Antibody Evaluation and Mutations of Antigenic Epitopes in the Spike Protein of the Porcine Epidemic Diarrhea Virus from Pig Farms with Repeated Intentional Exposure (Feedback).

The feedback strategy, or controlled exposure of pig herd to the porcine epidemic diarrhea virus (PEDV), significantly decreased losses during a severe outbreak in late 2013 in Taiwan. However, some pig farms still suffered from recurrent outbreaks. To evaluate the association between antibody titers and clinical manifestations, sera and colostra were analyzed from one pig farm that employed the feedback strategy. Furthermore, spike (S) gene full sequences from six positive samples of two farms with and without using feedback were compared to investigate the evolution of PEDV variants circulating in pig herds. The results in this study showed that high PEDV antibody titers do not correlate with the high rate of protection from PEDV infection. In addition, repeated feedback generated the emergence of PEDV variants with unique substitutions of N537S and Y561H in the COE domain and S769F in the SS6 epitopes. These mutations indicated the pathogenetic evolution of PEDV strains existing in the cycle of the feedback method. A very strict biosecurity practice to block the routes of pathogen transfer should be followed to achieve successful control of PEDV infections in pig herds.

Encephalitis induced by a newly discovered ruminant rhadinovirus in a free-living Formosan sambar deer (Rusa unicolor swinhoei).

We documented a case of a free-living Formosan sambar deer (Rusa unicolor swinhoei) infected with a newly discovered ruminant Rhadinovirus (RuRv). Non-purulent encephalitis was the primary histological lesion of the sambar deer. We conducted nested PCR to screen for herpesvirus using generic primers targeting the DNA polymerase gene. In addition, we found that DNA polymerase gene of the sambar deer RuRv was present in the macrophage distributed in the Virchow Robin space with histopathologic lesions by chromogenic in-situ hybridization (CISH). The phylogenetic analysis indicated a high similarity between the viral sequence isolated from fallow deer and our case. This result suggests the possibility of cross-species transmission from other exotic Cervidae reservoir to the Formosan sambar deer.

Pigeon circovirus infection in disqualified racing pigeons from Taiwan.

Pigeons (Columba livia) infected with pigeon circovirus (PiCV) have been reported worldwide. The present study diagnosed PiCV infection in tissue samples of disqualified racing pigeons in Taiwan, using molecular and microscopy diagnostics. Among the 164 dead pigeons examined, 96.95% (159/164) tested positive for PiCV. Severe histopathological lesions, with characteristic inclusions, were observed in various organs of the PiCV-infected pigeons. Multiglobular basophilic intranuclear and intracytoplasmic inclusion bodies were found in the bursa of Fabricius and non-lymphoid tissues. The present study identified, for the first time, the presence of inclusion bodies in the thyroid gland, oesophagus, gizzard, and in the third eyelid of circovirus-infected pigeons. The presence of inclusion bodies in the third eyelid and mucosa of the gizzard was confirmed by transmission electron microscopy. A high detection rate of PiCV and some severe lesions evident in disqualified racing pigeons, as well as PiCV sequences in this study were highly similar with those detected in European countries suggesting an epidemiological association possibly due to imported pigeons.

Filarial nematode infection in eclectus parrots (Eclectus roratus) in Taiwan.

A total of 166 psittacines belonging to 22 species were received by the Animal Hospital of National Pingtung University of Science & Technology (NPUST) from 2013 to 2015. Only eclectus parrots (Eclectus roratus) were identified as hosts for microfilariae. All eclectus parrots were adult birds and had been kept in Taiwan for more than three years. The relevance of filariae to eclectus parrots is evident as indicated by the 35.7% (5/14) infection rate. At necropsy, adult filarial nematodes 57-75 mm in length and 0.4-0.7 mm in width were found in the hepatic veins. The microfilariae were 170-230 µm in length. Histopathological examination confirmed that eggs and larvae were observed in the ovaries and uteri of female filariae. These nematodes were closely related to an unidentified Filaria sp. (KJ612514.1) as indicated by polymerase chain reaction (PCR) analysis and phylogenetic analysis of nucleotide sequences from 18S ribosomal DNA gene (18S rDNA), mitochondrial cytochrome c oxidase subunit 1 (COX1) gene, and internal transcribed spacers 1-5.8S ribosomal DNA gene (ITS 1-5.8S rDNA). However, structurally the filarial nematodes were similar to that of the Pelecitus sp. Eclectus parrot species are important pet birds and are highly traded, resulting in high uncertainty of the origin of the parasite infection. This study is the first of its kind to report the presence and potential impact of filarial nematode infection on eclectus parrots, suggesting parasite inspection prior to the international trade of these pet birds.

Development of a loop-mediated isothermal amplification method for rapid detection of pigeon circovirus.

There are no effective antiviral treatments for pigeon circovirus (PiCV); thus, rapid diagnosis is critical for effective control of the disease caused by this virus. The recent development of a novel LAMP technique that amplifies nucleic acids rapidly with high specificity and sensitivity under isothermal conditions has overcome some of the deficiencies of nucleic-acid-based diagnostic tests. We established a LAMP method for rapid detection of PiCV using two pairs of primers that were designed from PiCV and compared its sensitivity and specificity with that of PCR. Amplification by LAMP was optimal at 63 °C for 60 min. The detection limit was nearly 0.5 pg of PiCV DNA, making it ten times more sensitive than PCR. There was no cross-reaction with porcine circovirus type 2 (PCV2), pigeon Trichomonas gallinae, or pigeon herpesvirus (PHV) under the same conditions. The assay also successfully detected the pathogen DNA in the tissues of infected pigeons. This is the first report indicating that LAMP is a valuable, rapid method of detecting PiCV with high sensitivity and specificity.