Effects of oocytes exposure to bovine diarrhea viruses BVDV-1, BVDV-2 and Hobi-like virus on in vitro-produced bovine embryo development and viral infection

被引:11
作者
da Silva Cardoso Pinto, V. [1 ,3 ]
Alves, M. F. [2 ]
de Souza Nunes Martins, M. [1 ]
Basso, A. C. [2 ]
Tannura, J. H. [2 ]
Pontes, J. H. F. [2 ]
Santos Lima, M. [1 ]
Garcia da Silva, T. [1 ]
Okuda, L. H. [1 ]
Stefano, E. [1 ]
Romaldini, A. H. C. N. [1 ]
Arnold, D. R. [2 ]
Pituco, E. M. [1 ]
机构
[1] Inst Biol Sao Paulo, Lab Viroses Bovideos, Ave Conselheiro Rodrigues Alves 1-252, BR-04014900 Sao Paulo, SP, Brazil
[2] Vitro Brasil, Rodovia SP 340,Km 166, BR-13800970 Mogi Mirim, SP, Brazil
[3] Univ Sao Paulo, FMVZ, Fac Med Vet, BR-05508270 Sao Paulo, SP, Brazil
基金
巴西圣保罗研究基金会;
关键词
Oocyte infection; BVDV; IVP embryo development; TRANSMISSION;
D O I
10.1016/j.theriogenology.2017.04.028
中图分类号
Q [生物科学];
学科分类号
07 ; 0710 ; 09 ;
摘要
As production of in vitro (IVP) bovine embryos steadily increases, the sanitary risk associated with IVP embryos remains a concern. One of the greatest concerns is how BVDV may be transmitted through IVP embryos. The objective of this study was to evaluate the effects caused by BVDV-1, BVDV-2 and Hobi-like virus exposure during in vitro maturation on embryo development and viral infection. Abittior-derived oocytes were randomly assigned for in vitro maturation with serial concentrations of BVDV-1 (3.12 x 10(2) - 2.50 x 10(3) TCID50/100 mu L), BVDV-2 (6.25 x 10(1) - 5.20 x 10(2) TCID50/100 mu L) or Hobi-like virus (1.90 x 10(2) - 1.58 x 10(3) TCID50/100 mu L) for 22-24 h. After maturation, oocytes were fertilized and embryo cultured following standard in vitro procedures. Embryo development was evaluated and percentage of respective, positive BVDV degenerated and viable embryos were evaluated by RT-qPCR. No concentration of BVDV-1 altered embryo development as measured by cleavage and blastocyst rates, compared to negative control group. However 100% of degenerated embryos and 50-100% of viable embryos tested positive for BVDV-1, depending on the viral concentration. BVDV-2 exposed oocytes had higher cleavage rates than the negative control group (60.2-64.1% vs 49.8%; P = 0.003-0.032). However, no difference was detected for blastocyst rates. In aadition, 100% of degenerated embryos and 20-50% of viable embryos tested positive for BVDV-2. Hobi-like virus treated oocytes had reduced cleavage rates for the three highest viral concentrations (333-38.0% vs 49.8% for negative controls; P <= 0.001-0.014). Blastocyst rates were only reduced in the 7.9 x 10(2) Hobi-like virus concentration (6.9 +/- 0.9% vs 15.1 +/- 1.6%; P = 0.009), when calculated by oocyte number. 50-80% of degenerated embryos tested positive for Hobi-like virus. No viable embryos from the Hobi-like virus treated oocytes tested positive. These results suggest that IVP embryos from BVDV-1 and -2 infected oocytes develop normally, but carry the virus. However, Hobi-like virus infected oocytes had reduced cleavage and cause pre-implantation embryo loss, but viable embryos did not carry the virus. (C) 2017 Elsevier Inc. All rights reserved.
引用
收藏
页码:67 / 72
页数:6
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