Assessing and Prioritizing Zoonotic Diseases in Punjab, India: A One Health Approach.

Zoonotic diseases have a significant impact on both human and animal health globally. The present study was planned to prioritize the zoonoses in Punjab state of India. To develop a zoonotic disease prioritization scoring system, a comprehensive approach has been taken, including literature review, key person interviews with animal health experts (n = 12) and medical professionals (n = 7), and nine focus group discussions (FGDs) with veterinary academicians, medical professionals, and field veterinary doctors. The scoring system comprises of seven major criteria, each assigned a weightage score (ws): prevalence/incidence of the disease (ws = 0.20), severity of illnesses in humans (ws = 0.18), epidemic potential (ws = 0.16), socio-economic burden (ws = 0.16), availability of effective control and prevention measures (ws = 0.15), inter-sectoral collaborations (ws = 0.1), and bioterrorism potential (ws = 0.05). The finalized scoring system, accompanied by a list of 15 selected zoonotic diseases, was implemented among a group of 23 professionals engaged in zoonoses research (n = 7), animal health (n = 10), and medical health (n = 6) to determine their prioritization. The zoonotic diseases prioritized for the Punjab (India) included, Brucellosis (0.70) > Rabies (0.69) > Anthrax (0.64) > Leptospirosis (0.62) = Toxoplasmosis (0.62) = Highly Pathogenic Avian Influenza (HPAI) (0.62) > Bovine tuberculosis (0.61) > Q fever (0.60) > Cysticercosis (0.59) > Listeriosis (0.58) > Crimean-Congo haemorrhagic fever (CCHF) (0.57) > Japanese encephalitis (0.56) = Echinococcosis (0.56) > Dermatophytosis (0.53) > and Scrub typhus (0.48), respectively. Higher priority is suggested for endemic zoonoses (e.g., brucellosis and rabies) as compared to those with epidemic potential (e.g., CCHF, HPAI etc.) in Punjab. Results of the current study will help in the development of targeted control and prevention strategies for zoonotic diseases in Punjab and other geographical regions facing similar challenges.

Prevalence and distribution of Taenia solium cysticercosis in naturally infected pigs in Punjab, India.

BACKGROUND: Taenia solium (T. solium) cysticercosis remains a neglected zoonotic disease in India. The current study was planned to estimate the prevalence of T. solium porcine cysticercosis in the Punjab state of India, to compare this prevalence with the disease prevalence in pigs reared outside Punjab and to assess the distribution of the parasite in pig carcasses. METHODS: Two slaughter shops were selected in each of the 22 districts of Punjab. Pigs slaughtered on the day/s of inspection were post-mortem inspected to identify the presence of T. solium cysts. Estimated true prevalence was estimated by taking into account the diagnostic sensitivity (38%) and specificity (100%) of post-mortem inspection using the Rogan-Gladen estimator. Positive carcasses were purchased and brought to the laboratory to assess the tissue distribution of T. solium cysts and to conduct PCR targeting large subunit rRNA gene, internal transcribed spacer 1 gene, ITS1 gene and Cytochrome oxidase I gene. The selected PCR products were submitted for sequencing and phylogenetic analyses were performed. FINDINGS: We contacted 71 shop owners to achieve a sample of 44 shops for the study. We inspected 642 pigs reared in Punjab and 450 imported from other states at these slaughter shops. In addition, we sampled 40 pigs from an abattoir located in the state capital. Of the 642 pigs reared in Punjab, 9 had T. solium cysts with an apparent prevalence of 1·40% (95% CI: 0·74%, 2·64%) and the estimated true prevalence of 3.69% (95% CI: 1·95%, 6·95%). Pigs imported from outside the state had a significantly higher prevalence (odds ratio: 2·58; 95% CI: 1·12, 5·98; p-value: 0·026) as 15 of the 450 imported pigs were positive (apparent prevalence: 3.33%; 95% CI: 2.03%, 5.43%; estimated true prevalence: 8.77%; 95% CI: 5.34%, 14.28%). None of samples was positive from the pigs sampled at the abattoir in the state capital. The PCR confirmed T. solium cysts from all the 24 positive samples. We counted a median of 897 (range 526-1964) cysts per infected pig from the 19 infected pig carcasses inspected. The phylogenetic tree based on the alignment of partial cytochrome oxidase 1 sequences indicated all positive samples to be clustered with the T. solium Asian genotype. The analysis did not indicate the presence of T. asiatica in the slaughter pigs. CONCLUSIONS: Despite the underestimation of the prevalence due to missing mildly-infected carcasses, low participation and lack of representative sampling, the presence of heavily infected carcasses containing viable cysts, particularly those imported from outside the state, indicates that T. solium cysticercosis is an important food safety concern for pork consumers in Punjab, India. Measures should be taken to reduce the disease prevalence in pigs to reduce the disease burden in the public.

Prevalence and first molecular identification of Sarcocystis species in cattle and water buffaloes in India.

The importance of Sarcocystis hominis in causing zoonotic infections is well known. Recently, S. hominis like cysts have been reported from water buffalo in China. Previous studies indicate prevalence of Sarcocystis species in bovine populations in India but molecular evidence is required for proper species differentiation. We examined two hundred and ninety six cardiac tissue samples of Indian water buffaloes and cattle from northern and western parts of the country. Tissues were examined for Sarcocystis using intact cyst isolation method, pepsin acid digestion method and Sarcocystis 18S rRNA PCR. The combination of primers was used for 18S rRNA PCR amplification followed by sequencing. Twenty five representative samples were sent for sequencing and 19 readable sequences were obtained for phylogenetic analysis. Overall, the Sarcocystis cysts/zoites were recorded in 44% (95% CI 38-49%), 58% (95% CI 53-64%) and 68% (95% CI 63-73%) from both cattle and buffalo samples using intact cyst isolation, pepsin-HCl digestion method and conventional PCR, respectively. The results indicate that pepsin-HCl digestion method and conventional PCR are more sensitive than intact cyst isolation for detection of Sarcocystis species in tissue samples. The prevalence of Sarcocystis species was high in buffalo as compared to cattle intermediate hosts. Phylogenetic analysis indicated that more than one Sarcocystis species are circulating in cattle and water buffaloes in India. The results further indicate that experimental transmission studies are required to re-confirm the identities and host ranges of the Sarcocystis species in cattle and water buffaloes in India.

Prevalence and morphological characterisation of Echinococcus granulosus from north India.

The purpose of this study was to document the prevalence and to analyze morphological characteristics from hydatid cysts to test their suitability for strain identification. In the present study, 4,130 animals, including 278 cattle, 298 buffaloes, 760 sheep, 2,439 goat and 355 pigs were examined for the presence of hydatid cysts on post-mortem inspection at different slaughter houses/shops in northern India. Morphological characteristics from hydatid cysts were analyzed to test their suitability for strain identification. For statistical analysis, five variables were considered: number of hooks per rostellum, blade length of large and small hooks, and total length of large and small hooks. Principal component analysis was applied for analysis of morphological parameters. Out of a total of 4,130 animals examined, 66 were positive for hydatid cysts (prevalence 1.598 %). The prevalence of hydatid cysts was highest in cattle (5.39 %) followed by buffaloes (4.36 %), pigs (3.09 %), sheep (2.23 %) and goat (.41 %). The results indicate significant prevalence of hydatidosis in all the food producing animals and further that morphological analysis can also be used as a valid criterion for differentiation of different strains of E. granulosus particularly in developing countries where molecular studies could not be performed due to lack of infrastructure or financial constraints.