Investigation of the Hydrolytic Enzyme Activities of Candida Parapsilosis Isolated From Milk Samples of Bovine Mastitis by API ZYM and Molecular Method
Keywords:Candida parapsilosis, Bovine mastitis, Hydrolytic enzymes, Conventional PCR, API ZYM test.
Two hundred and fifty milk samples were collected from cows suffering from mastitis, these samples were
tested for mycotic mastitis using culture and conventional PCR method. The results showed 116 isolates of
Candida spp. were recovered with a prevalence of (46.4%),hence C. parapsilosis was the dominating species
15/116 (12.9%). Fifteen isolates (100%) belong C.parapsilosis was confirmed by PCR using a species
specific primer for the 18S rRNA gene. Some randomly chose isolates of C. parapsilosis were used for
detection of their hydrolytic enzymatic activity by API ZYM test and PCR. The method used to evaluate their
enzymatic activity was API ZYM (BioMerieux). The highest activity was observed for leucine arylamidase,
acid phosphatase, naphthol-AS-BI-phosphohydrolase and ?-glucosidase as 10/10 (100%) for each, followed
by esterase lipase (C8) 9/10 (90%), valine arylamidase and alkaline phosphatase 8/10 (80%) for both and
esterase (C4) 7/10 (70%). Conventional PCR was done for partial amplification of sterol esterase, alkaline
phosphates and alpha-glucosidase genes by specific primer sequences, The results of the PCR amplification
of these genes was 8/10 (80%) for sterol esterase gene. Concerning the alkaline phosphatase gene, it was
present in 10/10 (100%). Regarding alpha-glucosidase gene was present in 10/10 (100%). This work aims at
evaluating enzymatic activities of C. parapsilosis isolated from cow’s milk with mastitis.
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