[Another instance of sausage poisoning by Staphylococcus aureus haemolyticus].
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Humans↗
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Antimicrobial efficacy of a new antibiotic-loaded poly(hydroxybutyric-co-hydroxyvaleric acid) controlled release system.
OBJECTIVE: Failure of orthopaedic devices, mainly femoral hip replacements, due to infection is of increasing medical importance. There is a need for improved antibiotic delivery systems in the treatment of orthopaedic infections and here we have evaluated polyhydroxyalkanoate formulations for their suitability as a constant delivery system for gentamicin. METHODS: Gentamicin was incorporated in poly(hydroxybutyric-co-hydroxyvalerate) (PHBV) with 8% or 12% hydroxyvalerate (HV) content at 2:1 or 5:1 (weight to weight) ratio. In conjunction with an elution study, a scanning electron microscopy and a porosity study were carried out to explore physical characteristics of the complexes before and after the leaching effect. The antibacterial effectiveness of the complexes was analysed in a bacterial adhesion assay using clinical isolates of Staphylococcus haemolyticus and Staphylococcus aureus. In addition, the polymers were exposed to pooled human blood to test their biocompatibility in both static and dynamic environments. RESULTS: We have shown that increasing the HV content from 8% to 12% leads to a faster release of the integrated antibiotic. An increase in antibiotic content enhanced the homogeneity while decreasing the permeability of the complexes and reducing the release rate. A significant reduction in the number of the adherent S. aureus and gentamicin-resistant S. haemolyticus within a 48 h exposure to our formulations confirmed the effectiveness of the PHBV/gentamicin complexes. Finally, these formulations did not alter the haemodynamics of the pooled blood samples after an extended period of time. CONCLUSION: Taken together, the PHBV/gentamicin formulations may prove to be effective preventive therapeutic modalities in implant-related Staphylococcus infections.
Anti-Bacterial Agents↗
In vitro susceptibilities of four species of coagulase-negative staphylococci.
The in vitro susceptibilities of 260 strains of coagulase-negative staphylococci to penicillin G, oxacillin, nafcillin, methicillin, cephalothin, and seven non-beta-lactam antimicrobial agents were determined and compared with the susceptibilities of 54 strains of Staphylococcus aureus with known patterns of susceptibility. Penicillin G susceptibility for S. aureus, Staphylococcus epidermidis, Staphylococcus haemolyticus, and Staphylococcus hominis was readily determined by using beta-lactamase tests with induced cells and with a standardized microdilution test. MIC criteria for susceptibility used for S. aureus were applicable to the coagulase-negative species. Percentages of organisms susceptible were as follows: S. epidermidis, 7%; S. haemolyticus, 5%; and S. hominis, 47%. Oxacillin susceptibility for these four species was readily determined by using a modification of the microdilution test. MIC criteria for susceptibility used for S. aureus were applicable to S. haemolyticus and S. hominis, but alternate criteria were necessary for S. epidermidis. Percentages of organisms susceptible were as follows: S. epidermidis, 29%; S. haemolyticus, 36%; and S. hominis, 97%. Staphylococcus saprophyticus differed from the other staphylococcal species; all strains were beta-lactamase negative and were penicillin susceptible but had higher penicillin G MICs than did susceptible strains of the other species. There was total cross resistance among the penicillinase-resistant penicillins and cephalothin for the coagulase-negative staphylococci as well as for S. aureus; oxacillin MICs were more reliable than MICs of the other drugs or a standardized disk diffusion test for distinguishing resistant from susceptible strains. Vancomycin, rifampin, and ciprofloxacin were consistently active against all staphylococci. Erythromycin, clindamycin, gentamicin, and trimethoprim-sulfamethoxazole were more active against oxacillin-susceptible staphylococci than against oxacillin-resistant staphylococci.
Anti-Bacterial Agents↗
Coagulase negative Staphylococci as causative agents of urinary tract infections-prevalence and resistance status in IGIMS, Patna.
Previously considered solely as the laboratory contaminants and normal flora of skin in man, coagulase negative Staphylococci are now a major cause of nosocomial and opportunistic infections. This study was conducted at IGIMS, Patna to find out the coagulase negative Staphylococcus isolates from urine and their antimicrobial resistance. In a period of ten months, the relative frequency of main coagulase negative Staphylococci were as follows--Staphylococcus epidermidis-45.90%, Staphylococcus saprophyticus 34% and Staphylococcus haemolyticus-8.50%. others were 11.60%. most of the Staphylococcus saprophyticus was isolated from young female patients suffering from uncomplicated acute cystitis and Staphylococcus epidermidis was mainly from patients with indwelling catheters and complicated cases. Staphylococcus saprophyticus showed the highest sensitivities to all the antimicrobials whereas Staphylococcus haemolyticus had the highest resistance rates. 66.6% of staphylococcus epidermidis and 60% of staphylococcus haemolyticus were resistant to oxacillin, whereas only 10% of staphylococcus saprophyticus resistant and 90% were sensitive to it. As is in the emerging state, vancomycin resistance was very less but in future it may cause a major problem to treat these cases. So this area needs further exploration.
Adolescent↗
[Pathogenic properties of coagulase-negative staphylococci in experimental infections].
The frequency of persistence of three Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus saprophyticus strains, respectively, was studied in BALB/c mice at the 10th day of intraperitoneal (ip) challenge. 245 out of 416 mice survived after infections with four bacterial suspensions of different colony forming units (CFU) of each strain. Staphylococci persisted in 61 mice (24,9%). The main sites of persistence were the kidneys, while cocci were rarely isolated from the spleen and the liver. S. epidermidis persisted with a significantly higher rate than the other two species, because S. epidermidis in 28,8%, S. haemolyticus in 4,9%, and S. saprophyticus in 3,6% were reisolated from the organs of the respective infected and surviving animals. The organ persistence was proportional to the amount of bacteria injected. The persistence resulted in subacute microabscesses in the organs. Reisolates of persisting bacteria remained stable in phenotype and genotype concerning antibiotic resistance patterns and biochemical activities for the taxonomic implication, whereas cell surface properties characterizable with phage types altered considerably during persistence. It is concluded that cocci of all three Staphylococcus species are invasive and can persist to a certain extent in the organs of animals with normal immune system, too, after artificial inoculation into the peritoneum i. e. to the serosal surfaces.
Animals↗
Survey of methicillin-resistant coagulase-negative staphylococci isolated from the fingers of nursing students.
To clarify the state of methicillin-resistant coagulase-negative staphylococci (MRCNS) contamination in the hospital environment, we compared MRCNS isolated from the fingers of 40 nursing students who had not yet experienced clinical practice and 40 who had just completed clinical practice in the hospital. Fourteen MRCNS strains were detected in 13 students (32.5%) after clinical practice; Staphylococcus epidermidis in 9 students, Staphylococcus haemolyticus in 3, and Staphylococcus saprophyticus in 2. Drug sensitivity tests were performed, and the minimum inhibitory concentration (MIC) of penicillin-G (PCG) was more than 2 microg/ml in all strains, and that of ampicillin (ABPC) was more than 16 microg/ml in many strains. Only a few strains showed high MIC values for the other drugs tested. However, some Staphylococcus haemolyticus strains showed high MIC values for cefazolin (CEZ), arbekacin (ABK), gentamicin (GM), ofloxacin (OFLX), or imipenem (IPM). In all strains, the mecA gene was detected by polymerase chain reaction (PCR), and penicillin binding protein 2' (PBP2') was detected by the latex agglutination method. Methicillin-resistant Staphylococcus epidermidis (MRSE) isolated from the fingers of nursing students was compared with that isolated from blood culture specimens by arbitrarily primed (AP)-PCR analysis. The patterns obtained were different, a finding which excluded the presence of cross-infection. The present results show that basic preventive measures for cross-infection should be considered in the future, using such genetic analysis methods, so that MRCNS may not cause hospital infection.
Adult↗
An inexpensive and reliable method for routine identification of staphylococcal species.
The aim of this study was to develop a simple, reliable, and inexpensive in-house system for routine species identification of staphylococci in clinical practice. The system combines 15 key tests (including carbohydrate fermentation) performed in micro-well strips and antimicrobial disk diffusion susceptibility tests performed on standardised paper disk method antibiotic sensitivity medium agar. Twenty-eight staphylococcal reference strains belonging to 18 different species were correctly identified using this in-house system. A total of 291 clinical staphylococci isolates were evaluated with the in-house system and a conventional identification scheme. The in-house system identified 281 (96.6%) of these 291 isolates. Eleven different species were recognised. The five species most frequently identified were Staphylococcus epidermidis (48.6%), Staphylococcus aureus (27.8%), Staphylococcus haemolyticus (8.2%), Staphylococcus hominis (5.7%), and Staphylococcus warneri (5.3%). There was an agreement of 86.3% between the species identification obtained with the in-house system and the conventional identification scheme. All coagulase-negative isolates initially identified as species other than Staphylococcus epidermidis as well as indistinctly identified isolates were also evaluated with a commercial identification system. The agreement between species identification obtained with the in-house system and the commercial system for 101 identified isolates was 73%. Several isolates that were difficult to distinguish with the conventional scheme and/or the commercial system were identified with the aid of the antimicrobial susceptibility test included in the in-house system. The described test scheme should be of value for identification of clinically significant staphylococci species.
Agar↗
Failure of teicoplanin therapy in two neutropenic patients with staphylococcal septicemia who recovered after administration of vancomycin.
A report is given on two neutropenic patients with staphylococcal septicemia caused by Staphylococcus haemolyticus and Staphylococcus aureus (both strains methicillin-resistant) who failed to respond to therapy with teicoplanin. Both strains were resistant to teicoplanin (MIC 16 and 8 mg/l respectively), but remained sensitive to vancomycin (MIC 2 and 4 mg/l respectively). Replacement of teicoplanin with vancomycin led to full recovery of both patients and their discharge from hospital. These two cases emphasize the importance of clinical and microbiological monitoring of patients with staphylococcal septicemia, even when glycopeptides are used for treatment.
Adult↗
Speciation and antibiotic susceptibility patterns of coagulase-negative staphylococci.
During a six month period, 191 isolates of coagulase-negative staphylococci from blood, urine, cerebrospinal fluid and heart valves were identified to species level and tested for antimicrobial susceptibility. Seventy-one percent of isolates were Staphylococcus epidermidis, 8% Staphylococcus warneri, 7% Staphylococcus hominis, 7% Staphylococcus haemolyticus, 4% Staphylococcus capitis, 2% Staphylococcus saprophyticus and 1% Staphylococcus cohnii. Approximately 4% of isolates were felt to be associated with infection. Overall, 18% of isolates were susceptible to penicillin G, 61% oxacillin, 98% cephalothin, 98% cefamandole, 72% cefotaxime, 95% cefsulodin, 76% gentamicin, 64% clindamycin and 98% rifampicin. All isolates were susceptible to vancomycin. Vancomycin, rifampicin, cephalothin and cefamandole showed excellent activity against oxacillin-resistant isolates. With one exception, speciation was not helpful in determining whether or not an isolate was associated with infection.
Anti-Bacterial Agents↗
Glycopeptide resistance in Staphylococcus aureus: is it a real threat?
Following the discovery in 1992 that resistance to vancomycin could be transferred in vitro from a strain of Staphylococcus haemolyticus to Staphylococcus aureus, the threat that a similar event could happen in vivo has existed. In 1996, the threat became a reality in Japan followed by reports of low-level heteroresistant S. aureus from elsewhere in the world. However, the threat has not gone away; indeed it has become more intimidating within the past 2 years with the isolation of higher-level resistance to vancomycin transferred on a plasmid from Enterococcus faecalis. The mechanisms of resistance are different in each of the types of S. aureus with reduced susceptibility to the glycopeptides: in one hetero-vancomycin intermediate-susceptible S. aureus (hVISA) and vancomycin intermediate-susceptible S. aureus (VISA), cell wall biosynthesis is altered, hindering glycopeptide reaching its target, and in the other vancomycin-resistant S. aureus (VRSA) the target is altered. This review attempts to place into context the threat posed to patients by the appearance of strains of S. aureus more resistant to one of the key therapeutic agents used in our hospitals. It will do so by raising a number of important questions followed by the presentation of experimental evidence that may or may not provide answers that heighten or lower the threat.
Anti-Bacterial Agents↗
Invasive potential of bacterial isolates associated with subclinical bovine mastitis.
This work describes differences in the invasive ability of bacterial isolates associated with mastitis. Invasion ability was determined by the uptake and survival in a primary culture of bovine mammary epithelial cells (BMEC). BMEC were isolated from a healthy lactating cow and characterized by their morphology, immunostaining for cytokeratin and the detection of beta- and kappa-casein mRNAs. Ten bacterial isolates comprising the staphylococcal species Staphylococcus aureus (3), Staphylococcus epidermidis (1), Staphylococcus haemolyticus (1), Staphylococcus equorum (2), Staphylococcus xylosus (1) and Brevibacterium stationis (2) obtained from raw milk of cows with mastitis from backyard farms were assayed for their ability to invade BMEC. Only two S. aureus and one S. epidermidis isolates were able to invade BMEC, at similar levels to the S. aureus control strain ATCC 27543. In conclusion, using the in vitro model of infection used in this study, differences in bacterial invasion capability may be detected.
Animals↗
Coagulase-negative Staphylococci in Danish blood cultures: species distribution and antibiotic susceptibility.
The distribution and antibiotic susceptibility of coagulase-negative staphylococci (CoNS) isolated from blood cultures was examined in samples from hospitals covering most of Denmark. A total of 499 CoNS isolates were detected in 477 blood cultures from 340 patients and speciated as Staphylococcus epidermidis, 285; Staphylococcus hominis, 61; Staphylococcus haemolyticus, 43; Staphylococcus warneri, 12; Staphylococcus cohnii, 7; Staphylococcus saprophyticus, 4; Staphylococcus capitis, 2 and Staphylococcus lugdunensis, 1. Seventy-eight isolates could not be identified to species level and six were Micrococcus spp. In 108 (22.6%) blood culture sets, more than one CoNS strain were found, as detected by species identification, antibiogram and biotyping. Significantly more blood cultures from patients in university hospitals were drawn from central venous catheters. Comparing university and non-university hospitals, the overall antibiotic susceptibility among CoNS was only slightly different, except for methicillin and amikacin. The prevalence of methicillin-resistant strains was 35.1% in the university hospital strains vs. 25.3% in the non-university hospital strains. The overall prevalence of methicillin resistance was 32%. Great geographic variation in both species distribution and antibiotic resistance was observed. The high prevalence of S. epidermidis makes subtyping of this species important.
Blood↗
Activation of the human immunodeficiency virus long terminal repeat in THP-1 cells by a staphylococcal extracellular product.
Staphylococcal strains can release a factor that strongly activates the human immunodeficiency virus type 1 (HIV-1) long terminal repeat (LTR) in THP-1 cells transfected with the HIV-1 LTR-driven luciferase reporter gene (THP-1 LTRluc). The factor is present in the overnight culture fluid and is readily released from the organisms into aqueous medium by vigorous mixing. Staphylococcal extracellular material is a complex mixture of polysaccharide and protein containing peptidoglycan and teichoic acid, released in part by cell wall turnover. The importance of the carbohydrate component is emphasized by concanavalin A (Con A) inhibition of staphylococcal product-induced LTR activation but not of activation by phorbol 12-myristate 13-acetate or tumor necrosis factor. The effect of Con A was decreased or abolished by sugars in the order methyl alpha-D-mannopyranoside > methyl alpha-D-glucopyranoside > mannose > glucose = fructose > N-acetylglucosamine. Wheat germ agglutinin was less inhibitory than Con A; in this instance N-acetylglucosamine decreased inhibition, whereas methyl alpha-D-mannopyranoside or methyl alpha-D-glucopyranoside did not. The induction of luciferase activity in THP-1 LTRluc by the staphylococcal extracellular product also was inhibited by fetal bovine and normal human serum. A comparison of 31 staphylococcal isolates (9 Staphylococcus aureus, 11 Staphylococcus epidermidis, 2 Staphylococcus haemolyticus, 4 Staphylococcus hominis, 2 Staphylococcus capitis, 2 Staphylococcus warneri, 1 Staphylococcus saprophyticus) revealed wide variation in LTR activating activity that did not correlate closely with slime production. Our findings, using induction of luciferase in THP-1 LTRluc as a model for upregulation of HIV infection, raise the possibility that staphylococci, as well as certain other microorganisms, release carbohydrate-containing exopolymers, which can activate the HIV-1 LTR, thus influencing progression of HIV infection.
Animals↗
Relatedness of coagulase-negative staphylococci causing bacteremia in low-birthweight infants.
OBJECTIVE: To investigate coagulase-negative staphylococcus (CONS) causing bacteremia in a neonatal intensive care unit (NICU). DESIGN: A 14-month retrospective review of 47 infants in the NICU with CONS bacteremia was undertaken to determine CONS glycocalyx production, plasmid pattern, total DNA restriction fragment polymorphism, and clinical risk factors. RESULTS: The isolates included 32 Staphylococcus epidermidis, six Staphylococcus haemolyticus, four Staphylococcus warneri, four Staphylococcus saprophyticus, and one Staphylococcus hominis. Sixty-five percent of S epidermidis produced glycocalyx; other species did not. Oxacillin resistance (52%) and the antibiograms of the CONS were consistent with other units in the hospital. Five similar CONS plasmid patterns were found among 16 isolates; 31 isolates had unique patterns. Extractions of total DNA from these isolates were digested using HindIII, HaeIII, and BstEII. Those with similar restriction fragment length patterns could not linked as nosocomially transmitted among infants with bacteremia. CONCLUSION: Our observations suggest that multiple strains of CONS infect infants in the NICU who have similar risk factors. Although current infection control practices limit transmission of a pathogen, they do not prevent CONS bacteremias.
Bacteremia↗
Antibacterial activity of quinolones against coagulase-negative staphylococci and the quinolone resistance-determining region of the gyrA genes from six species.
Antibacterial activity of quinolones against three species of coagulase-negative staphylococci was investigated. Tosufloxacin and sparfloxacin exhibited potent activities against Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus saprophyticus compared with other quinolones tested. From the analysis of the DNA sequence in the quinolone resistance-determining region (QRDR), greater than 80% homology was recognized in coagulase-negative staphylococci. A series residue was conserved in all six species at the position corresponding to position 84 in Staphylococcus aureus.
Anti-Infective Agents↗
In vitro activity of quinupristin/dalfopristin, linezolid, telithromycin and comparator antimicrobial agents against 13 species of coagulase-negative staphylococci.
OBJECTIVES: To determine in vitro susceptibilities of a large series of speciated coagulase-negative staphylococci (CNS) against three new antibiotics, linezolid, quinupristin/dalfopristin and telithromycin. METHODS: Susceptibilities to three new antibiotics and oxacillin, vancomycin, clindamycin and erythromycin were determined by the agar dilution method, as described by the NCCLS. RESULTS: Resistance to linezolid was not observed in any isolates, although MIC90 values varied between species. Fifteen of 658 (2.3%) isolates were resistant to quinupristin/dalfopristin, but < 1% of the clinically most important isolates of Staphylococcus epidermidis, Staphylococcus haemolyticus and Staphylococcus hominis demonstrated resistance to this agent. Susceptibility to clindamycin correlated with susceptibility to quinupristin/dalfopristin; however, resistance to clindamycin did not predict quinupristin/dalfopristin resistance. Telithromycin was the least active of the new agents tested, showing activity similar to that of clindamycin. Susceptibility and resistance to clindamycin were predictive of susceptibility and resistance to telithromycin. CONCLUSION: Clindamycin susceptibility can be used as a surrogate marker for susceptibility to quinupristin/dalfopristin and telithromycin. Quinupristin/dalfopristin and linezolid show good activity against both mecA-positive and -negative CNS.
Acetamides↗
Identification and molecular characterization of mannitol salt positive, coagulase-negative staphylococci from nasal samples of medical personnel and students.
The identification of mannitol salt positive, coagulase-negative staphylococci (CNS) is often disregarded when Staphylococcus aureus is screened in clinical samples using mannitol salt agar. However, the emergence of CNS as important human pathogens has indicated that reliable methods for the identification of clinically significant CNS are of great importance in understanding the epidemiology of infections caused by them. The identification and molecular characterization of mannitol salt positive CNS from nasal samples of medical personnel and students is reported here. A total of 84 mannitol salt positive staphylococcal isolates were obtained from 240 nasal samples, of which 15 were CNS. The API STAPH system classified the CNS isolates into six species, and one-third of the isolates were identified with confidence levels of <80 %. 16S-23S rRNA intergenic spacer length polymorphism analysis (ITS-PCR) identified only two species (Staphylococcus haemolyticus and Staphylococcus saprophyticus). This identification was confirmed by antibiotyping, species-specific PCR and PFGE. The results from this study indicate that ITS-PCR is a potentially useful and reliable tool, enabling hospital laboratories to obtain rapid, full and accurate identification of CNS at the species level.
Bacterial Typing Techniques↗
[Prostate histopathology of NIH category IV prostatitis detected by sextant prostate needle biopsy from the patients with high prostatic specific antigen].
Asymptomatic prostatitis is classified as category IV in NIH classification of prostatitis syndrome (1999). No report concerning this category has been present. We investigated this category histopathologically and clinically, in order to clarify the histopathological distribution and its correlation to the clinical features, in this study. Among 785 patients who were suspected prostate cancer because of their high prostatic specific antigen (PSA) values and to have a sextant prostate needle biopsy was performed between January, 1996 and December, 2000, 88 patients (11.2%) were diagnosed as NIH category IV prostatitis (asymptomatic prostatitis). We observed all pathological specimens stained with Hematoxylin-Eosine, and classified them into subtypes according to the classification criteria for prostatitis defined by True et al. (1999). We also investigated the relationship between histopathological distribution and clinical features such as PSA values, PSA density, the incidence of pyuria or bacteriuria. In the histopathological study, grade distributions were 12.5% (11/88) in mild, 71.6% (63/88) in moderate, and 15.9% (14/88) in severe. Location distributions were 2.3% (2/88) in glandular, 68.2% (60/88) in periglandular, and 29.5% (26/88) in stromal. No relationship between these subtypes and clinical features was recognized statistically. However, 7 patients (7.95%) were diagnosed as prostate cancers, later. Pyuria was found in 29.1% (23/79). Bacteriuria was present in 14.3% (11/77). Isolated bacteria were 4 strains of Enterococcus faccalis, 2 strains of each of Pseudomonas aeruginosa and Staphylococcus aureus, and one strain of each of Escherichia coli, Klebsiella oxytoca, Enterobacter cloacae, Enterobacter aerogenes, Staphylococcus haemolyticus, and Staphylococcus epidermidis. Gram positive rod, and Candida sp. No relationship between these subtypes and bacterial species was recognized. These results indicated that the incidence of NIII category IV prostatits was not low without correlation to any clinical features. However, we should pay attention to the presence of prostate cancer, because a small number of the patients were diagnosed as prostate cancer, later.
Aged↗