Porcine epidemic diarrhea virus (PEDV) has been detected sporadically in Italy since the 1990s. We report the phylogenetic relationship of swine enteric coronaviruses collected in Italy during 2007–2014 and identify a drastic shift in PEDV strain variability and a new swine enteric coronavirus generated by recombination of transmissible gastroenteritis virus and PEDV.
Porcine epidemic diarrhea virus (PEDV) and Transmissible gastroenteritis virus (TGEV) (family Coronaviridae, genus Alphacoronavirus) are enveloped viruses that contain a single-stranded, positive-sense RNA genome of ≈28 kb. Infection with these viruses causes watery diarrhea, dehydration, and a high mortality rate among suckling pigs. Coronaviruses (CoVs) are prone to genetic evolution through accumulation of point mutations and homologous recombination among members of the same genus. Porcine respiratory coronavirus (PRCV), a mutant of TGEV, appeared in pigs in the 1980s. The spread of PRCV, which conserved most of the antigenic sites and causes cross-protection against TGEV, led to the gradual disappearance of TGEV. Newly emerging CoVs pose a potential threat to human and animal health because multiple human CoV infections have been derived from animal hosts. Emerging swine coronaviruses are of great concern to swine health because of the potential increase in viral pathogenesis.
In 1978, PEDV was first identified in Europe; subsequent reports occurred in many countries in Asia, including China, Japan, Korea, and Thailand. In 2010–2012, genetically different PEDV strains emerged, causing severe outbreaks in China. PEDV spread to the United States, Canada, and Mexico in 2013–2014, and genetically related strains were detected in South Korea and Taiwan. The PEDV outbreak caused large global economic losses to the swine industry. In Europe, a severe PEDV epidemic occurred in Italy during 2005–2006 (8), and in 2014–2015, PEDV was detected in Germany, France, and Belgium. These strains have a high nucleotide identity to PEDV strains that contain distinct insertions and deletions (INDELs) in the S gene (S-INDELs) from the United States). We report the detection and genetic characterization of swine enteric CoVs circulating in Italy during 2007–2014. We also report a recombinant TGEV and PEDV strain (identified as the species Swine enteric coronavirus [SeCoV]) circulating from June 2009 through 2012. Finally, we describe the phylogenetic relationship of the 2014 PEDV S-INDELs to the recent PEDV strains circulating in Europe.
During 2007–2014, most (92%) samples collected from the Po Valley in Italy were positive for PEDV by ELISA; only 72% were positive by pan-CoV PCR. However, because we were investigating the presence of PEDV or TGEV in samples with clinical signs of diarrhea, the high occurrence of PEDV may not reflect the actual prevalence of PEDV in Italy. The increased percentage of PEDV found in samples tested by ELISA, compared with the proportion found by PCR, may be explained by the number of ambiguous bases in the pan-CoV primers; the ambiguous bases severely reduce the efficiency of the reaction. The swine enteric CoV strains from Italy in our study, including the recombinant strain, were reported in pigs with mild clinical signs, indicating that PEDV and SeCoV have been circulating in Italy and likely throughout Europe for multiple years but were underestimated as a mild form of diarrhea.
To understand the evolution of PEDV in Italy, the partial RdRp, S, and M genes were sequenced from 18 samples and grouped in 3 different temporal clusters. Cluster I (2007–mid 2009) resembles the oldest PEDV strains; cluster II resembles a new TGEV and PEDV recombinant variant; and cluster III, identified from 2 pig farms in northern Italy in 2014, resembles the PEDV S-INDEL strains identified in Germany, France, Belgium, and the United States. The >99.3% nt identity of the S1 gene within cluster III and in previously identified strains could suggest the spread of the S-INDEL strain into Europe. However, directionality of spread cannot be determined because of a lack of global and temporal PEDV sequences.
Although our findings could indicate 3 introductions of PEDV in Italy, the results more likely suggest the high ability of natural recombination among CoVs and the continued emergence of novel CoVs with distinct pathogenic properties. Further investigation is needed to determine the ancestor of the SeCoV strain or to verify whether the recombinant virus was introduced in Italy. Recombinant SeCoV was probably generated in a country in which both PEDV and TGEV are endemic, but because the presence of these viruses in Europe is unclear and SeCoV has not been previously described, it is difficult to determine the parental strains and geographic spread of SeCoV. Future studies are required to describe the pathogenesis of SeCoV and its prevalence in other countries.
http://wwwnc.cdc.gov/eid/ Emerging Infectious Diseases
http://wwwnc.cdc.gov/eid/article/22/1/15-0544_article Original web page at Emerging Infectious Diseases