Ahead of Print -Gouleako and Herbert Viruses in Pigs, Republic of Korea, 2013 - Volume 20, Number 12—December 2014 - Emerging Infectious Disease journal - CDC
Volume 20, Number 12—December 2014
Gouleako and Herbert Viruses in Pigs, Republic of Korea, 2013
Several viruses in the family Bunyaviridae, such as severe fever thrombocytopenia syndrome virus, sandfly fever Naples virus, and La Crosse virus, cause vector-borne zoonotic problems (1–7). Recently, outbreaks of severe disease caused by Rift Valley fever virus and Schmallenberg virus produced abortion storms, resulting in a high mortality rate among newborn lambs and calves (4,8). Gouleako virus (GOLV) and Herbert virus (HEBV) have been isolated from mosquitoes (Culex spp.) trapped in Côte d'Ivoire (9,10); however, their infectivity or virulence have not been proven. Investigation of the cause of death of pigs in the Republic of Korea identified GOLV and HEBV infection.
In March 27, 2013, a piglet, ≈8 weeks of age, on a 150-sow farm in Gyeonggi, Republic of Korea, died after onset of high fever (40°C), wasting, respiratory disease, and diarrhea. The carcass was sent to the Department of Veterinary Medicine Virology Laboratory, Seoul National University, Seoul, Republic of Korea, for diagnostics. Necropsy and microscopic examinations revealed greenish lung tissue with lymphoid depletion, consistent with severe bronchopneumonia. Despite the presence of multiple clinical signs, the results of routine tests for major pathogens in pigs (e.g., porcine reproductive and respiratory syndrome virus, porcine circovirus type 2, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, Escherichia coli, Streptococcus spp., and Salmonella spp.) were negative.
To further explore cause of the death, we used the particle-associated nucleic acid –random PCR method (Technical Appendix[PDF - 1.19 MB - 6 pages]). Sequencing and BLAST analysis (http://www.ncbi.nlm.nih.gov/blast/Blast.cgi) of the agent-specific amplicon simultaneously detected 2 viruses in lung tissue RNA samples. One partial sequence had 100% identity with 63 nt of the GOLV strain F23/CI/2004 glycoprotein gene (GenBank accession no. FJ765411). Another sequence had 97% similarity with 66 nt of the HEBV strain F23-K4 RNA-dependent RNA polymerase (RdRp) gene (GenBank accession no. EF423168).
Results were validated with reverse transcription PCR (RT-PCR) (Technical Appendix[PDF - 1.19 MB - 6 pages]). We obtained partial sequences of 235 nt of GOLV and 324 nt of HEBV. These sequences had 97.1% and 96.9% similarity with GOLV and HEBV, respectively, previously isolated from mosquitoes (9,10). The sequences were registered as GenBank accession nos. KF361520 and KF361522 and designated as GOLV/P1 and HEBV/P1, respectively.
During March–May 2013, we received a total of 9 dead pigs from the same farm; they had displayed various clinical signs. We further screened these pigs for the presence of GOLV and HEBV by using the same primer sets (Technical Appendix[PDF - 1.19 MB - 6 pages]) selective for their glycoprotein andRdRp genes, respectively. The results showed that the pigs were infected with GOLV and HEBV at a prevalence of 83.3% and 100%, respectively, mostly in lung samples (Table 1). The sequences obtained from this assay were registered as GenBank accession nos. KF361521 and KF361523 and designated GOLV/P8 and HEBV/P9, respectively.
Because of the high rate of GOLV positivity, we conducted a histopathologic RNA in situ hybridization study (Technical Appendix[PDF - 1.19 MB - 6 pages]). Hybridization signal was positive in lung and lymph node tissues and negative in intestine and control tissues. Hybridization was strong in the cytoplasm of mononuclear cells (deep blue color) (Technical Appendix[PDF - 1.19 MB - 6 pages] Figure 1).
Mr Chung is a virologist at the Department of Veterinary Medicine Virology Lab, College of Veterinary Medicine, Seoul National University, Republic of Korea. His research interests include swine virology, transgenic pigs, endogenous retroviruses, viral enteritis of pigs, and viral diseases of animals.
We thank Su Hee Yun for expert technical assistance.
This study was supported by a grant (PJ009015) from BioGreen 21 Program, Republic of Korea.
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Suggested citation for this article: Chung HC, Nguyen VG, Goede D, Park CH, Kim AR, Moon HJ, et al. Gouleako and Herbert viruses in pigs, Republic of Korea, 2013. Emerg Infect Dis [Internet]. 2014 Dec [date cited]. http://dx.doi.org/10.3201/eid2012.131742