Document Detail


Evaluation of virus decontamination techniques for porcine embryos produced in vitro.
MedLine Citation:
PMID:  15910918     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
The objective of this study was to explore approaches to decontaminate embryos either contaminated naturally or under experimental conditions with different viruses. Embryos were obtained from in vitro maturation and fertilisation of porcine oocytes. After 7 days of development, morula and blastocyst stages were exposed for 1 h to the following viruses: encephalomyocarditis virus (EMCV), porcine circovirus type 2 (PCV2), porcine parvovirus (PPV), porcine reproductive and respiratory syndrome virus (PRRSV), and bovine viral diarrhea virus (BVDV) at an infectivity of 100 TCID50/mL. Embryos samples were treated with different washing procedures, which all included the following standard washing solutions: PBS+0.4% BSA (five times for 10 s), Hank's+0.25% trypsin (two times for 60-90 or 120-150 s, or one time of 5 min), Hank's+0.1 mg/mL DNase 1+20 U/mL RNase One (one time of 30 min) and PBS+0.4% BSA again (five times for 10s). Two new approaches were used to improve trypsin treatment, 0.1% hyaluronidase (one time for 5 min) instead of trypsin and a pre-incubation with oviductal cells. Therefore, in the first experiment, oocytes received standard maturation treatments and in the second, they were also co-incubated with oviductal cells for the last 3 h of maturation. The effectiveness of the different washing techniques in removing viruses was evaluated by polymerase chain reaction (PCR) analysis. In the first experiment, trypsin treatment did not eliminate PRRSV, PPV, PCV, and EMCV from contaminated embryos. Surprisingly, treatment with hyaluronidase eliminated all tested viruses. In the second experiment, all viruses tested were removed from the oocytes following the different enzymatic treatments. In conclusion, in vitro embryo decontamination was more effective following exposure to oviductal secretions and hyaluronidase eliminated more virions than trypsin in washing techniques.
Authors:
Mariève Bureau; Serge Dea; Marc-André Sirard
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Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Theriogenology     Volume:  63     ISSN:  0093-691X     ISO Abbreviation:  Theriogenology     Publication Date:  2005 Jun 
Date Detail:
Created Date:  2005-05-24     Completed Date:  2005-08-11     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  0421510     Medline TA:  Theriogenology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  2343-55     Citation Subset:  IM    
Affiliation:
Centre de Recherche en Biologie de la Reproduction, Département des Sciences Animales, Pavillon Paul-Comtois, Université Laval, Ste-Foy, Qué., Canada G1K 7P4.
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MeSH Terms
Descriptor/Qualifier:
Animals
Circovirus / genetics,  isolation & purification
Decontamination / methods*
Diarrhea Viruses, Bovine Viral / genetics,  isolation & purification
Embryo Culture Techniques / veterinary
Embryonic Development
Encephalomyocarditis virus / genetics,  isolation & purification
Fertilization in Vitro / veterinary*
Hyaluronoglucosaminidase
Parvovirus, Porcine / genetics,  isolation & purification
Porcine respiratory and reproductive syndrome virus / genetics,  isolation & purification
Swine / embryology*,  virology*
Trypsin
Chemical
Reg. No./Substance:
EC 3.2.1.35/Hyaluronoglucosaminidase; EC 3.4.21.4/Trypsin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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