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Original Article

AJVS. 2018; 58(1): 109-119


Molecular Study on Bacterial Resistance to Antibiotic with Special Reference to Plasmid Curing

Hala A. Abdellatif, Samy A. Khalil, Abdelhalim Hegazy.




Abstract

Abstract
Widespread use of antimicrobials in poultry farming here is a concern of multi-drug microbial resistance development that can potentially be transmitted to human pathogens even from non-pathogenic carrier strains. 200 samples were collected from 30 broilers flocks at private farm in Kafr El-Sheikh governorate. E. coli was isolated from 60 (30%) cases, P. aeruginosa was isolated from 30(15%) cases and S. aureus was isolated from 15 (7.5%) cases. Studying antibiotic sensitivity pattern of three bacterial strain revealed that isolates bacteria multiple drug (MDR) resistance was 17 isolates from E.coli with Average Multiple Antibiotic resistance index(MAR) index 0. 68, 8 isolates from P. aeruginosa with average MAR index 0.48 and 6 isolates from S. aureus with average MAR index 0.33. All MDR E.coli isolates were resistant to erythromycin while Ceftriaxone the most effective one; All MDR S. aureus isolates resistant AML (100%), while sensitive to OFX, LEV and Do (100%); All MDR P. aeruginosa isolates resistant AML, E and CTX (100%) while more sensitive to CN and LEV. ALL MDR isolates from three bacterial strain subjected to plasmid profiling, genomic extraction and PCR; plasmid profiling revealed that 14 (82%) isolates of E.coli, 3 (50%) isolates of S. aureus and 7 (87%) isolates of P. aeruginosa harboring plasmid with varying size and number; PCR detection for antibiotic resistance gene revealed ß-lactamase resistance gene was detected by amplifying 516 bp of bla TEM in 14 isolates (100%) and 6 isolates of E.coli and P. aeruginosa, respectively, while bla Z was detected in S. aureus by amplifying 173 bp in 5 isolates (100%); aminoglycosides resistance gene was detected by amplifying 484 bp of aadA1 in 13 isolates (93%) and 6 isolates (86%) of E.coli and P. aeruginosa, respectively, while aac(6')aph (2'') was detected in S. aureus by amplifying 491 bp in 4 isolates (80%). Quinolone resistance gene was detected by amplifying 417 bp of qnrS in 6 isolates (43%) and 4 isolates (57%) of E.coli and P. aeruginosa, respectively, while norA was detected in S. aureus by amplifying 620 bp in 5 isolates (100%). GC-MS analysis confirmed that cinnamoldhyde was the major constituents in cinnamon essential oil (57.15%) and the main constituents in clove oils were eugenol (45.27%). Antibacterial inhibition zone of Cinnamon oil 29.25, 34.75 and 28.25 mm in case of E. coli, P. aeruginosa and Staphylococcus aureus. MIC of cinnamon, clove oil and extract was determined. Treatment of MDR isolates of three bacterial strain with sub MIC of cinnamon and clove oil and extract and acridine organ, bacteria which showed change in phenotype were examined for antibiotic resistance gene by PCR, plasmid profiling and electron microscope.
Key words: Antibiotic resistance pattern, plasmid profile, Molecular identification, GC-MS, MIC, cinnamon, clove

Key words: Key words: Antibiotic resistance pattern, plasmid profile, Molecular identification, GC-MS, MIC, cinnamon, clove






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