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1.
Reproduction ; 158(1): 71-83, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-31013477

RESUMO

Beneficial effects of n-3 polyunsaturated fatty acid (PUFA) supplementation on dairy cow reproduction have been previously reported. The objectives of the present study were to assess whether n-3 PUFA supplementation would affect in vitro embryo production (IVP) after ovarian stimulation. Holstein cows received a diet with 1% dry matter supplementation of either n-3 PUFA (n = 18, microencapsulated fish oil) or a control, n-6 PUFA (n = 19, microencapsulated soy oil). Both plasma and follicular fluid FA composition showed integration of total PUFA through the diet. All cows underwent an IVP protocol consisting of ovarian stimulation, ultrasound-guided transvaginal oocyte retrieval (ovum pick-up, OPU, five per cow) followed by in vitro maturation, fertilisation and 7 days of embryo development. A tendency toward an increase in the blastocyst rate (diet effect, P = 0.0865) was observed in n-3 cows, with 49.6 ± 5.5% vs 42.3 ± 5.5% in control n-6 cows. A significant increase (diet effect, P = 0.0217) in the good-quality blastocyst rate (freezable blastocysts) was reported in n-3 cows (42.2 ± 7.7%) compared to control n-6 cows (32.7 ± 7.7%). A significant difference in lipid composition was shown in the oocytes recovered by OPU from n-3 and n-6 treated cows, by intact single-oocyte MALDI-TOF mass spectrometry. The 42 differentially abundant identified lipids were mainly involved in cell membrane structure. In conclusion, n-3 PUFA supplementation enhanced oocyte quality and modified their lipid composition. Further studies are necessary to investigate the potential link of these lipid modifications with enhanced oocyte quality.


Assuntos
Dieta/veterinária , Suplementos Nutricionais , Embrião de Mamíferos/citologia , Desenvolvimento Embrionário/efeitos dos fármacos , Ácidos Graxos Ômega-3/administração & dosagem , Oócitos/citologia , Indução da Ovulação/veterinária , Animais , Bovinos , Embrião de Mamíferos/efeitos dos fármacos , Feminino , Fertilização in vitro/veterinária , Oócitos/efeitos dos fármacos
2.
Theriogenology ; 86(6): 1482-1488, 2016 Oct 01.
Artigo em Inglês | MEDLINE | ID: mdl-27341774

RESUMO

The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from infected donor goats to healthy recipients and/or their offspring. Nevertheless, the detection of C. abortus DNA by polymerase chain reaction does not prove that the bacteria found was infectious. Further studies are required to investigate whether enzymatic and/or antibiotic treatment of caprine embryos infected by C. abortus would eliminate the bacteria from the ZP.


Assuntos
Infecções por Chlamydia/veterinária , Chlamydia , Transferência Embrionária/veterinária , Doenças das Cabras/embriologia , Doenças das Cabras/microbiologia , Animais , Chlamydia/genética , Chlamydia/isolamento & purificação , Infecções por Chlamydia/transmissão , DNA Bacteriano/análise , Embrião de Mamíferos/microbiologia , Cabras , Reação em Cadeia da Polimerase/veterinária , Zona Pelúcida/microbiologia
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