Detection of Japanese Encephalitis Virus in Culex Mosquitoes in Singapore.

Mosquito-borne flaviviruses are emerging pathogens of an increasing global public health concern because of their rapid increase in geographical range and the impact of climate change. Japanese encephalitis virus (JEV) and West Nile virus (WNV) are of concern because of the risk of reemergence and introduction by migratory birds. In Singapore, human WNV infection has never been reported and human JEV infection is rare. Four sentinel vector surveillance sites were established in Singapore to understand the potential risk posed by these viruses. Surveillance was carried out from August 2011 to December 2012 at Pulau Ubin, from March 2011 to March 2013 at an Avian Sanctuary (AS), from December 2010 from October 2012 at Murai Farmway, and from December 2010 to December 2013 at a nature reserve. The present study revealed active JEV transmission in Singapore through the detection of JEV genotype II in Culex tritaeniorhynchus collected from an Avian Sanctuary. Culex flavivirus (CxFV), similar to the Quang Binh virus isolated from Cx. tritaeniorhynchus in Vietnam and CxFV-LSFlaviV-A20-09 virus isolated in China, was also detected in Culex spp. (vishnui subgroup). No WNV was detected. This study demonstrates the important role that surveillance plays in public health and strongly suggests the circulation of JEV among wildlife in Singapore, despite the absence of reported human cases. A One Health approach involving surveillance, the collaboration between public health and wildlife managers, and control of mosquito populations remains the key measures in risk mitigation of JEV transmission in the enzootic cycle between birds and mosquitoes.

First Report of Co-invasion by the Reptile Nematode Ozolaimus megatyphlon (Nematoda: Pharyngodonidae) with Invasive Green Iguanas (Iguana iguana) in the Asia-Pacific.

PURPOSE: Co-invasion of naïve ecosystems by non-native parasites is a serious threat to global biodiversity, though such events are difficult to detect early in the invasion process. Green iguanas (Iguana iguana) are an emerging invasive species and have colonised several countries in the Asia-Pacific. A survey was undertaken to determine whether parasites of the green iguana had co-invaded naïve ecosystems with their introduced host. METHODS: Over a 10-month period, wild green iguanas were trapped and euthanised in Singapore. All animals were necropsied and sampled for parasites. Parasites were then identified morphologically and subsequently characterised molecularly at the cytochrome c oxidase I (COI) locus. RESULTS: The reptile nematode Ozolaimus megatyphlon was found in 38% of the sampled green iguanas, with burdens of 100 + worms in all infected animals. This represents the first recorded co-invasion of this species with wild green iguanas in the Asia-Pacific. Based on the molecular characterisation of the cytochrome c oxidase I (COI) locus, the first DNA barcode is provided for O. megatyphlon. CONCLUSION: For the first time, the reptile nematode Ozolaimus megatyphlon is shown to be invasive and to have colonised the Asia-Pacific region with its introduced host, the green iguana. The DNA barcode provided here will facilitate future monitoring programmes as O. megatyphlon invades new habitats and countries.

Serological evidence of continued Japanese encephalitis virus transmission in Singapore nearly three decades after end of pig farming.

BACKGROUND: Singapore used to report an annual average of 14 cases of Japanese encephalitis, but ever since the abolishment of pig farms in the early 1990s, the local incidence rate for Japanese encephalitis virus (JEV) infections has reduced drastically. Studies done in the early 2000s demonstrated the presence of JEV-specific antibodies in animals such as wild boars, dogs, chickens and goats on the offshore island and peripheral parts of the Singapore, indicative of prior JEV exposure. A JEV wildlife and sentinel chicken surveillance system was initiated in 2010 through to 2017 to study the animal host seroprofiles. RESULTS: A total of 12/371 (3.23%) of resident bird samples, 24/254 (9.45%) of migratory bird samples and 10/66 (15.16%) of wild boar samples were positive for the presence of JEV antibodies. Seroconversions in sentinel chickens were observed at two time points. Through this study, two sites with active transmission of JEV amongst avian or porcine hosts were identified. CONCLUSIONS: JEV transmission in animal hosts has continued despite the phasing out of pig farming nearly thirty years ago; however, the public health risk of transmission remains low. Environmental management for mosquito population remains key to keeping this risk low.