Daignosis of Staphylococcus aureus mastitis in bovine in Al-Najaf province by using Polymerase chain reaction (PCR)

This study was conducted to collect 388 milk samples from cows at different villages and townships in Al-Najaf province to examine about Staphylococcus aureus mastitis .CMT was used for subclinical mastitis screening ,212(54.6%) milk samples were mastitic .The molecular method (PCR assay) was used to detected the presence (glpF) gene in classically diagnosed S.aureus, which appeared that 38(92.6%) S.aureus mastitis as 13(32.5%) clinical and 25(14.5%) subclinical mastitis .There was high significant incidence of Staphylococcus aureus mastitis in left posterior udder quarter rather than others quarters. Key word: S.aureus, bovine mastitis, glpf gene Introduction Mastitis remains the most common disease of dairy cattle, causing the biggest economic losses to the dairy industry ( 1). S.aureus is among the most common etiologic agents of bovine mastitis ( 2 ) Staphylococcus aureus is a major pathogen in dairy cattle mastitis ( 3 , 4, 5) , it causes big financial/economic loss to the dairy industry worldwide, mainly due to reduced milk production and the need to discard contaminated milk (6,7 ).Iraq have many researches were done on mastitis in cows' herds for detection of causative agents, (8 ) were isolated just one isolate S.aureus from 29 cows suffered from acute mastitis in cows' herd in Al-Sulaymaniya governorate during two years (19781979),While (9) recorded the highest percentage to S.aureus isolates (36%). (10) had been revealed that S.aureus was isolated at 7.64% . (11) at Al-Nasir station of cows, She resulted that S.aureus mastitis was (28.73%) . (12) show the highest results of isolation of S.aureus from mastitic cows were (58%) , while (13) found the percentage of Staphylococcus aureus mastitis in cows in Ninevah governorate was 55% . Materials and methods Materials Cultures media:1. Blood agar base : 2.Nutrient agar: 3. Mannitol Salt Agar : 4. Brain Heart Infusion agar: 5. Nutrient Broth: 6A. Urea Agar: All media were prepared according to information's of manufactured company . Reagents : 1. Catalase reagent : According to (14) 2.Oxidase reagent:According to (15) 3.CaliforniaMastitisTest (CMT):It used for detection a subclinical mastitis (16) 4.Coagulase reagent ( rabbit plasma ): Bacton , Dickinson Company (Spain) 5. Gram Stain :It Prepared according to (15) . 6 . Urea solution (20 % ) : Commercial kits:The commercial kits used in the present study are shown in Table (1)and its appendices, as follow:Table (1): Commercial kits used in the present study AL-Qadisiya Journal of Vet.Med.Sci. Vol./12 No./2 2013 82 Source Types of kits No. Geneid/Korea DNA extraction Kit(1) 1 BIONEER/Korea Green master mix 2X Kit(2) 2 BIONEER/Korea Primers(3) 3 2. Green master mix consist of :DNA polymerase enzyme (Taq) 1