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PubMed1995

Evaluation of new agglutination test for identification of oxacillin-susceptible and oxacillin-resistant Staphylococcus aureus.

A new agglutination test (Monostaph +; Bionor, Skien, Norway) has been developed. This new agglutination test has been compared with two other agglutination tests for the identification of 128 isolates of Staphylococcus aureus and 82 coagulase-negative staphylococci. The sensitivities of both Monostaph + and Pastorex Staph-Plus were excellent (98.7 and 97.4%, respectively) in detection of oxacillin-resistant Staphylococcus aureus. The specificity was 96.4% (two Staphylococcus epidermidis isolates and one Staphylococcus hominis isolate were false positive).

Agglutination Tests↗

PubMed1988

Rate of environmental mastitis in quarters infected with Corynebacterium bovis and Staphylococcus species.

Rates of environmental streptococcal and coliform intramammary infections were compared among quarters uninfected and infected with either Corynebacterium bovis or Staphylococcus species. Rate of environmental streptococcal intramammary infections was 3.9 times greater in C. bovis-infected quarters than in uninfected quarters. Rate of environmental streptococcal infections was 2.6 times greater in quarters infected with Staphylococcus species than in uninfected quarters. Rate of coliform intramammary infections did not differ among quarters with differing bacteriological infection statuses. Quarters infected with either C. bovis or Staphylococcus species had higher milk SCC than did uninfected quarters. Intramammary infection with either Corynebacterium bovis or Staphylococcus species did not protect quarters against coliform infection. Rate of environmental streptococcal infections was enhanced in quarters infected with either C. bovis or Staphylococcus species.

Animals↗

PubMed2026

Antimicrobial resistance in Staphylococcus spp. isolated from sporotrichosis-affected cats in Brazil: Detection of MRSP and MRSA.

Recently, Brazil has experienced a zoonotic emergence of sporotrichosis. The associated cutaneous lesions are often extensive and slow to heal, thereby providing a gateway for opportunistic bacteria belonging to the normal skin microbiota. Among these, Staphylococcus spp. are of particular concern due to their high prevalence and notable levels of antimicrobial resistance. The objective of this study was to identify and characterize Staphylococcus spp. isolated from the cutaneous wounds of domestic cats undergoing treatment for sporotrichosis and exhibiting clinical signs of secondary bacterial infection. A total of 233 samples from 203 cats were analyzed. Staphylococcus spp. was isolated from 156 samples (67%), with S. aureus (42.3%) and S. felis (25.6%) being the most prevalent. Antimicrobial susceptibility testing revealed high levels of resistance to penicillin (51.9%), erythromycin (28.8%), and clindamycin (19.2%). In contrast, most isolates were susceptible to chloramphenicol (98%), ciprofloxacin (96.7%), and nitrofurantoin (93%). Multidrug-resistant strains were identified in 24% (38/156) of the isolates. Overall, 12 isolates (7.7%) were classified as methicillin-resistant staphylococci, including four methicillin-resistant S. pseudintermedius (MRSP) and one methicillin-resistant S. aureus (MRSA). To investigate the genetic profiles and epidemiological relationships of these isolates, all the MRSP and MRSA strains were subjected to whole-genome sequencing. Among the MRSP isolates, four sequence types (STs) were identified, including ST551, the founder of clonal complex (CC)551, which is commonly associated with infection in dogs. The MRSA isolate belonged to ST1176, a member of CC5, which is a globally prevalent lineage and is frequently associated with nosocomial infections in humans. This study demonstrates that Staphylococcus species, including methicillin-resistant isolates, are frequently present in the wounds of sporotrichosis-infected cats exhibiting clinical signs of secondary bacterial infection. The detection of MRSA and MRSP in a cat highlights an additional public health concern associated with feline sporotrichosis and further reinforces the growing concern regarding antimicrobial resistance in companion animals.

Animals↗

PubMed1999

Presence and expression of collagen adhesin gene (cna) and slime production in Staphylococcus aureus strains from orthopaedic prosthesis infections.

Prosthesis-associated infections still represent one of the most serious complications in the clinical use of biomaterials. The most frequent causes are Staphylococcus aureus and Staphylococcus epidermidis. Several studies have been devoted to identify adhesion mechanisms for these bacteria. Slime in particular has been extensively investigated. Recently, in Staphylococcus aureus species, considerable attention has been given to the host protein receptors that have been shown in in vitro assays to serve as substrates for bacterial adhesion. Collagen-rich tissues, as bone and cartilage, that are the preferential sites of staphylococcal infections, are also the tissues that harbour orthopaedic implants. These can be easily coated in vivo by collagen and thus become prone to adhesion of Staphylococci strains which carry the collagen adhesin gene (cna). In this study the frequency of cna was determined within a collection of 35 Staphylococcus aureus strains from orthopaedic prosthesis infections by a PCR method. Also the collagen-binding ability and slime forming capacity was evaluated. 29% of the strains were cna-positive and also able to bind collagen in vitro. 83% of the strains were slime forming. The results indicate that in the examined bacterial population slime-positive strains predominate over the cna-positive strains, with a striking association of the two adhesion mechanisms in cna-positive strains.

Adhesins, Bacterial↗

PubMed2006

Influence of disk separation distance on accuracy of the disk approximation test for detection of inducible clindamycin resistance in Staphylococcus spp.

We undertook this study to assess the accuracy of the clindamycin-erythromycin disk approximation test (D-test) for detection of inducible clindamycin resistance in Staphylococcus spp. One hundred sixty-three Staphylococcus aureus and 68 coagulase-negative Staphylococcus (CoNS) spp. which were erythromycin nonsusceptible but clindamycin susceptible were tested using the D-test performed at both 15-mm and 22-mm disk separations and compared with genotyping as the "gold standard." The rate of inducible clindamycin resistance was 96.3% for S. aureus and 33.8% for CoNS spp. The sensitivities of the D-tests performed at 15 mm and 22 mm were 100% and 87.7%, respectively, and specificities were 100% for both. The use of 22-mm disk separation for the D-test to detect inducible clindamycin resistance results in an unacceptably high very major error rate (12.3%). All isolates with false-negative results harbored the ermA gene, and the majority were methicillin-resistant Staphylococcus aureus. False-negative results were associated with smaller clindamycin zone sizes and double-edged zones. We recommend using a disk separation distance of </=15 mm. There is wide geographic variation in the rates of inducible clindamycin resistance, and each laboratory should determine the local rate before deciding whether to either perform the D-test routinely or else report that all erythromycin-resistant S. aureus isolates are also clindamycin resistant.

Anti-Bacterial Agents↗

PubMed1992

Prevalence of Staphylococcus species during the periparturient period in primiparous and multiparous cows.

During a 14-mo period, 77 multiparous and 36 primiparous cows were sampled to determine the prevalence of staphylococci during the periparturient period. Distal streak canal swabs were taken at 14 d prepartum, and foremilk was sampled the first 5 consecutive wk of lactation. Staphylococcus aureus was isolated from 7.6% of quarters of primiparous cows but from only .6% of quarters of multiparous cows at parturition. Prevalence in primiparous cows declined to 3.5% by the wk-1 sampling. Quarter prevalence of coagulase-negative Staphylococcus species prepartum, at parturition, and wk 1 to 5 in primiparous cows was 38.9, 27.8, 15.3, 14.6, 13.2, 15.3, and 14.6%, respectively. In multiparous cows, prevalence at these times was 50.3, 12.3, 6.2, 8.1, 10.7, 7.1, and 8.1%. Staphylococcus chromogenes was the predominant species isolated, accounting for over 50% of the staphylococci isolated at each sampling time. Results suggest that high prevalence of staphylococci isolated prepartum is a reflection of natural skin flora and that a higher postpartum prevalence of these organisms was observed in primiparous cows than in multiparous cows. These data suggest also that the peripartum heifer could be a source of Staphylococcus aureus in the lactating herd.

Animals↗

PubMed1988

[An analysis of incidents of Staphylococcus in Kashima Rosai Hospital (I)].

Isolation patterns and drug susceptibilities of Staphylococcus aureus and Staphylococcus epidermidis were investigated in Kashima Rosai Hospital, a local hospital with 300 beds opened in June, 1981. Our investigation covered 218 Staphylococcus strains isolated from various clinical materials in the Clinical Laboratory during a one year period in 1985. These isolated strains comprised of 8 species of genus Staphylococcus with S. aureus and S. epidermidis together accounted for 93.6% of all the clinically isolated Staphylococci. S. aureus was depicted in materials obtained from outpatients at a higher frequency than S. epidermidis, while the opposite was the case in materials obtained from inpatients. When distribution of these organisms were classified depending on clinical materials from which they were isolated, outpatient sources from which S. aureus were isolated at high frequencies were otorrhea and pus, while inpatient sources with high incidents of S. aureus isolation were sputum and pus. With regards to S. epidermidis, urine and pus from outpatients as well as from inpatients yielded this organism at high frequencies. Isolation frequency ratios in this hospital of methicillin-resistant S. aureus (MRSA) and S. epidermidis (MRSE) were 8.6% and 30.*%, respectively. These resistant strains were found from a large spectrum of clinical materials. Thus, these strains seemed to have established themselves more or less firmly among bacterial population in the hospital, hence alerting us the risk of hospital infections in compromised hosts. As to the coagulase typing of MRSA, type IV was the most frequent from outpatient sources whereas type II was the most frequent from inpatient sources including a large variety of clinical materials. The coagulase typing has been and rapid than the phage typing. We confirmed these in our investigation. Among beta-lactam antibiotics we tested against MRSA, flomoxef was found to be superior to the others. Cephalothin and cefamandole proved to be highly active against MRSE. Minocycline and vancomycin showed good activities against MRSE as well as MRSA.

Anti-Bacterial Agents↗

PubMed1980

Protective effect in the lactating bovine mammary gland induced by coagulase-negative staphylococci against experimental Staphylococcus aureus infections.

The susceptibility of quarters preinfected either experimentally with Staphylococcus epidermidis or Staphylococcus xylosus or naturally by other coagulase-negative staphylococci was measured after challenge with two different strains of Staphylococcus aureus. The proportion of infected control quarters (95%) was raised significantly higher than that of those preinfected by the coagulase-negative staphylococci (24.5%). Differences were observed in the frequency of superinfection of the quarters according to the strain of the preinfective coagulase-negative staphylococcus, duration of preinfection and somatic cell count at the time of challenge. The protective mechanism against Staph. aureus induced by preinfection with coagulase-negative staphylococci is discussed.

Animals↗

PubMed1994

Identification of coagulase-negative staphylococci and typing of Staphylococcus epidermidis by a 4 h micromethod.

A new 4 h micromethod (Minibact-S) for identification of coagulase-negative staphylococci (CNS) important in human medicine, Staphylococcus epidermidis, Staphylococcus hominis, Staphylococcus haemolyticus and Staphylococcus saprophyticus, has been investigated. The reproducibility for species identification was 100% and the reproducibility for the single reactions was 95%. When compared to a modified Kloos & Schleifer conventional identification method, discrepant identification was obtained with the Minibact-S in only 8 of 382 clinical and reference strains. The system has been used for typing and biotyping of 201 S. epidermidis strains, and gave a subdivision in 69 different types; 4 related types accounted for 91 (45%) of the isolates, whereas 44 types were represented by only 1 isolate. The discriminatory index was 0.82. In a comparison with other typing methods performed on 81 isolates from 15 patients Minibact-S gave about the same number of types as antibiograms, but in 20% of the patients extra types were obtained with Minibact-S alone compared to antibiogram alone and vice versa. The Minibact-S is of some value in the routine clinical microbiology laboratory as a first screening method for identification of coagulase-negative staphylococci and typing of S. epidermidis in combination with the antibiogram.

Coagulase↗

PubMed1998

A polymerase chain reaction (PCR) method for the identification of collagen adhesin gene (CNA) in Staphylococcus-induced prosthesis infections.

Staphylococci are well-recognized pathogens of foreign body-associated infections. The pathogenesis of such infections involves an initial step of contact between the colonizing bacterium and the biomaterial, with subsequent colony formation. Several studies have been devoted to identify adhesion mechanisms for these bacteria. Slime in particular has been extensively investigated. Recently, considerable attention has been given to the host protein receptors that have been shown in in vitro assays to serve as substrates for bacterial adherence. To determine the importance of the collagen adhesin as virulence factor in Staphylococcus-induced prosthesis infection, a simple and reliable method using a polymerase chain reaction (PCR) was devised to identify collagen adhesin gene (cna). By using lysates of ten strains from orthopedic prostheses (5 Staphylococcus aureus and 5 Staphylococcus epidermidis) and two 20-oligonucleotides as primers, a 192-bp region of the cna gene was amplified by PCR and detected by agarose gel electrophoresis. Results obtained by this method were in accordance with those obtained by the in vitro phenotypic characterization of binding ability to collagen of Staphylococcus strains.

Adhesins, Bacterial↗

PubMed2005

A multiplex PCR method for the detection of all five individual genes of ica locus in Staphylococcus epidermidis. A survey on 400 clinical isolates from prosthesis-associated infections.

In Staphylococcus epidermidis, ica locus encodes for the synthesis of a polysaccharide intercellular adhesin (slime or biofilm). A multiplex polymerase chain reaction (PCR) for the detection of the five individual genes of ica locus was developed, with the aim to probe the set of genes in a large collection of Staphylococcus epidermidis clinical isolates. Single representative fragments for icaR, icaA, icaD, icaB, and icaC genes were selected. Multiplex PCR was applied to two reference Staphylococcus epidermidis strains [the non-biofilm-forming ATCC 12228 and the biofilm-forming ATCC 35984 (RP62A)] and to 400 clinical isolates of Staphylococcus epidermidis from orthopedic prosthesis associated infections. The gene profile was compared with the phenotypic biofilm-forming ability, evaluated by means of an optimized Congo red agar (CRA) plate test. Among the clinical isolates, 228 (57%) turned out completely ica positive and were biofilm producing. Among the 172 non-biofilm-forming strains (43%), 164 (41%) were completely ica negative and 8 strains (2%) harbored all five ica genes. The ica locus thus proves to be a cluster of strictly linked genes, without any evidence of single gene deletion.

Biofilms↗

PubMed2001

Interaction of coagulase-negative Staphylococcus species with bovine mammary epithelial cells.

Three coagulase-negative Staphylococcus species (CNS) (Staphylococcus epidermidis, Staphylococcus xylosus and Staphylococcus hyicus), from the milk of cows with mastitis, were used to evaluate adherence to and internalization by bovine mammary epithelial cells, and to investigate involvement of host cell signal transduction and host cell cytoskeleton rearrangement on internalization of CNS. S. xylosus showed highest adherence and internalization values of the species evaluated. Host cell cytoskeleton polymerization and protein kinase (PK) phosphorylation were required for internalization of CNS. Both protein kinase C (PKC) and tyrosine kinase (TPK) pathways were involved, but internalization of S. xylosus occurred preferentially through epidermal growth factor TPK activity. S. epidermidis and S. hyicus seemed to exploit other TPK pathways. Results of this study showed that S. xylosus, S. hyicus and S. epidermidis were able to adhere and internalize bovine mammary cells in a process that appeared to be receptor(s) mediated and exploited host signal transduction and cytoskeleton to induce an uptake signal.

Animals↗

PubMed1987

Early evaluation of coagulase negative staphylococcus in blood samples of intensive care unit patients. A clinically uncertain judgement.

Of 2160 intensive care unit patients, 36 patients with positive blood cultures had coagulase-negative staphylococcus in one blood bottle, whereas the organism was present in two or more bottles in 38 cases. The groups were not significantly different in 27 clinical variables, obtained at the time of their first positive blood culture. There was also no significant difference in the antimicrobial sensitivities. No initial clinical data supported the classification of coagulase-negative staphylococcus as either pathogen or contaminant. When the 74 patients with blood culture positive coagulase-negative staphylococcus were compared with three "control groups" ("absent septicemia," "probable septicemia" and "proven septicemia") they were not different from those with "probable septicemia." A discriminant analysis was performed comparing patients with "absent septicemia" and with "proven septicemia" in an attempt to classify patients with isolates of coagulase-negative staphylococcus in one of these groups at an early stage. Patients with two or more positive blood cultures were not statistically classified more frequently as septicemic than patients with one blood bottle positive for this organism. However, patients categorized as septicemic had a significantly higher mortality (59%) than those classified as non-septicemic (35%) (p less than 0.05).

Coagulase↗

PubMed1982

Studies on multi-antibiotic resistant strains of Staphylococcus aureus.

Twenty-seven isolates of Staphylococcus aureus were found to be resistant to methicillin, nafcillin, tobramycin, gentamicin, amikacin, cefoxitin, clindamycin, erythromycin and chloramphenicol by disc diffusion testing and tube dilution studies; they were sensitive to cefamandole, cephalothin and vancomycin by disc testing. A discrepancy between minimum inhibitory and bactericidal concentrations was noted for the cephalosporins which was not appreciated on disc testing. All isolates were very sensitive to rifampin (minimum bactericidal concentration 0.3 mg/l). All isolates were susceptible to phage type 83A. A subpopulation of variant small-colony forms of Staphylococcus aureus was recovered when superinhibitory amounts of aminoglycosides and methicillin were used. The variant Staphylococcus aureus strains retained the same antibiotic susceptibility patterns and phage type as their parent strain of Staphylococcus aureus. Further studies are in progress concerning these isolates and their impact on the hospital flora at our institution.

Anti-Bacterial Agents↗

PubMed1989

Effect of amoxycillin and clavulanic acid, alone and in combination, on human polymorphonuclear leukocyte function against Staphylococcus aureus.

The effect of amoxycillin and clavulanic acid on the interaction in vitro of human polymorphonuclear leukocytes with Staphylococcus aureus was examined. The exposure of a non-penicillinase producing Staphylococcus aureus strain to one-fourth the MIC of amoxycillin or clavulanic acid alone significantly increased the uptake of both unopsonized and opsonized bacteria by human polymorphonuclear leukocytes. This effect was also observed when bacteria were exposed to one-fourth the MIC of different proportions of the combination of amoxycillin and clavulanic acid (4/1, 1/1, 1/8 and 1/32). When a penicillinase-producing Staphylococcus aureus strain was used, only clavulanic acid significantly increased the uptake of unopsonized bacteria. The production of superoxide radicals by human polymorphonuclear leukocytes was impaired only by the presence of high concentrations (100 mg/l) of both clavulanic acid or the combination amoxycillin/clavulanic acid (4/1). At this high concentration, however, amoxycillin/clavulanic acid (4/1) significantly increased the intracellular killing of Staphylococcus aureus.

Amoxicillin↗

PubMed1995

Frequency of toxic shock syndrome toxin- and enterotoxin-producing clinical isolates of Staphylococcus aureus.

In 183 clinical isolates of Staphylococcus aureus, toxic shock syndrome toxin was detected in 13.7%, enterotoxin A in 20.2%, enterotoxin B in 7.7%, enterotoxin C in 5.5% and enterotoxin D in 3.3%. Seventy-three (39.9%) of the strains were found to produce one or more toxins. Multiple secretion of toxins was rare (< 1% of all strains) except for the combinations enterotoxin A with toxic shock syndrome toxin and enterotoxin A with enterotoxin D. These combinations were observed at significantly higher frequencies (4.9% and 2.2%, respectively) than would have been expected from values calculated from respective individual frequencies (0.88% and 0.06%) (p < 0.0001). In comparison with Staphylococcus aureus strains isolated from miscellaneous material, Staphylococcus aureus blood culture isolates produced enterotoxin D in significantly larger amounts and at higher frequencies (p = 0.01 in both cases). These toxins might play an important pathogenic role in Staphylococcus aureus infections.

Bacterial Toxins↗

PubMed1991

Identification of Staphylococcus and Micrococcus species with the STAPHYtest system.

A collection of 216 well-characterized strains of Staphylococcus, Micrococcus and Stomatococcus was examined by a commercially available STAPHYtest system (Lachema, Brno, Czechoslovakia). The results of STAPHYtest agreed with those of conventional tests. The STAPHYtest permitted a clear-cut separation of Staphylococcus from Micrococcus and Stomatococcus strains and correctly identified 104 of 145 (72%) Staphylococcus strains after 24 h of incubation. However, it allowed the identification only of 19 of 29 validly published Staphylococcus species. The STAPHYtest proved to be a simple and rapid system for the separation of staphylococci from micrococci and for the identification of most frequent clinically significant staphylococci.

Bacteriological Techniques↗

PubMed2001

Clinical and microbiological characteristics of 28 patients with Staphylococcus schleiferi infection.

The aim of this study was to analyse the clinical and microbiological characteristics of a series of patients with infection by Staphylococcus schleiferi. Seventy-one isolates were recovered from 36 patients between January 1993 and June 1999 at a tertiary care centre in northern Spain. There were 28 patients with well-documented clinical data. Infection was more frequent in men (89.3%), and more than half of the patients had some degree of immunosuppression, mainly malignant neoplasms. Infection was nosocomial in 22 cases and community-acquired in the remaining cases. Staphylococcus schleiferi was frequently associated with wound infections, mainly surgical-site infections, although unusual types of infections were detected. Infection-related mortality was low. This study highlights the importance of careful identification of Staphylococcus schleiferi in the clinical microbiology laboratory. Due to the documented association of Staphylococcus schleiferi with clinical infections in humans, any isolates of this organism should be assumed to be pathogenic, unless proven otherwise.

Adult↗