Prasad Minakshi

Evidence of genetic reassortment between Indian isolate of bluetongue virus serotype 21 (BTV-21) and bluetongue virus serotype 16 (BTV-16)

The genome of bluetongue virus (BTV) consists of 10 segments. Of these seg-2 encoded VP2 is the major serotype determining protein, and seg-6 encoded VP5 protein enhances the protective neutralizing activity of VP2 protein inducing higher serotype specific antibody titer than the VP2 alone. Out of the twenty-six BTV serotypes found worldwide, 22 were reported from different states of India. These include serotype 21 which was recently isolated from Andhra Pradesh, and was involved in a severe outbreak of bluetongue in Indian native sheep. BTV21 (KMNO-7) and BTV16 were circulating at the same time. This co-circulation, along with the fact that the virus genome is segmented, provides an opportunity for these two isolates of different serotypes to simultaneously infect the same animal, and even the same cell or a same vector with the potential for generation of reassortant viruses. This study was carried out to provide some insights into the outbreak. We carried out full length sequencing of genome seg-2 and seg-6 of Indian isolates VJW64 (BTV16) and KMNO-7 (BTV21). Detailed phylogenetic analysis revealed that genome seg-6 of Indian isolate KMNO-7 (BTV21) clusters with isolates of BTV16 showing maximum nucleotide similarity of 97.6% with TUR/2000/02 isolate of BTV16, which is much more than it shows with any isolate of BTV21. KMNO-7 (BTV21) significantly diverged from original strain of BTV21, and is a reassortant strain having acquired seg-6 from an isolate of BTV16. This study provides some useful insights into the epidemiology of the bluetongue disease, and undermines serotyping on genome seg-6 basis.

Biomineralization Abilities of Cupriavidus Strain and Bacillus subtilis Strains In Vitro Isolated from Speleothems, Rani Cave, Chhattisgarh, India

ABSTRACT In vitro culture experiments using three bacterial strains CSJC1, CSJC2, and CSJC3 isolated from speleothems, Rani cave, Chhattisgarh, India, were studied to examine their biomineralization potential. These speleothems showed high microbial cell enumerations on nutrient agar and iron agar (9 × 104 CFU/g) followed by thiosulfate agar (7 × 104 CFU/g), and 60 diverse strains were isolated. The BLASTn sequence search of 16S rRNA sequences with the NCBI database to establish the identity of CSJC1, CSJC2, and CSJC3 strains yielded similarity scores of ≥99% with the respective organisms, and the strains were identified as CSJC1 – Bacillus sp., CSJC2 – Cupriavidus sp., CSJC3 – Bacillus sp. The phylogenetic analysis of CSJC2 strain suggests that it formed a separate major cluster with Cupriavidus sp. and Cupriavidus necator. The phylogenetic analysis of CSJC1 and CSJC3 strains revealed that it formed a major cluster with several strains of Bacillus sp. and Bacillus subtilis. The biominerals induced by Cupriavidus sp. CSJC2 strain imaged with an ultra high-resolution field emission scanning electron microscope (FE-SEM) were seen as calcified coccoid shells that transformed into calcified dumbbells. FE-SEM imaging of biominerals induced by B. subtilis CSJC1 and CSJC3 tested both on B4 media and sheep blood agar individually showed that the precipitates formed calcified dumbbells that were almost similar but not identical phenotypically, indicating that strain-specific morphologies and crystal formation is easier when Ca is present in the media. This is the first comprehensive report on the possible evidences about the role of Cupriavidus sp. in calcite precipitation isolated from speleothems in the Indian caves. These results allow us to postulate that the identified strains may have a role in the biogenic influences in mineral formations at Rani cave.

Nanodiagnostics: a new frontier for veterinary and medical sciences.

Infectious diseases are one of the greatest threats to animal and human population living in the developing world. These diseases have capacity to instigate in a small area and then open out very fast to the rest of the world and causing a heavy pandemic situation, for example; avian influenza pandemic. Such diseases infect large masses of population and may lead to loss of lives and also incur huge economic losses. Therefore, the best way to control these diseases is by diagnosing it at a very primary level and taking necessary precautionary measures so as to avoid the spread. Since last few years, the diagnostic approach has changed from tedious molecular biological techniques, to easy and rapid diagnostic techniques. Nanotechnology has extended the molecular diagnostics limit to nanoscale. These developed techniques do not require sophisticated laboratories and expert personnel, and hence are a cheap diagnostic approach. These assays can also be performed at the field level where the patient is present and get the results there itself. Hence, they are also called as pen side test or lab on chip diagnostic assays. The biological tests using nanotechnology become quicker, more flexible and more sensitive. These techniques have greatly influenced the diagnostic approach in the veterinary as well as medical field. Especially in the developing countries such as India, where the laboratory services are not Upendra Lambe 1 , Minakshi P 1, *, Basanti Brar 1 , Madhusudan Guray 1 , Ikbal 1 , Koushlesh Ranjan 2 , Nitish Bansal 1 , Sandip Kumar Khurana 3 and Manimegalai J 1

CANINE PARVOVIRUS- AN INSIGHT INTO DIAGNOSTIC ASPECT

Canine parvovirus (CPV) leads to an acute disease, characterized by hemorrhagic gastroenteritis, vomiting and myocarditis in dogs. The disease can affect dogs of any age but is fatal in pups. CPV has undergone genetic variations that have led to emergence of various CPV-2 antigenic variants such as 2a, 2b and 2c with replacement of the original CPV-2 circulating in the dog population. CPV genome is made up of 5.2 Kb nucleotides. Viral protein VP2 plays a very important role in determining antigenicity and host range specificity of CPV. The antigenicity as well as host range of CPV is determined by virus specific VP2 protein. That’s why the mutations that affect the VP2 gene are the main source of different antigenic variants. It spreads rapidly in the wild population of canines as well as domestic animals, infected feces serve as a main source of infection because the virus is shed in large quantity in the feces particularly 4 - 7 days post infection. The present review is focused on various

New Approaches for Improvement of Diva Vaccines and Their Companion Diagnostic Tests for Foot-and-Mouth Disease and Bluetongue Disease

Vaccination is one of the effective ways to control disease outbreak and virus circulation both in endemic and disease-free countries. Endemic countries follow vaccination annually as a prophylactic measure subsequently increasing antibody titre of animals. In disease-free countries, emergency vaccination with high PD50 vaccine is done. In both the cases vaccine capable of eliciting long-lasting immune response followed by a robust companion Differentiating Infected from Vaccinated Animals (DIVA) test is required.