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Kerala Veterinary and Animal Sciences University, Wayanad

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20211
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Subject: Veterinary Public Health × Author: MATHEW, BINSY × Thesis Type: Ph.D ×
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    ThesisItemOpen Access
    DETECTION OF DRUG RESISTANT AND BIOFILM FORMING FOODBORNE PATHOGENIC BACTERIA IN BROILER CHICKEN AND MITIGATION USING PLANT EXTRACTS
    (COLLEGE OF VETERINARY AND ANIMAL SCIENCES MANNUTHY, 2021-07-05) MATHEW, BINSY; Latha, C.
    The study was undertaken to detect the presence of drug resistant and biofilm forming food-borne pathogens with special reference to E. coli, Salmonella spp. and Campylobacter spp. from retail chicken sold in different parts of Kerala and study the effect of aqueous leaf extracts of Couroupita guianensis Aubl. and Annona muricata on genotypically confirmed isolates of the aforesaid organisms. The districts included in the study were Kozhikode and Palakkad representing northern part of Kerala, Thrissur and Ernakulam representing central part and the southern part represented by Kollam and Pathanamthitta. The prevalence of E. coli, Salmonella spp. and Campylobacter spp. was 54.17, 14.33 and 17.17 per cent, respectively. The positive isolates were subjected to molecular confirmation of virulence genes targeting eaeA, aggR and ipaH for E. coli by standardising a multiplex PCR; invA and spvC for Salmonella spp. using a duplex PCR and mapA and ceuE for Campylobacter spp. using a multiplex PCR. The eaeA, aggR and ipaH were detected in 84.31, 4.31 and 1.54 per cent isolates, respectively. Virulence genes of Salmonella spp. were detected in 100 (invA) and 4.65 (spvC) per cent of Salmonella spp. isolates. The mapA and ceuE genes were detected in 74.76 and 27.18 per cent of Campylobacter spp. isolates, respectively. The antibiotic resistance profiles of the virulent isolates against nine antibiotics: ampicillin, cefotaxime, tetracycline, trimethoprim, streptomycin, ciprofloxacin, nalidixic acid and chloramphenicol were tested for E. coli and Salmonella spp. In addition to these antibiotics susceptibility to erythromycin was also studied against Campylobacter spp. The highest per cent of resistance was detected to nalidixic acid in E. coli and to tetracycline in Salmonella spp. and Campylobacter spp. Multiple drug resistance was seen in 49.64, 25.58, 23.94 and 24 per cent for E. coli, Salmonella spp., C. jejuni and C. coli, respectively. Polymerase chain reaction was used to detect antibiotic resistance genes in antibiotic resistant isolates. Four mismatch amplification mutation assay (MAMA) PCR was standardised for detection of point mutations in gyrA and parC genes in quinolone resistant isolates. In E. coli isolates, gyrA mutation was detected in 64.41 per cent of ciprofloxacin resistant isolates. In Salmonella spp cent per cent of the isolates showed the presence of blaTEM and catA genes. Among the Campylobacter spp. isolates, tetA gene was detected in 80.43 per cent of C. jejuni isolates and cent per cent of C. coli isolates. Biofilm forming ability was detected in all the three organisms. None of C. jejuni and C. coli showed strong biofilm production. The effect of four types of plant extracts: C. guianensis (hot and cold) and A. muricata (hot and cold) was studied on genotypically confirmed resistant isolates. The cold extract of C. guianensis was found to show maximum antibacterial activity in all the three drug resistant organisms. The detection of multidrug resistant foodborne pathogens with biofilm forming ability is a potential threat to human health. Nevertheless, mitigation using plant extracts seems to be a ray of hope in the present scenario of antimicrobial resistance in foodborne pathogens. In addition to the use of plant extracts, a multifaceted one health approach can effectively address the issue of antimicrobial resistance