Evaluation of a simplified key for the identification of coagulase-positive Staphylococcus isolated from bovine mastitis (original) (raw)
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2013
In order to determine the prevalence of S. aureus in clinical and subclinical bovine mastitis as well as the relationship between the presence of protein A gene and the udder inflammatory response . A total of 400 quarter milk samples were collected from 100 lactating Friesian cows at Ismailia Governorate . The percentage of clinical mastitis was ( 19.5%.), while the percentage of subclinical mastitis according to C.M.T was ( 42.85%) . The percentage of S.aureus in clinical mastitis was (28.2%) , while in in subclinical mastitis was (22.46% ).The identification of S,aureus was confirmed by PCR amplification of species-specific parts of the gene encoding the 16S rRNA. Twenty nine S.aureus isolates were subjected to PCR for detection of X region of protein A (spA) gene,10 isolates from clinical mastitis and 19 isolates subclinical mastitiC milk specimens (+++ve CMT and ++ve CMT) were (100%) positive . Results indicate that there was a positive relationship between the presence of prot...
Brazilian Journal of Microbiology, 2001
Contamination of fresh milk with Staphylococcus aureus was assessed comparatively through routine phenotypic (coagulase tube test and coagulase slide test) and genotypic (PCR) screening of 128 S. aureus strains isolated from 555 milk samples. These samples were collected from 362 cows with subclinical mastitis, hosted in different dairy herds at various locations of the Northern and Northeastern rural areas of the State of Rio de Janeiro, 39.7% of which were CMT-positive. All S. aureus isolates tested positive for the presence of the coagulase gene by PCR and the isolates could be grouped into four distinct classes according to the size of the PCR product. The strains also yielded variable results when assayed with coagulase test. Taken together, these data indicate the existence of extensive polymorphism at the coagulase gene locus in the genus Staphylococcus and exemplifies the extent of molecular and phenotypic heterogeneity associated with the strains circulating in rural herds.
Comparative Clinical Pathology, 2010
The aim of this study was the genotypical characterization of 58 Staphylococcus aureus isolates from nine dairy herds in the Tabriz and Urmia regions of east and west Azerbaijan provinces, Iran. In this study, 58 S. aureus isolates from 370 milk samples from cows with clinical and subclinical mastitis were analyzed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis of the aroA gene. Amplification of the aroA gene resulted in a single amplicon with a size of approximately 1,153 bp from all 58 isolates of S. aureus. To obtain the RFLP patterns of the isolates, the PCR products were digested with TaqI restriction enzyme and the fragments separated by gel electrophoresis. Four distinct RFLP patterns were observed among the studied isolates. Three out of four detected genotypes had the same RFLP patterns (A, B, and N) as reported by previous studies. The fourth newly detected genotype in this study was named H. Genotypes A and B were the most frequent, being observed in 24 (41.38%) and 29 (50%) isolates, respectively. Genotypes N and H comprised 1.7% and 6.9% of all isolates, respectively. With the exception of the RFLP pattern N, which was observed only in the Tabriz region, all other patterns were found in both Tabriz and Urmia regions. The results demonstrate that strain variations of S. aureus could occur within and between herds and also between different regions, although a few genotypes of S. aureus were predominant in bovine mastitis. This study also indicated that PCR amplification of the aroA gene is specific for S. aureus identification.
Molecular subtyping of Staphylococcus aureus isolates from cases of bovine mastitis in Brazil
Veterinary Microbiology, 1999
Sixty-six isolates of Staphylococcusaureus obtained from milk samples of dairy cows suffering from subclinical mastitis in southern Brazil were analysed by five different molecular typing methods. These included the analysis of plasmid profiles, the analysis of coagulase (coa) gene polymorphisms by PCR amplification of the 3′ terminal region of the coa gene, the PCR-based detection of polymorphisms in the X region of the protein A gene (spa), the PCR-directed analysis of variations in the spacer region between 16S and 23S rRNA, and the comparison of pulsed-field gel electrophoretically separated genomic SmaI fragment patterns. The molecular typing methods were supplemented with the biochemical characterization of the isolates and the determination of their in-vitro susceptibility to 14 different antibiotics. All genotypic and phenotypic typing methods were analyzed for their ability to discriminate between the isolates. Macrorestriction analysis proved to be the most discriminatory single method (D = 0.96) followed by rRNA spacer typing (D = 0.85), coa PCR (D = 0.82), and spa PCR (D = 0.80).
Indian Journal of Comparative Microbiology, Immunology and Infectious Diseases, 2016
To understand the prevalence and characteristics of Staphylococci causing goat mastitis in Algeria, mastitis milk samples were collected and cultured on both blood agar and mannitol salt 2 agar medium. Staphylococci isolates were identified by API-Staph gallery and tested for antibiotic susceptibility by disk diffusion method. DNA microarrays analysis was performed on S. aureus strains in order to detect virulence factors, including toxins, and to assign the isolates to theirs MLST clonal complexes. Overall, 86/266 (32.3%) goats suffered from mastitis, whose 32.5% (28/86) infected with Staphylococci. Most of strains were susceptible to many antibiotics, except for penicillin, tetracycline and fusidic acid, and the resistance incidence of which were 21.4%, 25%, and 10.7% respectively. Only twelve (42.8%) isolates were resistant to utmost two antibiotics and bi-drug resistance was associated to four Coagulase negative Staphylococci (CNS) strains. Nine different species were identified in CNS isolates while S. aureus strains were clustered within CC130-agr3-[lukF-P83, lukM+] (four isolates; 50%), CC6-agr1 (three isolates; 37.5%) and one agr4 non-typables cluster. Enterotoxins genes were found in 62.5% of the S. aureus strains. Of which sec, sel were the most prevalent (62.5%: n=5) followed by sea (12.5%: n=1). CC130 showed edinB, tst and tetK (2/4) genes and CC6 were positive to cna and fosB genes. The current investigation provide the data for prevalence of Staphylococci in goats in Tizi-Ouzou region and S. aureus characterization which will help in tracking evolution of epidemic strains and their control methods.
Characterization ofStaphylococcus aureusstrains involved in human and bovine mastitis
Fems Immunology and Medical Microbiology, 2011
Staphylococcus aureus is one of the main etiological agents of mastitis in different mammalian species. At present, it is unknown whether strains isolated from human mastitis cases share phenotypic properties and genetic background with those obtained from animal mastitis cases. Therefore, the objective of this study was to characterize S. aureus strains isolated from women with lactational mastitis and to compare them with the strains responsible for bovine mastitis and noninfectious strains. All the strains were genotyped by both pulsed field gel electrophoresis and multilocus sequence typing and submitted to a characterization scheme that included diverse assays related to pathogenic potential and antibiotic resistance. Apart from siderophore production, no significant association was observed between the strains from bovine and human mastitis. Statistical differences between human-and bovine-mastitis-associated strains were detected for some traits and virulence determinants, such as the presence of prophages and cna and hlb genes, which were more frequently found within the bovine group. On the contrary, resistance to penicillin was significantly higher among strains isolated from human lactational mastitis, probably related to the common presence of the blaZ gene. A high genetic diversity was found among the strains involved in mastitis in breastfeeding women.
Journal of Veterinary Diagnostic Investigation, 2009
Staphylococcus aureus is one of the most important etiologic agents of ovine mastitis. To develop effective control measures for mastitis, it is important to type S. aureus strains that have considerable genetic heterogeneity. In the current study, 47 S. aureus strains isolated from ovine mastitis were typed by polymerase chain reaction (PCR) based on coagulase ( coa) and protein A ( spa) polymorphisms and by pulsed-field gel electrophoresis (PFGE). Eight different coa types and 4 spa types were identified by PCR. While the most prevalent coa type was CG2 (42.56%), the spa types S4 and S1 were the most commonly observed (44.68% and 38.29%, respectively). Nineteen different pulsotypes were identified, and 12 of these were represented by a single isolate. Pulsotypes J and K were predominant and each represented 9 isolates (19.14%). All isolates belonging to J and K pulsotypes were CG2. Although all 9 isolates belonging to the J pulsotype were S4, all isolates in the K pulsotype were S...