Phytohemagglutinin improves the development and ultrastructure of in vitro-cultured goat (Capra hircus) preantral follicles

The objective this study was to determine the effect of phytohemagglutinin (PHA) on survival, growth and gene expression in caprine secondary follicles cultured in vitro. Secondary follicles (∼0.2 mm) were isolated from the cortex of caprine ovaries and cultured individually for 6 days in α-MEM+ supplemented with PHA (0, 1, 10, 50, 100, or 200 µg/mL). After 6 days of culture, follicle diameter and survival, antrum formation, ultrastructure and expression of mRNA for FSH receptors (FSH-R), proliferating cell nuclear antigen (PCNA), and neuronal nitric oxide synthase were determined. All treatments maintained follicular survival [α-MEM+ (94.59%); 1 µg/mL PHA (96.43%); 10 µg/mL PHA (84.85%); 50 µg/mL PHA (85.29%); 100 µg/mL PHA (88.57%), and 200 µg/mL PHA (87.50)], but the presence of 10 µg/mL PHA in the culture medium increased the antrum formation rate (21.21%) when compared with control (5.41%, P < 0.05) and ensured the maintenance of oocyte and granulosa cell ultrastructures after 6 days of culture. The expression of mRNA for FSH-R (2.7 ± 0.1) and PCNA (4.4 ± 0.2) was also significantly increased in follicles cultured with 10 µg/mL PHA in relation to those cultured in α-MEM+ (1.0 ± 0.1). In conclusion, supplementation of culture medium with 10 µg/mL PHA maintains the follicular viability and ultrastructure, and promotes the formation of antral cavity after 6 days of culture in vitro.

INIBIÇÃO DOS LENTIVÍRUS DE PEQUENOS RUMINANTES POR DROGAS ANTIVIRAIS

ABSTRACT Inhibitors of the reverse transcriptase and protease enzymes were evaluated for their inhibitory effect on the replication of caprine arthritis encephalitis virus (CAEV) strain CAEV Cork and of the Maedi-Visna virus (MVV) strain K1514 cultured in fibroblastic cells. The drugs lamivudine, didanosine, stavudine, zidovudine, efavirenz, atazanavir and lopinavir/ritonavir were used. The highest concentration used for lamivudine, stavudine, zidovudine and efavirenz was 500 ?M, for atazanavir it was 50 ?M and 5.0 ?M for lopinavir/r and didanosine. The in vitro antiviral activity was investigated by evaluating the cell viability by the MTT method and testing for inhibition of cytopathic effects (CPE) of the virus. The replication of the virus was not completely inhibited by the protease inhibitors atazanavir and lopinavir/r in the test for CPE, while the others drugs showed significant antiviral efficacy in different concentrations. The protease inhibitors together with the efavirenz did not show antiviral activity when they were assessed by the reduced MTT technique. These data showed that the reverse transcriptase inhibitor drugs lamivudine, didanosine, stavudine and zidovudine were effective in the in vitro inhibition of small ruminant lentivirus.

RESUMO Inibidores da enzima transcriptase reversa e da protease foram avaliados quanto ao seu efeito inibitório na replicação do Vírus da Artrite Encefalite Caprina (CAEV) cepa CAEV Cork e do vírus Maedi-Visna (MVV) cepa K1514 cultivados em células fibroblásticas de caprinos. Os fármacos utilizados foram: lamivudina, didanosina, estavudina, zidovudina, efavirenz, atazanavir e lopinavir/ritonavir. A maior concentração utilizada para lamivudina, estavudina, zidovudina e efavirenz foi 500 ?M, para atazanavir foi 50 ?M e 5,0 ?M para lopinavir/r e didanosina. A atividade antiviral in vitro foi pesquisada por meio da avaliação da viabilidade celular através da redução do MTT e pela pesquisa de inibição dos efeitos citopáticos (CPE) dos vírus. A replicação dos vírus só não foi completamente bloqueada pelos inibidores de protease (IP) atazanavir e lopinavir/r enquanto os demais apresentaram eficácia antiviral significativa em diferentes concentrações. Os IP juntamente com o efavirenz, não mostraram atividade antiviral quando foram avaliados pela técnica de redução do MTT. Esses dados indicam que os fármacos inibidores da transcriptase reversa lamivudina, didanosina, estavudina e zidovudina são eficazes na inibição in vitro dos lentivírus de pequenos ruminantes.

Evaluation of induced molting methods on the livability and reproductive system regression of Japanese quails (Coturnix japonica)

This study aimed at evaluating the alternative method of zinc oxide and fasting to induce molt in Japanese quails. A total number of 190 48-week-old quails was used. They were at end of laying cycle, and presented low egg production. Quails molted by zinc oxide (Z) were fed a diet containing 25,000 ppm of zinc oxide, and received water ad libitum. Quails treated by fasting (F) received no feed and a day of water restriction. The treatment period was determined by the experimental level of body weight loss (BWL). Birds were submitted to different levels of BWL in order to analyze reproductive system regression (ovary + oviduct), and livability. The following groups were established according to their BWL: Control (untreated quails); F25 (25% BWL by F); F35 (35% BWL by F); Z25 (25% BWL by Z), and Z35 (35% BWL by Z). Z25, Z35, and F35 presented no significant differences in reproductive system weights after molting; however, their weights were lower than F. Z, Z, and F presented the following livability: 97.5, 72.5, and 90%. Japanese quails treated by the alternative method of zinc oxide, presenting body weight loss of 25%, showed low mortality rate, and adequate regression of the reproductive organs.

Evaluation of induced molting methods on the livability and reproductive system regression of Japanese quails (Coturnix japonica)

This study aimed at evaluating the alternative method of zinc oxide and fasting to induce molt in Japanese quails. A total number of 190 48-week-old quails was used. They were at end of laying cycle, and presented low egg production. Quails molted by zinc oxide (Z) were fed a diet containing 25,000 ppm of zinc oxide, and received water ad libitum. Quails treated by fasting (F) received no feed and a day of water restriction. The treatment period was determined by the experimental level of body weight loss (BWL). Birds were submitted to different levels of BWL in order to analyze reproductive system regression (ovary + oviduct), and livability. The following groups were established according to their BWL: Control (untreated quails); F25 (25% BWL by F); F35 (35% BWL by F); Z25 (25% BWL by Z), and Z35 (35% BWL by Z). Z25, Z35, and F35 presented no significant differences in reproductive system weights after molting; however, their weights were lower than F. Z, Z, and F presented the following livability: 97.5, 72.5, and 90%. Japanese quails treated by the alternative method of zinc oxide, presenting body weight loss of 25%, showed low mortality rate, and adequate regression of the reproductive organs.