Porcine astrovirus (PAstV) is a small, non-enveloped virus of the genus Mamastrovirus within the family Astroviridae. Its genome consists of a positive-sense, single-stranded RNA of approximately 6.4–7.3 kb in length. PAstV displays considerable genetic diversity and is presently categorized into five distinct genotypes. While primarily recognized as an enteropathogen linked to varying degrees of gastroenteritis in pigs, its precise role in inducing diarrhea remains uncertain, as it has been detected in both diarrheic and healthy pigs with a similar prevalence. In addition to gastrointestinal disease, PAstV has recently been detected in pigs with respiratory and nervous disorders. The current review provides a comprehensive summary of the published literature on the PAstV genome, viral replication, epidemiology, geographical distribution, pathogenesis, clinical manifestations, diagnostic methods, co-infections, and probable cross-species transmission. Furthermore, it underscores the gaps in the current understanding, serving as a valuable reference for future investigations into the PAstV.

In India, despite the documented presence of porcine parvoviruses (PPV1 and PPV3), only a limited number of studies reveal the genetic and evolutionary characteristics of PPV1, with no report on PPV3 genetic analysis. The VP2 protein, which plays a pivotal role as a critical antigenic domain and encodes the structural capsid protein of PPVs, serves as the focal point in this investigation. We delved into the genetic characterization of highly immunogenic VP2 gene fragments in 84 samples for PPV1 and PPV3 isolates, which were found to have a prevalence of 14.28

Introduction:Bluetongue virus (BTV) is a species of genus Orbivirus belonging to the Sedoreoviridae family. Bluetongue (BT) is endemic in India and responsible for causing significant economic losses to livestock farmers. In India, antibodies to BTV serotype 24 (BTV-24) have been reported in 2005; it was first isolated in 2010, and it caused several outbreaks in sheep during 2012-2014. The in vivo studies investigating the pathogenetic potential of various BTV serotypes in the susceptible host sheep are scarce. Furthermore, detailed investigations to elucidate the pathogenetic mechanisms of BTV-24 under experimental conditions in sheep are not available. Because of its impact on the livestock economy, the present study was undertaken for the first time to explore the infection kinetics, pathology, pathogenesis, and immune responses against the Indian isolate of BTV-24 in sheep under experimental conditions. Methods:Six native sheep were infected intradermally with BTV-24 at 106 TCID50/mL concentration, and six sheep were inoculated with uninfected cell culture fluid. Animals were euthanized at 4, 7, 11, 16, 45, and 60 days post-inoculation (DPI). The sequential pathology, BTV localization by immunohistochemistry, BTV quantification by quantitative PCR (qPCR), immune cell kinetics [CD4+ and CD8+ T lymphocytes in peripheral blood mononuclear cells (PBMCs), prescapular lymph node (PSLN), and spleen] by fluorescence-activated cell sorting (FACS), and cytokine estimation by qRT-PCR were studied. Results:The BTV-24-infected animals showed pyrexia, conjunctival and oral mucosal congestion, cyanosis of tongue, serous to catarrhal nasal discharge, and viremia. Gross pathological lesions were observed in the lymph nodes, lungs, and kidneys, with the lymph nodes being enlarged, edematous, and hemorrhagic. Subintimal hemorrhage at the base of the pulmonary artery (pathognomonic lesion of BT) was observed at 7 DPI. Histopathological lesions were prominent in lymph nodes, spleen, heart, lungs, and cerebral endothelium. Severe hemosiderosis in spleen, and hemorrhages and hyalinization of tunica media in pulmonary artery at 7 DPI were observed. Development of clinical signs and gross and histopathological lesions in BTV-24-infected animals emphasized the moderate progression of disease and enhanced virulence of the serotype. Humoral immune response was significantly high at 5, 11, 16, 21, 45, and 60 DPI. Cell-mediated immune response-like kinetics of CD4+ and CD8+ T lymphocytes showed a sharp decline during the early stage and an increase of CD8+ T lymphocytes during later stages of infection. BTV antigen was detected consistently in tongue, thymus, trapezius muscle, heart, and pulmonary artery by immunohistochemistry and qPCR. Significant changes in the levels of cytokines [interferon-alpha (IFN-α), IFN-β, IFN-γ, interleukin-2 (IL-2), IL-12, and tumor necrosis factor-alpha (TNF-α)] and upregulated expression of apoptotic markers, B-cell lymphoma-2 (Bcl-2), and caspase-3 in the spleen and lymph nodes were correlated with peak viremia. Conclusion:The results of this study can be used to formulate effective preventive and control measures and to develop a suitable vaccine against BTV-24 to minimize economic losses.

Neonatal calf diarrhea is a serious health issue in dairy calves with significant economic losses. The immature gut immunity and the inadequate passive transfer of immunity in neonates favour the transmission and replication of the bovine rotavirus A (RVA) and the apicomplexan Cryptosporidium spp., and others, causing significant morbidity and mortality. To prevent such losses against these major pathogens, the efficacy of two-strain probiotics, namely Lacticaseibacillus (L.) rhamnosus strain NCDC 610 and Bifidobacterium (B.) bifidum strain BbVK3, were explored in a laboratory mouse model. A total of 120 seven-day-old Swiss albino suckling mice were grouped into six: Gr I (pups gavaged with RVA, 106.5TCID50/ml @ 100 μl/pup on 0-day), Gr II (gavaged with C. parvum oocysts @ 105 on 0-day), Gr III (pups infected with RVA on 0-day followed by C. parvum oocysts @ 105 after 36 h of infection), Gr IV (pups pre-treated with10 μl of cell-free supernatant of two-strain probiotic mix (L. rhamnosus strain NCDC 610 @ 108 CFU/ml and B. bifidum strain BbVK3 @ 108 CFU/ml) from 2 days prior to mixed infection till 7 day post-infection with RVA and C. parvum), Gr V (two-strain probiotic mix from day one to day 7 post-mixed infections), and Gr VI (sterile PBS). The mice were observed daily for the clinical signs of diarrhea and 4 mice from each group were sacrificed on 3, 5, 7, 11, and 16 day post-infection for sample collection. The samples were analyzed for the counting of the C. parvum oocyst in the feces, gross and histopathological lesions in the ileum, RVA antigen in the distal ileum by immunohistochemistry, cellular apoptosis by TUNEL assay, bacterial loads in the ileocaecal and rectal contents by culturing, RVA quantification and expression levels of TLR-2, TLR-3, TNF-α, MUC-2, INF-γ, IL-12, TGF-β, and IL-10 genes in distal ileum by the Real time-PCR, and the sIgA in fecal contents and serum by ELISA. The prophylactic group (Gr IV) showed reduced severity and duration of diarrhea, reduced severity of intestinal lesions and apoptosis, reduced viral load and oocyst count, reduced levels of TNF-α, and higher Il-10, and TGF-β levels, higher levels of innate immune response (higher levels of INF-γ, TLR-3, TLR-2, and reduced levels of Il-12), and up-regulation of MUC-2 genes as compared to infected (Grs 1, II, III) and therapeutic (Gr V) groups. The study showed promising prophylactic potential of the two-strain probiotic mix of L. rhamnosus strain NCDC 610 and B. bifidum strain BbVK3 against the mixed infections of rotavirus A and C. parvum.

Detailed spatiotemporal pathogenesis data for virulent Indian Goatpox virus (GTPV) isolates in the natural host remain limited. This study characterized systemic dissemination and tissue tropism of the GTPV Mukteshwar strain following experimental intradermal inoculation (106 SRD50) in goats. Sixteen GTPV-seronegative goats (12-21 months) were monitored longitudinally, with replicated serial necropsies conducted from 3 to 35 days post-infection (dpi). Viral dissemination was quantified by real-time PCR targeting the RPO147 gene, and tissue lesions and antigen localization were evaluated by gross pathology, histopathology, immunohistochemistry, and special staining. Infected goats developed fever, lymphadenopathy, and characteristic cutaneous lesions after an incubation period of 3-11 days. Primary replication was detected at the inoculation site by 3 dpi, followed by viremia between 5 and 14 dpi, with peak blood viral loads of 1.64-3.55 log10 DNA copies/μL during 8-11 dpi. Systemic dissemination involved skin, lymphoid organs, respiratory and gastrointestinal tissues, urinary system, and male reproductive organs, while central nervous system tissues remained negative. Viral shedding occurred predominantly via conjunctival and nasal secretions between 8 and 21 dpi, peaking at 5.04 and 4.24 log10 DNA copies/μL, respectively, at 11 dpi, with intermittent urinary detection reaching 2.11 log10 DNA copies/μL at 14 dpi. Persistent antigen and/or viral DNA detection in respiratory and gastrointestinal tissues beyond clinical recovery indicates tissue-specific viral maintenance. Defined viremia peaks and persistence windows establish a kinetic framework for systemic dissemination and provide quantitative benchmarks for vaccine efficacy assessment, surveillance, and control strategies in endemic regions.

Bluetongue disease remains a global economic threat, necessitating the development of rapid, high-precision diagnostics. The genome of the Bluetongue virus (BTV) encodes seven structural proteins, VP1–VP7. Among these proteins, VP7 is the major immunodominant structural protein conserved across BTV serotypes and is therefore targeted for the development of serogroup-specific immunodiagnostic assays. This study aimed to identify an immunodominant VP7 epitope recognized by BTV-specific monoclonal antibodies (MAbs) and to evaluate its utility in a peptide-based ELISA. The gene encoding VP7 protein were expressed in three overlapping fragments in a prokaryotic system and analyzed for its reactivity with the two MAbs by Western blot and indirect ELISA. By dissecting the VP7 gene into overlapping recombinant fragments, we identified the binding domain is located in Fragment II (27 kDa) region. However, linear monomeric peptides designed from this region failed to replicate the protein’s native antigenicity and did not show any reactivity with the MAbs. To overcome this, we designed a chimeric peptide bridging the junction of Fragments II and III and harnessed Multiple Antigenic Peptide (MAP) technology to present the epitope in a four-armed dendrimeric scaffold. An indirect ELISA based on the MAP antigen was subsequently evaluated using field serum which demonstrated diagnostic sensitivity of 92.3

The river ecosystems provide habitats and source of water for a number of species including humans. The uncontrolled accumulation of pollutants in the aquatic environment enhances the development of antibiotic-resistant bacteria and genes. Water samples were collected seasonally from different sites of Gomti and Ganga River. Bacteria were isolated by plating on nutrient agar supplemented with individual antibiotics (100 µg/ml) to select the resistant strains. These isolates were subsequently tested for cross-resistance to other antibiotics using the disc diffusion method. PCR was performed to detect selected ARGs. The enumeration of microbial population of Gomti River, the tetracycline-resistant bacteria comprised 38

Wars have devastating effects on all the components of the One Health approach: humans, animals, and ecosystems. Wars and the resulting migratory waves massively disrupt normal animal health services and surveillance. Among other consequences, they adversely impact the early detection, prevention, and control of animal diseases. Uncontrolled movement of animals or their undisposed carcasses, the destruction of wildlife habitats, and the increased interface between humans, wildlife, and domestic animals contribute to uncontrolled transmission and spread of zoonotic pathogens from animals to humans. In the last millennium, zoonotic diseases such as the "Black Death" were triggered by devastating wars and led to the deaths of a large fraction of the human population. However, also recent and ongoing wars carry the risk of an uncontrollable increase in zoonotic diseases. The most significant zoonotic diseases reported during the recent wars are African swine fever, highly pathogenic avian influenza, rabies, leptospirosis, and brucellosis, as well as foodborne and waterborne zoonotic diseases. Indeed, alarming rates of infections by antimicrobial-resistant pathogens such as Mycobacterium tuberculosis go along with wars, as seen in the current Ukraine-Russia conflict. Considering human migration, foodborne and waterborne zoonotic diseases are key health threats for refugees due to the consumption of unsafe food, lack of safe water, and disruption of the water supply and sanitation system. This review summarizes the potential factors and some data associated with the increased risk of zoonotic disease emergence and transmission during recent and ongoing conflicts.

Early pregnancy diagnosis in buffaloes enhances reproductive efficiency and dairy production. This study aimed to develop and validate a duplex TaqMan-based RT-qPCR assay using two interferon-stimulated genes (ISGs), ISG15 and LGALS3BP, for early pregnancy diagnosis in buffaloes. Whole blood samples were collected from artificially inseminated buffaloes before (day 0) and after (days 20, 25 and 40) artificial insemination (AI). On 40 days, post-insemination ultrasonography was performed to categorize animals as pregnant or non-pregnant. RNA from peripheral blood mononuclear cells (PBMCs) was isolated and converted into cDNA. A duplex TaqMan-based RT-qPCR assay was developed to predict pregnancy based on the expression of ISG15 and LGALS3BP in PBMCs. Gene expression was evaluated in a test dataset at various time points. The assay's performance was validated using two additional datasets, under which blood samples were collected randomly at 20 ± 2 days to evaluate its utility for predicting pregnancy. Colour Doppler ultrasonography was utilized to confirm pregnancy at 40 days post-insemination for all the animals. Significantly different abundance of transcripts of ISG15 and LGALS3BP, in pregnant buffaloes, was observed on day 20 post-insemination. A statistically significant fold change (p < 0.05) of ISG15 and LGALS3BP transcripts was observed between pregnant and non-pregnant buffaloes. The receiver operating characteristic curve of validation datasets demonstrated the AUC = 0.95 and AUC = 0.90. The negative predictive value and positive predictive value range from 90% to 95% and 75% to 85%. In conclusion, the developed duplex RT-qPCR-based assay demonstrates high sensitivity and specificity in detecting early pregnancy in buffaloes.

This study introduces an assay combining real-time qPCR and high-resolution melting (HRM) analysis for the detection of African Swine Fever Virus (ASFV). The assay utilizes two subunits of RNA polymerases (RNAP1 or NP1450 and RNAP6 or C147L) gene-targeting primers. Initially, the assay was developed on plasmids containing synthetic gene constructs and later validated on clinical samples obtained from field outbreaks. The assay demonstrated high specificity, as evidenced by the absence of amplification from closely related viruses such as porcine parvovirus type 1 (PPV1), porcine parvovirus type 2 (PPV2), and porcine circovirus (PCV). Comparative analysis between qPCR-HRM assay and TaqMan real-time PCR showed similar detection limits for both methods. The assay showed an accuracy of 100% compared to the World Organisation for Animal Health (WOAH)-recommended assay when tested on 25 representative clinical samples. In silico computational analysis revealed that the identified primer sets can detect all the ASFV genotypes circulating globally. Notably, this assay offers an advantage of differentiating strain with single base change (single nucleotide polymorphisms; SNP) using HRM peaks. The study revealed that utilizing real-time PCR with HRM is a cost-effective method of early detection of ASFV in field. This provides an alternative method for swiftly, simply, and accurately detecting ASFV with high specificity and sensitivity.

Diarrheal diseases, particularly those caused by rotaviruses, pose a significant health threat, especially among children, and cause huge economic losses to the pig industry in the form of high morbidity, mortality, and stunted growth. Rotavirus A (RVA) remains the predominant viral agent for severe diarrheal episodes, contributing to high hospitalization and mortality rates in India. RVA’s high genetic diversity is attributed to frequent reassortment and mutations. This study aims to characterize the VP4, VP6, VP7, and NSP4 genes of RVA in stool samples collected from children and piglets in and around Bareilly, Uttar Pradesh, India. A total of 300 samples, including 100 from children and 200 from piglets, were screened for the detection of double-stranded RNA of RVA using ribonucleic acid-polyacrylamide gel electrophoresis (RNA-PAGE) and reverse transcription polymerase chain reaction (RT-PCR). Results revealed the RVA incidence, particularly in winter (end of November to beginning of February), aligning with observed seasonal trends. Among the 32 Rotavirus (RV)-positive samples from children, 21 (65.63%) were detected by RNA-PAGE, whereas 28 (87.5%) were identified by RT-PCR. Whereas, of the 80 RT-PCR positive samples from piglets, only 51 (63.75%) were detected by RNA-PAGE, indicating the superiority of RT-PCR. Molecular analysis identified the prevalent genotypes in human strains as G1, G2, G3, and P[8], whereas G9P[13]-I5-E1 dominated among piglets in a single farm outbreak. The findings underscore the critical need for continuous surveillance to monitor evolving RV genotypes from both humans and piglets, enabling the identification of new strains of RVA and subsequent modification of vaccination strategies to reduce RVA’s impact in India.

Pig farming plays a vital role in economic growth. Unfortunately, the swine farming sector of India has experienced a decline, partially due to several viral and bacterial infections. These diseases have direct impact on animal health, productivity, mortality rates, and economic returns. Viral gastroenteritis, caused by enteric viruses, is a severe condition that primarily results in infection in neonatal animals. Many enteric viruses are responsible for viral gastroenteritis amongst which Porcine astrovirus (PAstV) and Porcine kobuvirus (PKV) are the emerging ones. Current diagnostic approaches are serological tests, conventional PCR, and other molecular diagnostics, which are reliable but take time and are expensive for high throughput screening. To overcome these challenges a duplex RT-PCR assay was developed for identification of PAstV and PKV in pigs to boost up the intensity of diagnosis. This approach allowed rapid and inexpensive testing, which is particularly useful for the swine population. A duplex PCR assay showed sensitivity to a reliable range of concentrations for PAstV at an LOD of 2.74 ng and for PKV at 30 fg. This developed assay also found to be specific and did not detect other porcine viruses like PCV and PPV. The applicability of assay was also done for screening (n=50) pig samples collected from different regions of Punjab, India, and the developed nucleo-diagnostic identified both viruses (PAstV and PKV) at a single stance and thus enhanced control measures that would reduce losses occasioned by these diseases in the swine industry.

Rotaviruses are widely acknowledged as the prime cause of severe gastroenteritis in children globally especially in under developed countries. This contemporary study presents data on the prevalence and genotypic distribution of human group A rotaviruses across various regions of India. 217 fecal samples were collected entirely from children and adults over 5 years of age from the Western (n=30), Southern (n=98), and North-Eastern Hill (NEH) (n=89) regions/zones of India and tested for the presence of rotaviruses. The RNA extracted from these stool samples was analyzed using Polyacrylamide Gel Electrophoresis (PAGE) and Reverse Transcription Polymerase Chain Reaction (RT-PCR). Rotaviruses were identified in 20.27% of the samples through PAGE and/or RT-PCR, with the highest detection rate of 41.37% observed in the Western region, followed by 17.34% in the Southern zone/ region, and 18.98% in the NEH region. A higher prevalence of 27.38% was observed in males than female children (14.06%), especially during winter (November to March) (34.84%) followed by April to June (15.62%). A total of fifteen samples were successfully typed for the G genotype, and six for the P genotype, through amplification and subsequent sequencing of VP4 and VP7gene products. The prevalent G genotypes detected were G1, succeeded by G12 and G11, with one sample each also identifying G2 and G10. Amid P genotypes, P[6] was the most prevailing genotype, while P[4] and P[8] were each recognised in a single sample. The dominant genotype was G1P[6], with one sample showing a G1P[4] combination, and two samples exhibiting G11P[6] and G11P[8] combinations, which are rarely reported in India.

In recent times, the understanding of the human microbiome and its impact on health and disease has undergone a paradigm shift, leading to ground-breaking discoveries in the field of probiotics. Probiotics, live microorganisms known for conferring health benefits when administered adequately, have garnered significant interest for their potential to modulate the immune system's response to viral infections in both humans and animals. The emergence of enteric and respiratory viruses as significant global health threats has prompted intensive research efforts to identify novel therapeutic strategies. Traditional antiviral therapies often face challenges such as drug resistance, limited efficacy and adverse effects, underscoring the urgent need for alternative approaches. In this context, probiotics have emerged as a promising avenue for the prevention and treatment of viral infections due to their ability to modulate the host immune response, enhance mucosal barrier function and exert direct antiviral effects. This review aims to provide a comprehensive overview of the therapeutic landscape of probiotics against enteric and respiratory viruses. Based on latest findings from preclinical and clinical studies, we have explored the mechanisms underlying the antiviral activity of probiotics and their potential role in mitigating viral infections. Furthermore, promising avenues for harnessing probiotics as adjunctive or standalone interventions against enteric and respiratory viral infections have been discussed here.

Porcine Astrovirus (PAstV) is a positive-sense single-stranded RNA virus, responsible for gastrointestinal diseases in swine populations across the globe. There are sparse reports regarding diagnostic platforms for the detection of PAstV in the porcine population. The present study reported a nested PCR (nPCR) assay for the identification of PAstV in faecal specimens of pigs in Punjab, India. The technique was developed using outer and inner sets of primers targeting the RNA-dependent RNA polymerase (RdRp) gene of the virus. The nPCR was standardized and optimized, including sensitivity and specificity. The sensitivity of the assay was found to be 225 femtograms. The technique was specific and did not amplify with certain other viruses i.e., Porcine Kobuvirus (PKV) and Bovine Rotavirus (BoRV). The applicability of the nPCR assay was further assessed on faecal samples (n=50) collected from field conditions comprising diarrhoeic (n=39) and non-diarrhoeic (n=11) animals. The nPCR detected only 12 [diarrhoeic (n=8) & non-diarrhoeic (n=4)] out of 50 samples tested as positive. This work demonstrated that nPCR assay can be effectively used as a rapid, specific, and sensitive method for routine molecular screening and/or diagnosis of PAstV in swine population.

Lumpy Skin Disease Virus (LSDV), a Capripoxvirus of significant veterinary and economic importance, has been reported to manipulate host cellular processes, including autophagy, to enhance its replication and persistence. However, the precise mechanisms by which LSDV interacts with autophagy remain unclear. This study investigates the effect of LSDV infection on host autophagic pathways in MDBK cells, focusing on autophagic flux modulation and its implications for viral replication. Western blot and immunofluorescence analyses demonstrated that LSDV does not robustly induce autophagy but interferes with autophagic flux by suppressing lysosomal degradation, as indicated by reduced LAMP2 expression at later stages of infection. Pharmacological modulation of autophagy using activators (Rapamycin, Torin 2) and inhibitors (Bafilomycin A1, MRT68921) revealed that increased autophagy enhanced LSDV replication, whereas inhibition of autophagy significantly impaired viral propagation, underscoring the virus's reliance on autophagic processes for efficient replication. Notably, cells infected with BEI-inactivated LSDV exhibited significantly higher LC3B II accumulation than those infected with live LSDV, suggesting that active viral replication is required for autophagy modulation. Additionally, dose-dependent analysis revealed that LSDV-mediated suppression of LC3B II was most prominent at higher viral titers, further confirming the virus's capacity to regulate host autophagic responses. These findings suggest that LSDV strategically modulates autophagy by maintaining basal autophagic levels while suppressing lysosomal degradation, allowing for optimal viral replication. Understanding the interplay between LSDV and autophagy provides novel insights into its pathogenesis and identifies potential antiviral targets to mitigate LSDV infections in cattle.

This book provides immunopathology of animals, their etiology, clinical signs, diagnosis and treatment with the suitable references.