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Antibacterial activity of human cervical mucus.

The antimicrobial property of human uterine cervical mucus was tested in three groups of women. Healthy women, using no contraception, women using an intrauterine device and women receiving hormonal treatment for contraception. Cervical mucus was taken on the 10th, 14th, 18th and 22nd day of the menstrual cycle. Cervical mucus had a strong inhibitory effect on the growth of Micrococcus lysodeicticus in all three groups. The strength of the inhibitory effect on the other microorganisms were in the following order: Staphylococcus albus, Staphylococcus aureus, Proteus mirabilis, Escherichia coli, Candida albicans, Streptococcus haemolyticus, Streptococcus faecalis. Use of an intrauterine device did not affect the antimicrobial effect of cervical mucus. The use of hormonal contraceptive canceled the antimicrobial effect on the series of microorganisms, with the exception of M. lysodeicticus. The maximum inhibitory effect occurred on the 14th day and declined toward the end of the menstrual cycle.

Adult↗

Resident aerobic microbiota of the adult human nasal cavity.

Recent evidence strongly suggests that the microbiota of the nasal cavity plays a crucial role in determining the reaction patterns of the mucosal and systemic immune system. However, little is known about the normal microbiota of the nasal cavity. The purpose of this study was to determine the microbiota in different parts of the nasal cavity and to develop and evaluate methods for this purpose. Samples were collected from 10 healthy adults by nasal washes and by swabbing of the mucosa through a sterile introduction device. Both methods gave results that were quantitatively and qualitatively reproducible, and revealed significant differences in the density of the nasal microbiota between individuals. The study revealed absence of gram-negative bacteria that are regular members of the commensal microbiota of the pharynx. Likewise, viridans type streptococci were sparsely represented. The nasal microbiota was dominated by species of the genera Corynebacterium, Aureobacterium, Rhodococcus, and Staphylococcus, including S. epidermis, S. capitis, S. hominis, S. haemolyticus, S. lugdunensis and S. warneri. These studies show that the microbiota of the nasal cavity of adults is strikingly different from that of the pharynx, and that the nasal cavity is a primary habitat for several species of diphtheroids recognized as opportunistic pathogens. Under special circumstances, single species, including IgA1 protease-producing bacteria, may become predominant in a restricted area of the nasal mucosa.

Actinomycetales↗

Antimicrobial susceptibility of coagulase-negative staphylococci and characterization of isolates with reduced susceptibility to glycopeptides.

The antimicrobial susceptibility of 239 coagulase-negative staphylococci (CNS) isolates consecutively collected from blood culture in patients admitted in a 600-bed teaching hospital was evaluated. The isolates were identified to the species level by conventional methods and the MicroScan Positive Combo Panel type 6 system, and their susceptibility to vancomycin, teicoplanin, and oxacillin were tested by agar dilution, disk diffusion, and MicroScan-WalkAway system. The species distribution was as follows: Staphylococcus epidermidis 120 (50.2%), S. hominis 29 (12.1%), S. haemolyticus 24 (10.0%), S. cohnii 14 (5.9%), and isolates from other CNS species 52 (21.8%). The percentage of resistance to oxacillin was 74.5% by agar dilution. The highest percentages of oxacillin resistance were found among S. haemolyticus (95.8%) and S. epidermidis (80.8%). Teicoplanin resistance (MIC > or = 32 micrograms/mL) was detected in five S. haemolyticus isolates, whereas intermediate resistance (MIC = 16 micrograms/mL) was detected in nine strains. These isolates with reduced susceptibility to teicoplanin were resistant to oxacillin, but remained susceptible to vancomycin (MIC < or = 4 micrograms/mL). Two isolates, one S. haemolyticus and one S. epidermidis, showed a vancomycin MIC of 8 micrograms/mL, and both MicroScan and disk diffusion methods classified these isolates as susceptible. Our results showed that glycopeptide resistance is emerging among CNS isolates in our institution and the disk diffusion method may not detect isolates with decreased susceptibility to these antimicrobial agents.

Adult↗

Synergistic hemolysis exhibited by species of staphylococci.

The synergistic hemolysis reactions of 61 reference strains and 189 clinical isolates representing 17 species of staphylococci were examined on plates of Trypticase soy blood agar (BBL Microbiology Systems, Cockeysville, Md.). Some or all of the strains of Staphylococcus aureus, S. epidermidis, S. capitis, S. cohnii, S. haemolyticus, S. hyicus, S. simulans, S. warneri, and S. xylosus produced a delta-hemolysin that gave synergistic, complete hemolysis of washed human, sheep, and ox blood cells in an area of beta-lysin activity from strains of S. aureus and S. intermedius. Strains of the same nine species were positive with a commercial beta-lysin paper disk designed for presumptive identification of group B streptococci; most of these strains also gave synergistic, complete hemolysis with exotoxin from a strain of Corynebacterium pseudotuberculosis. None of the strains of S. auricularis, S. carnosus, S. caseolyticus, S. hominis, S. intermedius, S. saprophyticus, S. sciuri, or S. lentus were positive by any of these tests for synergistic hemolysis. These results indicate that a synergistic hemolysis test could prove very useful for differentiating these species; they also suggest that one role of some of these organisms in human infections could be that of a synergist. Further studies of synergism may clarify the clinical significance of these results.

Antimicrobial Cationic Peptides↗

Distribution and antimicrobial susceptibility of coagulase-negative staphylococci from skin lesions.

The distribution and antimicrobial susceptibility of coagulase-negative staphylococci from skin lesions were investigated. Staphylococcus epidermidis was found on all areas of the body, whereas S. capitis, S. haemolyticus and S. hominis were mainly found on the face/head or arm/leg. The distribution of coagulase-negative staphylococci in skin lesions and at the same location on normal skin was similar. Staphylococcus lugdunensis was the most susceptible to the nine tested antimicrobials (benzylpenicillin, ampicillin, piperacillin, cefazolin, erythromycin, minocycline, gentamicin, vancomycin and ofloxacin) and S. epidermidis the least susceptible. S. haemolyticus also showed low susceptibility to all nine antimicrobials. Low susceptibility to penicillins may be explained by beta-lactamase production. The existence of coagulase-negative staphylococci, especially concerning their potential pathogenicity and multiple drug resistance, should not be neglected.

Adolescent↗

[Oropharyngeal bacterial flora in asthmatic and health subjects].

MATERIAL AND METHODS: It was carried out cultivation of oozing pharyngeal to 248 fellows, 128 of them were asthmatic and 120 seemingly healthy. From group of asthmatic 55.46% they correspond to the masculine sex and 44.54% to the feminine; 72.97% they present more than two sharp squares of asthma per month. RESULTS: In the cultivation of 107 asthmatic the presence of germs is reported like: Staphylococcus aureus positive 43.56% (p = NS), the Streptococcus betahemolytic 72.97% (p = 0.001), Candida albicans 70.58% (p = 0.004), Streptococcus alpha haemolyticus 61.42% (p = 0.008). CONCLUSIONS: When they exist more than two germs in a same individual frequently the Staphylococcus aureus is detected with the Streptococcus beta hemolytic 42.10% or with the Candida albicans 31.57%. We proposes to magnify the problem to which the patients face in order to order a medication or examine of laboratory with opportunity and when we hoped to obtain the good benefit for the health of the population.

Adolescent↗

Polynucleotide sequence divergence among some coagulase-negative staphylococci.

The degree of binding was determined, at both exacting and non-exacting temperatures, between deoxyribonucleic acid (DNA) preparations from coagulase-negative staphylococci and carbon (14C) labelled DNA from nine reference strains, Staphylococcus capitis LK499, S. cohnii GH137, S. epidermidis GH37, S. haemolyticus SM131, S. hominis DM122, S. saprophyticus CCM883, S. simulans SM128, S. warneri AW25 and S. xylosus KL162. The close similarity in nucleotide sequences found between five of the six representatives received as S. capitis, S. hominis and S. warneri indicate that these organisms are separated into too many species. In contrast, little homology was found between the pairs of strains representing S. cohnii and S. haemolyticus, respectively. The divergence indices of the nine reference strains were calculated and the relationship between them expressed in a sorted similarity matrix and dendrogram based upon the average linkage algorithm. The pattern of nucleotide heterogeneity was found by plotting the relative DNA bound at the exacting temperature against the divergence indices. The coagulase-negative staphylococci exhibit extensive genetic diversity due mainly to random mutation.

DNA, Bacterial↗

Predictive values of species' identifications from the Vitek Gram-Positive Identification Card using clinical isolates of coagulase-negative staphylococci.

The Gram-Positive Identification Card (GPI, bioMerieux Vitek) was compared to conventional tests for identification of 616 clinical isolates of coagulase-negative staphylococci. All tests were inoculated with subcultures from single isolated colonies. The predictive values of GPI species' identifications (the number of the system's correct calls divided by the total number of calls for each species) were 100% (12 of 12) for Staphylococcus capitis, 100% (79 of 79) for S saprophyticus, 98.4% (246 of 250) for S epidermidis and 96.0% (120 of 125) for S haemolyticus, but only 64.5% (69 of 107) for the other species analyzed. When an infrequently encountered Staphylococcus species is named using the GPI, the identification should be confirmed using additional tests.

Bacteremia↗

The pattern of micro-organisms and the efficacy of new macrolide in acute lower respiratory tract infections.

Lower respiratory tract infection (LRTI) is one of the major health problems in developing countries such as Indonesia. According to the National Household Health Survey conducted by the Ministry of Health in 1992, LRTIs still rank fourth as the main cause of death in Indonesia. The problem of LRTIs could be simply managed as long as the causative organism can be identified and the proper antibiotic known. In some occasions, it is not quite so easy to identify the causative micro-organism, especially in lower tract infections. There are several methods of obtaining specimens from LRTIs for cultures. The easiest, most simple way is to collect expectorated sputum. Unfortunately, because of the high rate of contamination by upper respiratory tract flora, this method is not reliable. Recognizing the difficulties with routine expectorated sputum cultures, two alternative approaches have been suggested. One approach is to bypass potential expectorated sputum 'contaminants' in the oropharynx by transtracheal aspiration or transthoracic aspiration. The second approach is to modify the usual technique of processing expectorated sputum by either washing techniques or by quantitative cultures. Azithromycin and clarithromycin are chemically related to macrolide erithromycin. Both antibiotics retain the traditional macrolide spectrum of activity against gram-positive and atypical pneumonia pathogens, while demonstrating improved activity against gram-negative bacteria. The American Thoracic Society (ATS) recommended the use of macrolide for outpatients with community-acquired pneumonia, without comorbidity and 60 years of age or younger. A total of 34 outpatients with acute LRTIs were open-comparative, randomly allocated to treatment with the new macrolide in Persahabatan Hospital, Jakarta, 1996. The purposes of this study were: (i) to identify the causative micro-organisms; and (ii) to evaluate the clinical efficacy of the new macrolide in these infections. Azithromycin 500 mg was given orally once a day for 3 days and was administered 1 h before or 2 h after every meal. Clarithromycin 500 mg was given orally every 12 h for 10 days. The diagnosis of the patients were: 16 with pneumonia, 10 with acute bronchitis and 8 with acute exacerbation of chronic bronchitis. In this study of 34 patients, the sputum specimens were washed with N acetylcysteine before culture and we could only detect micro-organisms in one patient. Before treatment, we found 47 strains in 33 (97.05%) patients and after treatment we found five strains. From serological examination, only four (11.76%) atypical bacterial were detected. The most frequently found microorganisms were 23 strains of Klebsiella pneumoniae (40.42%), 10 of Streptococcus alpha haemolyticus (21.26%), five of Streptococcus pneumoniae (10.63%) and five of Staphylococcus aureus (10.63%). The atypical bacterial were: two Legionella pneumophila, one Mycoplasma pneumoniae and one Chlamydia pneumoniae. The clinical efficacy of new macrolides were 100% and the bacteriological responses with eradication of 94.12% vs 70.59% of isolates in the azithromycin and clarithromycin groups are shown in Table 1. There were no adverse reactions detected in the two treatment groups until the end of the study.

Administration, Oral↗

Pulsed-field gel electrophoresis of the genomic restriction fragments of coagulase-negative staphylococci.

The genomes of 47 coagulase-negative staphylococcal strains assigned to different species were analysed by pulsed-field electrophoresis. The strains were clustered on the basis of their similarity in the SmaI restriction patterns into various groups, each group consisting of the type strain and the strains whose SmaI restriction patterns were similar to that of the type of strain. The SmaI restriction groups seem to correspond to the following species: Staphylococcus warneri, S. hominis, S. xylosus, S. lugdunensis, S. kloosii, S. haemolyticus, S. lentus, S. cohnii, S. equorum, S. chromogenes, S. saprophyticus, S. simulans, S. carnosus, S. capitis and S. auricularis. The species S. sciuri, S. caseolyticus, S. gallinarum, S. epidermidis and S. schleiferi were represented only by their type strains and showed no similarity in their SmaI restriction patterns neither to each other nor to all the other species investigated here. Thus, the classification of coagulase-negative staphylococcal strains into the above species seems to be confirmed also by genome restriction analysis carried out by pulsed-field gel electrophoresis.

DNA Restriction Enzymes↗

The ica operon and biofilm production in coagulase-negative Staphylococci associated with carriage and disease in a neonatal intensive care unit.

Coagulase-negative staphylococci (CoNS) are a major cause of sepsis in the neonatal intensive care unit (NICU). We evaluated the hypothesis that the ica operon and biofilm production are associated with CoNS disease in this setting. CoNS associated with bacteremia or blood culture contamination and from the skin of infants with CoNS bacteremia or healthy controls were obtained during a prospective case-control study on a busy NICU. A total of 180 strains were identified, of which 122 (68%) were Staphylococcus epidermidis and the remainder were S. capitis (n = 29), S. haemolyticus (n = 11), S. hominis (n = 9), S. warneri (n = 8), and S. auricularis (n = 1). The presence of the genes icaA, icaB, icaC, and icaD was determined by PCR, and biofilm production was examined using qualitative (Congo red agar [CRA]) and quantitative (microtiter plate) techniques. There were no significant differences in the presence of the ica operon or CRA positivity among the four groups of strains. However, quantitative biofilm production was significantly greater in strains isolated from either the blood or the skin of neonates with S. epidermidis bacteremia. We conclude that the quantity of biofilm produced may be associated with the ability to cause CoNS infection. This conclusion suggests that the regulation of biofilm expression may play a central role in the disease process.

Bacteremia↗

Variation in the expression of cell envelope proteins of coagulase-negative staphylococci cultured under iron-restricted conditions in human peritoneal dialysate.

Strains of coagulase-negative staphylococci (CNS), including Staphylococcus epidermidis, S. hominis, S. warnerii, S. simulans, S. capitis, S. haemolyticus and S. saprophyticus, were isolated from patients with continuous-ambulatory-peritoneal-dialysis-related peritonitis. The cell wall and cytoplasmic membrane protein profiles of CNS strains cultured in either nutrient broth (NB) or pooled human peritoneal dialysate (HPD) were compared. Some interspecies variation in both the wall and membrane protein profiles was noted. However, the cell wall protein profiles of HPD-grown CNS strains differed markedly from those cultured in NB. Growth in HPD resulted in a marked reduction in the total number of cell-wall-associated proteins but up to three antigenically related proteins in the 40-56 kDa range, depending on the species, predominated. Growth in HPD also resulted in the induction of two iron-repressible cytoplasmic membrane proteins (IRMPs) of 32 and 36 kDa in S. epidermidis. Other CNS strains only appeared to express a single IRMP, which varied in molecular mass from 32 to 36 kDa. Whilst the IRMP in these CNS strains showed considerable antigenic homology with the 32 kDa IRMP, the S. warneri IRMP showed cross-reactivity with both the 32 and 36 kDa IRMPs of S. epidermidis. Immunoblotting experiments revealed that whilst the CNS cell wall proteins were poorly immunogenic, the IRMPs were the immunodominant CNS protein antigens, reacting strongly with antibodies present in HPD. This finding provides evidence to suggest that the IRMPs are expressed in vivo during infection.

Antibodies, Bacterial↗

Cloning and restriction analysis of DNA conferring new quinolone antimicrobial agent resistance from Staphylococcus aureus and other coagulase-negative Staphylococcus species.

DNA conferring resistance to new quinolone antimicrobial agents (NQR) in clinical isolates of Staphylococcus aureus and coagulase-negative staphylococci (CNS), S. epidermidis and S. haemolyticus, was analyzed after cloning in Escherichia coli. The NQR phenotypes were expressed in E. coli at lower levels. NQR gene(s)-carring HindIII fragments were very similar to each other among S. aureus or CNS, although the S. aureus fragments differed from the CNS fragments in terms of the NQR phenotypes and restriction endonuclease maps. The data suggest the possibility that the NQR genes have disseminated in evolutionarily distinct routes among S. aureus and CNS.

4-Quinolones↗

Reduction in the colonization of central venous cannulae by mupirocin.

In an in-vitro simulation of an intravascular cannula enclosed in a fibrin sheath, 0.03 mg1(-1) of mupirocin prevented significant colonization [greater than 15 colony forming units (cfu)] by two clinical isolates of Staphylococcus epidermidis and one each of S. saprophyticus, S. hominis and S. haemolyticus. This suggests that in vivo, where protein binding of mupirocin is 95-97%, 1 mg 1(-1) of mupirocin at the cannula surface would be required to prevent colonization. These results support the findings of our previously published prospective controlled trial, in which mupirocin applied to the insertion sites of 172 internal jugular cannulae reduced the rate of colonization of cannula tips to 5%, compared with 25% for the 186 controls (P less than 0.001). Of the 46 colonized cannula tips from 110 control patients, the same species was isolated from the skin of the insertion site in 67% and from the lumen flush in only 15%. Analysed by patient, mupirocin reduced the proportion of patients with colonized tips from 17% to 3% after 24 h of infection, and from 35 to 10% after 48 h (P = 0.002). The use of agar containing charcoal, as a mupirocin neutralizer, and the incubation of tip-culture plates flooded with the Oxford staphylococcus, gave no evidence of carry over of mupirocin onto cannulae removed from mupirocin-treated patients.

Administration, Topical↗

Distribution of Staphylococcus species among human clinical specimens and emended description of Staphylococcus caprae.

By DNA-DNA hybridization on microplates, we identified 1,230 strains of staphylococci from human clinical specimens and determined the distribution of species. The 10 Staphylococcus species isolated most often were S. epidermidis (31.3%), S. aureus (23.3%), S. haemolyticus (12.2%), S. caprae (10.7%), S. simulans (4.4%), S. hominis (4.0%), S. capitis (3.9%), S. saprophyticus (3.6%), S. warneri (2.2%), and S. lugdunensis (1.3%). From these results, we realized that S. caprae strains were widely distributed in human clinical specimens. The description in Bergey's Manual of Systematic Bacteriology indicates that no strains of S. caprae produce acid from fructose and mannitol, but all our S. caprae strains produced acid from fructose and mannitol. Consequently, many strains of S. caprae isolated from human clinical specimens have been misidentified as S. haemolyticus or S. hominis by conventional biochemical tests. In this paper, we propose an emended description of S. caprae.

Bacterial Typing Techniques↗

Methicillin-resistant coagulase-negative staphylococci isolated from healthy horses in Japan.

OBJECTIVE: To determine patterns of methicillin-resistant staphylococci isolated from apparently healthy horses. SAMPLE POPULATION: 44 horses from 8 riding clubs in Japan. PROCEDURE: Methicill in-resistant staphylococci were isolated from the skin or nares, using a selective medium containing a beta-(symboric) lactam antibiotic, ceftizoxime. Clonality of isolates was determined by use of pulsed-field gel electrophoresis. Detection of mecA, mecl, and mecR1 genes was accomplished by use of polymerase chain reactions. RESULT: Of the 44 horses, 13 (29.5%) yielded 15 isolates of methicillin-resistant staphylococci. The 15 isolates were identified as 6 species (Staphylococcus epidermidis, S lentus, S saprophyticus, S xylosus, S sciuri, and S haemolyticus). However, methicillin-resistant S aureus was seldom isolated. Each isolate contained the mecA gene and had a high resistance to beta-lactam antibiotics. Some isolates also were resistant to other antibiotics such as erythromycin and kanamycin. CONCLUSIONS AND CLINICAL RELEVANCE: Methicillin-resistant coagulase-negative staphylococci that were highly resistant to various antibiotics were isolated from apparently healthy horses in Japan. These organisms must be considered a potential threat to horses and veterinarians who care for them.

Animals↗

Comparative performances of six agglutination kits assessed by using typical and atypical strains of Staphylococcus aureus.

Six commercial agglutination tests designed for the identification of Staphylococcus aureus were compared by using a strain collection which included 512 staphylococci representing 33 species (318 isolates of Staphylococcus aureus [including 144 oxacillin resistant], 46 S. epidermidis isolates, 15 S. haemolyticus isolates, 12 S. saprophyticus isolates, 29 S. schleiferi isolates, 30 S. lugdunensis isolates, and 62 other coagulase-negative staphylococci). This group also included a proportion of strains with unusual phenotypes (e.g., 19 coagulase-negative S. aureus isolates, 26 clumping factor-negative S. aureus isolates, and 4 S. aureus isolates each with a double deficiency). The overall sensitivity for identification of typical and atypical S. aureus was high with the Staphaurex Plus test (Murex Biotech) (99.7%), the Pastorex Staph Plus test (Sanofi Diagnostics Pasteur) (99.7%), and the Slidex Staph Plus test (bioMérieux) (100%). The overall rate of specificity was affected by the unusual inclusion in this study of a high proportion of non-S. aureus species, such as S. lugdunensis and S. schleiferi, which express a clumping factor and therefore produce a positive result with the agglutination tests.

Agglutination Tests↗

Antimicrobial susceptibility of staphylococci isolated from otitis externa in dogs.

Samples were obtained from 65 unmedicated adult dogs, processed for isolation of Staphylococcus species and tested for susceptibility to penicillin G, gentamicin, oxacillin, tetracycline, trimethoprim-sulphamethoxazole, streptomycin, ampicillin and rifampin. Forty-four isolates were obtained, which represents 67.7% of samples. Coagulase-negative species were most commonly found, and the most frequently isolated staphylococcus species were Staph. epidermidis and Staph. aureus. Other species, such as Staph. simulans, Staph. haemolyticus, Staph. saprophyticus and Staph. intermedius were also isolated. Resistance to antibiotics was frequently observed, with 90.9% of the isolates showing resistance to at least one drug. The most active antimicrobial agents against staphylococci isolated from otitis externa of dogs were rifampin and oxacillin. Multidrug resistance was a common finding, and one strain of Staph. haemolyticus species, was resistant to all tested antimicrobial agents. Resistance to three or more different drugs was a common finding, observed in 16 strains (36.4%) of both coagulase-positive and coagulase-negative staphylococci. This study highlights the emergence of cases of otitis externa determined by coagulase-negative staphylococcus strains and once more emphasizes the need for bacterial culture with species identification and susceptibility testing of swab specimens from the ear canal in order to choose appropriate antimicrobial agents.

Animals↗