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SETSCI - Volume 4 (9) (2019)
ISAS WINTER-2019 (HSS) - 4th International Symposium on Innovative Approaches in Health and Sports Sciences, Samsun, Turkey, Nov 22, 2019

Pseudomonas aeruginosa detection methods from fish samples
Belgin Sırıken1, Veli Öz2, Ceren Başkan3*
1Ondokuz Mayıs University, Samsun, Turkey
2Ondokuz Mayıs University, Samsun, Turkey
3Amasya University, Amasya, Turkey
* Corresponding author: cerennyavuz@hotmail.com
Published Date: 2019-12-23   |   Page (s): 141-145   |    160     5
https://doi.org/10.36287/setsci.4.9.083

ABSTRACT A total of 70 fish samples were randomly purchased from different butchers and supermarkets brands in Samsun Province, Turkey in 2018, and analysed for present of Pseudomonas (P.) aeruginosa. For that purpose, P.  aeruginosa isolation was carried out in conventional culture technique; briefly, under aseptic condition 10 g fish samples were transferred into a sterile polyethylene bag and 90 ml of peptone water (PW-Oxoid CM 00099) broth was added. The mixture was homogenized and prepared decimal dilution up to 10-6. Following that, the broth was plated onto Pseudomonas CN Selective Agar [Oxoid SR 102E, suppl. Pseudomonas Agar base-(Oxoid CM 0559)] (EN ISO 13720) using spread plate technique and the plates was incubated aerobically for 24-48 h at 37 °C. After the incubation, up to five susceptible colonies grown on the Pseudomonas CN Selective Agar were subcultured onto Tryptone Soya Agar plates (TSA-Oxoid-CM0131-L21). The presumptive P. aeruginosa colonies were tested with the Gram staining, oxidase (Oxoid BR 64) and catalase test. In addition, the colonies were streaked onto Endo Agar Base (Oxoid, CM0479, suppl. BR0050), for confirmation of the isolates for being P. aeruginosa at molecular levels, two types of genes were detected in the isolates- oprL and PA-SS (16 S rDNA) genes using PCR assay.  In conclusion, we obtained 100 Pseudomonas spp. isolates. Of those, 65 isolates were identified as a P. aeruginosa using classing culture technique. However, 30 out of 65 isolates were confirmed at molecular levels. Between two genes regions, although only oprL gene detected in the 30 isolates, PA-SS (16 S rDNA) gene was not detected in any of the isolates. 
KEYWORDS Pseudomonas aeruginosa, fish, oprL gene, PA-SS (16 S rDNA) gene
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