas matrizes com 60 semanas, sendo dependente do crescimento do jejuno. A presença do alimento no lúmen teve influência na utilização das reservas do saco vitelino apenas nas matrizes com 30 semanas de idade. Os resultados deste experimento mostraram que a idade da matriz é importante fator no desenvolvimento do trato gastrointestinal do embrião, sendo fator relevante no crescimento pós-ecloão dos pintos.

Influence of egg pre-storage heating period and storage length on incubation results

This experiment aimed at evaluating the influence of different heating times of settable eggs of Cobb 500® broiler breeders before submitting them to different storage periods on egg weight loss, embryo mortality, and hatchability. A total number of 1,980 eggs were distributed in a completely randomized experimental design with a 3 x 3 factorial arrangement, comprising nine treatments with 22 replicates of 10 eggs each. The following factors were analyzed: pre-storage heating periods (0, 6, 12 hours at 36.92°C) and storage periods (4, 9, 14 days at 12.06°C). After storage, eggs were incubated under usual conditions, and were transferred to the hatcher at 442 hours of incubation. Eggs were weighed before heating, incubation, and transference to determine weight loss. Partial hatchability was determined at 480 hours, and total hatchability at 498 hours of incubation. Embryo mortality was determined in non-hatched eggs. It was concluded that heating eggs for six hour before storage improves incubation results as it decreases incubation length and late embryo mortality, therefore its use can be indicated in commercial operations. Storing eggs for 14 days and pre-heating for 14 days and pre-heating for 12 hours severely impair incubation results, and therefore are not recommended.

Influence of egg pre-storage heating period and storage length on the digestive tract of newly-hatched broiler chicks

An experiment was carried out to evaluate the effect of different heating times of settable eggs of Cobb 500® broiler breeders before submitting them to different storage periods on body weight, digestive tract organ weights, and intestinal mucosa morphology of newly-hatched chicks. Settable eggs were distributed in a completely randomized experimental design with a 3 x 3 factorial arrangement: pre-storage heating periods (0, 6, 12 hours at 36.92°C) and storage periods (4, 9, 14 days at 12.06°C). Body weight and relative weights of the yolk sac, heart, liver, proventriculus+gizzard, and intestinal segments were measured in chicks hatching at 480 and 498 hours of incubation. Villi height, width and perimeter, and crypt depth (ìm) were measured in duodenal histological sections. It was concluded that pre-storage heating for six hours of eggs stored for four or nine days increases small intestine weight of newly-hatched chicks, but does not influence the morphology of the duodenal mucosa. Pre-storage heating for 12 hours negatively influences body weight and duodenal mucosa development, and therefore this practice is not recommended. Storage length does not have consistent effect on body weight and development of the gastrointestinal tract.

Cultura de embrião e indução de brotos in vitro para micropropagação do pinhão-manso

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Sperm Organelle Morphologic Abnormalities: Contributing Factors and Effects on Intracytoplasmic Sperm Injection Cycles Outcomes

OBJECTIVE To (1) analyze possible relationships between motile sperm organelle morphology examination (MSOME) and sperm chromatin status, aneuploidy incidence, and patient's age; (2) determine the effects of sperm morphologic abnormalities on intracytoplasmic sperm injection (ICSI) outcomes; and (3) identify the benefits of intracytoplasmic morphologically selected sperm injection (IMSI) in patients with high DNA fragmentation rate.METHODS The study was performed in 50 patients undergoing ICSI cycles. The MSOME, sperm DNA fragmentation, and sperm aneuploidy incidence were performed in 200 sperm cells of each patient. Regression models were used to assess the relationships among sperm morphology and sperm aneuploidy, sperm DNA fragmentation, patient's age, and ICSI outcomes. In cycles with patients showing a high incidence of DNA fragmentation, oocytes were split into 2 groups according to the sperm selection method: Standard-ICSI (n = 82) and IMSI (n = 79). Fertilization and high-quality embryo rates were compared between the groups.RESULTS A close relationship between sperm DNA fragmentation and the presence of vacuoles in the MSOME was noted. The patient's age was correlated to the presence of vacuoles. No correlation between sperm aneuploidy and IMSI was observed. Vacuolated cells were negatively correlated with fertilization, pregnancy, and implantation. In patients with a high incidence of sperm DNA fragmentation, fertilization and high-quality embryo rates were similar when comparing IMSI and Standard-ICSI.CONCLUSIONS Our data demonstrate a correlation between paternal age and the incidence of nuclear vacuoles, as well as an effect of large and small vacuoles on late embryo development. UROLOGY 78: 786-791, 2011. (C) 2011 Elsevier B.V.

Efeito das células do cumulus e cisteamina durante o cultivo de maturação in vitro de oócitos bovinos sobre a maturação nuclear e aquisição da competência para desenvolvimento embrionário

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Targeted Nucleotide Exchange in Bovine Myostatin Gene

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Developmental anatomy and morphology of the ovule and seed of Heliconia (Heliconiaceae, Zingiberales)

The developmental anatomy and morphology of the ovule and seed in several species of Heliconia were investigated as part of an embryological study of the Heliconiaceae and to provide a better understanding of their relationships with the other families of the Zingiberales. Heliconia species have an ovule primordium with an outer integument of both dermal and subdermal origin. The archesporial cell is divided into a megasporocyte and a single parietal cell, which in turn are divided only anticlinally to form a single parietal layer, disintegrating later during gametogenesis. The embryo sac was fully developed prior to anthesis. In the developing seed, the endosperm was nuclear, with wall formation in the globular stage; a nucellar pad was observed during embryo development, but later became compressed. The ripe fruit contained seeds enveloped by a lignified endocarp that formed the pyrenes, with each pyrene having an operculum at the basal end; the embryo was considered to be differentiated. Most of these characteristics are shared with other Zingiberales, although the derivation of the operculum from the funicle and the formation of the main mechanical layer by the endocarp are unique to the Heliconiaceae.

Os histricomorfos sul-americanos: uma análise comparativa do desenvolvimento embriológico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Efeito da utilização de diferentes diluidores para a produção in vitro de embriões bovinos

Objetivou-se avaliar as características morfológica e funcional do sêmen bovino congelado comparando-se a eficácia de dois diferentes diluidores. O ejaculado de quatro touros foi dividido em duas partes iguais, uma submetida ao diluidor Tris e gema de ovo (A) e outra ao diluidor à base de lecitina de soja (Andromed®) (B). No experimento I, cinco palhetas dos diluidores A e B de cada touro foram descongeladas e avaliadas quanto à motilidade, vigor, concentração, morfologia espermática e teste de termor-resistência lento. Foram feitas, ainda, avaliação da integridade de membranas, por meio da associação das sondas iodeto de propídio, isotiocionato de fluoresceína - Pisum sativum e carbocianina catiônica lipofílica, e avaliação funcional da membrana plasmática com teste hiposmótico. A avaliação da integridade da cromatina foi realizada pelo método de coloração com laranja de acridina. No experimento II, o sêmen com os diferentes diluidores foi utilizado na fecundação in vitro, sendo observadas taxas de clivagem e desenvolvimento embrionário in vitro. em relação aos resultados obtidos, apenas a porcentagem de espermatozoides no sêmen congelado foi discretamente maior com o diluidor A, concluindo-se que o diluidor composto por lecitina de soja pode substituir o composto por Tris e gema de ovo, respeitando-se as variações individuais de cada touro utilizado no presente experimento.

Aplicação da ultrassonografia colorida doppler em programas de transferência de embriões equinos

Os programas comerciais de transferência de embriões em éguas existem por mais de três décadas e são hoje uma das biotécnias mais utilizadas na reprodução assistida de equinos. O exame ultrassonográfico nos períodos pré e pós-cobertura de doadoras de embriões, assim como a avaliação de receptoras no momento da inovulação é de vital importância para o êxito de um programa de transferência de embriões. A ultrassonografia Doppler é uma técnica não-invasiva que permite a avaliação em tempo real da hemodinâmica do trato reprodutivo de animais de grande porte. Por fornecer detalhes anatômicos e informações imediatas sobre a fisiologia do fluxo sanguíneo de tecidos e órgãos, o exame doppler permite a avaliação do potencial ovulatório de folículos e do status funcional de corpo lúteo e útero, além de ser uma técnica auxiliar para o diagnóstico de distúrbios na hemodinâmica do sistema reprodutivo. Alterações na perfusão sanguínea dos futuros folículos dominantes são detectáveis em diferentes fases de seu desenvolvimento, como na divergência folicular e no período pré-ovulatório. A partir dessas informações, é possível determinar o princípio da atividade sexual e o momento ideal para o início de tratamentos superovulatórios e indutores de ovulação, assim como o momento mais apropriado para a realização de coberturas de éguas doadoras de embriões. A avaliação através do modo-Doppler do corpo lúteo e útero de éguas receptoras é também um instrumento auxiliar para a seleção de animais com perfil sérico de progesterona e ambiente uterino adequados para a sobrevivência do embrião e manutenção da gestação. Essa técnica pode ser útil ainda ao se avaliar a interação concepto-maternal. Apesar da aplicabilidade da tecnologia doppler dentro de programas de transferência de embriões, novos estudos visando a determinar padrões de normalidade e posterior caracterização de distúrbios de fluxo sanguíneo de trato reprodutivo ainda se fazem necessários.

Fecundity in Portunus spinimanus Latreille, 1819 (Crustacea, Brachyura, Portunidae) from Ubatuba, São Paulo, Brazil.

The object of this work was to study the conditions of embryo development and egg production in Portunus spinimanus Latreille, 1819 at Ubatuba region of Brazil. The number of eggs laid by 21 females in a range of size was determined. Eggs at 3 differ ent stages of embryonic development were measured P. spinimanus fecundity varied between 188,065 and 682,992, with an average of 429,676 +/- 157,203, for animals with a carapace width ranging from 56.2 to 86.6 mm. Between the initial and final stages of development there was an increase of 55.1% in egg diameter. The relatively high fecundity of P. spinimanus may be a strategy of the species faced with a high mortality rate in the larval phase; however this hypothesis should be tested.

Influence of sire breed (Bos indicus versus Bos taurus) and interval from slaughter to oocyte aspiration on heat stress tolerance of in vitro-produced bovine embryos

Based on in vitro experiments, Bos indicus embryos were more resistant to heat stress (HS) than Bos taurus embryos. To increase knowledge regarding differences between Bos indicus and Bos taurus in resistance to HS, the primary objective of this study was to determine if tolerance to HS is due to the breed, origin of the oocyte, sperm, or both. Additionally, the influence of the interval between ovary acquisition (in the abattoir) and oocyte aspiration in the laboratory, on early embryo development was ascertained. Oocytes were collected from Nelore and Holstein cows in an abattoir; 4.0 or 6.5 h later, oocytes were aspired in the laboratory, and then matured and fertilized using semen from Nelore (N), Gir (GIR), or Holstein (H) bulls. Ninety-six h post insemination (hpi), embryos with >= 16 cells were divided in two groups: control and HS. In the control group, embryos were cultured at 39 degrees C, whereas in the HS group, embryos were subjected to 41 degrees C for 12 h, and then returned to 39 degrees C. Rates of cleavage, and formation of morula and blastocysts were higher (P < 0.05) for oocytes aspirated at 4.0 versus 6.5 h after ovaries were acquired. Heat stress decreased rates of blastocyst formation for all breeds (N X N; H x H; and H X GIR) and in both time intervals (4.0 and 6.5 h). However, N X N had higher cleavage rate (P < 0.05) in both time intervals when compared with H X H and H X GIR. In addition, Nelore oocytes fertilized with Nelore semen (N X N) had higher blastocyst yields (P < 0.05) in the control and HS group, when compared with the other two breeds (H X H and H X GIR). We concluded that the breed of origin of the oocyte was more important than that of the sperm for development of thermotolerance, because bull breed did not influence embryo development after HS, and in vitro early embryonic development was impaired by increasing (from 4 to 6.5 h) the interval between ovary acquisition and oocyte aspiration. (C) 2011 Elsevier B.V. All rights reserved.