Isolation and characterization of lumpy skin disease virus from cattle in India.

Lumpy skin disease (LSD) has devastating economic impact. During the last decade, LSD had spread to climatically new and previously disease-free countries, which also includes its recent emergence in the Indian subcontinent (2019). This study deals with the LSD outbreak(s) from cattle in Ranchi (India). Virus was isolated from the scabs (skin lesions) in the primary goat kidney cells. Phylogenetic analysis based on nucleotide sequencing of LSD virus (LSDV) ORF011, ORF012 and ORF036 suggested that the isolated virus (LSDV/Bos taurus-tc/India/2019/Ranchi) is closely related to Kenyan LSDV strains. Further, we adapted the isolated virus in Vero cells. Infection of the isolated LSDV to Vero cells did not produce cytopathic effect (CPE) until the 4th blind passage, but upon adaptation, it produced high viral titres in the cultured cells. The kinetics of viral DNA synthesis and one-step growth curve analysis suggested that Vero cell-adapted LSDV initiates synthesizing its genome at ~24 hours post-infection (hpi) with a peak level at ~96 hpi whereas evidence of progeny virus particles was observed at 36-48 hours (h) with a peak titre at ~120 h. To the best of our knowledge, this study describes the first successful isolation of LSDV in India, besides providing insights into the life cycle Vero cell-adapted LSDV.

Improvement in fluoride remediation technology using GIS based mapping of fluoride contaminated groundwater and microbe assisted phytoremediation.

Fluoride (F) in groundwater is a major issue of water pollution. Geo-statistical analysis of groundwater quality in Newai Tehsil, (India) has been done in order to identify the possible spatial distribution of water quality parameters and to assess the spatial dependence of water properties with the help of principal component analysis (PCA) structure. Two types of maps (spatial map and principal component map) of groundwater quality have been developed. A field experiment was conducted to investigate the effect of different Fluoride (F) concentration combined with Pseudomonas fluorescens (P.F) on Prosopis juliflora plant. The field design was used as completely randomized block design with three replicates. Study revealed that parameters were found to be positively and highly correlated with principal component. Low and high values (with their acceptable limit) have also been displayed over the each spatial map. Plants treated with P. fluorescens showed the highest F uptake in root, shoot and leaves tissues were 33.14, 19.41 and 15.15 mg kg-1 after 120 days, respectively. Both total bioaccumulation factor (BF) and translocation factor (TF) were obtained above one i.e., 1.06 and 1.04, this confirmed the high accumulation and translocation of F in plant tissues. The F uptake efficiency of plant was enhanced to 67.7% and plant biomass was increased upto 57.03%. According to the available literature, this is the first spatial field study for the remediation of F polluted soil through P. fluorescens. The present study will be beneficial for researchers working towards further improvement of F phytoremediation technology. Also, GIS based study can be very useful for decision maker's exploration of groundwater to understand the potential of present research work on fluoride contamination.

Isolation, identification and characterization of a Peste des Petits Ruminants virus from an outbreak in Nanakpur, India.

A Peste des Petits Ruminants virus (PPRV) was isolated from an outbreak that occurred in sheep and goats in Nanakpur village of Mathura District in Uttar Pradesh (India). Based on hemagglutination of chicken red blood cells (rbcs), cytopathic effect similar to that caused by the Morbilliviruses in Vero cells, and amplification and sequence analysis of the viral nucleoprotein (N) gene, the identity of the virus was confirmed as PPRV and named PPRV/C. hircus-tc/India/2012/Nanakpur1 (in short PPRV/Nkp1/2012). However, based on its poor neutralization with monoclonal antibodies, escape detection by commercial ELISA, and unsuccessful amplification of the hemagglutinin (H) and the fusion (F) genes by several pairs of published PCR primers it was concluded that the PPRV/Nkp1/2012 may not be closely related to lineage type IV PPR viruses believed to be present in the Indian subcontinent. A plaque assay for titration of the PPRV was developed for the first time. The virus was plaque purified and its growth characteristics were studied in the African green monkey kidney (Vero) cells and baby hamster kidney (BHK-21) cells. In a one-step growth curve analysis it was concluded that the duration of the PPRV life cycle is 6-8h, an uncharacterized part of PPRV replication. These findings provide information for devising control strategies against PPR in India by choosing a homologous candidate vaccine prototype.