Abstract
Severe climate change and global warming may impact significantly on vector-borne disease including Japanese encephalitis (JE) infection in human and animals. Thus, veterinary authority requires large quantity of diagnostic tools to survey vector-borne diseases. New producing method having a relation with JE antigen is needed to substitute conventional sucrose-acetone extraction method using suckling mouse. So, we developed new manufacturing method using polyethylene glycol (PEG) precipitation. Japanese encephalitis virus (JEV) was propagated in roller bottle containing Vero cell and inactivated with two kinds of inactivating reagents. Viability of the supernatant of bulk containing antigen was checked using Vero cell after inactivation. The supernatant did not show hemagglutination (HA) activity with goose erythrocytes. The antigen inactivated by binary ethylenimine (BEI) and concentrated by PEG precipitation method was found to be 2048 HA, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer. The antigen prepared from mice brain using sucrose-acetone extraction method showed 256 HA titer. This BEI inactivation method does not evoke animal welfare problem and can replace the conventional method that required biological hazardous reagents and suckling mice in preparing HA antigen. This new BEI inactivation method was safe in producing HA antigen against JEV in laboratory and can reduce environmental contamination of acetone.
Highlights
Climate change through recent global warming and extreme weather may have significantly impacted the transmission of vector-borne diseases such as Japanese encephalitis virus (JEV), West Nile virus (WNV), and tick-borne encephalitis virus (TBEV), as rapid weather and climate changes can directly or indirectly affect migratory birds and mosquitoes
The antigen inactivated by binary ethylenimine (BEI) and concentrated by polyethylene glycol (PEG) precipitation method was found to be 2048 HA, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer
The antigen inactivated with 0.001 M BEI and concentrated by PEG precipitation method showed 2048 HA titer, but the antigen inactivated by 0.3% formaldehyde solution and concentrated by PEG precipitation method did not show HA titer (Table 1)
Summary
Climate change through recent global warming and extreme weather may have significantly impacted the transmission of vector-borne diseases such as Japanese encephalitis virus (JEV), West Nile virus (WNV), and tick-borne encephalitis virus (TBEV), as rapid weather and climate changes can directly or indirectly affect migratory birds and mosquitoes. JEV can lead to infection in several animals such as reptiles, pigs, horses, chickens, breeding birds such as herons. Fattening pigs bitten by mosquitoes holding JEV show viremia for a short period of time with subclinical signs. Pregnant sows with JEV infection cause still birth, mummified fetus, weaken piglets showing neurological symptoms. Horse infected with JEV naturally shows lack of appetite, fever, neurological symptoms and pass away in the long run [6]. Since 2001, sero-surverveillance for JEV has been carried out over 9000 pigs every year
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