Streptococcus agalactiae mastitis of bovine detection by Polymerase Chain Reaction (PCR) test in AL-Diwanyia province

This study was conducted for exam 348 milk samples from (clinically mastitic and other healthy cows) in many areas in AL-Diwanyia province by using CMT and bacteriological testing , which appeared that (64.9%) as percentage of mastitis ( clinically 15.9% , subclinically 84.0% ) Streptococcus agalactiae mastitis 13.2% ( 26.6% clinically , 73.3 % subclinicaly) diagnose by PCR assay by using specific primer (16SrRNA). Streptococcus agalactiae (30 isolates) after classical methods applied for streptococcus agalactiae identification (86 isolates). Introduction Mastitis is one of the most prevalent and most costly production diseases affecting the dairy cattle industry worldwide (1). In dairy cows streptococcus agalactiae is a major cause of mastitis , which is currently considered the economically most important disease affecting the dairy industry (2 , 3) .In Iraq many researches,(4) found that streptococcal mastitis (15.6%) , (5) was isolate streptococcus agalactiae in Iraq from three cow stations for breeding cow as (39.75%) .In the Dar es Salaam region of Tanzana, The streptococcus agalactiae mastitis was 15.4% ( 6).In Ahavaz ,Iran the most bacteria isolated from diary herd in this area were Streptococcus agalactiae (20 %) (7).Prevalence of sub-clinical mastitis and bacterial etiology in the West Littoral Region of Uruguay were: Staphylococcus aureus, Streptococcus agalactiae, (62.8%, 11.3% ) respectively (8) . Materials and Methods Materials: 1.Blood agar base. 2. Nutrient agar. 3. Edward agar. 4. Nutrient Broth. 5DNA Extraction kits . All media were prepared according to company instructions . Reagents : 1Catalase reagent : Prepared according to (9) . 2California Mastitis Test (CMT): AlSyria for veterinary preparation (Syria) according to (10). 3Coagulase reagent ( rabbit plasma ): Bacton , Dickinson Company (Spain) according to ( 11). 4Gram Stain :Prepare according to (11) . Commercial kits: The commercial kits used in the present study are shown in Tables (1,2,3). and its appendices, as follow:Table (1): Commercial kits used in the present study. Source Types of kits No. Geneid / Korea DNA extraction Kit(1) 1 BIONEER/Korea Green master mix 2X Kit(2) 2 BIONEER/Korea Primers (forward and reverse ) 3 AL-Qadisiya Journal of Vet.Med.Sci. Vol./12 No./2 2013 ______________________________________________________________________________ 102 Table (2) : DNA extraction Kit consist of : Lyses cells solutions:-Nuclei lyses solution 1 DNA precipitation alcohols:-Isopropanol alcohol -Ethanol alcohol 2 DNA rehydration solution 3 Table (3) : Green master mix consist of : DNA polymerase enzyme (Taq) 1