| Summary: | 碩士 === 國立嘉義大學 === 獸醫學系研究所 === 96 === Mastitis often cause economic loss of dairy-related industry and staphylococcus areus is main infectious pathogen. We developed a rapid multiplex PCR identification method to suit to dairy goat, including 16S rRNA, clfA, nuc and mecA gene, and take together with traditional microbiology and commercial API kit to upgrade total efficiency of identification. During August, 2006 to December, 2007, we other isolated 86 strains of Staphylococcus areus and susceptible to methicillin, 181 strains of Staphylococcus sp., 183 strains of Escherichia coli and 967 strains of other species in 3,427 samples of fresh milk collected from 20 middle-southern Taiwan dairy herds. We divided 86 strains identified as MSSA into eight biotypes, 34 antibiograms, and 7 coa-PCR-RFLP types. The frequence of maintypes differed among typing mwthods. When efficacy of all typing methods was analyzed by Simpson’s index, index was 0.9929, 0.9800 and 0.9989, separately for each method, easicially coa-PCR-RFLP method was best, with 16 used antibiotics antibiotic test resistance was of 86 strains of MSSA was analyzed by disk-diffusion test which is significant in anti-drug analysis. The prevalence of antibiotic had 72.09 % (68/86) for Oxytetracyclin, 69.77 % (60/86) for Tetracycline, 63.95 % (55/86) for Vancomycin, 56.98 % (49/86) for penicillin G and 56.98 (49/86) for Ampicillin. In addition, all strains were susceptible to Methicillin, Cephalothin, Cefuroxine and Bacitracin. To understand the genes responsible for antibiotic resistance, mecA and tet were examind. 86 strains of MSSA were negative to mecA, 7 strains for tetK, 52 strains for tetM, 1 strain containing tetK and tetM In addition, enterotoxin genes including sea, seb, sec, sed and see was analyzed in 86 strains of MSSA. We sec was detected in 25 strains of MSSA, and the major group in Taiwan milky-goat. In summary, we established a set of fast-diagnostic Multiplex PCR method to detect Staphylococcal suitable for fresh milk sample. Further more, result of typing, anti-drug analysis and incidence of enterotoxin to staphylococcus areus could supply a accordance to dairy-relate industry in epidemiology.
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