Characterisation of coagulase positive Staphylococcus species isolated from bovine mastitis using protein and plasmid patterns (original) (raw)
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The objective of this work was to study the protein patterns, plasmid profiles, and antibiotic susceptibility of Staphylococcus intermedius and Staphylococcus aureus isolates originating from mastitic mammary glands of dairy cattle in different parts of Konya province. A total of 114 Staphylococcus species were isolated and identified by conventional bacteriological methods from bovine mastitis. Of the total isolates 77 were identified as S. aureus and 37 as S. intermedius. Intra-and inter-species diversities in the coagulase-positive staphylococci were investigated by analysis of whole-cell protein profiles using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Plasmid profiling also demonstrated that 75 S. aureus isolates and 36 S. intermedius isolates contained plasmid. In addition, 88.3% of S. aureus and 59.4% of S. intermedius isolates were resistant to penicillin. Sixty-six of the 77 S. aureus isolates were also resistant to amoxicillin+clavulanic acid (85.7%). The corresponding number for S. intermedius was 17 (45.9%). Only 1 S. aureus isolate was resistant to danofloxacin. One of each of the Staphylococcus isolates was resistant to methicillin. Results from the study showed that the susceptibility of S. intermedius isolates to antibiotics used widely in mastitis therapy is a matter of concern.
A total of 572 California Mastitis Test (CMT) positive milk samples were collected from 423 lactating cows on 18 private farms in the Middle Western Anatolia. Coagulase-negative staphylococci colonies and CNS species identification was performed based on conventional biochemical techniques and using the API Staph test. Slime production was detected by Congo Red Agar (CRA) method. The antibiotic susceptibility was determined according to the National Committee for Clinical Laboratory Standards guidelines (NCCLS). A total of 67 (11.7%) coagulase-negative staphylococci (CNS) were isolated from CMT positive milk samples. In total, 11 CNS species: S. epidermidis (n=18), S. simulans (n=14), S. warneri (n=10), S. hominis (n=5), S. chromogenes (n=4), S. caprae (n=4), S. xylosus (n=3), S. haemolyticus (n=3), S. hyicus (n=3), S. cohnii (n=2), and S. capitis (n=1) were identified. The most commonly identified CNS species were Staphylococcus epidermidis (26.8%) and Staphylococcus simulans (20.8%) followed by Staphylococcus warneri (14.9%). Out of 67 CNS isolates, slime production was found in 37 (55.2%) CNS strains. CNS isolates were the most resistance to trimethoprim+sulphamethoxazole (76.2%), erythromycin (73.2%), oxacillin and ampicillin (70.2%) followed by penicillin (58.3%), gentamicin (53.8%), tetracycline (52.3%), vancomycin (51.8%), ciprofloxacin (26.9%), cefoxitim (23.9%), and cephalothin (13.5%). These results indicate that CNS species are resistant at high rates to the beta-lactam antibiotics which are intensively used in the prevention and treatment of mastitis without any antibiotic susceptibility test in the Middle Western of Turkey.
2017
The purpose of the study was to isolate Staphylococcus aureus from bovine subclinical mastitis, determine their antibiotic susceptibilities and investigate the coagulase gene polymorphism by using a PCR-based restriction fragment length polymorphism (RFLP) method. Milk samples from 463 CMT positive udders from 237 cows cultured. The antimicrobial susceptibility of the isolates were determined by disc diffusion method. A total of 82 out of the 83 isolates (98.8%) were found to be resistant at least one out of the 16 antibiotics studied. In this experiment 53 isolates (63.8%) were found to be resistant to penicillin; 52 (62.67%) to trimethoprim/sulphamethoxazole; 51 (61.5%) to ampicillin; 40 (48.2%) to erytromycin; 29 (34.9%) to tetracycline; 18 (21.6%) to ciprofloxacin, 16 (19.3%) to clindamycin, 13 (15.6%) to chloramphenicol; 8 (9.6%) to gentamicin; 5 (6.0%) to cefoxitin; 4 (4.9%) to vancomycin; 3 (3.6%) to cephalotin; 2 (2.4%) nafcillin; one (1.2%) to oxacillin and one to (1.2%) fu...
Veterinary World
The present study was planned to determine the in vitro antibiotic susceptibility of coagulase-negative Staphylococci (CNS) strains isolated from clinical and subclinical cases of mastitis in dairy cows. Antibiotic sensitivity profile will be helpful to recommend early therapy at the field level prior to availability of CST results. Materials and Methods: The milk samples from cases of clinical mastitis received in Mastitis Laboratory, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana and those of subclinical mastitis collected during routine screening of state dairy farms, were subjected to microbial culture. Identification of CNS organisms was done by standard biochemical tests. Antibiotic sensitivity testing, based on 30 antibiotics belonging to 12 groups, was done on 58 randomly selected CNS isolates (clinical isolates: 41, subclinical isolates: 17). Results: Isolates were highly susceptible to chloramphenicol (98.3%), gentamicin (93.1%), streptomycin (91.4%), linezolid (91.4%), ceftixozime (87.9%), cloxacillin (86.2%), clotrimazole (86.2%), bacitracin (86.2%), enrofloxacin (84.5%) and ceftrioxone + tazobactum (70.7%), while resistance was observed against amoxicillin (77.6%), penicillin (75.9%), ampicillin (74.1%) and cefoperazone (51.7%). Overall, isolates from clinical cases of mastitis had a higher resistance than subclinical isolates. Conclusion: CNS isolates were susceptible to chloramphenicol, gentamicin and streptomycin, while higher resistance was recorded against routinely used penicillin group.
Journal of Dairy Science, 2014
The aims of this study were to determine the prevalence and antibiotic resistance of Staphylococcus aureus isolated from bovine clinical mastitis in Varamin, Tehran Province, Iran. All of the isolated Staph. aureus were identified by morphology and culture and confirmed using the API Staph identification system (bioMérieux, Marcy-l'Étoile, France). Antibiotic resistance genes were detected by PCR with oligonucleotide primers specific for each gene. Staphylococcus aureus was recovered from 43 of 207 (20.1%) bovine clinical milk samples. Using disk diffusion, methicillin-resistant Staph. aureus was detected in 5 of 43 (11.6%) samples. The pathogen showed high resistance against penicillin G (86%) and tetracycline (76.7%). The blaZ (penicillin) (86%), tetM (tetracycline), and ermC (erythromycin) genes (39.5% each) were the most prevalent antibiotic resistance genes. The findings of this study are useful for designing specific control programs for bovine clinical mastitis caused by Staph. aureus in this region of Iran.
Hassan W. H.a,*, Hatem M. E.b, Elnwary H. A.c, Sediek S. H.c, 2016
In the current study, a total of 20 and 78 milk samples were collected from animals showed signs of clinical and subclinical mastitis, for isolation and identification of different causative pathogens in some dairy farms of Beni-Suef Governorate, and for investigation of in vitro sensitivity. The recovered microorganisms were Staphylococcus species (n=79; 80.61%), Enterococcus spp. (n=28; 28.57%), CAMP negative Streptococci, Pseudomonas aeruginosa (n=7; 7.14%), E. coli (n=3; 3.06%) and Proteus vulgaris (n=1; 1.02%). Antibiogram profile for S. aureus showed that the most effective drug was vancomycin and the least was penicillin. Trials were done to detect biofilm production for recovered isolates of S. aureus (n=23) by the use of a phenotypic method (Congo red agar, CRA) and genotypic methods through determination of some biofilm related genes using PCR. All recovered S. aureus isolates were seeded on the CRA media to detect the biofilm forming ability. It has been found that all tested isolates showed a biofilm forming ability either strong (13; 56.52%) or intermediate (10; 43.48%). The detection of some biofilm associated genes (icaA, icaD and bap genes) using polymerase chain reaction revealed that two (10.53%) isolates out of 19 were negative for all tested genes, 16 (84.21%) isolates harbored both icaA and icaD gene, while only one (5.26%) isolate had all tested genes.
Medical Laboratory Journal, 2017
Background and Objectives: Mastitis caused by Staphylococcus aureus is a widely distributed disease in cattle, goats and sheep. The infection is often subclinical in cattle, leading to reduced milk production and quality, but acute catarrhal or even gangrenous inflammation may also occur. The aim of this study was to investigate resistance of S. aureus isolates from bovine subclinical mastitis to a number of antibiotics. Methods: Milk samples were collected from 120 cows in different regions of Tabriz, Iran. Milk samples were cultured and bacteriological identification was performed. Antimicrobial susceptibility of the isolates was assessed by determining minimum inhibitory concentration. Plasmid DNA was extracted by an alkaline lysis method. Results: The highest frequency of resistance was observed against gentamicin (100%) and βlactam antibiotics including amoxicillin (96%), ampicillin (40%) and penicillin (96%). The isolates mostly contained large plasmids, which might harbor acquired antibiotic resistance. Conclusion: The results confirm the high frequency of antibiotic resistance among staphylococci isolated from bovine subclinical mastitis.
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.
3,000 half-udder milk samples from 8 goats flocks reared in Sardinia were collected and analysed for mastitis pathogens. From 469 positive samples, 421 Coagulase Negative Staphylococci, 25 S. aureus, 4 Micrococcus spp, and 21 other species were isolated and identified. Four of the strains were not identified. Antibiotic susceptibility tests were carried out on 25 S. aureus and 75 CNS randomly selected strains. Minimum Inhibitory Concentrations (MICs) were determined by using the broth micro-dilution method according to the guidelines and standards of The Clinical and Laboratory Standards Institute (CLSI). The MIC values were compared to breakpoints of ampicillin (AMP), cephalothin (KF), cefoperazone (CFP), ceftriaxone (CRO), cloxacillin (OB), kanamycin (K), novobiocin (NV), ofloxacin (OFX), oxytetracycline (OT) and vancomycin (VA). More than 96 % or over of the strains were susceptible to all the antimicrobial agents tested, with the exception of AMP (70.0%), K (74.0%), OT (92.0%) and OFX (94.0%). The MIC90 for these antimicrobial agents were, respectively, 4.0, 0.25, 4.0, 8.0,1.0, 64.0, 0.5,1.0, 2.0 and 4 ug/ml. Multiple resistance was detected in 8 strains.