cDNA cloning, structural analysis, SNP detection and tissue expression profile of the IGF1 gene in Malabari and Attappady Black goats of India.

Insulin-like growth factor 1 (IGF1) plays an important role in growth, reproduction, foetal development and cell proliferation. The present study was conducted to clone and sequence the full-length coding sequence of the caprine IGF1 gene from Attappady Black and Malabari breeds, two indigenous goat breeds of south India, to analyse its structure, and to ascertain the relative abundance of IGF1 mRNA in different tissues. The caprine IGF1 cDNA (GenBank accession nos: KJ549851 and KJ549852) contained a 465-bp open reading frame encoding IGF1 protein with 154 amino acid residues. A novel SNP was detected in the 3'UTR region, g.931A>G. Genotyping was performed in 277 goats from the two genetic groups using the PCR-single strand conformational polymorphism (SSCP) and two genotypes, AA and AG were observed at this locus. IGF1 is a secretary pathway protein with 49 amino acid-long signal peptide with 19 phosphorylation sites. Caprine IGF1 amino acid sequence was 83-99% identical to other species with highest identity with the ruminants. Relative expression of IGF1 was highest in uterus and liver (P < 0.05), followed by oviduct and muscle. This work provided an important experimental basis for further research on the functions of IGF1 in goats.

Utility of a rapid immunochromatographic strip test in detecting canine parvovirus infection compared with polymerase chain reaction.

AIM: The present study was undertaken to detect the presence of canine parvovirus (CPV) in fecal samples of diarrheic dogs by conventional polymerase chain reaction (PCR) and immunochromatographic (IC) strip test and to compare the diagnostic potential of these tests. MATERIALS AND METHODS: A total of 50 fecal samples collected from diarrheic dogs suspected for CPV infection were subjected to PCR using CPV-555 primer amplifying the gene coding for the VP1 protein. These samples were also tested by IC strip test using a commercial rapid Ag test kit. The results were statistically analyzed using McNemar test. RESULTS: A total of 22 samples (44%) were detected as positive by PCR, which yielded a specific amplicon of 583 bp. In IC strip test, 18 (36%) samples were found to be positive. The sensitivity of the test as compared to PCR was found to be 72.22% and specificity was 92.86%. Positive predictive value and negative predictive value of IC strip test was found to be 88.89% and 81.25%, respectively. Statistical analysis of the results of PCR and IC assay using McNemar test revealed no significant difference (p>0.05). CONCLUSION: The IC strip test could be employed as a rapid field level diagnostic tool for the diagnosis of canine parvoviral diarrhea.

Biofilm formation of Pasteurella multocida on bentonite clay.

BACKGROUND AND OBJECTIVES: Biofilms are structural communities of bacterial cells enshrined in a self produced polymeric matrix. The studies on biofilm formation of Pasteurella multocida have become imperative since it is a respiratory pathogen and its biofilm mode could possibly be one of its virulence factors for survival inside a host. The present study describes a biofilm assay for P. multocida on inert hydrophilic material called bentonite clay. MATERIALS AND METHODS: The potential of the organism to form in vitro biofilm was assessed by growing the organism under nutrient restriction along with the inert substrate bentonite clay, which will provide a surface for attachment. For quantification of biofilm, plate count by the spread plate method was employed. Capsule production of the attached bacteria was demonstrated by light microscopic examination following Maneval staining and capsular polysaccharide estimation was done using standard procedures. RESULTS AND CONCLUSION: The biofilm formation peaked on the third day of incubation (1.54 ×10(6) cfu/g of bentonite clay) while the planktonic cells were found to be at a maximum on day one post inoculation (8.10 ×10(8) cfu/ml of the broth). Maneval staining of late logarithmic phase biofilm cultures revealed large aggregates of bacterial cells, bacteria appearing as chains or as a meshwork. The capsular polysaccharide estimation of biofilm cells revealed a 3.25 times increase over the planktonic bacteria. The biofilm cells cultured on solid media also produced some exclusive colony morphotypes.

Outbreaks of salmonellosis in three different poultry farms of Kerala, India.

Avian salmonellosis is an important disease causing serious impediment to the development of poultry industry especially in developing countries of Asia and Africa. Since no "effective" immunoprophylactic measures are available for the disease till date, strict biosecurity is the only alternative to preclude the disease. For formulating the control measures, an understanding of the epidemiology of the disease, proper diagnosis and identification of the causative agent is quintessential. This report sheds light on three different outbreaks of salmonellosis in three different farms in Kerala (India) describing the disease diagnosis, antibiotic resistance and the suggested control measures. All the three isolates were revealed to be Salmonella gallinarum and were resistant to at least three of the antimicrobial agents tested.