Abstract
Porcine epidemic diarrhea virus (PEDV), which is the causative agent of porcine epidemic diarrhea in China and other countries, is responsible for serious economic losses in the pork industry. Inactivated PEDV vaccine plays a key role in controlling the prevalence of PEDV. However, consistently low viral titers are obtained during the propagation of PEDV in vitro; this represents a challenge to molecular analyses of the virus and vaccine development. In this study, we successfully isolated a PEDV isolate (strain NJ) from clinical samples collected during a recent outbreak of diarrhea in piglets in China, using porcine intestinal epithelial cells (IEC). We found that the isolate was better adapted to growth in IECs than in Vero cells, and the titer of the IEC cultures was 104.5 TCID50/0.1 mL at passage 45. Mutations in the S protein increased with the viral passage and the mutations tended towards attenuation. Viral challenge showed that the survival of IEC-adapted cultures was higher at the 45th passage than at the 5th passage. The use of IECs to isolate and propagate PEDV provides an effective approach for laboratory-based diagnosis of PEDV, as well as studies of the epidemiological characteristics and molecular biology of this virus.
Highlights
Porcine epidemic diarrhea (PED), which is caused by the porcine epidemic diarrhea virus (PEDV), is an acute and highly contagious enteric viral disease in nursing pigs
Porcine epidemic diarrhea virus (PEDV) was isolated from PEDV‐positive samples collected from pig farms in China, using Vero
Vero cell cultures were negative for the M gene after two passages (Figure 1A), and no cytopathic effect (CPE) were cells and intestinal epithelial cell (IEC), in which a severe outbreak of acute diarrhea had been reported in sucking piglets
Summary
Porcine epidemic diarrhea (PED), which is caused by the porcine epidemic diarrhea virus (PEDV), is an acute and highly contagious enteric viral disease in nursing pigs. PED is characterized by vomiting and lethal watery diarrhea; and is a global problem, especially in many swine-producing countries [1,2,3,4,5,6,7]. PED was first reported in feeder pigs and fattening swine in the United Kingdom in. In China, PED outbreaks have occurred infrequently with only sporadic incidents [9,10]. In late 2010, a remarkable increase in PED outbreaks was reported in the pork-producing provinces [11,12]. In 2014, an outbreak of severe acute diarrhea, with high morbidity and mortality, occurred in sucking piglets in Nanjing, China. Herds vaccinated with the CV777-inactivated vaccine were infected
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