Bacteriological and genetic study of the bacteria Campylobacter spp locally isolated

  • Noor Monim Al Shammari Department of Biology, College of Education Pure Sciences, Diyala University
  • Adnan Neama Al Azawy Department of Biology, College of Education Pure Sciences, Diyala University
  • Ziad Tariq Sedrah Department of Animal Production , College of Agriculture , Diyala University
Keywords: Campylobacter ( C.jejuni,, C.coli,C. hyointestinalis, , diarrhea for children

Abstract

The study is conducted in the Graduate Studies Lab. In the College of Pure Sciences. The researcher compiled (123) samples from various sources comprising of (30) raw milk samples, (30) local white cheese samples, (30) local cream samples compiled from local markets in Baqubah city during the period of time from (1/11/2015 to 28/2/2016), and (33) samples of feces from children with diarrhea (watery, mucoid, bloody) of ages between (3 months to 5 years) in Al-Batoul Teaching Hospital for Maternity and Children.

Results of the study exposed that (33) samples, with a ratio of (26.82%), showed negative growth to bacterial plantation, and (90) samples, with a rate of (73.17%) mirrored positive growth to bacterial plantation. Moreover, studied phenotypic characteristics and attributes biochemical bacterial isolates, it has indicated diagnostic results to be (65) isolation of them, and by (65) samples with a ratio of (52.84%) were Campylobacter bacteria as the ratio of isolated bacteria from raw milk reached up to (15) samples with a rate of (50%), (7) samples from local white cheese with a rate of (23.33%) and (8) samples from local cream in a ratio of (26.66%). the ratios of isolated bacteria from feces samples were; (3) samples of bloody sustenance (9.90%), (10) samples of mucoid sustenance (30.30%) and (20) watery samples (60.60%). The bacteria were also diagnosed by means of micrography which showed its negativity and positivity towards Gram stein; (90,90%) were negative to Gram stain, while (9.90%) were positive to Gram stain.

Biochemical and physiological examinations were executed and isolates were oxidase-positive, catalase, urease test, Hippurate hydrolysis test, and growth test within (37 c-42 c) temperatures as well as their tolerance of salinity with concentrations between (1.5 %-3.5%). Isolates were sensitive to Nalidixic acid and resistant to Cephalothin.

Moreover, total DNA extraction process of the Campylobacter bacterial samples was executed via delineating extraction kit (Mini DNA Bacteria Kit) which is equipped by Pioneer Co. the extracted DNA purity ranged between (2-1.8) regarding all selected samples. Next to that, the polymerase chain reaction (PCR) was done with positive growth bacteria on selective media (Skirrow-Preston) and non-selective media (Blood agar, Chocolate agar, Macconkey agar) via the delineation of the specialized primers kit which target the quantitative sequence of cadF gene with a molecular weight of 400 base pairs. The doubling outputs were deported on the Agoaros gel with a concentration of 1%. One package was observed to appear in all tracks in the gel at the same level regarding the gene .The results of the PCR with a use of specialized primer for this gene showed that from (12) samples, (7) of them contained this gene with a rate of (58.33%) and at the same molecular weight of 400 base pairs, while the other (5) samples did not contain this gene with a rate of (41.66%). 

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Published
2019-04-17
Section
Articles