ABSTRACT
Background & objectives: Timely intervention is needed to minimize the economic losses of vector-borne bovine anaplasmosis which can be possible by the isothermal amplification assay. Methods: Anaplasma marginale in the cattle of south Gujarat, India was detected in the PCR and LAMP by amplifying the fragment of msp5 gene. The PCR product was digested with EcoRI, and sequenced to confirm its pathogen specific detection. Results: Species specific PCR observed a band of 457 bp of msp5 DNA following 1% agarose gel electrophoresis. Positive LAMP reaction turned into yellow colour while negative sample depicted original pink colour. A detection limit of PCR and LAMP was up to 10-6 and 10-8 of the original genomic DNA of A. marginale, respectively. A single cut site of EcoRI was observed in the PCR product. Current msp5 DNA sequences of A. marginale (MW538962 and MW538961) showed 100% homology with the published sequences. Monophyletic lineage type relationship was observed with high bootstrap proportion among the msp5 DNA sequences of A. marginale in the phylogram. Prevalence rate of A. marginale was significantly higher (p<0.05) in the PCR [43/280 (15.36%)] and LAMP [62/280 (22.14%)] than the microscopic technique [17/280 (6.07%)]. Diagnostic sensitivity, specificity, positive and negative predictive values at 95% CI for LAMP assay with respect to PCR were 93.02%, 90.72%, 64.52% and 98.62%, respectively. Interpretation & conclusion: Thus LAMP can be a practical alternative to the PCR for the diagnosis of A. marginale infection in the cattle even in field condition
ABSTRACT
Bacillus cereus incorporates the most important group of endospore-forming micro organism and can cause emetic and diarrheal food poisoning. A total of 42 B. cereus strains isolated from marketed raw chicken meat and human subjects swab samples were assessed by a triplex and multiplex PCR for the presence of enterotoxin genes. The detection rate of nheB, hblA, hblD, cytK, nheA,CER, hblC and entFM enterotoxin genes among all B. cereus strains was 83.33%, 80.95%, 69.04%, 21.42%, 47.61%, 0%, 61.90%, and 92.85% respectively. Enterotoxigenic profiles were determined in enterotoxin-producing strains showed 19 different patterns. The results offer essential information on toxin genes prevalence and toxigenic profiles of B. cereus from sources of origin. The present study was taken into consideration about extreme fitness danger for public health and insuring extra ability in difficulty to food safety amongst all B. cereus group members. Also, there may be need for extensive and continuous tracking of food products embracing both emetic toxin and enterotoxin genes.
ABSTRACT
The study was designed to isolate predominance contamination of Pseudomonas aeruginosa in marketed raw buffalo milk (n=122) samples, collected from private dairy farms from different places of south Gujarat, India. Pre-enrichment of 1 ml of each sample was done with inoculation in 9 ml tryptone soya broth and incubated at 37ºC for 24 hrs. A loopful of culture was taken from broth and streaked on selective Pseudomonas agar F plates and incubated at 37ºC for 24 hrs, after completion of incubation period, the colonies characteristics were studied and further confirmed by various biochemical tests and found 14 samples contaminated with P. aeruginosa, were further more biochemical testes are used and give positive results with IMViC, Motility test, catalase and sugar fermentation confirm at 37ºC for 24 hrs incubation. All biochemically conformed isolates were further subjected for molecular characterization and were also tested for antimicrobial susceptibility by using various antibiotics discs such as vancomycin, penicillin, tylosin, cefixime, chloramphenicol, kanamycin, ceftriaxone, cefixime, tetracycline, streptomycin, ciprofloxacin, enrofloxacin, oxytetracycline and gentamicin, which has shown multi drug resistant ranging from seven to nine antimicrobials and Multiple Antibiotic Resistance (MAR) index ranges from 0.50 to 0.64.The isolates of P. aeruginosa in the present study are extremely resistant to vancomycin, penicillin, tylosin, cefixime, chloramphenicol and maximum sensitive to ciprofloxacin and enrofloxacin followed by gentamicin. Further statistical analysis of antibiotics wise zone diameter interpretative standard (mm) reveled susceptibility phenotypes under significant of difference at P≤0.05 in one way ANOVA using Duncan’s multiple range test and found ciprofloxacin having maximum sensitivity among antibiotic tested and it could be considered as a drug of choice for controlling P. aeruginosa mediated animal and human infections in the studied regions for insuring food safety as well.