Molecular characterization of a novel variant of infectious bronchitis virus from field outbreaks in backyard chicken population of North East India.

Infectious bronchitis virus (IBV) causes considerable economic impacts on global poultry production. Since its emergence in early 1930, IBV continues to evolve and now exists in a wide range of antigenically and genetically distinct variants, that makes the prevention and the control of the disease both complex and challenging. Although IBV has been reported regularly from different corner of India, information about the molecular epidemiology of circulating strain in relation to clinical form of the disease is not available. We have studied the clinico-pathology and confirmed eight distinct field outbreaks of the disease from poultry population of Mizoram, India. The clinical disease in affected birds resulted sever pathological lesions involving respiratory, gastrointestinal, and urinary system together. The complete S1 nucleotide sequences and protein analyses have revealed a distinct variant of genotype I-IBV (GI), designated as GI-24 circulating in India. The S1 protein of the field strains displayed unique additional eighteen amino acids at C terminal end when compared with M41strain. Comparison of the S1 protein among all the 27 lineages of GI revealed five mutations that are exclusive to only the Indian strains. All the field strains have also possessed the amino acid mutations at highly variable region 2 (HVR2) of S1 receptor-binding domain (RBD) that are considered characteristic of nephropathogenic strains. The circulating GI-24 strains displayed potency for a wide range of tropism from respiratory epithelium to GIT and urinary system. This study provides insight on recently emerging IBV outbreaks in NER, India, which might be causing huge economic losses to the poultry farmers in the region.

Emergence of a novel genotype of class II New Castle Disease virus in North Eastern States of India.

Outbreaks of New Castle Disease from three north eastern states of India were confirmed by clinico-pathological examination followed by reverse transcription-PCR detection of F gene of ND Virus (NDV). Irrespective of vaccination, the outbreaks resulted 90-100% mortality in the affected flocks. The analysis of fusion protein sequences from ten field isolates revealed them as the velogenic or highly virulent strain. Phylogenetic analyses based on the complete F gene nucleotide sequences of the isolates have characterized only one of the isolate (OK149201) in the genotype XIII.2.2. The rest of the nine isolates are depicted in a distinct monophyletic group with average nucleotide distances from the other 20 genotypes ranged from 10.90 - 20.70. The nine isolates were further divided into two sub branches with the bootstrap support value of 100% at the nodes that define the two subgroups with an average evolutionary nucleotide distance of 6.00between the isolates in the two subgroups. As per the recommendation put forth in recently updated unified phylogenetic classification system for NDV, our findings clearly indicates emergence of a novel genotype of class II NDV in the biodiversity hot spot region of NER, India. The isolates in the newly identified genotype is designated with next available Roman numerals XXII. Further, the two subgroups within the genotype are designated as XXII.1 and XXII.2.

Delta variant (B.1.617.2) of SARS-CoV-2: Mutations, impact, challenges and possible solutions.

Since commencement of COVID-19 pandemic, several SARS-CoV-2 variants have emerged amid containment efforts via vaccination. The Delta variant (B.1.617.2), discovered in October 2020, was designated as a VOC by the WHO on May 11, 2021. The enhanced transmissibility of Delta variant has been associated with critical mutations such as D614G, L452R, P681R, and T478K in the S-protein. The increased affinity of the S-protein and ACE2 has been postulated as a key reason for decreased vaccine efficacy. As per evidence, the Delta variant possesses increased transmissibility and decreased vaccine efficacy compared to other VOCs like Alpha and Beta. This has led to concerns regarding the acquisition of novel mutations in the Delta variant and outbreaks in vulnerable communities, including vaccinated people. In this mini-review of Delta variant, we have explained its evolution and characteristics, the impact of spike mutations on infectivity and immune evasion, and measures to combat future outbreaks.

Molecular detection and characterization of highly pathogenic porcine reproductive and respiratory syndrome virus from a natural outbreak in wild pigs, Mizoram, India.

This study reports for the first time a natural outbreak of highly pathogenic porcine reproductive and respiratory syndrome (HP-PRRS) caused by HP-PRRS virus (HP-PRRSV) in wild pigs characterized by sudden onset of depression, anorexia, respiratory distress, and high fever. The disease has caused severe haemorrhagic pneumonia, haemorrhagic lymphadenitis, enlarged spleen with areas of infarction, and petechial haemorrhages on the myocardium and on the surface of kidneys. HP-PRRSV was detected in representative tissue samples by reverse transcription-PCR, and the field strain was isolated in the MA104 cell line. The phylogenetic analyses based on the whole genome sequences and nucleotide sequences of open reading frame 5 (ORF5) gene showed close grouping with the subtype IV of lineage 8/8.7 of PRRSV II, which represents the HP-PRRSV strains that predominate in the pig population of China since 2010. The amino acid sequence analysis of the ORF5 gene revealed the replacement of leucine (L) at position 39 to isoleucine (I) in the primary neutralizing epitope. Among the four potential N glycosylation sites, the N34 was mutated and found to be restricted to only three N glycosylation sites. The present findings have indicated that HP-PRRSV can cause fatal outbreaks and may emerge as a major threat to the wild pig population.

Molecular detection and characterization of African swine fever virus from field outbreaks in domestic pigs, Mizoram, India.

African swine fever (ASF) is a devastating haemorrhagic disease of domestic pigs, which can cause mortality up to 100%. Sudden mortality in pigs following an acute course of systemic disease was investigated in Mizoram state of India and confirmed the outbreak as ASF. Affected pigs suffered from severe depression, high fever, bloody diarrhoea, cutaneous haemorrhages and showed haemorrhagic lesions in visceral organs. The outbreak was confirmed by detection of p72, p54 and the central variable region of B602L genes by PCR in representative tissue samples collected from dead pigs. The nucleotide and phylogenetic analyses of p72, p54 and B602L characterized the ASFV as genotype II. Interestingly, the analysis of B602L gene has revealed that the ASFV from Mizoram state of India is more closely linked to the Eurasian ASFV strains isolated prior to 2014 and discriminated the Indian strains in two separate groups indicating that the source of origin for the Mizoram outbreak could be different from that of the other states of India.

Effect of dietary phytobiotic mixture on growth performance, nutrient utilization, and immunity in weaned piglets.

This study investigated the effects of dietary phytobiotic mixture on growth performance, blood profiles, immune response, and fecal microorganisms in weaned piglets. Twenty four weaned crossbred piglets were equally divided into four groups in a completely randomized design. The animals in 4 groups were fed a basal diet added with (1) no antibiotics and phytobiotics (CON), (2) bacitracin (0.5 g/kg; AB), (3) a blend of Cinnamomum zeylanicum and Trachyspermum copticum essential oils (0.3 g/kg and 0.4 g/kg, respectively; EO), and (4) plant extracts (PEO) of Mikania micrantha and Garcinia lanceifolia (2.8 g/kg and 1.4 g/kg, respectively) and C. zeylanicum and T. copticum essential oils (0.3 g/kg and 0.4 g/kg, respectively). Inclusion of AB, EO, and PEO did not affect final body weight, average daily gain, feed intake, feed efficiency, and nutrient digestibility. Compared with the CON, serum protein profiles were not affected, but a few lipid profiles were improved, particularly cholesterol, low-density lipoprotein, and high-density lipoprotein in the EO and PEO groups. Lymphocyte proliferation index and concentrations of IgG and IgA and TNF-α were not affected by any treatments. The concentrations of IgM increased (P = 0.04) at 28 days and tended to increase (P = 0.10) at 56 days in the EO group. Serum IL-1ß levels decreased on days 28 and 56 in the EO and PEO groups. Fecal Lactobacilli population generally increased (P < 0.01) in the AB, EO, and PEO groups compared with the CON. Fecal enterobacterial numbers were always greater for AB than for CON, EO, or PEO, but enterobacterial populations were sometimes lower in the EO group than the CON group. In conclusion, dietary EO or PEO has no effect on the growth performance, but it may improve a few lipid profiles, immune responses, and fecal microbial populations in piglets.

Classical Swine Fever Virus Biology, Clinicopathology, Diagnosis, Vaccines and a Meta-Analysis of Prevalence: A Review from the Indian Perspective.

: Classical swine fever (CSF) is an economically significant, multi-systemic, highly contagious viral disease of swine world over. The disease is notifiable to the World Organization for Animal Health (OIE) due to its enormous consequences on porcine health and the pig industry. In India, the pig population is 9.06 million and contributes around 1.7% of the total livestock population. The pig industry is not well organized and is mostly concentrated in the eastern and northeastern states of the country (~40% of the country's population). Since the first suspected CSF outbreak in India during 1944, a large number of outbreaks have been reported across the country, and CSF has acquired an endemic status. As of date, there is a scarcity of comprehensive information on CSF from India. Therefore, in this review, we undertook a systematic review to compile and evaluate the prevalence and genetic diversity of the CSF virus situation in the porcine population from India, targeting particular virus genes sequence analysis, published reports on prevalence, pathology, and updates on indigenous diagnostics and vaccines. The CSF virus (CSFV) is genetically diverse, and at least three phylogenetic groups are circulating throughout the world. In India, though genotype 1.1 predominates, recently published reports point toward increasing evidence of co-circulation of sub-genotype 2.2 followed by 2.1. Sequence identities and phylogenetic analysis of Indian CSFV reveal high genetic divergence among circulating strains. In the meta-analysis random-effects model, the estimated overall CSF prevalence was 35.4%, encompassing data from both antigen and antibody tests, and region-wise sub-group analysis indicated variable incidence from 25% in the southern to nearly 40% in the central zone, eastern, and northeastern regions. A country-wide immunization approach, along with other control measures, has been implemented to reduce the disease incidence and eliminate the virus in time to come.

Genetic Characterization of Infectious Bursal Disease Viruses from Field Outbreaks of the North East Region of India.

In recent years, acute severe outbreaks of infectious bursal disease (IBD) are frequently observed in commercial chicken populations of the North East Region (NER) of India, resulting in huge economic loses to poultry farmers. Field outbreaks of IBD in 30 different poultry farms in the NER were confirmed by clinicopathologic examination and reverse transcriptase PCR. A total of 10 isolates of IBD virus (IBDV) from these outbreaks were characterized by the genetic analysis of VP1 and the hypervariable region of the VP2 gene. Nucleotide sequences, deduced amino acid sequences, and phylogenetic analysis of both VP2 and VP1 genes revealed two genetically diverse strains of very virulent IBDV (vvIBDV) and one intermediate strain circulating in the NER. These isolates differ at nucleotide and amino acid levels from vvIBDV isolates of mainland India and are clustered in distinctly separate groups in the phylogenetic tree. Six of the isolates revealed a unique combination of vvIBDV amino acid signatures in the VP2 gene (A222, I256, I294), while bearing the non-vvIBDV amino acid signatures of the VP1 gene (146E, 147G, 242D), but they are clearly classified as vvIBDV in a phylogenetic analysis of both genes. Interestingly, one of the isolates showed 99% sequence homology with attenuated vaccine strains in the VP2 gene and clustered together. This study demonstrates the diversity of IBDVs in India and document for the first time the possible involvement of attenuated vaccine strains in the epidemiology of IBD in India.