Epidemiology, genetic characterization and population structure analysis of lumpy skin disease virus from North India.

Lumpy skin disease (LSD) is a transboundary, viral disease of cattle with a significant economic impact on the livestock industry. This study describes the epidemiological investigations of outbreaks of LSD that occurred in 2022 in three Indian states viz., Haryana, Himachal Pradesh and Rajasthan and the genetic characterization of Lumpy skin disease viruses (LSDVs). Also, the population structure analysis of LSDVs was carried out. Out of 138 scab samples tested from suspected cattle for LSDV, 106 were found positive. Of these, nine representative scab samples were further genetically characterized. Phylogenetic analysis based on the P32 and EEV gene sequences depicted that the LSDV strains of the present study had nucleotide identity of 100% and 99.10-99.45%, respectively with the LSDV/2019 field strains of India. Multiple sequence alignment of the EEV glycoprotein gene sequences revealed nucleotide polymorphisms at three positions viz., G178A, G253A and A459G in circulating LSDV/2022 field strains. Based on the median joining network analysis of the EEV gene, 12 haplotypes were identified among the LSDV populations. Population structure analysis corresponding to the EEV gene revealed high haplotype (0.8486 ± 0.026) and low nucleotide diversities (0.00636 ± 0.0006) and negative values for neutrality indices, indicating a high number of closely related haplotypes and the studied population may have undergone a recent expansion. The findings will help in understanding the distribution and dynamics of LSDV populations, which will prove pivotal in designing and implementing effective management and control strategies.

Therapeutics and prophylactic efficacy of novel lytic Escherichia phage vB_EcoS_PJ16 against multidrug-resistant avian pathogenic E. coli using in vivo study.

Avian pathogenic Escherichia coli (APEC) is the causative agent of avian colibacillosis, which causes significant economic losses to the poultry industry. The growing resistance of bacteria to antibiotics is a major global public health concern. However, there is limited data on the efficacy of phage therapy in effectively controlling and treating APEC infections. In this study, a novel lytic Escherichia phage, vB_EcoS_PJ16, was isolated from poultry farm wastewater and characterized in both in vitro and in vivo conditions. Transmission electron microscopy analysis revealed the presence of an icosahedral head and a long non-contractile tail, classifying the phage under the Caudoviricetes class. Host range determination showed that Escherichia phage vB_EcoS_PJ16 exhibited lytic activity against multiple strains of pathogenic E. coli, while no significant signs of lysis for Klebsiella pneumoniae, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus. Biophysical characterization revealed that the isolated phage was sturdy, as it remained viable for up to 300 days at temperatures of 30 °C, 37 °C, and 42 °C and for up to 24 h at pH 5 to 11, with only minor changes in titer. Kinetic analysis at multiplicity of infection (MOI) 0.1 showed a latency period of about 20 min and a burst size of 26.5 phage particles per infected cell for phage vB_EcoS_PJ16. Whole genome sequencing unveiled that the phage vB_EcoS_PJ16 genome consists of a double-stranded linear DNA molecule with 57,756 bp and a GC content of 43.58%. The Escherichia phage vB_EcoS_PJ16 genome consisted of 98 predicted putative ORFs, with no transfer RNA identified in the genome. Among these 98 genes, 34 genes were predicted to have known functions. A significant reduction in APEC viability was observed at MOI 100 during in vitro bacterial challenge tests conducted at different MOIs (0.01, 1, and 100). In vivo oral evaluation of the isolated phage to limit E. coli infections in day-old chicks indicated a decrease in mortality within both the therapeutic (20%) and prophylactic (30%) groups, when compared to the control group. The findings of this study contribute to our current knowledge of Escherichia phages and suggest a potentially effective role of phages in the therapeutic and prophylactic control of antibiotic-resistant APEC strains.

Identification and assessment of stress and associated stressors among veterinary students in India using a cross-sectional questionnaire survey.

Background: Veterinary education, is a rigorous professional training program, which exposes students to significant academic and non-academic pressures. The identification of stressors and stress levels among veterinary students mighty help the designing and implementation of coping strategies to protect the students' mental health. Methods: A 44-item based cross-sectional questionnaire survey was prepared and disseminated among veterinary students in India to identify the stressors responsible, measure the amount of stress, and relate stress to characteristics like gender, degree year, and family income. A total of n = 611 veterinary students across 14 states including 27 colleges/universities participated in the study. The collected data was evaluated for sampling adequacy, construct validity, and reliability using a set of statistical tests. Results: The analysis revealed high sampling adequacy with a KMO value of 0.957 and a highly significant anti-image correlation (p < 0.001). The principal component analysis generated six factors or subscales which effectively explained 51.98% of the variance in the data, depicting high construct validity. The Cronbach's alpha value of 0.957 revealed high internal consistency for the questionnaire. Analysis revealed more than 94% of pupils under stress, with levels ranging from moderate to severe. Academic-related stressor (95.58%) was the leading cause of overall stress in the present study followed by inter- and intrapersonal and career related-stressors (93.12%) and exams and evaluation-related stressor (90.99%). In comparison to male students, female students reported significantly higher levels of overall stress, academic stress, and intrapersonal and interpersonal stress (p < 0.001) using Chi-square. The students from lower-income families experienced significantly higher overall stress as well as stress due to family responsibilities (p < 0.001). The first-year undergraduate students reported significantly higher (p < 0.001) stress due to family responsibilities-related stressors whereas second-year students due to social activities-related stressors. The hierarchal regression model predicted that gender, family income, academic-related stressors, inter- and intrapersonal and career-related stressors, and social activities-related stressors can be employed to evaluate overall stress among students, as they ensured the maximum variance in the data (p < 0.001). Conclusions: To the best of our knowledge, this is the first Indian study to identify stressors, quantify associated stress and predict major attributes to be targeted in future studies for veterinary students.

Detection of Leptospira in urine of apparently healthy dogs by quantitative polymerase chain reaction in Haryana, India.

BACKGROUND AND AIM: Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira. The organism can spread through the urine of infected animals, which can get into water or soil and can survive there for weeks to months. The study was undertaken to detect the pathogenic Leptospira in healthy dogs' urinary shedding by real-time polymerase chain reaction (qPCR). MATERIALS AND METHODS: Leptospirosis is a zoonotic disease caused by bacteria of the genus Leptospira. To detect the pathogenic Leptospira organisms in dogs' urinary shedding, 239 urine samples were collected from healthy dogs from April 2018 to March 2019 from different areas of Haryana. All the urine samples were processed for DNA extraction and qPCR technique was used to detect the presence of Leptospira. RESULTS: Out of 239 urine samples of dogs, none of the samples resulted in the detection of DNA of pathogenic Leptospira organisms. CONCLUSION: The present study indicated low risk of transmission of Leptospira organisms from dogs' urine to human beings in Haryana.

Emerging status of anaplasmosis in cattle in Hisar.

AIM: Anaplasma marginale is a rickettsial pathogen responsible for progressive anemia in ruminants leading to huge economic losses. The objectives of this cross-sectional study were to determine the prevalence of anaplasmosis and therapeutic evaluation of traditional line of treatment. MATERIALS AND METHODS: A total of 168 cattle presented to Teaching Veterinary Clinical Complex, Lala Lajpat Rai University of Veterinary & Animal Sciences, Hisar during the period of 3 months (July-September, 2014) with history of fever, anorexia, reduced milk yield and tick infestation were analyzed for prevalence of hemoprotozoan diseases using classical giemsa stained thin blood smear parasitological method. RESULTS: Out of these 168 animals, 7 (4.17%) were found to be suffering from anaplasmosis on the basis of presence of dense, rounded, intra-erythrocytic bodies situated on or near the margin of the erythrocytes. Overall prevalence of theileriosis and babesiosis were found to be 42.9% and 1.8%, respectively. Level of parasitemia was noticed to be 1.2%, 0.8% and 0.9% in babesiosis, theileriosis, and anaplasmosis, respectively. The most marked and common clinical signs reported in all the cases were severe anemia (hemoglobin=3-6 g/dl) and history of fever, followed by normal body temperature. Following treatment with oxytetracycline parenterally along with supportive therapy out of seven cases six got recovered without any side-effects. CONCLUSION: The current study indicates the emerging status of anaplasmosis in this part of the country as during the past few years there are very few reports showing the prevalence of clinical cases of anaplasmosis. Treatment with oxytetracycline yielded excellent result showing recovery in most of the clinical cases.