RESUMO
In this study, we investigated the effects of melatonin supplementation in the culture medium and blastocoel fluid removal (BFR) before vitrification on the quality and viability of in vitro-derived bovine embryos. After fertilization, presumptive zygotes were assigned to one of the following treatments: control, in vitro standard culture (IVC) medium; IVC + M10-9, IVC medium supplemented 10-9 M melatonin; or IVC + M10-9 BFR, IVC medium supplemented with 10-9 M melatonin plus BFR on day 7 (D7) of culture. D7 blastocysts were vitrified by the Cryotop method and, after 5 mo of storage, were warmed and incubated for an additional 72 h. The re-expansion rate was evaluated after 2 and 24 h, and the hatching rate was evaluated after 24, 48, and 72 h. At 72 h, the total number of cells (TNC); number of apoptotic cells (NAC); and expression of genes related to oxidative stress (HSPA5), cell metabolism (SLC2A3), cell repair (MSH6), placentation (KRT8 and PLAC8), and implantation (FOSL1) were assessed in the blastocysts. Less than 30% of the control blastocysts re-expanded until 2 h, whereas more than 85% of the IVC + M10-9 and IVC + M10-9 BFR blastocysts re-expanded (P < 0.05). The hatching rate of IVC + M10-9 BFR blastocysts increased at all time points (P < 0.05), reaching 66.8% at 72 h of incubation. The TNC was similar among treatments (P > 0.05), regardless of vitrification/warming and re-cultivation. The NAC:TNC was smaller for melatonin-treated blastocysts (P < 0.05). BFR increased HSPA5 (P = 0.0118) expression and did not affect SLC2A3, MSH6, KRT8, and FOSL1 expression (P > 0.05). In conclusion, melatonin (10-9 M) supplementation in the culture medium and BFR on D7 of culture increased the hatching rate 24, 48, and 72 h after warming of the vitrified embryos, indicating an improvement in cryotolerance.
Assuntos
Melatonina , Animais , Blastocisto , Bovinos , Criopreservação/veterinária , Suplementos Nutricionais , Técnicas de Cultura Embrionária/veterinária , Feminino , Fertilização in vitro/veterinária , Melatonina/farmacologia , Gravidez , VitrificaçãoRESUMO
Eighteen enzyme systems were examined in Ilex paraguariensis St. Hil. using starch gel electrophoresis. Seven out of 12 active isozyme systems revealed one or more polymorphic loci (PGI, GOT, MR, G-6PDH, MDH, NDH, and 6-PGDH). However, the segregation and linkage analyses were performed only for PGI, GOT, G-6PDH and 6-PGDH systems. Gene segregation at these loci was regular, except for a few trees that showed segregation distortion. Weak linkage disequilibrium between loci was detected, but it was not enough to influence the multilocus estimate.
Dezoito sistemas enzimáticos foram analisados em Ilex paraguariensis, utilizando eletroforese em gel de amido. Sete dos 12 sistemas que apresentaram atividade enzimática revelaram um ou mais locos polimórficos (PGI, GOT, MR, G-6PDH, MDH, NDH e 6-PGDH). Entretanto, as análises de segregação e desequilíbrio de ligação foram realizadas somente nos sistemas PGI, GOT, G-6PDH e 6-PGDH. A segregação destes locos foi regular, exceto para algumas árvores, que apresentaram distorções de segregação. Foram detectados indícios de desequilíbrio de ligação entre alguns locos, mas que não devem influenciar as estimativas multilocos.
RESUMO
Os termos pseudo-hermafrodita e intersexo são utilizados para se referir a animais com traços sexuais ambíguos. O diagnostico de pseudo-hermafrodita é baseado nos vestígios de gônadas e aspecto fenotípico do trato reprodutivo. Os pseudo-hermafroditas masculinos são aqueles que possuem somente testículo. Este trabalho relata um caso de pseudo-hermafrodita masculino em um suíno, cujo aparelho reprodutor foi colhido na linha de abate de um frigorífico. O animal, com idade de 150 dias e peso aproximado de 100 kg, possuía órgãos genitais exteriores femininos. O material coletado foi analisado macro e microscopicamente. Na análise macroscópica, verificou-se a presença de uma vulva anormal, ainda que a vagina e o útero (cérvix, corpo e cornos) apresentassem anatomia topográfica normal. As gônadas eram morfologicamente compatíveis com testículos, havendo também a presença dos epidídimos e do plexo pampiniforme. No exame histopatológico, confirmou-se que as gônadas apresentavam morfologia compatível com a de testículos, apresentando alterações degenerativas. Com base nos achados macroscópicos e microscópicos, concluiu-se que o animal era um pseudo-hermafrodita masculino.
The terms hermaphrodite and intersex are used to refer to animals with ambiguous sexual traits. The diagnostic of pseudo-hermaphrodites must be based on survey of the gonads and in the phenotypic aspect of the reproductive trait. The male pseudo-hermaphrodites have only testicle. This report describes a case of a male pseudo-hermaphrodite in a pig, whose reproductive organs were collected in a slaughterhouse. The pig was 150 days old, weighed nearly 100 kg, and showed external genitalia compatible with a female. The organs were evaluated grossly and microscopically. Grossly, the vulva showed abnormal morphology, but the vagina and the uterus (cervix, body, and horns) showed normal topographic anatomy. The gonads were morphologically compatible with testes with epididymis and pampiniform plexuses present. Histological examination of the gonads confirmed the identification of testes showing degenerative changes. Based on the gross and microscopic findings, the animal was diagnosed as a male pseudo-hermaphrodite.