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Background and Aim: Hepatitis E virus (HEV) infection formerly and predominantly occurred in rural areas. However, it has recently been spread to urban and peri-urban areas. This study aimed to estimate the seroprevalence of HEV in pigs collected from urban and rural areas in Bali. The potential of the pig farmers’ risk level for being exposed to HEV and the virus transmitted to them in association with their pig-rearing practices was also assessed. Materials and Methods: A total of 183 pigs from 68 herds were sampled in this study, with 91 pigs collected from Denpasar as the representative samples of urban areas and 92 pigs from Karangasem Regency as the representative samples from rural areas. Sera from the sampled pigs were collected and immunoglobulin G antibodies against HEV were detected using a commercial enzyme-linked immunosorbent assay. A questionnaire was prepared for interviewing the farmers. Bivariable and multivariable logistic regression analyses were performed to identify the putative factors associated with seropositivity. Meanwhile, the potential risk-incurring practices of the farmers for HEV being transmitted to them from their pig-rearing practices were assessed by scoring their responses from the interview. Results: Overall, 23.5% (43/183) (95% confidence interval [CI]: 17.6–30.3) pig sera tested were detected to have the antibodies against HEV. Among 68 pig herds, 36.8% (25) (95% CI: 25.4–49.3) of them had antibodies in at least one pig sampled from each herd. Pigs sampled from Karangasem were 5 times (Odds ratio [OR] 5.34, 95% CI: 2.27–13.54, p < 0.001) more likely to be seropositive than pigs collected from Denpasar. However, no difference was found in the seropositivity to HEV in pig herds between Denpasar and Karangasem (p = 0.05). In assessing the pig rearing management factors, pig farmers from Denpasar were 3 times (OR 3.0, 95% CI: 1.07–8.52, p = 0.05) more likely to rear pigs for economic investment compared to the farmers from Karangasem. Regarding anticipating pig diseases that can be transmitted to humans, farmers from Denpasar were 6 times (OR 5.72, 95% CI: 1.48–26.7, p = 0.0074) more likely to anticipate zoonotic diseases compared to the farmers from Karangasem. Similarly, pig farmers from Denpasar were 3 times (OR 3.29, 95% CI: 1.08–10.23, p = 0.035) more likely to anticipate pig diseases that could be transmitted to humans than the farmers from Karangasem. Pig farmers from Denpasar had 4 times the odds (OR 4.49, 95% CI: 1.11–18.19, p = 0.03) of washing their hands after going to the pigpens compared to the farmers from Karangasem. All the participants were categorized as being at high risk of HEV exposure and transmission. Conclusion: IgG antibodies against HEV were detected among pigs reared in rural areas of Karangasem and those reared in urban areas of Denpasar. This suggests that the risk of HEV exposure and transmission in these areas is not negligible. To minimize the risk, public education on zoonotic diseases, including HEV infection, transmission, and prevention, needs to be implemented and particularly targeted to local pig farmers.

Background and Aim: Despite the endemicity of Japanese encephalitis virus (JEV) in humans and animals in the Province of Bali, Indonesia, there is little data on whether seroconversion to the virus occurs in pigs, JEV genotypes circulating, and it’s potential mosquito vectors in the area. The aims of this study were to (i) Determine whether JEV infection in Balinese pigs occurs before reaching their sexual maturity, (ii) identify the genotypes of circulating JEV, and (iii) identify potential JEV mosquito vectors at the study sites in urban and peri-urban areas of Bali. Materials and Methods: Sixteen 1-week-old Landrace piglets from two different sows were housed in Denpasar. Similarly, 18 one-week-old mixed-breed piglets of two different sows were housed in Badung Regency. The piglets were bled every 1 to 4 weeks for up to 24 weeks. Serum samples from the 11 piglets were tested for antibodies against JEV, and seroconversion-suspected sera were titrated using an enzyme-linked immunosorbent assay. Blood of seroconverted sera from pigs were tested using polymerase chain reaction (PCR) to detect the genetic sequence of JEV. The mosquitoes in the sentinels were trapped throughout the study period to identify the potential mosquito vectors of JEV. Results: Antibodies were detected in most of the selected piglets’ sera from weeks 1 to 24 of their age. However, sera of pig B9 collected from the sentinel setting in Badung Regency showed a four-fold increase in antibody titer from week 4 to week 8, indicating seroconversion. PCR testing of blood from B9 (pooled blood sample collected from week 5 to week 8) identified JEV nucleic acids, which were phylogenetically classified as belonging to the JEV genotype III. Meanwhile, 1271 of two genera of mosquitoes, Anopheles spp. and Culex spp. were trapped in the pig sentinels. Conclusion: JEV seroconversion likely occurs before the pig reaches sexual maturity in Badung Regency. Sequence data indicate that JEV genotype III is circulating in the pig sentinel setting in the regency; however, circulating genotypes need to be clarified through increased surveillance. Meanwhile, Culex spp. and most likely Culex quinquefasciatus and Anopheles spp. were the dominant mosquitoes present in the study sites set in the urban area of Denpasar and peri-urban areas of Badung, Bali, indicating that these are likely vectors in spread of JEV in the region.

Background and Aim: Japanese encephalitis (JE) is a zoonotic infectious inflammatory brain disease caused by the JE virus (JEV). Considerable research into the seroprevalence of JE in domestic animals has been conducted, but there have been no reports of its occurrence in wild animals, including long-tailed macaques (Macaca fascicularis). This study aimed to estimate the seroprevalence of JEV infection and its determinants in long-tailed macaques in Bali and the prevalence of mosquito vectors. Materials and Methods: Blood samples (3 mL) were collected from a population of M. fascicularis (92 heads) inhabiting a small forest with irrigated rice field nearby (wetland area) in Ubud, Gianyar, and from two populations in dryland areas with no wet rice field (Uluwatu, Badung, and Nusa Penida, Bali Province, Indonesia). The collected sera were tested for antibodies against JEV using a commercially available enzyme-linked immunosorbent assay kit (qualitative monkey JE Immunoglobulin G antibody kit). The seropositivity of the antibodies was then compared based on different variables, namely, habitat type, age, and sex. Results: The seroprevalence of the JEV antibodies in all the samples tested was found to be 41.3%. The seropositivity of the monkey serum samples collected from the wetland area was 46.4%, which was higher than the seropositivity of the sera samples collected from the dried field areas (1.25%). Monkey sera collected from the wetland areas were 6.1 times (odds ratio [OR]: 6.1; 95% confidence interval [CI]: 0.71-51.5, p>0.05) more likely to be seropositive compared to the monkey sera collected from the dried field areas. Meanwhile, female monkeys were 1.79 times (OR: 1.79; 95% CI: 0.76-4.21; p>0.05) more likely to be seropositive to JEV than males. Similarly, juvenile monkeys were 2.38 times (OR: 2.38; 95% CI: 0.98-5.79); p>0.05) more likely to be seropositive against the JEV than adult monkeys. However, none of these differences achieved statistical significance. Regarding the JEV mosquito vector collection, more Culex mosquitoes were found in the samples from the wetland areas than from the dried field areas. Conclusion: The study confirms the existence of JEV infection in long-tailed macaques in Bali. There were patterned seropositivity differences based on habitat, age, and sex of the monkeys, but these were not significant. The possibility of monkeys as a JEV reservoir and the presence of the mosquitoes as the JEV vector are suggested but require more study to confirm.

Aim: This study aimed to prepare binary ethylenimine (BEI)-inactivated virulent Newcastle disease virus (NDV) vaccine and to examine their ability to induce a protective antibody response in commercial chickens. Materials and Methods: A virulent NDV field isolate Gianyar-1/AK/2014 was propagated in chicken-embryonated eggs and was then inactivated with BEI at a concentration of 4 mM. Three groups of chickens with low-level (2 log2 hemagglutination inhibition [HI] units) maternally derived antibodies against NDV were then immunized with the BEI-inactivated vaccine. A commercial live vaccine (LaSota strain) was used as positive control, and phosphate-buffered saline (PBS) was used as negative control. A challenge experiment with a virulent NDV of Tabanan-1/ARP/2017 was performed at 3 weeks post-vaccination. Results: At 2 weeks post-immunization, the mean titers of antibodies against NDV in serum samples of chickens immunized with 0.2 mL of BEI-inactivated NDV (Group I), with live commercial NDV vaccine (Group II) and with PBS (Group III) were 3±0.94 log2 HI units, 4.9±0.99 log2 HI unit, and 0.0±0.0 HI units, respectively. At week 3 post-immunization, the mean titers of the antibodies for the three groups were 5±1.09 log2 HI units, 6.9±0.32 log2 HI units, and 0.00 HI units, respectively. The antibody titer induced by inactivated NDV Gianyar-1/AK/2014 isolates examined at 2 and 3 weeks post-vaccination was still at a significantly (p

Research was conducted to detect APMV-1 infection by examining microscopic lesions of chicken suspected ND and confirming the causative agent with serological and molecular assay. Samples obtained from commercial and back yard farm in 9 regencies and city of Bali Province were tested by rapid test for AIV antigen detection. AI negative samples were necropsied, then brain, lungs, and intestines were collected for histopathological examination. Samples tissue of brain, lung, spleen, and intestine were taken aseptically for viral isolation and amplification. Infected allantoic fluid was collected and tested by hemagglutination assay (HA) and hemagglutination inhibition (HI) test to prove APMV-1 serologically. Viral ribonucleic acid was isolated and subsequently reverse transcribed by reverse trasncription reaction followed by amplification by polymerase chain reaction to multiply the cDNA. Microscopically, perivascular cuffing (20%), endoteliosis (75%), and gliosis (75%) were found in the brain. In the lung, an interstitialis pneumonia (50%), lobar pneumonia (5%), and proliferation of pneumosit type 2 (100%) were observed. The most prominent intestinal lesions were catarrhal enteritis (75%) and hemorrhagic necrotizing enteritis (10%). Confirmation of the 20 isolates obtained in this study both serologically and molecularly were positive APMV-1. Moreover PCR results showed that 80% of its amplicon showed a single band and 20% still require some optimizations to get single good bands.

virus was isolated from suspected Newcastle disease (ND) chicken in backyard farm at Sibang Village, Badung Regency, Bali Province. The isolates were then propagated and confirmed for NDV serologically. RNA isolation was performed by standard Trizol method. Phylogenetic tree analysis of NP gene (nt1020-1561) and HN gene (nt7019-7754) were performed using sequence of Badung-02/AK/14 and selected NDV strains from GenBank. Based on the NP gene sequence, the newly isolate closely related with other NDV strains belong to genotype VII that previously isolated such as Banjarmasin-010/10, Bali-1/07, and Cockatoo/90 with genetic distance 0.2%, 12.6%, and 18.0% respectively. The genetic distance with LaSota/46 virus (genotype II) is 19.6%. Based on HN gene sequence, genetic distance of the Badung-02/AK/14 with other viruses belong to genotype VII such as Banjarmasin-010/10, Bali-1/07, and Cockatoo/90 are 0.4%, 3.7%, and 4.2% respectively. The genetic distance with LaSota/46 virus (genotype II) is 9.0%. There were no difference between the result of nucleotide and amino acid sequence analysis, both in NP and HN gene (P0.05).