Production and characterization of biologicals for disease diagnosis and pathological evaluation of elephant endotheliotropic herpesvirus (EEHV).

Elephant endotheliotropic herpesviruses (EEHV) belong to the family Herpesviridae and cause a highly fatal hemorrhagic infection in elephants. EEHV poses a global threat to the already endangered elephant population. Since EEHV is a non-cultivable virus, there is a scarcity of specific diagnostics, therapeutics, and vaccines. In this study, our objective was to develop biologicals for diagnosis and pathological studies against the most prevalent EEHV1A/1B. We expressed two truncated fragments of the DNA polymerase, glycoprotein B (gB), and glycoprotein (gL) of EEHV in the prokaryotic system. Hyperimmune serum against the purified antigens was raised in rabbits and guinea pigs. We validated the reactivity of this hyperimmune serum using western blotting, ELISA, and immune-histochemistry on known positive infected tissues. Samples collected from 270 animals across various states in India were evaluated with these biologicals. The raised antibodies successfully demonstrated virus in immune-cytochemistry. Additionally, all known positive samples consistently exhibited significant inhibition in the OD values when used in the competitive format of ELISA across all four antigens when compared to the serum collected from known negative animals. An apparent sero-prevalence of 10 % was observed in the randomly collected samples. In summary, our study successfully developed and validated biologicals that will be invaluable for EEHV diagnosis and control.

High mortality in free-ranging wild boars associated with African swine fever virus in India.

The present study focuses on the pathological and molecular characterization of African swine fever virus (ASFV) associated with an outbreak in wild boars in two national parks in southern India in 2022-2023. Significant mortality was observed among free-ranging wild boars at Bandipur National Park, Karnataka, and Mudumalai National Park, Tamil Nadu. Extensive combing operations were undertaken in both national parks, spanning an area of around 100 km2, originating from the reported epicenter, to estimate the mortality rate. Recovered carcasses were pathologically examined, and ASFV isolates was genetically characterized. Our findings suggested spillover infection of ASFV from nearby domestic pigs, and the virus was equally pathogenic in wild boars and domestic pigs. ASFV intrusion was reported in the Northeastern region of the country, which borders China and Myanmar, whereas the current outbreak is very distantly located, in southern India. Molecular data will help in tracing the spread of the virus in the country.

Development and evaluation of recombinant gD protein based ELISA for sero-surveillance of BoHV-1 in India.

Bovine herpes virus-1 (BoHV-1) is responsible for production losses through decreased milk yields, abortions, infertility, and trade restrictions in the bovine population. The disease is endemic in many countries including India. As the virus harbors a unique feature of latency animals once infected with the virus remain sero-positive for lifetime and can re-excrete the virus when exposed to stressful conditions. Hence, identification and culling of infected animals is only the means to minimize infection-associated losses. In this study, an economical indigenous assay for the detection of BoHV-1 specific antibodies was developed to cater to the huge bovine population of the country. The viral structural gD protein, expressed in the prokaryotic system was used for optimization of an indirect ELISA for bovines followed by statistical validation of the assay. The diagnostic sensitivity and specificity of the indirect ELISA were 82.9% and 91.3% respectively. Systematically collected serum samples representing organized, unorganized and breeding farms of India were tested with the indigenously developed assay for further validation.

An observational study on carbapenem-resistant Enterobacterales (CRE) colonisation and subsequent risk of infection in an adult intensive care unit (ICU) at a tertiary care hospital in India.

Background: Carbapenem-resistant Enterobacterales (CRE) are a global health problem with a growing prevalence. India has a high prevalence of CRE. CRE infections are difficult to treat, and are associated with significant morbidity and mortality. Colonisation is generally a prerequisite for infection and the prevention of CRE colonisation is key to the prevention of CRE infection. Objectives: To determine the prevalence of CRE colonisation and subsequent infections in an adult intensive care unit (ICU) in India. Methods: We conducted a prospective observational study in which perirectal swabs were obtained along with relevant clinical details of consenting adult patients upon ICU admission between January 2019 and August 2020. Rectal screening was performed using MacConkey agar plates with ertapenem disks and further identification was performed using conventional microbiological techniques. Ertapenem minimum inhibitory concentration (MIC) was determined using an epsillometer (E) test. The modified carbapenem inactivation (mCIM) test and EDTA carbapenem inactivation test (eCIM) were performed to confirm carbapenem resistance using the Clinical Laboratory Standards Institute (CLSI) 2020 guidelines. Results: 192 ICU patients were screened for CRE. 37 patients were found to be colonised with CRE. Klebsiella pneumoniae (N=25; 67.6%) was the most frequent CRE isolate, followed by Escherichia coli (N=11; 29.7%) and one Enterobacter species (N=1; 2.7%). 89.2% (33/37) patients developed CRE infection. Pneumonia was the most common CRE infection identified in 12/33 (36.4%) patients.during the hospital stay. The median duration of hospital stay was longer (17 days) for CRE colonised compared to CRE non-colonised patients (9 days) (P<0.001). Death occurred in 27 % (N=10/37) of CRE-colonised patients during the hospital admission. Conclusion: CRE colonisation is associated with high risk of subsequent CRE infection and longer ICU and hospital admission.

Refractory giardiasis in a child with steroid-resistant nephrotic syndrome.

Giardiasis is an infection of the small intestine caused by the protozoan parasite Giardia intestinalis and one of the most common parasitic intestinal diseases in humans worldwide. It mainly manifests as a self-limited illness in the case of immunocompetent patients and usually does not require treatment. However, immunodeficiency is a risk factor for the onset of severe Giardia infection. In this report, a case of recurrent giardiasis refractory to nitroimidazole therapy is presented. A 7-year-old male patient with steroid-resistant nephrotic syndrome came to our hospital because of chronic diarrhoea. The patient was on long-term immunosuppressive therapy. Microscopic examination of stool showed a significant number of trophozoites and cysts of G. intestinalis. Treatment with metronidazole for longer duration than recommended has failed to clear the parasite in the present case.

Cryptosporidiosis in India and the World: A Review.

Cryptosporidiosis is caused by infection with a coccidian parasite belonging to the genus Cryptosporidium. Initially, human cryptosporidiosis was believed to be caused only by one species, but since the advent of molecular studies, 15 more species have been discovered to cause this infection. Among them, Cryptosporidium hominis and Cryptosporidium parvum are the most common species involved. This mainly affects children and causes diarrhea in most cases. It is mainly diagnosed by microscopy, especially in low-middle-income countries. This review covers the epidemiology, life cycle, risk factors, clinical manifestations, different diagnostic methods and treatment of this disease.

A retrospective study showing a high rate of seropositivity against SARS-CoV-2 in wild felines in India.

We report a high rate of seropositivity against SARS-CoV-2 in wild felines in India. Seropositivity was determined by microneutralization and plaque reduction neutralization assays in captive Asiatic lions, leopards, and Bengal tigers. The rate of seropositivity was positively correlated with that of the incidence in humans, suggesting the occurrence of large spillover events.

Pathological and molecular studies on elephant endotheliotropic herpesvirus haemorrhagic disease among captive and free-range Asian elephants in India.

In the present research pathology and molecular diagnosis of elephant endotheliotropic herpes virus-haemorrhagic disease (EEHV-HD) among Asian elephants was studied. Out of 76 cases, 20 were positive for EEHV infection in PANPOL and POL1 based semi-nested PCR. Out of 20 samples, 10 samples were fatal cases of EEHV-HD while 10 were of either subclinical or latent infection. Acute onset haemorrhagic disease with EEHV-HD had anorexia, facial and neck swelling, cyanotic buccal mucosa and tongue, nasal and ocular discharge, and colic. The hallmark of gross finding in all cases were severe haemorrhagic lesions in the internal organs viz. cyanosis of tongue with multifocal petechial haemorrhages, diffuse epicardial and endocardial haemorrhages, swollen liver (rounded edges) with parenchymal haemorrhages, serosal and mucosal haemorrhages in gastrointestinal tract, congested kidneys with corticomedullary haemorrhages, highly congested meninges, and brain capillaries with haemorrhages. Microscopic findings in all the cases had severe vascular changes in the visceral organs. Microthrombi was present in the vasculature of tongue, heart, lung, liver, kidney, and brain. The endothelial lining of most of the blood vessels were swollen with apoptotic changes. Amphophilic to basophilic intranuclear inclusion bodies were observed in the endothelial cells. Immunostaining using anti-EEHV DNAPOL hyperimmune sera revealed intense positive signals in the endothelium of blood vessels and their walls. Quantification of viral load in necropsy tissue samples revealed highest in the heart (7.4 × 106/µg of sample) and least in the brain (9 × 103/µg of sample). The PCR amplicons from EEHV1 specific genes (POL1(U38) and TER were subjected to partial genome sequencing which had 99.9% similarity with the EEHV1A subtype. It was concluded that Asian elephants in India are latently infected for EEHV1 and in all the fatal EEHV-HD cases, EEHV1A subtype was the causative agent with characteristic pathomorphological changes in visceral organs.

Detection of Antibodies against Toxoplasma from Human Serum Sample using ELISA.

BACKGROUND: Toxoplasmosis is a common worldwide zoonotic infection affecting warm blooded animals and humans caused by Toxoplasma gondii. Clinical features range from mildfebrile illness and lymphadenopathy in the immunocompetent host to encephalitisin the immunosuppressed host (E.g. HIV infected patients). Congenital infection can also occur. For effective control and treatment of toxoplasmosis, accurate detection of T. gondii infection is important. OBJECTIVE: In this study, ELISA detecting anti-toxoplasma antibodies IgM and IgG has been used for diagnosing Toxoplasmosis in patients. METHODS: Enzyme-linked immunosorbentassay (ELISA) was carried out in serum samples collected from 40 patients to detect anti-toxoplasma IgG and IgM antibodies as a part of work up in suspected cases. Relevant clinical history was also taken. RESULTS: Of the total 40 samples taken, only one sample came positive for IgM and 9 came positive for IgG antibody. All patients who were seropositivefor T. gondii antibodies had HIV infection. Five patients were in the age group between 30 to 40 years. CONCLUSION: From the limited data available in this study, it may be recommended to screen for T. gondii antibodies in HIV patients.

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.

Influences of a novel cylindrical solar dryer on farmer's income and its impact on environment.

Poor grain drying facilities, along with the burden of drying cost, bound the farmers to market their produce soon after harvest. Thus, this research paper intends to study the influences of a novel cylindrical solar-assisted dryer on farmers' income and its impact on the environment. The paper also presents the design and fabrication of a drying system for maize cobs using non-conventional solar energy. The performance of the solar-assisted drying system was also investigated for drying of yellow dent maize cobs. As a very energy-intensive post-harvest process, drying consumes a lot of electricity, which is usually provided by conventional energy. Here, solar dryers are the perfect solution in terms of efficiency, uniform drying of agricultural food products, less drying time, increased marketability of agricultural products, and reducing the load on farmer's pocket for drying. With a high internal rate of return of 66 percent, the designed dryer proved to be technically and economically viable. Compared to open sun drying, the solar drying system produced better quality and drying time results. Compared to other models, the Midilli model fits the experimental maize drying data better, with a coefficient of determination of R2 = 0.89729. Energy analysis inferred savings of 1352.97 kWh electrical energy and 128.18 liters of diesel fuel plus a reduction of 1.22 t CO2 per annum can be achieved by using this dryer.