Outbreak of porcine epidemic diarrhea in suckling piglets, China.
Article Type: Letter to the editor
Subject: Diarrhea (Health aspects)
Epidemics (Thailand)
Epidemics (China)
Epidemics (Health aspects)
Authors: Sun, Rui-Qin
Cai, Ru-Jian
Chen, Ya-Qiang
Liang, Peng-Shuai
Chen, De-Kun
Song, Chang-Xu
Pub Date: 01/01/2012
Publication: Name: Emerging Infectious Diseases Publisher: U.S. National Center for Infectious Diseases Audience: Academic; Professional Format: Magazine/Journal Subject: Health Copyright: COPYRIGHT 2012 U.S. National Center for Infectious Diseases ISSN: 1080-6040
Issue: Date: Jan, 2012 Source Volume: 18 Source Issue: 1
Geographic: Geographic Scope: Thailand; China Geographic Code: 9THAI Thailand; 9CHIN China
Accession Number: 277344822
Full Text: To the Editor: Beginning in October 2010, porcine epidemic diarrhea (PED), caused by a coronaviral infection affecting pigs, emerged in China in an outbreak characterized by high mortality rates among suckling piglets. The outbreak overwhelmed >10 provinces in southern China, and >1,000,000 piglets died. This outbreak was distinguished by -100% illness among piglets after birth (predominantly within 7 days and sometimes within only a few hours) and death rates of 80%-100% (online Technical Appendix Table 1, wwwnc. cdc.gov/EID/pdfs/11-1259-Techapp. pdf). Few sows or boars showed any clinical signs during the outbreak, which is not consistent with a recent report from Thailand (1). In that outbreak during late 2007, pigs of all ages were affected, exhibiting different degrees of diarrhea and no appetite. We characterized the genetic variation of the PED virus (PEDV) that caused a large-scale outbreak in China during 2010-2011 and compared it with viruses in other outbreaks. We also report a possible novel transmission pathway for PEDV.

A total of 177 samples (intestine, stool, and maternal milk) were collected from pigs from different farms who had diarrhea; 100% of farms had [greater than or equal to] 1 porcine sample positive for PEDV. A total of 125/177 porcine samples were confirmed as positive for PEDV by reverse transcription PCR using primers as described (2). PEDV was detected in 105 (82.0%) of 128 fecal samples and 20 (40.8%) of 49 sow milk samples. Piglets infected with PEDV showed mild hemorrhage, undigested curdled milk in the stomach, and thin-walled intestines with severe mucosal atrophy and foamy fluid (data not shown).

The spike (S) gene of the family Coronaviridae has a high degree of variation and can induce neutralizing antibody (3). Reverse transcription PCR products of the 651-bp partial S gene of PEDV and the deduced amino acid sequences were aligned by using ClustalW (www.genome. jp/tools/clustalw), and a neighborjoining tree with 1,000 bootstraps was constructed. Sequences of the S genes from this outbreak were 99.1%-100.0% homologous and had 88.7%-98.9% nt identity with all reference strains (online Technical Appendix Table 2), 98.5%-98.9% with Thailand strains, and 94.5%-95.1% with vaccine strain CV777. The partial S gene deduced amino acid sequences were compared and also showed a high degree of homology (98.0% 100.0%); they had 85.3%-98.7% identity with all reference strains listed in online Technical Appendix Table 2, 98.0%-98.7% with Thailand strains, and 93.3%-94.7% with vaccine strain CV777 (data not shown).

Phylogenetic analysis indicated that the PEDV in the China outbreak was different from foreign and other domestic strains on the basis of the reported partial S gene sequences. All new strains were clustered in the same branch, close to the cluster of Thailand strains, and far from the cluster of vaccine strain CV777 (Figure).

In the China outbreak, PEDV caused severe diarrheal disease in piglets; heavy economic losses in many provinces resulted, despite use of commercial vaccines (inactivated transmissible gastroenteritis [TGEV H] and porcine epidemic diarrhea [CV777]). To determine why the vaccines showed poor efficacy, we investigated evolution of the virus. Comparison of amino acid sequences from isolates from the outbreak and from the CV777 vaccine strain showed 9 amino acid mutations of fragments containing major hydrophilic regions: 16 (L[right arrow]H), 18 (S[right arrow]G), 22 (V[right arrow]I), 44 (T[right arrow]S), 89 (G[right arrow]S), 100 (A[right arrow]E), 107 (L[right arrow]F), 130 (I[right arrow]V) and 160 (I[right arrow]F) (online Technical Appendix Figure, panel A). Three of these 9 mutations were at positions 16, 18, and 22 in the isolates from China; they influenced the hydrophobicity of the S protein as compared with that for CV777 (online Technical Appendix Figure, panel B).

[FIGURE OMITTED]

Phylogenic analysis showed that strain CV777 did not cluster with current common strains and showed considerable genetic distance from them. Isolates in the outbreak in China had only a minor nucleotide sequence variation from the Thailand isolates, indicating that the virus has a high genetic relatedness to the Southeast Asia strain. However, previous studies showed that isolates from Europe, South Korea, and China were serologically identical to the prototype CV777 strain (1,4).

To our knowledge, fecal-oral transmission is probably the main or only route of PEDV transmission (5-7). In our study, if a fecal sample from a sick piglet was found to be positive for PEDV, we also collected and studied milk from its mother. These results showed that PEDV was present in sow milk (online Technical Appendix Table 3), but the detection rate was lower for these samples (40.8%) than for the fecal samples (82.0%).

On the basis of these results, we hypothesize that sow milk could represent a possible (and potentially major) route for the vertical transmission of PEDV from sow to suckling piglet. This hypothesis could be indirectly verified by our field observation that piglet death rates decreased as a result of fostering (data not shown). Our findings show that PEDV was identified not only in fecal samples from sick piglets, as expected, but also in the milk of the sow, which suggests vertical transmission of the virus.

Acknowledgments

We thank the technicians from the pig farms for assistance in sample collection and Bin Wu for assistance in figure preparation. We also thank the Guangdong Province Pig Industry Innovation Projects for their support.

References

(1.) Puranaveja S, Poolperm P, Lertwatcharasarakul P, Kesdaengsakonwut S, Boonsoongnern A, Urairong K, et al. Chinese-like strain of porcine epidemic diarrhea virus, Thailand. Emerg Infect Dis. 2009;15:1112-5. doi:10.3201/ eid1507.081256

(2.) Kim SY, Song DS, Park BK. Differential detection of transmissible gastroenteritis virus and porcine epidemic diarrhea virus by duplex RT-PCR. J Vet Diagn Invest. 2001;13:516-20. doi:10.1177/104063870101300611

(3.) Jimenez G, Correa I, Melgosa MP, Bullido MJ, Enjuanes L. Critical epitopes in transmissible gastroenteritis virus neutralization. J Virol. 1986;60:131-9.

(4.) Pospischil A, Hess RG, Bachmann PA. Light microscopy and ultrahistology of intestinal changes in pigs infected with epizootic virus diarrhoea (EVD): comparison with transmissible gastroenteritis (TGE) virus and porcine rotavirus infections. Zentralbl Vet erinarmed B. 1981;28:564-77. doi:10. 1111/j.1439-0450.1981.tb01774.x

(5.) Riley S. Large-scale spatial-transmission models of infectious disease. Science. 2007;316:1298-301. doi:10.1126/science.1134695

(6.) Utiger A, Tobler K, Bridgen A, Ackermann M. Identification of the membrane protein of porcine epidemic diarrhea virus. Virus Genes. 1995;10:137-48. doi:10.1007/BF01702594

(7.) Turgeon DC, Morin M, Jolette J, Higgins R, Marsolais G, DiFranco E. Coronavirus-like particles associated with diarrhea in baby pigs in Quebec. Can Vet J. 1980;21:100-xxiii.

Rui-Qin Sun, Ru-Jian Cai, Ya-Qiang Chen, Peng-Shuai Liang, De-Kun Chen, and Chang-Xu Song

Author affiliations: Guangdong Academy of Agricultural Sciences Veterinary Medical Institute, Guangzhou, China (R.-Q. Sun, R.-J. Cai, C.-X. Song); Northwest A &F University, Xi'an, Shanxi, China (R.-Q. Sun, D.-K. Chen); Inner Mongolia Agriculture University, Huhhot, Inner Mongolia, China (Y.-Q. Chen); and Henan Agriculture University, Zhengzhou, Henan, China (P.-S. Liang)

DOI: http://dx.doi.org/10.3201/eid1801.111259

Address for correspondence: Chang-Xu Song, Veterinary Medicine Institute, Guangdong Academy of Agricultural Sciences, Guangzhou, People's Republic of China; email: cxsong2004@163.com
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