PMID- 15910918
OWN - NLM
STAT- MEDLINE
DCOM- 20050811
LR  - 20191210
IS  - 0093-691X (Print)
IS  - 0093-691X (Linking)
VI  - 63
IP  - 9
DP  - 2005 Jun
TI  - Evaluation of virus decontamination techniques for porcine embryos produced in 
      vitro.
PG  - 2343-55
AB  - 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.
FAU - Bureau, Marieve
AU  - Bureau M
AD  - Centre de Recherche en Biologie de la Reproduction, Departement des Sciences 
      Animales, Pavillon Paul-Comtois, Universite Laval, Ste-Foy, Que., Canada G1K 7P4.
FAU - Dea, Serge
AU  - Dea S
FAU - Sirard, Marc-Andre
AU  - Sirard MA
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Theriogenology
JT  - Theriogenology
JID - 0421510
RN  - EC 3.2.1.35 (Hyaluronoglucosaminidase)
RN  - EC 3.4.21.4 (Trypsin)
SB  - IM
MH  - Animals
MH  - Circovirus/genetics/isolation & purification
MH  - Decontamination/*methods
MH  - Diarrhea Viruses, Bovine Viral/genetics/isolation & purification
MH  - Embryo Culture Techniques/veterinary
MH  - Embryonic Development
MH  - Encephalomyocarditis virus/genetics/isolation & purification
MH  - Fertilization in Vitro/*veterinary
MH  - Hyaluronoglucosaminidase
MH  - Parvovirus, Porcine/genetics/isolation & purification
MH  - Porcine respiratory and reproductive syndrome virus/genetics/isolation & 
      purification
MH  - Swine/*embryology/*virology
MH  - Trypsin
EDAT- 2005/05/25 09:00
MHDA- 2005/08/12 09:00
CRDT- 2005/05/25 09:00
PHST- 2001/11/20 00:00 [received]
PHST- 2004/02/23 00:00 [revised]
PHST- 2004/05/03 00:00 [accepted]
PHST- 2005/05/25 09:00 [pubmed]
PHST- 2005/08/12 09:00 [medline]
PHST- 2005/05/25 09:00 [entrez]
AID - S0093-691X(04)00381-4 [pii]
AID - 10.1016/j.theriogenology.2004.05.034 [doi]
PST - ppublish
SO  - Theriogenology. 2005 Jun;63(9):2343-55. doi: 
      10.1016/j.theriogenology.2004.05.034.