Detection and molecular characterization of porcine enterovirus G15 and teschovirus from India.

Porcine enterovirus G (EV-G) and teschovirus (PTV) generally cause asymptomatic infections. Although both viruses have been reported from various countries, they are rarely detected from India. To detect these viruses in Western India, fecal samples (n = 26) of diarrheic piglets aged below three months from private pig farms near Pune (Maharashtra) were collected. The samples were screened by reverse transcription-polymerase chain reaction using conserved enterovirus specific primers from 5' untranslated region. For genetic characterization of detected EV-G strain, nearly complete genome, and for PTV, partial VP1 gene were sequenced. EV-G strain showed the highest identity in a VP1 gene at nucleotide (78.61%) and amino acid (88.65%) level with EV-G15, prototype strain. However, its complete genome was homologous with the nucleotide (78.38% identity) and amino acid (91.24% identity) level to Ishi-Ka2 strain (LC316832), unassigned EV-G genotype detected from Japan. The nearly complete genome of EV-G15 consisted of 7398 nucleotides excluding the poly(A) tail and has an open reading frame that encodes a 2170 amino acid polyprotein. Genetic analysis of the partial VP1 gene of teschovirus identified porcine teschovirus 4 (PTV-4) and putative PTV-17 genotype. To the best of our knowledge, this is the first report on nearly full genome characterization of EV-G15, and detection of PTV-4 and putative PTV-17 genotypes from India. Further, detection and characterization of porcine enteroviruses are needed for a comprehensive understanding of their genetic diversity and their association with symptomatic infections from other geographical regions of India.

Molecular characterization of unusual G10P[33], G6P[14] genomic constellations of group A rotavirus and evidence of zooanthroponosis in bovines.

Group A rotaviruses (RVA) are a major cause of diarrhea in neonatal calves and children. The present study examined G/P combinations and genetic characteristics of RVAs in diarrheic bovine calves in Western India. RVAs were detected in 27 samples (17.64%) with a predominance of G10P[11] (51.85%), followed by previously unreported genomic constellations, G6P[14] (14.81%), and, G6P[4] (7.40%) and G10P[33] (3.70%). Sequencing and phylogenetic analysis revealed circulation of G10 (Lineage-5), G6 (Lineage-2), P[11] (Lineage-3), P[14] (proposed Lineage-8) and P[4] (Lineage-3) genotypes. The predominant G10P[11] strains were typical bovine strains and exhibited genotypic homogeneity. The rare, G10P[33] strain, had VP7 and VP4 genes of bovine origin but, a resemblance of the VP6 gene with simian strain indicated possible reassortment between bovine and simian (SA11-like) strains. The VP6 and VP7 genes of two rare strains, G6P[14] and G6P[4], were identical to those of bovine stains, but the VP4 was closely related to those of the human-bovine like and human strains, respectively. Additionally, in the VP4 gene phylogenetic tree, Indian P[14] strains constituted a closely related genetic cluster distinct from the other P[14] strains. Hence Lineage-8 was proposed for them. These findings indicated that bovines could serve as a source for anthropozoonotic transmission of G6P[14] strains while zooanthroponotic transmission followed by reassortment with human strain gave rise to G6P[4] strains. The observations of a present study reinforce the potential of rotaviruses to cross the host-species barrier and undergo reassortant to increase genetic diversity which, necessitates their continuous surveillance for development and optimization of prevention strategies against zoonotic RVAs.

Development of a DNA vaccine for chicken infectious anemia and its immunogenicity studies using high mobility group box 1 protein as a novel immunoadjuvant indicated induction of promising protective immune responses.

Chicken infectious anaemia (CIA) is an economically important and emerging poultry disease reported worldwide. Current CIA vaccines have limitations like, the inability of the virus to grow to high titres in embryos/cell cultures, possession of residual pathogenicity and a risk of reversion to virulence. In the present study, a DNA vaccine, encoding chicken infectious anaemia virus (CIAV) VP1 and VP2 genes, was developed and co-administered with truncated chicken high mobility group box 1 (HMGB1ΔC) protein in young chicks for the evaluation of vaccine immune response. CIAV VP1 and VP2 genes were cloned in pTARGET while HMGB1ΔC in PET32b vector. In vitro expression of these gene constructs was evaluated by Western blotting. Further, recombinant HMGB1ΔC was evaluated for its biological activity. The CIAV DNA vaccine administration in specific pathogen free chicks resulted in moderately protective ELISA antibody titres in the range of 4322.87 ± 359.72 to 8288.19 ± 136.38, increased CD8(+) cells, and a higher titre was observed by co-administration of novel adjuvant (HMGB1ΔC) and booster immunizations. The use of vaccine with adjuvant showed achieving antibody titres nearly 8500, titre considered as highly protective, which indicates that co-immunization of HMGB1ΔC may have a strong adjuvant activity on CIAV DNA vaccine induced immune responses. The able potential of HMGB1 protein holding strong adjuvant activity could be exploited further with trials with vaccines for other important pathogens for achieving the required protective immune responses.

Tuberculosis in Birds: Insights into the Mycobacterium avium Infections.

Tuberculosis, a List B disease of World Organization for Animal Health, caused by M. avium or M. genavense predominantly affects poultry and pet or captive birds. Clinical manifestations in birds include emaciation, depression and diarrhea along with marked atrophy of breast muscle. Unlike tuberculosis in animals and man, lesions in lungs are rare. Tubercular nodules can be seen in liver, spleen, intestine and bone marrow. Granulomatous lesion without calcification is a prominent feature. The disease is a rarity in organized poultry sector due to improved farm practices, but occurs in zoo aviaries. Molecular techniques like polymerase chain reaction combined with restriction fragment length polymorphism and gene probes aid in rapid identification and characterization of mycobacteria subspecies, and overcome disadvantages of conventional methods which are slow, labour intensive and may at times fail to produce precise results. M. avium subsp. avium with genotype IS901+ and IS1245+ causes infections in animals and human beings too. The bacterium causes sensitivity in cattle to the tuberculin test. The paper discusses in brief the M. avium infection in birds, its importance in a zoonotic perspective, and outlines conventional and novel strategies for its diagnosis, prevention and eradication in domestic/pet birds and humans alike.