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Relationship of Staphylococcal isolates in a Moroccan hospital by comparing phenotypical and genotypical tests.

The relationship of Staphylococcus isolates was determined among a collection of 26 clinical strains at the Centre Hospitalier Universitaire de Rabat. These isolates originated principally from blood culture and wounds. In order to affirm the clonal origins of these isolates, six phenotype (biotype, anti-biotype, serotype, phage type), and genotype (random amplified polymorphic DNA, pulsed field gel electrophoresis) methods were used. Biotyping, anti-biotyping, phage and serotyping were generally not sufficient while many isolates remained non-phage typeable. Random amplified polymorphic DNA analysis used in epidemiological typing seemed suitable for S. epidermidis and S. haemolyticus. However, rigorous standardization will be needed to assure reliable results. Pulsed field gel electrophoresis discriminated more efficiently than random amplification polymorphic DNA analysis. This study attests to the suitability of two or more methods in combination for typing Staphylococcus isolates.

Bacterial Typing Techniques↗

Relatedness among coagulase-negative staphylococci: deoxyribonucleic acid reassociation and comparative immunological studies.

DNA-DNA-homology values were determined under restrictive to relaxed reassociation conditions with type strains and some additional strains of coagulase-negative staphylococci belonging to ten different species. The immunological relationship of the catalases present in the type strains of these species was also determined by applying double immunodiffusion and microcomplement fixation. The results of these studies support the previous proposal to subdivide the coagulase-negative staphylococci into at least ten separate species. However, it is evident that some of the species are more closely realted than others and can form species groups. According to the results presented in this study, the coagulase-negative staphylococci can be combined into five species groups: The Staphylococcus saprophyticus group is composed of S. saprophyticus, S. xylosus and S. cohnii. The S. epidermidis group comprises S. epidermis, S. capitis and S. warneri. The S. hominis group which exhibits a significant relationship to S. epidermidis includes S. hominis and S. haemolyticus. The species group S. sciuri consists of S. sciuri ssp. sciuri and S. sciuri ssp. lentus and the species group S. simulans is presently represented by the corresponding single species.

Catalase↗

[Virulence factors of coagulase-negative Staphylococcus of human and bovine origin].

Virulence factors such as hydrophobicity of cell surface, capsule formation and slime production were investigated in 393 CNS strains, 190 isolated from bovine mastitis and 203 from human infections; in addition a selected number of CNS species were experimentally infused into the teat cistern of lactating rabbits for pathogenicity tests. Most human (95%) and bovine (82.6%) strains showed cell hydrophobicity properties, however only 11.3% and 7.9% were capsulated strains, respectively. Slime production was detected in 10.5% of bovine strains but only in 0.9% of human strains. Using the lactating rabbit model it was shown that S. chromogenes, S. epidermidis, S. saprophyticus and S. haemolyticus were the most pathogenic species.

Animals↗

The investigation of Staphylococcus aureus and coagulase-negative staphylococci nasal carriage among patients undergoing haemodialysis.

The frequency of nasal staphylococcal colonization among haemodialysed patients was investigated. The swabs were collected in 1998 and 2004 from 28 and 43 patients, respectively. Staphylococcus aureus colonization rates were 57.1% and 27.9% in 1998 and 2004, respectively. Twenty-six coagulase-negative staphylococci (CNS) isolates were cultured: S. epidermidis (21), S. lugdunensis (2), single S. haemolyticus, S. warneri, and S. capitits isolates. One S. aureus and 10 CNS isolates were methicillin resistant. The methicillin-resistant S. aureus (MRSA) was resistant to beta-lactams, tetracycline, and harbored the pvl gene encoding the Panton-Valentine leukocidin. The decrease in S. aureus colonization at 6-year interval was observed. The presence of the pvl gene and a favorable antibiotic susceptibility pattern of the MRSA suggest that the isolate was a member of community-acquired MRSA (CA-MRSA). Concluding, screening of haemodialysed patients for staphylococcal colonization accompanied by characterization of cultured isolates is important to understand its epidemiology and to develop infection prevention measures and treatment strategies.

Carrier State↗

[Drug sensitivity and teicoplanin resistance of coagulase-negative Staphylococcus isolated from blood culture].

Drug sensitivity and teicoplanin (TEIC) resistance of coagulase-negative Staphylococcus spp (CNS) isolated from blood culture in 10 years between 1992 and 2001 were investigated, and epidemiological analysis was performed using the survey of clinical backgrounds. Regarding the resistance rates of CNS to various drugs, no isolate was resistant to vancomycin (VCM), arbekacin (ABK), and linezolid (LZD), but the minimum inhibitory concentration (MIC) of TEIC was 16 micrograms/ml or higher in 10.3% of isolates. The isolates include 56 S. epidermidis strains, 1 S. capitis strain and 4 S. haemolyticus strains. As for other drugs, the minocycline (MINO) resistance rate was relatively low (6.1%), and methicillin-resistant CNS (MRCNS) accounted for 67.1%. The resistance rate was high for many drugs between 1995 and 1997. MIC50/90 of drugs considered to be anti-Staphylococcus were 1/2 microgram/ml (VCM), 1/16 microgram/ml (TEIC), 0.25/1 microgram/ml (ABK) and 1/2 microgram/ml (LZD), showing that both MIC 50 and MIC90 of ABK were the lowest. Regarding cross-resistance to TEIC, the resistant group (MIC: 16 micrograms/ml or higher) exhibited cross-resistance to all drugs excluding MINO. Particularly, 90.2% of the TEIC resistant group was MRCNS. TEIC-resistant strain was most frequently (30.8%) detected in the pediatric ward. In comparison of clinical backgrounds of the TEIC-resistant and TEIC-sensitive groups, there were no significant differences in age, use of IVH, or prognosis, but a significant difference in detection of TEIC-resistant strains was noted in patients treated with the 4 th cepharosporins and carbapenems (p < 0.01). This survey clarified the presence of TEIC-resistant CNS strains. We will continue the survey to monitor resistance conversion. Investigation of the involvement of TEIC-resistant CNS strains in hospital infection and therapeutic course is also necessary.

Blood↗

Subspecies discrimination of staphylococci from revision arthroplasties by ribotyping.

Sixty-six cultures of Staphylococcus spp. were obtained from bone and tissue samples collected from 37 patients during revision arthroplasties and were speciated and ribotyped to assess strain diversity in each species. There were 10 ribotypes among 51 isolates of S. epidermidis, three among three isolates of S. capitis, two among four isolates of S. aureus and two among two isolates of S. simulans. One ribotype was found among each of: two isolates of S. warneri; two isolates of S. haemolyticus and single isolates of S. cohnii and S. saprophyticus. Low molecular weight bands of ribotype patterns characterized one or two related species whereas high molecular weight bands were useful for distinguishing types within species. Specimens from 17 patients yielded more than one isolate of Staphylococcus spp. In 13 of these patients the isolates were representatives of a single species but in only eight did ribotyping show the isolates to be identical. The findings of multiple species and ribotypes from samples taken from the same patient may have implications for understanding the nature of infection in revision arthroplasty and for antibiotic therapy.

Bacterial Typing Techniques↗

Distribution of staphylococcal species on clinically healthy cats.

Among 827 isolates derived from 113 clinically healthy cats, 12 species of staphylococci were identified. Staphylococci were isolated from each cat and from 54.9% of the anatomic sites evaluated. A mode of 6 (range = 2 to 11) of the 11 anatomic sites evaluated per cat yielded staphylococci. A mode of 8 (range = 2 to 12) isolates were found per cat. Staphylococcus simulans was the most isolated (43.9% of total) coagulase-negative species. Moreover, S simulans was the most isolated species from each of the 11 sites evaluated and, except for the mouth and haircoat, comprised greater than 50% of the isolates from each site. Staphylococcus intermedius was the most isolated (13.5% of the total) coagulase-positive species. Three other species (S epidermidis, S xylosus, and S aureus) comprised 32.2% of the isolates, and 7 species (S haemolyticus, S hominis, S hyicus, S capitis, S warneri, and S saprophyticus) comprised 10.4% of the isolates. Six species (S intermedius [96 of 112 isolates], S haemolyticus [20 of 22], S sciuri [17 of 18], S warneri [10 of 13], S hyicus [10 of 10], and S capitis [7 of 8]) were isolated primarily from household cats. Only 1 species, S xylosus (75 of 87), was isolated primarily from cattery cats. Haircoat specimens (n = 452) yielded 508 isolates (61.4% of the total) distributed among all 12 staphylococcal species and included greater than 50% of the isolates of all species other than S simulans and S sciuri. A more heterogeneous population of staphylococci was isolated from household cats than was isolated from cattery cats.

Anal Canal↗

[Clinical significance of coagulase-negative staphylococci isolated from neonates].

OBJECTIVE: To evaluate the clinical significance of coagulase-negative staphylococci (CNS) isolated from newborns' infections at Neonatal Unit of Hospital das Clínicas da Faculdade de Medicina de Botucatu. METHODS: The CNS strains isolated were identified and classified as clinically significant and contaminant, based on a series of clinical and laboratory data obtained from patients who stayed in the Neonatal Unit. The following data were analyzed: risk factors for infections, clinical evolution, abnormal blood cell counts and/or C-reactive protein and antibiotic therapy. RESULTS: Among the 117 CNS strains isolated, 60 (51.3%) were classified as significant and 57 (48.7%) as contaminant. Among the 54 infants infected by CNS, 43 (79.6%) presented very low birthweight (<1,500 g). Most of the infants infected by CNS were submitted to two or more invasive procedures (77.8%), including use of catheter (88.9%), parenteral nutrition (64.8%) and mechanical ventilation (61.1%). Staphylococcus epidermidis was the most frequently isolated species (77.8%) and more often associated with infection (86.7%) than with contamination (68.4%). Other species of CNS, including two strains of S. haemolyticus, three strains of S. lugdunensis, one strain of S. simulans, one strain of S. warneri and one strain of S. xylosus were also isolated from infants with clinical evidence of pneumonia, necrotizing enterocolitis and sepsis. CONCLUSIONS: Most newborns infected by CNS presented important risk factors for infection onset, including birthweight <1,500 g, foreign body presence and previous use of antibiotics. The identification of CNS species constitutes a useful marker of infection, since S. epidermidis was the species more frequently associated with infection.

English Abstract↗

Immunological detection of penicillin-binding protein 2' in clinical isolates of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.

The additional penicillin-binding protein (PBP 2') that is important in determining intrinsic resistance in methicillin-resistant strains of Staphylococcus aureus (MRSA) has been detected immunologically in strains from a variety of world-wide locations. This additional protein has also been definitively identified both immunologically and as a PBP in methicillin-resistant strains of S. epidermidis (MRSE). The assay described is rapid, specific and sensitive and has been used to detect PBP 2' in S. haemolyticus but not in beta-lactam resistant Streptococci.

Acyltransferases↗

Incidence, adherence, and antibiotic resistance of coagulase-negative Staphylococcus species causing human disease.

Fifty-two isolates of coagulase-negative Staphylococcus species recovered from the blood or intravenous catheters of patients with clinically significant disease were compared to 60 similar isolates from patients who were presumably colonized. All isolates were identified and evaluated for ability to adhere to smooth surfaces, and resistance to anti-staphylococcal penicillins. S. epidermidis, S. hominis, and S. haemolyticus were the most frequently occurring species, representing 65%, 15%, and 10%, respectively, of disease isolates and 57%, 25%, and 8% of colonizers. The seven other species recovered accounted for only 10% of the total in both groups. Differences in isolation rates of each species within the two groups were not significant and were reflective of their reported incidence in the normal flora. All species of coagulase-negative Staphylococcus (except S. capitis and S. cohnii, which were isolated in very small numbers) were capable of adhering to smooth surfaces. S. hominis disease isolates were all capable of adherence, and the difference between the disease isolates and colonizers was statistically significant (p less than 0.02). This was not true for any other species that was analyzed nor for all isolates considered as a whole. Resistance to anti-staphylococcal penicillins was documented for all coagulase-negative Staphylococcus species, and was more frequent in S. epidermidis disease isolates than colonizers (p less than 0.05). No correlation was found between resistance to antistaphylococcal penicillins and ability to adhere.

Adhesiveness↗

Characterization of staphylococci associated with clinical and subclinical bovine mastitis.

Attempts were made to identify 900 species of staphylococci or micrococci recovered from samples of bovine milk examined for mastitis pathogens. The presence and identity of haemolysins was recorded together with results of disc diffusion antibiotic sensitivity tests. The occurrence of clinical mastitis was also noted and somatic cell counts (SCC) were performed on milk samples which were normal in appearance. Eight hundred and thirty-one coagulase positive staphylococci were obtained, of which 810 were S. aureus and 21 were S. intermedius. Of 65 coagulase negative staphylococci the species of 19 could not be determined by the identification systems used. The remainder were identified as S. hyicus sub sp. hyicus (1), S. hyicus sub sp. chromogenes (19), S. haemolyticus (17), S. hominis (3), S. epidermidis (4), S. capitis (1) and either S. hominis or S. warneri (1). Four other isolates could not clearly be assigned to the genus Staphylococcus or Micrococcus and were designated irregular strains. No micrococci were identified. The presence of alpha, beta, or delta haemolysins occurring singly or in various combinations was identified in 98.3% of coagulase positive staphylococci and in 60% of coagulase negative staphylococci. Epsilon haemolysin was detected in 47.6% of the coagulase negative staphylococci and in 9.5% of S. intermedius. All staphylococci were sensitive to tetracycline (30 microg), novobiocin (1.6 microg), nafcillin (30 microg), methicillin (10 microg) and cephalothin (30 microg) and variable numbers of each species were sensitive to penicillin (2 iu) and streptomycin (10 microg). One non-identified species of coagulase negative staphylococcus was sensitive to erythromycin (0.4 microg) the remaining staphylococci were resistant. Each of the four irregular strains was sensitive to erythromycin and novobiocin. Clinical mastitis was associated with 30.6% of coagulase positive staphylococci, 15.3% of coagulase negative staphylococci, and two of the four irregular strains (50%). Subclinical mastitis as determined by SCC of 500 x 10(3) or greater was associated with 92.7% of coagulase positive and 37.5% of coagulase negative staphylococci.

Journal Article↗

Novobiocin resistance and virulence of strains of Staphylococcus saprophyticus isolated from urine and skin.

A method was developed to study virulence of coagulase-negative staphylococci. Our results showed that coagulase-negative staphylococci injected into adult mice by the intracerebral route did not give rise to lethal infections, whereas mice aged 2 days were much more susceptible. Novobiocin-resistant strains of Staphylococcus saprophyticus were more virulent than strains of Staphylococcus epidermidis. Strains of S. saprophyticus biotype 3 of Baird-Parker's classification varied in virulence according to novobiocin sensitivity. In the classification of Kloos and Schleifer, S. saprophyticus biotype 3 can be subdivided into four distinct staphylococcal species, namely S. saprophyticus , S.cohnii, S.haemolyticus and S.warneri. S. chonii and S. saprophyticus were equally virulent for mice aged 2 days, but novobiocinsensitive S. haemolyticus was less virulent. On epidemiological grounds, however, it would seem that S. saprophyticus has some undefined advantage in invading the urinary tract.

Animals↗

[Phenotypic characteristics of coagulase-negative Staphylococci colonizing pleural drains in patients with lung cancer after thoracic surgery].

Coagulase-negative staphylococci (CNS) represent an important group of etiologic agents of infections associated with plastic biomaterials, e.g. drains. In the present paper 33 strains of CNS were characterized. All of them were isolated from fluid of pleural drains in patients with lung cancer after pulmonary resection under conditions of antimicrobial prophylaxis. The most frequently isolated species were Staphylococcus epidermidis and S. warneri. The majority of CNS strains showed ability to produce slime and possessed hydrophobic properties of cell surface. Strains of CNS resistant to penicillin and oxacillin, but sensitive to amoxicillin/clavulanate were isolated most frequently. Only two methicillin-resistant strains, belonging to S. haemolyticus, were found. The obtained data indicate that CNS strains colonizing pleural drains had potential ability to adhere to smooth surfaces. Most of isolated strains were susceptible to antibiotics and chemotherapeutics routinely used in staphylococcal infections.

Anti-Infective Agents↗

Evaluation of a mannitol-salt-oxacillin-tellurite medium for the isolation of methicillin-resistant Staphylococcus aureus from contaminated sources.

A mannitol-salt medium supplemented with 0.002% tellurite (MSOT) supported the growth of 104 of 109 strains of methicillin-resistant Staphylococcus aureus (MRSA) while suppressing 74 of 102 strains of mannitol-positive, methicillin-resistant coagulase-negative staphylococci. MSOT was equally or more sensitive and more differential than a similar formulation without tellurite for the isolation of MRSA from the skin and nares of hospitalized patients. Only S. haemolyticus produced dark gray-to-black colonies on MSOT similar to those of MRSA. MSOT is a useful adjunct to other nonselective culture formulations for the isolation of MRSA from contaminated sources.

Axilla↗

A Staphylococcus aureus-specific oligonucleotide probe derived from 16S rRNA gene sequences.

Genes encoding 16S rRNA were sequenced from 16 species of Staphylococcus. Sequence analysis highlighted a potential Staph. aureus-specific region and a complementary oligonucleotide probe was synthesized and its specificity tested. Northern blotting indicated molecular specificity, and dot blots to RNA from Staph. aureus, Staph. capitis, Staph. caprae, Staph. carnosus, Staph. caseolyticus, Staph. cohnii, Staph. epidermidis, Staph. gallinarum, Staph. haemolyticus, Staph. hominis, Staph. hyicus, Staph. saprophyticus, Staph. sciuri, Staph. simulans, Staph. warneri and Staph. xylosus indicated species-specificity.

Base Composition↗

In vitro antistaphylococcal activities of two investigative fluoroquinolones, CI-960 and WIN 57273, compared with those of ciprofloxacin, mupirocin (pseudomonic acid), and peptide-class antimicrobial agents.

By using broth microdilution, 373 clinical isolates of staphylococci were studied to determine their susceptibilities to CI-960, WIN 57273, ciprofloxacin, mupirocin, vancomycin, teicoplanin, and ramoplanin. Test strains comprised 179 strains of Staphylococcus aureus and 194 strains of coagulase-negative species. Strains of S. aureus were susceptible to CI-960, which had a mode MIC of 0.032 micrograms/ml and an MIC for 90% of the strains of 2 micrograms/ml. CI-960 was equally active against methicillin-susceptible and -resistant S. aureus strains as well as ciprofloxacin-resistant strains. Similarly, WIN 57273 was highly active, with a mode MIC of 0.008 micrograms/ml and an MIC for 90% of the strains of 1 micrograms/ml. No cross-resistance to CI-960 and WIN 57273 among ciprofloxacin-resistant strains was detected. Mupirocin was four- to eightfold more active than ramoplanin, vancomycin, and teicoplanin. With regard to coagulase-negative staphylococci, CI-960 and WIN 57273 were the most active of the test compounds, inhibiting all strains at 0.5 and 1 micrograms/ml, respectively. Against the same strains, mupirocin was fourfold more active than ramoplanin and eightfold more active than vancomycin. Five strains of S. haemolyticus were found to be resistant to ciprofloxacin, while resistance to teicoplanin was found among strains of S. epidermidis, S. haemolyticus, S. hominis, S. saprophyticus, S. simulans, S. warneri, and S. xylosus.

Anti-Infective Agents↗

Antimicrobial resistance and production of biofilms in clinical isolates of coagulase-negative Staphylococcus strains.

Coagulase-negative Staphylococcus (CNS) strains are frequently associated with bacteremia and hospital-acquired infections. 293 CNS strains were isolated from 744 samples from a dialysis center in S. M. de Tucumán, Argentina, from hemocultures, catheters and urine and identified as S. epidermidis, S. haemolyticus, S. saprophyticus, S. hominis and S. cohnii. 13 antibiotics were tested for antibacterial resistance. 75% of S. saprophyticus, 66% of S. epidermidis and 57% of S. haemolyticus was resistant to erythromycin and 50% of S. haemolyticus was resistant to ciprofloxacin. OXA resistance was found in 43% of S. haemolyticus. Presence of PBP 2a in OXA-R strains was confirmed with the modified agglutination assay (MRSA) and presence of the mecA gene. 15 strains with intermediate halos for vancomycin and teicoplanin showed a MIC in solid and liquid medium <or=1 microg/ml and <or=4 microg/ml respectively, which means that they were susceptible. 30% of the CNS strains produced biofilms: 42% of S. haemolyticus and 40% of S. epidermidis. 76% of the S. epidermidis and 59% of the S. haemolyticus biofilm-producing strains were isolated from catheters. The disc-diffusion technique together with the agglutination assay proved to be fast and reliable, which is of essential importance in intensive care units. Rapid identification of CNS strains, detection of resistance to methicillin and biofilm production are decisive for a prompt and appropriate antimicrobial therapy and limited use of inappropriate glycopeptides.

Anti-Bacterial Agents↗

Species identification & methicillin resistance of coagulase negative staphylococci from clinical specimens.

A total of 75 strains of coagulase negative staphylococci were isolated in pure culture from different specimens from patients suffering from various deep seated staphylococcal infections undergoing treatment between November, 1985 and December, 1986. Using the Baird Parker classification system, Staphylococcus epidermidis was the commonest isolate found (60%) of which biotype 1 was the most frequent (51.1%). Twenty seven (36%) untypable strains could be typed using Kloos and Schleifer's classification system. These strains were identified as Staph. cohnii (10), Staph. hominis (8), Staph. capitis (4), Staph. haemolyticus (3), Staph. simulans (1), and Staph. warneri (1). Three Staph. saprophyticus strains were isolated from patients of urinary tract infection. Methicillin resistant strains (14.6%) were isolated mainly from patients of meningitis, urinary tract infections and endocarditis. All these strains were sensitive to vancomycin. Coagulase negative staphylococci thus can cause a number of human infections and should no longer be regarded as harmless commensals.

Humans↗