Characterization of infectious laryngotracheitis virus isolates from laying hens during 2019-2020 outbreaks in Tamil Nadu, India.

Infectious laryngotracheitis (ILT) is an acute respiratory disease in chickens that is a serious threat to poultry-producing countries worldwide. In the present study, we isolated and characterized infectious laryngotracheitis (ILTV) virus isolates by sequencing and restriction fragment length polymorphism analysis of PCR-amplified products (PCR-RFLP). A total of 26 ILTV outbreaks were investigated that occurred between 2019 and 2020 in flocks that had not been vaccinated against ILTV. ILTV was isolated by cultivating tracheal samples in embryonated chicken eggs, which showed multiple opaque pock lesions and thickening of the chorioallantoic membrane after 120 hours of infection. The ILTV isolates were identified and characterized by PCR and sequencing a portion of the ICP4 and TK genes. Phylogenetic analysis based on the ICP4 region showed that the sequences clustered with chicken-embryo-origin vaccine-like strains. Sequence analysis of the ICP4 region differentiated chicken-embryo-origin (CEO), tissue-culture-origin (TCO), and field ILTV strains, with significant differences in nucleotide and amino acid sequences. Furthermore, PCR-RFLP analysis of the TK gene showed that the patterns were identical to those obtained with low-virulence and vaccine strains. In conclusion, sequencing of a portion of the ICP4 region of ILTV allowed differentiation of ILTV field, CEO, and TCO vaccine strains. In this study, CEO-vaccine-like strains were found to be the cause of ILTV outbreaks between 2019 and 2020 in Tamil Nadu in southern India.

Optimization and Characterization of PHA (SCL-SCL) Copolymer by Indigenous Bacillus thuringiensis A102 Strain for Biomedical Applications.

PHA is one of the leading commercially important bio-polyesteric compounds piled up as an intracellular lipid-based energy storage compound by numerous microorganisms. An indigenous Gram-positive bacterium isolated from fire ant (Solenopsis invicta) has known to potentially accumulate PHA. Various nutritional elements like carbon, nitrogen, phosphate and C/N ratio were optimized. The indigenous B.t.A102 strain grown in optimized RC medium yielded PHA of about 3.25 g/L. The extracted polymer was characterized by NMR, GC-MS, X-ray diffraction and thermal analysis via TGA & DTA. The characterized PHA was used to prepare scaffold using the solvent casting method. The non-toxic nature of the composite material was evaluated on NIH/3T3 fibroblast cell lines using different staining (like Giemsa staining, AO/EB dual staining, neutral red staining) techniques and cell viability assay. This paper dealt with the optimization of the media components that increase PHA production and primary in vitro testing for its possible application as wound dressing materials.

Dermatophilus congolensis infection in sheep and goats in Delta region of Tamil Nadu.

AIM: The study was conducted to isolate and identify Dermatophilus congolensis (DC) using conventional and molecular diagnostic techniques in scab materials collected from skin infections of sheep and goats in the Delta region of Tamil Nadu. MATERIALS AND METHODS: A total of 20 scab samples collected from 18 goats and 2 sheep from Nagapattinam, Thanjavur, and Tiruvarur districts of Tamil Nadu. Smears were made from softened scab materials and stained by either Gram's or Giemsa staining. Isolation was attempted on blood agar plates, and colonies were stained by Gram's staining for morphological identification. Identification was also done by biochemical tests and confirmed by 16S rRNA polymerase chain reaction (PCR), followed by sequencing and phylogenetic analysis of the amplified product. RESULTS: The peculiar laddering arrangement of coccoid forms in stained smears prepared from scab materials revealed the presence of DC. Isolated colonies from scab materials of sheep and goats on bovine blood agar plate were small, hemolytic, rough, adherent, and bright orange-yellow in color, but some colonies were white to cream color. Gram-staining of cultured organisms revealed Gram-positive branching filaments with various disintegration stages of organisms. 16S rRNA PCR yielded 500 bp amplicon specific for DC. Sequence analysis of a sheep DC isolate showed 99-100% sequence homology with other DC isolates available in NCBI database, and phylogenetic tree showed a close cluster with DC isolates of Congo, Nigeria, and Angola of Africa. Genes for virulence factors such as serine protease and alkaline ceramidase could not be detected by PCR in any of the DC strains isolated of this study. CONCLUSION: The presence of dermatophilosis in Tamil Nadu was established from this study.