coa types and antimicrobial resistance profile of Staphylococcus aureus isolates from cases of bovine mastitis

The objective of the study was to determine the interrelationship between coa types of Staphylococcus aureus isolates from bovine mastitis cases and their susceptibility patterns to commonly used antimicrobial agents. In this study, a total of 63 S. aureus isolates obtained from 387 clinical and subclinical bovine mastitis milk specimens were examined for antimicrobial susceptibility against various classes of antibiotics and coa typing according to the size of the PCR product. Overall, five resistance profiles, nomenclatured PA, PAAc, PAE, PAAcE, and PAAcT according to their resistance and five different coa types, numbered I to V, were noted among the isolates studied. The most coa types were type I (38.1% of the 63 isolates) and type III (36.51%) and indicates that a few coagulase gene types of S. aureus are responsible for the majority of bovine mastitis cases. Resistance to penicillin, ampicillin, and amoxicillin/clavulanic acid were most common, respectively, with 63 (100%), 63 (100%), and 41 (65.1%) isolates and appeared to be higher than those usually reported. No resistance was detected for trimethoprim and gentamicin. Furthermore, a good relationship between phenotypic antibiogram and the polymorphism of coagulase gene was found in this study. Predominant coa types I and III were significantly associated with resistance profiles PAAcT and PAAcE, respectively. These two predominant types also showed high antibiotic resistance characteristic in comparison with rare types. As a result, this study illustrated a tendency for isolates from different coa types to have specific antimicrobic susceptibility profiles, presumably because of genetic diversity between isolates and/or selection force through the use of drugs.     

Molecular basis of antimicrobial drug resistance in Staphylococcus aureus isolates recovered from young healthy carriers in Spain

The frequency and genetic bases of antimicrobial drug resistance was determined for 111 Staphylococcus aureus recovered from young healthy carriers in a Spanish region. Resistances to ampicillin (84.7%), kanamycin (27%), erythromycin (25.2%), clindamycin (22.5%), tetracycline (11.7%), amikacin and tobramycin (6.3% each), gentamicin (5.4%), chloramphenicol (2.7%), ciprofloxacin (0.9%; MIC 4 μg/ml), moxifloxacin (0.9%) and mupirocin (0.9%; MIC 60 μg/ml) were found, and all were susceptible to methicillin (MSSA). Nearly 50% of the isolates were resistant to one antibiotic, 30% to two, 15.3% to three and 1.8% to four, while only 6.3% remained fully susceptible. A total of 31 profiles were found. For each phenotypic resistance, at least one gene accounting for it was identified. The detected genes were blaZ; erm(A)-erm(B)-erm(C)-msr(A)-msr(B)-lnu(A), aphA-aadE-sat4-aacA + aphD-aadD, tet(K), cat, and qacA/B, for resistance to ampicillin, macrolides and/or lincosamides, aminoglycosides, tetracycline, chloramphenicol, and quaternary ammonium compounds, respectively. In all isolates carrying cat genes, in all except one of the isolates positive for tet(K), and in most isolates with blaZ, erm(C), msr(A), or msr(B), the gene(s) mapped on resistance plasmids, which were detected in 69.2% of the resistant isolates (65% of the total). The S. aureus from young healthy carriers analysed in the present study do not constitute a reservoir of MRSA, but they represent a repository of multiple determinants conferring resistance to “old” antimicrobials. Some of these have still clinical applications and, considering the increasing resistance to recently introduced antimicrobials, none of them can be disregarded.     

烧伤病房金黄色葡萄球菌分离株的分子分型及其耐药性

BACKGROUND: Methicillin-resistant Staphylococcus aureus (MRSA) infection had been a global problem up to 1980s, and it has become a leading pathogen giving rise to nosocomial infections now. OBJECTIVE: To determine the molecular types and drug susceptibilities of Staphylococcus aureus prevailed in burn ward, and to provide a basis for preventing and controlling MRSA intections. METHODS: A total of 53 Staphylococcus aureus strains were collected from the burn ward in the Urumqi General Hospital of Lanzhou Military Region of Chinese PLA. These MRSA strains were identified by PCR and cefoxitin disc diffusion test, and all MRSA strains were typed by spa, SCCmec and MLST typing. In the meanwhile, antibiotic susceptibilities of 17 kinds of drugs, such as oxacillin, to Staphylococcus aureus were also determined, and drug resistance of different types of Staphylococcus aureus especially MRSA, was analyzed. RESULTS AND CONCLUSION:Among 53 Staphylococcus aureus strains, 43 were identified as MRSA, containing determined for amplification of meoA (n=41) and positive for cefoxitin disc diffusion test (n=2). Three SCCmec types, four spa types, and three ST types were found. The major predominant clone was ST239-MRSA-III-t030 (90.7%), with highest resistant to oxacillin and other nine antibiotics. In conclusion, the higher MRSA isolation rate from the burn ward, and ST239-MRSA-III-t030, as the predominant clone, presents with an outbreak in the burn ward and stronger resistance to many different families of antibiotics.       

DNA microarray genotyping and virulence and antimicrobial resistance gene profiling of methicillin-resistant Staphylococcus aureus bloodstream isolates from renal patients

Thirty-six methicillin-resistant Staphylococcus aureus (MRSA) bloodstream isolates from renal patients were genetically characterized by DNA microarray analysis and spa typing. The isolates were highly clonal, belonging mainly to ST22-MRSA-IV. The immune evasion and enterotoxin gene clusters were found in 29/36 (80%) and 33/36 (92%) isolates, respectively.     

Antibiotic resistance in Staphylococcus aureus and Staphylococcus epidermidis clinical isolates from implant orthopedic infections

In the last decade the rising phenomenon of resistance to most common antibiotic drugs among staphylococcal clinical isolates has been a reason for serious concern and alarm. The present study investigated the prevalence of antibiotic resistance within a large microbial collection including 530 clinical strains of S. aureus and 408 strains of S. epidermidis to a panel of 16 different drugs. All strains were isolated from orthopedic infections, either associated or non-associated with implant materials. Interestingly, our data show that the profile of the prevalence of antibiotic resistance within the two species of pathogens is extremely similar for the vast majority of the drugs screened. The only statistically significant variations in prevalence concerned, in order of relevance, the following 5 out of 16 antibiotics: sulfamethoxazole (in combination with trimethoprim), erythromycin, and, to a lesser extent, oxacillin, imipenem, and clindamycin. In the case of Staphylococcus aureus, the isolates associated to implant materials were found more frequently resistant to all 4 aminoglycosides screened as well as to ciprofloxacin.     

Genotyping of clinical methicillin-susceptible Staphylococcus aureus isolates in a Dutch teaching hospital

Methicillin-susceptible Staphylococcus aureus isolates, recovered from 204 patients in our hospital in a 22-month period, were characterized by pulsed-field gel electrophoresis. Among the multiple S. aureus types six clonal lineages dominated, comprising isolates from 158 patients. Despite the limited genetic variation, cross-transmission was made plausible only sporadically.     

An international multicenter study of antimicrobial consumption and resistance in Staphylococcus aureus isolates from 15 hospitals in 14 countries

Antibiotic consumption during 1996 was measured in 15 large hospitals from 14 countries and 3000 consecutive Staphylococcus aureus samples were collected, allowing calculation of local resistance rates and typing of isolates. Antibiotic consumption data were converted to defined daily doses (DDD), and similar antibiotics were grouped if they belonged to the same therapeutic subgroup. Variations in hospital size were corrected by using DDD per 1000 bed-days. The total antibiotic consumption in the 15 hospitals varied between 296 DDD/1000 bed-days and 1108 DDD/1000 bed-days. Differences in the usage of therapeutical subgroups of antimicrobials varied significantly between hospitals. A positive correlation was found between S. aureus resistance to methicillin (MRSA) and consumption of beta-lactam combinations, between resistance to quinolones and consumption of beta-lactam combinations and carbapenems and resistance to aminoglycosides and consumption of beta-lactam combinations. The consumption of beta-lactamase-sensitive antibiotics was negatively correlated to resistance to methicillin, quinolones, and aminoglycosides. Usage of the different antimicrobial therapeutical subgroups was also correlated. Consumption of beta-lactamase-sensitive antibiotics (penicillin) was positively correlated to consumption of beta-lactamase-resistant penicillins and negatively correlated to consumption of carbapenems, quinolones, and glycopeptides, whereas consumption of cephalosporins was positively correlated to consumption of aminoglycosides, quinolones, and glycopeptides. In this study of hospitals with MRSA prevalence of between 0% and 63%, significant correlations were found between resistance and consumption of antimicrobials. These findings support the importance of antimicrobial consumption on resistance. An accompanying paper addresses the issue of antibiotic resistance and clonality of isolates.     

Genetic diversity and antimicrobial resistance of Staphylococcus aureus from recurrent tonsillitis in children

The aim of this study was to analyze the prevalence of Staphylococcus aureus in the tonsils of children subjected tonsillectomy due to recurrent tonsilitis and to determine the spa types of the pathogens, carriage of virulence genes and antimicrobial resistance profiles. The study included 73 tonsillectomized children. Bacteria, including S. aureus were isolated from tonsillar surface prior to tonsillectomy, recovered from tonsillar core at the time of the surgery, and from posterior pharynx 2–4 weeks after the procedure. Staphylococcus aureus isolates were compared by spa typing, tested for antimicrobial susceptibility and for the presence of superantigenic toxin genes (sea-seu, eta, etb, tst, lukS/lukF-PV) by multiplex polymerase chain reaction. Seventy-three patients (mean 7.1 ± 4.1 years, 61.6% male) were assessed. The most commonly isolated bacteria were S. aureus. The largest proportion of staphylococcal isolates originated from tonsillar core (63%), followed by tonsillar surface (45.1%) and posterior pharynx in tonsillectomized children (18.2%, p = 0.007). Five (6.3%) isolates were identified as MRSA (mecA-positive). Up to 67.5% of the isolates synthesized penicillinases (blaZ-positive isolates), and 8.8% displayed MLS_B resistance. The superantigenic toxin genes were detected in more than half of examined isolates (56.3%). spa types t091, t084, and t002, and clonal complexes (CCs) CC7, CC45, and CC30 turned out to be most common. Staphylococcus aureus associated with RT in children showed pathogenicity potential and considerable genetic diversity, and no clones were found to be specific for this condition although further studies are needed.     

Antibiotic resistance patterns of mecA-positive Staphylococcus aureus isolates from clinical specimens and nasal carriage

The present study demonstrates that the nasal carriage rate of Staphylococcus aureus was 40% in Jordanian healthy young adult population, and 19% of nasal S. aureus and 57% of clinical isolates over the same period were resistant to oxacillin (MRSA), respectively. The mecA gene was detected in all MRSA isolates in both groups. Most of MRSA isolates were multiresistant to three antibiotic classes (beta-lactams, aminoglycosides, macrolides-lincosamides). This result suggests a serious problem may be encountered in treatment of staphylococcal infections in Jordan.     

An international multicenter study of antimicrobial resistance and typing of hospital Staphylococcus aureus isolates from 21 laboratories in 19 countries or states

During 1996, 4065 consecutive Staphylococcus aureus strains from different patients were collected in 21 worldwide hospital laboratories. The strains, their resistance pattern, and hospital demographic data were forwarded to Statens Serum Institut where the strains were typed and data analyzed. Resistance patterns varied by region and resistance to other antibiotics than methicillin were mainly related to the occurrence of methicillin resistance, except for mupirocin, rifampicin, and fusidic acid. Methicillin-resistant S. aureus (MRSA) occurred with low levels in hospitals in Northern Europe (<1%), increasing levels in middle-European countries, United States, New Zealand, and Australia (6-22%), and very high levels in Southern European countries as well as in parts of the United States, Asia, and South Africa (28-63%). MRSA found in large hospitals were more resistant to other antibiotics than MRSA found in smaller hospitals serviced by the same laboratory. No difference in resistance levels was seen for methicillin-susceptible S. aureus (MSSA) isolated in large or small hospitals. Intensive Care Units had the highest level of MRSA. Strains from the lower respiratory tract showed the highest resistance levels and blood isolates the lowest. A dominating MRSA clone was found in hospitals with an MRSA frequency of more than 10%. Pulsed-field gel electrophoresis (PFGE) typing recognized several of these clones as international epidemic MRSA (E-MRSA). All MSSA isolates were phage typed (typeability 85.4%) and divided in seven major phage patterns. Isolates of all patterns were found in all hospitals except one, indicating that the MSSA seldom represented the spread of clones within the hospital. The comparison should evaluate the prevalence of community-acquired MRSA and identify internationally E-MRSA. The present study gives a snapshot of the MRSA situation, but it is important to build up a continuous national and international surveillance, because MRSA is a global socioeconomic problem. Global infection control procedures, including rational antibiotic use, should be agreed on. The accompanying paper will address the issue of antibiotic consumption and MRSA.     

Novel and uncommon antimicrobial resistance genes in livestock-associated methicillin-resistant Staphylococcus aureus

Livestock-associated methicillin-resistant Staphylococcus aureus (LA-MRSA) isolates have been the subject of numerous studies during recent years. The characterization of such isolates has usually also included the determination of their resistance phenotypes and associated resistance genotypes. Analysis of the resistance genes present in LA-MRSA isolates has revealed a number of genes commonly found in S. aureus and coagulase-negative staphylococci of humans and animals. In addition, novel resistance genes and/or resistance genes that have been rarely detected in staphylococci so far have been encountered. These include the phenicol exporter gene fexA, the multiresistance gene cfr, the tetracycline resistance gene tet(L), the trimethoprim resistance gene dfrK, the macrolide–lincosamide–streptogramin B resistance gene erm(T), the lincosamide–streptogramin A–pleuromutilin resistance genes vga(C) and vga(E), and the apramycin resistance gene apmA. Most of these genes were located on multiresistance plasmids in LA-MRSA. The co-localization of these resistance genes with other resistance genes enables their co-selection and persistence. LA-MRSA can therefore act as a donor and a recipient of antimicrobial resistance genes within the Gram-positive gene pool.     

Prevalence of inducible clindamycin resistance among community- and hospital-associated Staphylococcus aureus isolates

Methicillin-resistant Staphylococcus aureus (MRSA) infections have become common among both hospitalized and nonhospitalized patients. Optimal outpatient therapy for MRSA infections has yet to be determined, but this matter is complicated by the possibility of inducible macrolide-lincosamide-streptogramin B resistance (MLSBi). We studied the prevalence of MLSBi in community- and hospital-associated S. aureus isolates and the prevalence of community-associated MRSA (CA-MRSA) and identified clinical predictors of CA-MRSA and MLSBi. Among 402 S. aureus isolates, the overall prevalence of MLSBi was 52%, with 50% of MRSA and 60% of methicillin-susceptible S. aureus isolates exhibiting MLSBi. CA-MRSA represented 14% of all isolates and had a lower prevalence of MLSBi than hospital-associated MRSA (33% versus 55%). The presence of skin or soft-tissue infection was predictive for CA-MRSA, and the presence of a comorbidity was predictive for MLSBi. Due to the low prevalence of MLSBi among CA-MRSA isolates, clindamycin remains a useful option for outpatient therapy.     

Comparison of tests to detect oxacillin resistance in Staphylococcus intermedius, Staphylococcus schleiferi, and Staphylococcus aureus isolates from canine hosts

Multiple tests were compared to the reference standard PBP2a latex agglutination test for detection of mecA-mediated oxacillin resistance in canine staphylococci. Cefoxitin disk diffusion, using breakpoints for human isolates of coagulase-negative Staphylococcus spp., had low sensitivity for detection of oxacillin resistance in members of the Staphylococcus intermedius group.     

Testing for induction of clindamycin resistance in erythromycin-resistant isolates of Staphylococcus aureus

Disk diffusion and broth microdilution (BMD) were used to perform clindamycin (CLI) induction testing on 128 selected nonduplicate isolates of Staphylococcus aureus. Disk diffusion testing involved placing CLI and erythromycin (ERY) disks approximately 12 mm apart (measured edge to edge) on a Mueller-Hinton agar plate that had been inoculated with an S. aureus isolate; the plate was then incubated for 16 to 18 h. Two distinct induction phenotypes (labeled D and D(+)) and four noninduction phenotypes (designated as negative [Neg], hazy D zone [HD], resistant [R], and susceptible [S]) were observed in disk diffusion results. A clear, D-shaped zone of inhibition around the CLI disk was designated as the D phenotype and was observed for 21 isolates while a D-shaped zone containing inner colonies growing up to the CLI disk was designated as D(+) (17 isolates). In addition, 10 isolates were CLI susceptible and ERY resistant but were not inducible and showed no blunting of the CLI zone (Neg phenotype). Isolates that were CLI and ERY resistant (constitutive macrolide-lincosamide-streptogramin B resistance) demonstrated either a double zone of inhibition with an inner ring of reduced growth up to the edge of the disks (HD phenotype; 33 isolates) or solid growth around the CLI and ERY disks (R phenotype; 16 isolates). Finally, 31 isolates were susceptible by disk testing to both CLI and ERY (S phenotype). PCR results showed that isolates with a D phenotype harbored ermA, isolates with a D(+) phenotype contained either ermC (16 isolates) or ermA and ermC (one isolate), and all 10 isolates with a Neg phenotype contained msrA. All isolates with an HD or R phenotype harbored at least one erm gene. Isolates showing the D(+) phenotype by disk diffusion were also detected by BMD using a variety of CLI and ERY concentrations; however, isolates with the D phenotype were more difficult to detect by BMD and will likely require optimization of ERY and CLI concentrations in multilaboratory studies to ensure adequate sensitivity. Thus, at present, disk diffusion is the preferred method for testing S. aureus isolates for inducible CLI resistance.     

Immunoblots, antimicrobial resistance, and bacteriophage typing of oxacillin-resistant Staphylococcus aureus.

An immunoblotting system was developed for typing of oxacillin-resistant Staphylococcus aureus. Clinical isolates recovered during a 40-month period at a single institution were evaluated with this typing scheme. Results were compared with susceptibility patterns and with bacteriophage typing results for 100 clinical isolates and with plasmid fingerprints for 14 isolates. Immunoblotting was found to be a useful method with good reproducibility that distinguished seven major groups of patient isolates that were clinically and epidemiologically related. Susceptibility patterns showed specific correlations with other typing results but were inferior to immunoblotting and phage typing for differentiating major groups of organisms. Plasmid profiles failed to distinguish two major groups that were readily identified by immunoblots and phage typing. There was evidence of increasing antimicrobial resistance of endemic hospital strains. Immunoblotting correlated well with phage typing, offered an alternative method for typing isolates that could not be typed by phage typing, and was superior to susceptibility testing and plasmid profiles for distinguishing different groups of oxacillin-resistant S. aureus at our institution.     

Molecular characterization of resistance to mupirocin in methicillin-susceptible and -resistant isolates of Staphylococcus aureus from nasal samples

A total of 15 of 101 (14.8%) nasal methicillin-resistant Staphylococcus aureus (MRSA) isolates exhibited mupirocin resistance (Mup(r)) compared with 1 of 154 (0.6%) methicillin-susceptible Staphylococcus aureus isolates. A total of 14 (93%) isolates exhibiting high-level Mup(r) belonged to a single clone. Horizontal plasmid transfer and transmission of Mup(r) strains contribute to a high incidence of Mup(r) MRSA at our institution.     

Molecular and phenotypic typing of Staphylococcus aureus isolates from rabbit meat

Twenty-six Staphylococcus aureus isolates were recovered from rabbit carcasses and cuts during a period of seven months. Samples from 51 different animals, flocks and farms were obtained from five establishments in four Spanish provinces. To determine their diversity and possible origin, isolates were typed by three molecular and three phenotypic methods. PFGE, with the highest discrimination index (D=0.966), identified 19 patterns (more than one band difference) and 10 types (more than three band differences). Based on > or = 90% similarity, RAPD (D=0.877) produced nine patterns while ribotyping (D=0.786) produced seven types. On the basis of biotyping (D=0.644), 11 isolates belonged to human ecovars and 15 to the non-host-specific crystal violet type C (NHS CV:C) biotypes. By direct phage typing (D=0.761), 17 isolates were lysed by human phages into groups II (8 isolates), III (5 isolates), I/III (2 isolates) and V (2 isolates). The overall resistance to antimicrobials (D=0.783) was 76.9%, with most isolates being resistant to tetracycline (61.5%) and penicillin G (26.9%). PFGE showed that samples from each processing plant carried different S. aureus types, some of them persisting over time. There also was evidence of interestablishment dissemination of genetically related clones, most of them belonging to the PFGE type A and phenotype "NHS CV:C biotypes-3A/3C/55/71 phage type", which is highly virulent for European commercial rabbitries. The ubiquity of the virulent phenotype, as well as the high incidence of resistance to antibiotics with application in human medicine, is a matter of concern in public and animal health.     

Characterisation of methicillin-resistant Staphylococcus aureus isolates from dogs and their owners

Ten methicillin-resistant Staphylococcus aureus (MRSA) isolates from healthy owners and their pets were characterised by susceptibility testing, staphylococcal chromosome cassette (SCC)mec and agr typing, and detection of the Panton-Valentine leukocidin (PVL) genes. Two human and three dog isolates harbouring SCCmec type III appeared to be of hospital origin. The five remaining isolates carried SCCmec type IV, with three being multidrug-resistant. One type IV isolate was PVL-positive and a prototypic agr type 3, typified by strain MW2. This is the first report of this type in association with nasal carriage. Drug resistance may be increasing among community isolates of MRSA.