Spillover of Newcastle disease virus to Himalayan Griffon vulture: a possible food-based transmission.

The Newcastle disease virus (NDV) affects wild and domesticated bird species, including commercial poultry. Although the diversity of NDV in domestic chickens is well documented, limited information is available about Newcastle disease (ND) outbreaks in other bird species. We report an annotated sequence of NDV/Vulture/Borjuri/01/22, an avirulent strain of NDV reported from Borjuri, Northeast India, in Himalayan Griffon vulture. The complete genome is 15,186 bases long with a fusion protein (F) cleavage site 112GRQGR↓L117. The phylogenetic analysis based on the F protein gene and the whole genome sequence revealed that the isolate from the vulture belongs to genotype II, sharing significant homology with vaccine strain LaSota. The study highlights the possible spillover of the virus from domestic to wild species through the food chain.

Thermostability and Immunogenicity of Genotype II Avian Orthoavulavirus (AOaV-1) Isolates from Duck (Anas platyrhynchos) and Parrot (Eclectusroratus).

Newcastle disease (ND) is a highly contagious viral disease of poultry causing significant economic losses worldwide. Vaccination is considered the most reliable approach to curb the economic menace that is ND, but the thermolabile nature of Newcastle disease virus (NDV) vaccination poses a significant threat to its protective efficacy. This study aimed to profile the thermostability of NDV isolates from duck (As/Km/19/44) and parrot (As/WB/19/91) and evaluate their immunogenic potential in chicks. Fusion protein cleavage site (FPCS) and phylogenetic analysis demonstrated the lentogenic nature of both the isolates/strains and classified them as class II genotype II NDV. The characterized NDV isolates were adapted in specific-pathogen-free (SPF) chicks by serially passaging. Biological pathogenicity assessment of chicken-adapted As/Km/19/44 (PSD44C) and As/WB/19/91 (PSP91C) revealed both the isolates to be avirulent with a mean death time (MDT) of more than 90 h and an intracerebral pathogenicity index (ICPI) ranging from 0.2 to 0.4. Both of the NDV isolates displayed varied thermostability profiles. PSD44C was the most thermostable strain as compared to PSP91C and the commercially available LaSota vaccine strain. The immunogenicity of PSD44C and LaSota was significantly higher than PSP91C. Based on these results, it is concluded that NDV isolate PSD44C is more thermostable and immunogenic when administered intraocularly without any adverse effects. Therefore, PSD44C is suitable for further research and vaccine development.

Consideration of genetic variation and evolutionary history in future conservation of Indian one-horned rhinoceros (Rhinoceros unicornis).

BACKGROUND: The extant members of the Asian rhinos have experienced severe population and range declines since Pleistocene through a combination of natural and anthropogenic factors. The one-horned rhino is the only Asian species recovered from such conditions but most of the extant populations are reaching carrying capacity. India currently harbours ~ 83% of the global wild one-horned rhino populations distributed across seven protected areas. Recent assessments recommend reintroduction-based conservation approaches for the species, and implementation of such efforts would greatly benefit from detailed genetic assessments and evolutionary history of these populations. Using mitochondrial data, we investigated the phylogeography, divergence and demographic history of one-horned rhinos across its Indian range. RESULTS: We report the first complete mitogenome from all the extant Indian wild one-horned rhino populations (n = 16 individuals). Further, we identified all polymorphic sites and assessed rhino phylogeography (2531 bp mtDNA, n = 111 individuals) across India. Results showed 30 haplotypes distributed as three distinct genetic clades (Fst value 0.68-1) corresponding to the states of Assam (n = 28 haplotypes), West Bengal and Uttar Pradesh (both monomorphic). The reintroduced population of Uttar Pradesh showed maternal signatures of Chitwan National Park, Nepal. Mitochondrial phylogenomics suggests one-horned rhino diverged from its recent common ancestors ~ 950 Kya and different populations (Assam, West Bengal and Uttar Pradesh/Nepal) coalesce at ~ 190-50 Kya, corroborating with the paleobiogeography history of the Indian subcontinent. Further, the demography analyses indicated historical decline in female effective population size ~ 300-200 Kya followed by increasing trends during ~ 110-60 Kya. CONCLUSION: The phylogeography and phylogenomic outcomes suggest recognition of three 'Evolutionary Significant Units (ESUs)' in Indian rhino. With ongoing genetic isolation of the current populations, future management efforts should focus on identifying genetically variable founder animals and consider periodic supplementation events while planning future rhino reintroduction programs in India. Such well-informed, multidisciplinary approach will be the only way to ensure evolutionary, ecological and demographic stability of the species across its range.

Isolation of genotype VII avian orthoavulavirus serotype 1 from barn owl from Northeast India.

Newcastle disease virus (NDV) affects commercial poultry as well as other avian species in the wild and in captivity. Although the diversity of NDV in domestic chickens has been well understood, little light has been shed on NDV outbreaks in other avian species. We provide an annotated sequence of NDV/Owl/Guwahati/01/20, a virulent strain of NDV isolated from barn owls in captivity from Guwahati in Northeast India. The complete genome is 15,192 base pairs long with a fusion protein (F) cleavage site 112KRQKR↓F117. The isolate showed 97.67% identity with its closest match, another highly virulent strain from Indonesia isolated from vaccinated commercial chickens; however, they differ in the F cleavage site. The NDV isolate from the owl shares 83.02% and 81.88% identity with the vaccine strains R2B and LaSota, respectively. Phylogenetic analysis with F gene as well as whole-genome nucleotide sequence reveals that the NDV isolate from owl belongs to genotype VII, subgenotype VII.2, and differs significantly from all other isolates of NDV from India.

Gut microbiome architecture of wild greater one-horned rhinoceros:a vulnerable species from Kaziranga National Park, India.

Rhinoceros unicornis, also known as the greater one-horned rhinoceros (GoHR), is a vulnerable wildlife species found in the Indian subcontinent with an estimated global population of 3582, of which an estimated 2995 resides in India. The Kaziranga National Park of Assam is the home to ~80.56% of the GoH population in India. Recent advances in genetics and microbial studies underscored the importance of gut microbial symbiosis as a crucial factor for host metabolic health and environmental interaction, particularly for higher mammals. Alteration of the normal microbiome can also be an indicator of chronic disease and infection. Freshly voided dung samples from nine dung heaps of free ranging or wild GoH rhinoceros were collected from Kaziranga National Park for mapping the gut microbial architecture through 16S-metagenomic approach. In our sample, the GoH gut harbours 168.8±12.55 (SE) bacteria-specific OTUs belonging to 21 phyla of which the gram-negative Proteobacteria is the most abundant phyla. Other abundant phylas found in the GoH gut are Firmicutes and Bacteroidetes. Although the GoH rhinoceros gut can utilize fibrous plant by microbial fermentation, the aerobic, nonfermenting Acinetobacter (20.7%), Stenotrophomonas (17.8%) and Brevundimonas (9.1%) constitute about 50% of all identified genus. Functional prediction of the GoH microbiome reveals that>50% of the bacteria present are involved in metabolism followed by cellular processes and information processing. A significant proportion (>1%) are associated with different diseases. In summary, our study characterized bacterial communities of nine wild GoH to identify some unique features and its implication in disease and survival of GoH.

Evidence of Transmission of Goatpox between Domestic Goats and Wild Himalayan Goral (Naemorhedus goral) in Arunachal Pradesh, India.

Examination of carcasses of Himalayan goral (Naemorhedus goral) revealed nodular, pox-like eruptions in the skin. Similar disease was also seen in domestic goats (Capra aegagrus hircus) in the same area. Goatpox virus was identified as the etiology of the disease in both cases, with probable transmission between the species.

Listeriosis in a peri-urban area: Cultural and molecular characterization of Listeria monocytogenes isolated from encephalitic goats.

BACKGROUND AND AIM: Listeriosis in food animals bears a significant threat to human health. Detailed investigations into the cause facilitate proper management of the disease. This study reports the cultural, pathological, and molecular characterization of Listeria monocytogenes isolated from encephalitic goats from peri-urban Guwahati, Assam. MATERIALS AND METHODS: Out of nine suspected samples, five positive isolates of L. monocytogenes were subjected to bacteriological, biochemical, and molecular tests. The genus and species-specific L. monocytogenes 16S rRNA and prs genes were amplified by polymerase chain reaction (PCR) to yield 1200 and 370 bp sized products, respectively. The encephalitic form of the disease was characterized by circling movement, high fever, and terminal recumbence. RESULTS: All the five isolates were confirmed to be L. monocytogenes based on PCR amplification of genus and species-specific 16S rRNA and prs gene products. The isolates were sensitive to ciprofloxacin, oxytetracycline (OTC), and norfloxacin, but resistant to doxycycline and erythromycin. A high dose of OTC was used in a goat at the early stage of clinical symptom and the animal recovered clinically. CONCLUSION: Listeriosis in goats could pose a significant public health threat as the meat (occasionally milk) or meat products from goats are widely consumed by the people of Assam. Understanding the molecular epidemiological aspects of L. monocytogenes infections of food animal species should, therefore, be the priority in this part of the country.

Emergence of oriental theileriosis in cattle and its transmission through Rhipicephalus (Boophilus) microplus in Assam, India.

AIM: The aim of the present study was to investigate the presence of Theileria in blood samples of crossbred and indigenous adult cows raised under unorganized small scale farming system in a Babesia and Anaplasma endemic geographical area from Assam, India and to see its transmission through Rhipicephalus (Boophilus) microplus ticks. MATERIALS AND METHODS: For the present study, 57 clinical cases of cattle suspected to be of hemoparasitic infections were taken into consideration. The parasites were identified based on morphology in giemsa stained blood smear followed by polymerase chain reaction (PCR). Sera samples were tested for T. annulata antibodies in plate and Dot-ELISA. PCR was also conducted in eggs of Rhipicephalus (Boophilus) microplus tick collected from a Theileria orientalis positive animal. RESULTS: PCR amplified 1124, 776, and 160 bp DNA fragments of B. bigemina (64.91%), T. orientalis (21.05%) and A. marginale (14.03%), respectively. This assay further conducted in 12 T. orientalis positive blood samples with primers of Buffeli, Chitose, and Ikeda variants of T. orientalis showed 3 samples positive to Ikeda type and none for Buffeli and Chitose. Babesia bovis and Theileria annulata specific primers also did not amplify any fragment during the PCR assay of the blood samples. Further, all sera samples tested negative to T. annulata antibodies in Plate and Dot-ELISA. PCR conducted in eggs of R (B).microplus tick collected from a T. orientalis positive animal revealed presence of the parasite DNA. Gradual improvement in physical condition leading to complete recovery in 10 out of 12 T. orientalis infected clinical cases treated with buparvaquone(at 2.5mg/kg.b.wt I/M) was the feedback obtained from field veterinarians and the cattle owners. CONCLUSION: The present investigation represents the first report of occurrence of T. orientalis in cattle of Assam with involvement of pathogenic Ikeda strain in clinical outbreaks and its possible natural transmission by R (B). microplus through the transovarian mode.