The stability of porcine rotavirus in feces.
Rotaviruses are known as major causal agents of diarrhea in humans and animals. They affect young animals in intensive rearing and cause great economic losses. This study evaluated the infectivity of porcine rotavirus maintained for 32 months at approximately 10 degrees C in the original stool specimens. Thirty stool specimens of 1-4-week-old piglets from breeding farms located in the southwest of the State of Parana were selected for this study. They were randomly chosen from stool samples positive for rotavirus RNA by polyacrylamide gel electrophoresis (PAGE) at the time of collection. The thirty stool samples maintained for 32 months were re-tested by PAGE and 11 out of 30 were still positive showing physical integrity of the eleven segments of viral RNA. In order to demonstrate the maintenance of viral infectivity processed fecal homogenates were inoculated in MA-104 cell cultures. After an average of three blind passages 5 out of 11 samples demonstrated cytopathic effect similar to that of a simian rotavirus (SA-11) used as positive control. To confirm these findings an immunofluorescence test was performed and typical cytoplasmatic granular fluorescence was observed. Electron microscopy of stool samples showed that most of the virus particles were single-shelled and some were found to be in advanced state of degradation. The viral nucleic acid extracted from six fecal specimens out of those that showed physical integrity of rotavirus RNA by PAGE were also amplified when submitted to RT-PCR demonstrating stability of viral RNA. We therefore concluded that porcine rotavirus infectivity is maintained for a long period of time in stool specimens at low temperature.
Ramos
A P
AP
Departamento de Microbiologia, CCB/UEL, Londrina, Parana, Brazil.
Stefanelli
C C
CC
Linhares
R E
RE
de Brito
B G
BG
Santos
N
N
Gouvea
V
V
de Cassia Lima
R
R
Nozawa
C
C
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D005243:Feces / Q000821:virology*; D019084:Fluorescent Antibody Technique, Indirect / Q000662:veterinary; D008854:Microscopy, Electron / Q000662:veterinary; D012367:RNA, Viral / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D012401:Rotavirus / Q000235:genetics / Q000302:isolation & purification* / Q000648:ultrastructure; D012400:Rotavirus Infections / Q000662:veterinary* / Q000821:virology; D013552:Swine; D013553:Swine Diseases / Q000821:virology*
D012367:RNA, Viral