Investigation of the presence of Aspergillus Flavus in mastitis milk with PCR

Abstract

In this study, milk samples were taken in sterile 20 ml containers from 90 cows with clinical mastitis in various enterprises around Aydın province between May-September 2017. The presence of Aspergillus flavus M1 toxin in milk samples was investigated by ELISA. Aflatoxin M1 ELISA results were compared with positive controls with 0 ppt, 5 ppt, 15 ppt, 30 ppt, 60 ppt and 100 ppt. For the 88 samples examined; 5 ppt in 7 (8,0 %), 15 ppt in 12 (13,6 %), 100 ppt in 32 (36,4 %) and 0 ppt in 37 (42,0 %) aflatoxin M1 was detected. According to these results, 32 (36,4 %) AFM1 positivity was detected in the European Union countries over the limit (50 ppt) accepted for raw milk. In our study, DNA obtained from 88 mastitis milk samples examined with Aflatoxin M1 ELISA kit was subjected to Aspergillus flavus species specific PCR process. As a result of PCR; Aflatoxin M1 at a level of 0 ppt in 7 (36,8 %), 5 ppt in 1 (% 5,2), 15 ppt in 2 (10,5 %) and 100 ppt in 9 (47,5 %) ELISA kit toxin presence was determined. As a result, 32 samples of 100 ppt AFM1 positivity were detected by ELISA and 9 samples 100 ppt positivity was detected by PCR in our study. The results of the ELISA test revealed that more positive results were formed due to cross-reactions, and the PCR method with FLA gene primers was more reliable for diagnosis.