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Molecular Epidemiology of Campylobacter Species Isolated from Meat Source in Lahore - Pakistan

Thesis Info

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Author

Nisar, Muhammad

Program

PhD

Institute

University of Veterinary and Animal Sciences

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Epidemiology & Public Health

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/12823/1/Muhammad_Nisar_Epidemiology_%26_PH_2018_UVAS_Lahore_12.09.2018.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676724873512

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Thermotolerant Campylobacter, one of the well-known bacterial causes of foodborne illnesses in humans, worldwide. These bacteria are gram-negative short rods which grow under microaerobic conditions and are non-spore forming. Campylobacteriosis is characterized by diarrhea, abdominal pain, cramps and fever. The diarrhea may be associated with blood and can be accompanied by vomiting/nausea. Digestive tract of all warm blooded animal is a significant reservoir for Campylobacter species. Campylobacter causes huge economic losses to a country by restricting trade in international market for meat and meat products. A total of 600 meat samples were collected from three different species (i.e., Beef (n=200), mutton (n=200) and poultry (n=200)) during September 2014 to February 2015 from ten administrative towns of district Lahore (Pakistan). From each town 20 samples of each species were collected through convenience sampling method thus from each town 60 samples and from 10 towns 600 samples were collected for this study. Data regarding factors associated with Campylobacter were collected through a pre-designed questionnaire. The collected samples were transported at 4˚C to laboratory of Epidemiology & Public Health department for further processing. The ISO 10272-1:2006 (E) was followed for the conventional microbiological isolation and confirmation of Campylobacter species. DNA was extracted with the help of Qiagen kit and DNA was quantified by Nanodrop C2000 (Thermo Scientific). The isolated strains (in 20% glycerol) and the extracted DNA were stored at -80˚C and -20˚C, respectively. Real Time and multiplex PCRs were optimized for molecular detection and identification of Campylobacter species in meat. Sangar method was used for sequencing of the genes and phylogenetic trees were generated with the help of Mega 6.0 software. The isolated strains were characterized by MLST (Multi Locus Sequence Typing) according to protocol described by pubmlst.org.com. The identified sequences were processed, entered and queried to PubMLST data base for comparison with already existing Campylobacter species alleles to obtain specific allelic numbers and sequence types (STs) for our isolates (http://campylobacter.mlst.net/). Antibiotic susceptibility tests for selected isolates were performed according to the Kirby-Bauer method. Chi square test was applied to see the association of various factors with Campylobacter strains circulating in meat sources in Lahore. A total of 125 Campylobacter isolates were recovered from 600 meat samples. The percentage prevalence was 20.8%. Campylobacter isolates were frequently present in poultry meat, followed by mutton and beef with 29%, 18% and 15.5%, respectively. Campylobacter jejuni was isolated from 93 meat samples and C. coli from 32 samples. The prevalence of Campylobacter in poultry was 29% (58/200). In these positive samples C. jejuni was frequently isolated (38/58) as well as C. coli (20/58). From mutton out of 200 samples, 36 were found positive for Campylobacter species. Campylobacter jejuni was isolated from 69.4% (25/36), C. coli 30.6% (11/36). A total of 31 samples were positive for Campylobacter isolates. The contribution Campylobacter jejuni and Campylobacter coli in beef was 61.2 % (19/31) and 38.8% (12/31), respectively. MLST analysis of 3 Campylobacter jejuni and 3 Campylobacter coli isolates yielded 2 different STs (i.e., 5 ST 1769 belongs to 828 CCs and 1 ST 2282 belongs to 206 CC). The highest resistance was observed against enrofloxacin 79.2% (99/125), followed by tylosin 77.6% (97/125), ciprofloxacin and amoxicillin 71.2% (89/125), colistin 69.6% (87/125), neomycin 32.8% (41/125), nalidixic acid 31.2% (39/125), gentamicin 25.6% (32/125) and doxycycline 8.8% (11/125). Thus doxycycline (91.2%) was found drug of choice followed by gentamicin (74.4%), nalidixic acid (68.2%) and neomycin (59.2%).
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