Unveiling genotypic diversity of Theileria orientalis in lethal outbreaks among bovines in Karnataka, India.

Theileria orientalis, the causal agent of oriental theileriosis, is known to cause mild disease in cattle and buffalo across the world. Recently, different genotypes of T. orientalis have emerged as pathogenic, causing high reported morbidity in cattle. This study focuses on investigating three suspected outbreaks of oriental theileriosis that resulted in fatalities among crossbred and indigenous bulls in Karnataka, India. Examination of blood smears revealed the presence of T. orientalis piroplasms within erythrocytes. The genetic characterization of T. orientalis was conducted by targeting specific markers, including the mpsp gene, p23 gene, and ribosomal DNA markers (18S rRNA gene, ITS-1, and ITS-2). Analysis based on the 18S rRNA gene unveiled the presence of both Type A and Type E genotypes of T. orientalis in the outbreaks. The mpsp gene-based analysis identified genotype 7 of T. orientalis in crossbred cows, whereas genotype 1 (Chitose B) was found to be present in indigenous bulls. Haplotype network analysis based on the mpsp gene revealed the presence of 39 distinct haplotypes within the 12 defined genotypes of T. orientalis with a high haplotype diversity of 0.9545 ± 0.017. Hematological and biochemical analysis revealed a decrease in calcium, hemoglobin levels, red blood cell counts, and phosphorus. This study constitutes the initial documentation of a clinical outbreak of oriental theileriosis in indigenous bulls with genotype 1 (Chitose 1B). Substantial epidemiological investigations are imperative to gain a comprehensive understanding of the geographical distribution of distinct genotypes and the diverse clinical manifestations of the disease across various hosts.

Development of dipstick enzyme linked immunosorbent assay for on-site sero-diagnosis of Japanese encephalitis in swine.

Japanese encephalitis (JE) is an important viral zoonotic disease in Asia, especially in rural and suburban areas where rice cultivation and pig farming coexist. Pigs serve as a suitable sentinel model, the surveillance of which could predict a potential JE outbreak in human population in the immediate vicinity. However, existing diagnostics like ELISA and VNT require sophisticated laboratory facilities which are more often not available in field conditions. In the present study, we aimed at developing recombinant non-structural (NS1) protein-based dipstick IgG ELISA as an on-site assay for sero-diagnosis of JE in swine. The assay was standardized by optimizing various parameters and the following conditions were found to be ideal including 1 µg of rNS1 protein in carbonate buffer per strip of nitrocellulose membrane comb; bovine serum albumin as blocking agent at 4 °C overnight; serum dilution of 1:10 and conjugate dilution of 1:5000 in skimmed milk powder. Relative diagnostic sensitivity and specificity of dipstick IgG ELISA was 100% and 92.9%, respectively. The dipstick assay was validated in three laboratories as per OIE guidelines. The storage life of dipstick was up to 7 months at 4 °C. The assay is easy to perform and the results can be interpreted with visual observation that precludes the need for absorbance reading equipment. The standardized dipstick assay was found promising for screening swine serum samples in field conditions. Timely detection of JE virus in swine will aid in predicting the outbreak in humans and thus in taking suitable preventive and control measures.

Serological evidence of Japanese encephalitis virus infection in pigs in a low human incidence state, Goa, India.

Japanese encephalitis (JE) is a mosquito-borne zoonosis caused by Japanese encephalitis virus (JEV). It is a leading cause of encephalitis in humans especially children in Asia. Aquatic wading birds are the reservoirs and pigs serve as amplifying hosts for JEV. Humans and horses are dead-end hosts. JE is endemic in several states of India. Goa, a small state on the west coast of India, had witnessed JE outbreaks in the past and as on date human JE cases are reported sporadically. Although human JE cases are well documented in Goa, the status of JEV exposure of pigs has not been well documented. Hence the present study was undertaken with an objective of identifying JEV exposure in the pig population of Goa state in the light of declining human JE cases. To achieve the objective, between January 2017 and May 2019, serum samples from 666 pigs were screened using enzyme-linked immunosorbent assay (ELISA) for the detection of anti-JEV IgG. The apparent prevalence of anti-JEV IgG in pigs was found to be 7.1 % (95 % confidence interval 5.3 %-9.3 %) and true prevalence was 4.6 % (95 % confidence interval 2.7 %-7.1 %). The seroprevalence of JE recorded in pigs of Goa state was low compared to other endemic states in India, which may also be one of the reasons for the lower prevalence of human JE cases in Goa state. Univariate analysis revealed that the age of the pigs and district did not significantly influence the JE seroprevalence in pigs of Goa state. However, in multivariable logistic regression, the North Goa district was found to significantly (p = 0.017) influence the JE seroprevalence in pigs. The study identified that JEV is still circulating in the Goan pig population and hence constant vigil is required to monitor the intensity of JEV circulation in pigs. Besides forewarning possible human outbreaks in the locality, evidence of JEV exposure in pig population provides valuable data on the magnitude and extent of geographical spread.