Isolation of porcine epidemic diarrhea virus in porcine cell cultures and experimental infection of pigs of different ages.
This paper describes the isolation of porcine epidemic diarrhea (PED) virus in Vero and porcine cell cultures, and the influence of age on disease in experimental infection. PED virus was isolated from the small intestine of piglets inoculated with PED samples and cultured in Vero, porcine bladder and kidney cells propagated in collagen-coated tissue culture plates in maintenance medium (MM) containing trypsin. In porcine bladder and kidney cell cultures inoculated with isolated PED virus, cytopathic effects (CPE) including cell fusion were detected. Specific brilliant fluorescence was observed in the cytoplasm of these cells. Two- and 7-day old, and 2-, 4-, 8- and 12-week old specific pathogen-free (SPF) pigs were orally inoculated with PED virus isolated from an outbreak. All 2- and 7-day old pigs inoculated developed severe watery diarrhea from post-inoculation day (PID) 1 and died between PID 3 and 4. Although three of five 2-week old pigs developed diarrhea on PID 1-4, they eventually recovered. In the 4-week old group, three of five pigs had mild diarrhea for 1-2 days. None of the 8- and 12-week old pigs showed any clinical signs. Antibodies against PED virus were detected in all surviving pigs by virus neutralization (VN) test and immunofluorescence assay (IFA). Therefore, there is an age-dependent resistance to pathogenic PED virus infection in pigs.
Shibata
I
I
Zen-noh Institute of Animal Health, 7 Ohja-machi, Sakura, Chiba, Japan. shibatai@zk.zennoh.or.jp
Tsuda
T
T
Mori
M
M
Ono
M
M
Sueyoshi
M
M
Uruno
K
K
D000367:Age Factors; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000032:analysis; D002478:Cells, Cultured; D002522:Chlorocebus aethiops; D003332:Coronaviridae / Q000276:immunology / Q000302:isolation & purification*; D003333:Coronaviridae Infections / Q000276:immunology / Q000662:veterinary* / Q000821:virology; D003588:Cytopathogenic Effect, Viral; D003967:Diarrhea / Q000662:veterinary / Q000821:virology; D005455:Fluorescent Antibody Technique / Q000662:veterinary; D007150:Immunohistochemistry; D007421:Intestine, Small / Q000821:virology; D007668:Kidney / Q000166:cytology / Q000821:virology; D009500:Neutralization Tests / Q000662:veterinary; D013047:Specific Pathogen-Free Organisms; D013552:Swine; D013553:Swine Diseases / Q000276:immunology / Q000821:virology*; D001743:Urinary Bladder / Q000166:cytology / Q000821:virology; D014709:Vero Cells
D000914:Antibodies, Viral; D000956:Antigens, Viral