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[Immunosuppressive effect of cyclophosphamide in experimental coagulase-negative staphylococcal infection].

BALB/c mice were given 200 mg/kg cyclophosphamide intraperitoneally. Three days later, mice were infected intraperitoneally with Staphylococcus epidermidis, Staphylococcus haemolyticus, Staphylococcus saprophyticus at 4 inocula ranging between 2 x and 20 x 10(8) CFU/ml suspended in dextran microcarrier solution. Controls were treated only with bacteria. Lethality rates and organ persistence of cocci were significantly higher, and more peritoneal abscesses and adhesions developed in the mice pretreated with cyclophosphamide than in the untreated controls, regardless of the species of Staphylococcus injected. Splenomegaly was also more pronounced indicating a probably enhanced compensatory reactivity of the immune system liberated from suppression 13 days after the administration of cyclophosphamide. Our results show that cyclophosphamide treatment increases the susceptibility of mice to infection by coagulase-negative staphylococci and it is also responsible for a more severe course of the diseases provoked.

Animals↗

Bacteria in surface infections of neonates.

A bacteriological work on surface infections was done among live births (study group I) and neonates admitted in hospital (study group II). Out of 134 cases of conjunctivitis in group I Gram-negative bacilli predominated (48.5%) with Escherichia coli accounting for 29 (14.9%) cases, Klebsiella species 15 (11.2%) cases, Citrobacter freundii 3 (2.2%) cases, Pseudomons aeruginosa 18 (13.4%) cases and Aeromonas hydrophila 3 (2.2%) amongst pure isolates (73.9%). Gonococcus was noted in 2 (1.5%) cases. In group II, 41.7% were Staphylococcus aureus in pure growth (75%), compared to only 9.0% in group I. Skin infections were caused by both Staphylococcus aureus and Staphylococcus epidermidis. Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa were the principal insolates from umbilical sepsis. Pseudomonas aeruginosa was isolated as pure growth from local site of noma neonatorum. Anaerobic cultures were negative in all except in 2 cases of umbilical sepsis with tetanus neonatorum revealing Clostridium tetani which however proved to be non-toxigenic. Blood cultures were positive in 4 out of 14 cases bearing 50% correlation with bacteria from surface infections. A source study established partial correlation with the cases of pseudomonas conjunctivitis. Phage typing of Staphylococcus aureus and biochemical typing failed to detect any definite marker of clinical entities, except that the skin infections were caused by group III phages predominantly (65.0%).

Bacteriophage Typing↗

Quantitation of skin bacteria: lethality of the wash solution used to remove bacteria.

A widely used technique for the quantitative removal of bacteria from the skin uses a detergent, Triton X-100 (p, t-octylphenoxynonaethoxyethanol), to remove and suspend the bacteria. We determined the half-life for the survival of five common skin bacteria suspended in the solution. The shortest-lived was Streptococcus pyogenes with a half-life of 0.9 hours. All of the others (Propionibacterium acnes, Staphylococcus epidermidis, Staphylococcus aureus, and Klebsiella pneumoniae) had half-lives of 1.5 hours or longer. K. pneumoniae, the only Gram-negative species tested, had a half-life of more than 30 hours. Thus, a one hour delay in plating and incubation of samples suspended in this detergent solution inhibits quantitation of most species tested.

Bacteria↗

[Bacteriological study of chronic sinusitis].

OBJECTIVE: To explore the characteristics of bacteria isolated from patients with chronic sinusitis (CR) and antibiotic-resistance. METHODS: The purulent discharges taken from the maxillary sinus and posterior ethmoid sinus of 76 patients during endoscopic sinus surgery were cultured for both bacteria and fungi, then the antimicrobial susceptibility test and beta-lactamase-producing bacteria (beta-LPB) were detected. In the control group, the discharges from the nasal cavity from 10 healthy adults were also cultured for bacteria. RESULTS: Twenty species, including Staphylococcus epidermidis, Escherichia coli, Staphylococcus aureus, alpha-Hemolytic streptococcus and Streptococcus pneumoniae, were cultured from the testing group. The total positive rate of bacteria was 81.8%. The detectable rates of aerobe, anaerobe, gram-positive bacteria and gram-negative bacteria were 83.4%, 16.6%, 56.3% and 43.7%, respectively. The fungi was only detected in 1.4% patients. The antimicrobial susceptibility test demonstrated that Imipenem, Cefotaxime and Amoxicillin-clavulanate were good in vitro activity to aerobic pathogens and Metronidazole, Imipenem and Chloramphenicol was good to anaerobic pathogens. The beta-LPB was detected in 37.7% strains in which Staphylococcus epidermidis, Staphylococcus aureus, alpha-Hemolytic streptococcus and Branhamella catarrhalis were more frequent. Five species including Staphylococcus epidermidis and Escherichia coli etc were cultured from the control group. There was no significant difference of distribution of the same bacteria between the control group and the testing group (P > 0.05). CONCLUSIONS: The pathogenic bacteria of CR mostly involve aerobes. The antibiotics-resistance of these pathogen is serious. The bacterial infection plays no leading role in CR, so the need of antibiotics should not be excessively emphasized. Before treatment by antibiotics, the antimicrobial susceptibility test should be performed. The broad-spectrum antibiotics containing beta-lactamase inhibitor such as Cefotaxime and Amoxicillin-clavulanate is recommended in condition that the test can not be finished.

Adolescent↗

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↗

Microbiological profile of a shipboard environment and the flora on contact lenses of seamen.

PURPOSE: The purpose of this study was to culture and identify the spectrum of organisms (and their sensitivities) that contaminate the extended wear contact lenses of seamen in their working environment. A secondary aim was to identify the most appropriate first-line antibiotic regimen to be used on seamen who sustain contact lens-related corneal keratitis on board ship. METHODS: Twenty pairs of contact lenses of 20 seamen in one group and 24 pairs in another group wearing contact lenses were collected at the end of 1 week of extended wear. Groups one and two differed only in the way lenses were stored prior to culturing. All contact lenses were then brought to the microbiological lab within 24 hours for culture and sensitivity testing. RESULTS: The most common contaminants on the contact lenses in this study were Staphylococcus epidermidis and Staphylococcus aureus, organisms resident on the normal eyelids. Enterobacterand Pseudomonas species, present in a shipboard environment, were also identified as contaminants on the contact lenses. The organisms cultured from the contact lenses correlated well with those of normal lid flora as well as from the ship environment and are probably derived from these sources. Storage in saline-containing preservatives yielded significantly less positive bacterial cultures from the contact lenses in our study. All bacterial isolates were sensitive to ciprofloxacin whereas several bacteria resistant to cefazolin and gentamicin were identified. CONCLUSION: The most common contaminants on the contact lenses in this study were Staphylococcus epidermidis and Staphylococcus aureus, organisms resident on the normal eyelids. Enterobacter and Pseudomonas species, which are normally present in a shipboard environment, were also identified as contaminants on the contact lenses. Ciprofloxacin is effective against all organisms identified as contaminants on the contact lenses in this study. Of all the antibiotics tested, it is probably the most suitable agent against contact lensrelated keratitis under such shipboard circumstances and is thus recommended in these situations.

Anti-Infective Agents↗

Coagulase-negative staphylococci in air samples from operating theatres.

The aim of our study was to recognize coagulase-negative staphylococci (CNS) in the air of operating theatres. Out of the identification of 449 isolates, the most frequent species were Staphylococcus epidermidis and Staphylococcus haemolyticus. The strains were adherent to glass in 52.3%. Most of the S. epidermidis showed adherent growth, while the majority of the S. haemolyticus failed to adhere. The disk-diffusion antibiotic sensitivity tests showed great differences in sensitivity to penicillin, tetracycline and erythromycin between adherence-positive and negative isolates. On the whole, the species S. haemolyticus proved to be much more resistant than S. epidermidis. Staphylococcus warneri was the most, while S. haemolyticus was the least sensitive to phages.

Air Microbiology↗

[Distribution of bacteria and analysis of their sensibility to antibiotics in patients with hospital-acquired pneumonia].

OBJECTIVES: This study was explore the distribution of the bacteria and their sensibility to antibiotics in hospital-acquired pneumonia. METHODS: One hundred and ninety-six bacterium species were collected in patients with the hospital-acquired pneumonia to make sputum culture. The sensibility of the bacteria to antibiotics were examined by KB paper method and the minimal-inhibitory-concentration by gel double multiple dilute method. RESULTS: Most of the G- bacteria were pseudomonas aeruginosa (30%) and klebsiella bacillus (22%). Most of the G+ bacteria were staphylococcus epidermidis (14%) and staphylococcus aureus (12%). G- bacteria were sensitive to impienem(98%), cefoperazone(90%), ceftriaxone(90%), leftazidime(92%), ciprofloxacin(90%), and amikacin(89%). The sensibility of vancomycin to G+ bacteria was 100%. CONCLUSIONS: The pseudomonas aeruginosa, klebsiella bacillus, staphylococcus epidermidis, and staphylococcus aureus are the most important bacteria in patients with hospital-acquired pneumonia. Imipenem, cefoperazone, ceftriazone, leftazidime, ciprofloxacin, amikacin, and vancomycin are effective antibiotics for treating hospital-acquired pneumonia.

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↗

Comparative assessment of antibiotic susceptibility of coagulase-negative staphylococci in biofilm versus planktonic culture as assessed by bacterial enumeration or rapid XTT colorimetry.

OBJECTIVES: To quantitatively compare the antibiotic susceptibility of biofilms formed by the coagulase-negative staphylococci (CoNS) Staphylococcus epidermidis and Staphylococcus haemolyticus with the susceptibility of planktonic cultures. METHODS: Several CoNS strains were grown planktonically or as biofilms to determine the effect of the mode of growth on the level of susceptibility to antibiotics with different mechanisms of action. The utility of a new, rapid colorimetric method that is based on the reduction of a tetrazolium salt (XTT) to measure cell viability was tested by comparison with standard bacterial enumeration techniques. A 6 h kinetic study was performed using dicloxacillin, cefazolin, vancomycin, tetracycline and rifampicin at the peak serum concentration of each antibiotic. RESULTS: In planktonic cells, inhibitors of cell wall synthesis were highly effective over a 3 h period. Biofilms were much less susceptible than planktonic cultures to all antibiotics tested, particularly inhibitors of cell wall synthesis. The susceptibility to inhibitors of protein and RNA synthesis was affected by the biofilm phenotype to a lesser degree. Standard bacterial enumeration techniques and the XTT method produced equivalent results both in biofilms and planktonic assays. CONCLUSIONS: This study provides a more accurate comparison between the antibiotic susceptibilities of planktonic versus biofilm populations, because the cell densities in the two populations were similar and because we measured the concentration required to inhibit bacterial metabolism rather than to eradicate the entire bacterial population. While the biofilm phenotype is highly resistant to antibiotics that target cell wall synthesis, it is fairly susceptible to antibiotics that target RNA and protein synthesis.

Anti-Bacterial Agents↗

Antimicrobial-resistance and enterotoxin-encoding genes among staphylococci isolated from expressed human breast milk.

Resistance traits and the presence of enterotoxin-encoding genes were investigated in staphylococcus isolates obtained from expressed human breast milk. A total of 54 staphylococcal isolates identified as Staphylococcus epidermidis (53.6 %), Staphylococcus warneri (20.4 %), Staphylococcus haemolyticus (13 %) and Staphylococcus aureus (13 %) were investigated. By using a disc-diffusion method, higher rates of resistance, including intermediate resistance, were observed for penicillin (87 %) and erythromycin (59.3 %). All strains were susceptible to clindamycin and vancomycin. Minimal inhibitory concentration (MIC) was determined by a macrodilution method for four clinically relevant antimicrobial drugs. High rates of resistance or intermediate resistance were observed for erythromycin, gentamicin and oxacillin. Additionally, three isolates showed reduced susceptibility to vancomycin (MIC, 8 microg ml(-1)). Genetic determinants of resistance were detected by using PCR and the results showed good correlation with the macrodilution tests. Moreover, in four staphylococcus isolates, the presence of enterotoxin-encoding genes (seg, seh and sea) was identified. The results demonstrated that expressed human breast milk can be a reservoir of multiresistant staphylococci that may also harbour important virulent determinants.

Anti-Bacterial Agents↗

[Two stages reimplantation for infection after knee arthroplasty. Apropos of a series of 29 cases].

PURPOSE OF THE STUDY: The purpose of this work was to precise diagnosis and treatment of infected total knee arthroplasty with two stage reimplantation. MATERIAL: 29 infected total knee arthroplasties were operated between 1984 and 1994 and included in this study (mean F.U. 3.5 Y). There were 20 females and 9 males, mean age 70 (46-83). The original arthroplasty was done for OA in 28 patients, RA in one. The arthroplasties were: UHK 2, Bicompartmental 2, Tricompartmental 19. 20 TKA were cementless. 14 patients showed one or several risk factors. Infection was diagnosed in 1 of 2 ways: preoperative aspiration or culture of surgical specimen. There were 12 staphylococcus epidermidis, 8 staphylococcus aureus, streptococcus (n = 2) acinetobacter (n = 2), peptococcus (n = 1) pseudomonas (n = 1), gemella morbidellum (n = 1). 6 were non identified. METHOD: The protocol for two stage reimplantation began with components and cement removal. A synovectomy was performed. The knee cavity was filled with antibiotic cement spacer and the wound was closed. The leg was placed in a splint. All patients underwend a continued antibiotic therapy, specific in 20 cases with isolated organisms. A total knee arthroplasty was performed, using a total posterior cruciate substituting prosthesis, 6 to 8 weeks after components removal (2-24). All patients received parenteral antibiotics after reimplantation for not less than 2 months (2-6). RESULTS: Infection was eradicated in 24 cases, 22 in one time, 2 bad second debridement. At last follow-up the average Hungerford score was 75.6/100, the average Knee society knee score was 80 and the average functional score was 70. Mean range of flexion was 95 degrees. 6 patients had recurrent infection and poor result. They underwent arthrodesis. 5 of the 6 patients had solid mature fusion at last follow-up. DISCUSSION: The results of two stage reimplantation for infected total knee replacement showed that this is the method of choice for infection treatment and acceptable function restoration. As other authors, we get a good success rate (82 per cent). Functional result was better with identified microorganisms, but we did not find any correlation with organisms type or infection length. Punction and bone scanning are of great help for diagnosis in difficult chronical cases. Organism identification is fundamental for infection duration. Staphylococcus epidermidis was the most frequent identified organism. New procedures using articulated cement spacer may improve functional results.

Aged↗

Staphylococcus epidermidis-fibronectin binding and its inhibition by heparin.

Staphylococcus epidermidis is able to adhere onto biomaterials and to cause implant infections. Recently, host matrix proteins, which in vivo cover the implants, have been indicated as substrates for adhesion by specific bacterial adhesins. Here, the binding of S. epidermidis to fibronectin, a main protein of the extracellular matrix, and the effect of heparin on this interaction were studied by dynamic force spectroscopy (DFS). Novelties are that S. epidermidis strains analysed by DFS were clinical isolates from prosthesis-associated infections, genotyped and phenotyped for their adhesion properties to fibronectin and examined as living cells. Thus, fibronectin-binding staphylococci adhered to the fibronectin-coated substratum and formed a continuous layer assuring their contact with the fibronectin-coated cantilever tip during the approach-retraction cycles of the DFS measurements. Results show that only a single molecular binding site of fibronectin is involved in the interaction with S. epidermidis, that it takes place at the domain near the C-terminus and that it is specifically inhibited by heparin.

Bacterial Adhesion↗

Immune response to Staphylococcus epidermidis-induced endophthalmitis in a rabbit model.

Although Staphylococcus epidermidis is the most common cause of postoperative pseudophakic endophthalmitis, little is known about the immune response to S. epidermidis-induced endophthalmitis. Using a rabbit model, the immune response to an intravitreal injection of 7000 S. epidermidis (group 1) or 30,000 S. epidermidis (group 2) organisms was investigated. Clinical evaluations showed that rabbits in group 2 had a more severe inflammatory reaction in the conjunctiva, cornea, iris, and vitreous than those in group 1. The inflammatory reaction in group 1 largely resolved by day 30; group 2 continued to show a severe inflammatory response. Histopathologic findings correlated with clinical findings, with rabbits in group 2 showing a more severe inflammatory reaction in both the anterior and posterior segments of the globe. Positive vitreous cultures for S. epidermidis were present in rabbits in group 1 on days 3, 7, 10, 14, and 21 but not thereafter. However, group 2 had higher vitreous colony counts at days 3, 7, and 14 and negative vitreous cultures thereafter. Neither group showed delayed hypersensitivity to S. epidermidis antigens (evaluated by skin tests). Serum immunoglobulin (Ig) G antibody levels to phenol-inactivated S. epidermidis and glycerol teichoic acid (GTA) increased progressively, reached a peak at days 10-14, and then declined in both groups. Serum IgA antibody levels to these antigens were not detected. Group 2 had a more prolonged IgG antibody response in vitreous and aqueous than group 1. Tear fluid showed the weakest IgG and IgA antibody response to S. epidermidis and GTA. S. epidermidis-induced endophthalmitis was associated with a humoral but not a delayed hypersensitivity response to this organism.

Animals↗

Alcohol increases hemolysis by staphylococci.

It was recently found that alcohols can confer hemolytic properties on certain species of yeast. Here, it is reported that alcohol can promote hemolysis by various species of staphylococci, including strains of Staphylococcus aureus, Staphylococcus epidermidis and Staphylococcus hominis. In order to study this novel phenomenon in S. aureus and S. epidermidis, strains that exhibit this phenomenon (e.g. S. aureus 8325-4, COL, SH1000, S. epidermidis), as compared with strains that exhibited little alcohol-enhanced hemolysis (e.g. S. aureus 8325-4 DeltaTRAP, RN6911) were examined. Both ethanol and n-butanol caused upregulation of the virulence regulator-RNAIII, with a concomitant increase in the production of alpha, beta and gamma-hemolysins in strain 8325-4. In S. aureus COL and SH1000, there was an increase in RNAIII but no change in transcription levels of alpha, beta and gamma hemolysins. Staphylococcus epidermidis stain sofi exhibited increased RNAIII and beta hemolysin production. Staphylococcus aureus mutant strains (8325-4 DeltaTRAP and RN6911) showed no change in the transcription level of the RNAIII regulator and the above hemolysins. Increased hemolysis in S. aureus COL, SH1000 and mutant strains may be caused by other hemolysins (not regulated by RNAIII) or through other mechanisms such as hyperoxidation or cytotoxic lipids.

1-Butanol↗

Effect of a Staphylococcus epidermidis-extracted slime factor on human natural killer cell activity.

A slime factor produced by Staphylococcus epidermidis was a complex glycoconjugate extracted by the phenol extraction method. The potential stimulatory or inhibitory capacity of the phenol-extracted slime (PES) was tested on human natural killer cell cytotoxic activity. Various concentrations of the PES preparation were incubated with the effector cells 30 min before and during the assay period. The PES factor inhibited natural killer cell cytotoxic activity at a concentration of 250 micrograms/ml and at higher concentrations (p < 0.05). The inhibition of natural killer cell cytotoxic activity may probably be related to the complex composition of the slime substance.

Glycoconjugates↗

Immunopathologic features of Staphylococcus epidermidis-induced endophthalmitis in the rat.

PURPOSE: To investigate the clinical, histopathologic and immunologic responses to Staphylococcus epidermidis endophthalmitis in a rat model. METHODS: Experimental rats received an intravitreal injection of viable S. epidermidis (7000 organisms), while control rats received sterile saline. The clinical scores, cellular infiltrate in vitreous, and levels of serum and vitreous IgM, IgG and IgA to glycerol teichoic acid (GTA), the major antigenic determinant of S. epidermidis cell wall, were all measured from day 1 to day 30 after injection. RESULTS: The ocular inflammation was largely resolved by day 14. The red reflex was abolished in 50% of rats between days 3 and 10. The bacteria were cleared from the vitreous by day 7. In vitreous, the neutrophils peaked at day 1 and decreased by day 7, and plasma cells were seen between days 1 and 3. Presence of B cells (CD45+/CD3-) was confirmed by flow cytometric analysis of pooled vitreous humor. IgM and IgG but not IgA antibodies to GTA were found in vitreous of injected eyes. The peak of anti-GTA IgM was observed in vitreous of S. epidermidis-infected rats on day 1 and declined by day 7. In contrast to vitreous antibodies, serum anti-GTA IgM antibodies were significantly elevated throughout the course of S. epidermidis endophthalmitis. A weak IgG but no IgA response were observed in serum. Anti-GTA antibodies were also found in low level in normal sera but not in normal vitreous. CONCLUSIONS: The vitreous antibodies may be involved in neutrophil-mediated opsonophagocytosis leading to 'spontaneous sterility' of the bacteria, and may play a role in the immunopathogenesis of staphylococcal endophthalmitis in the rat.

Animals↗

Removal of infected dual chambered transvenous pacemaker and implantation of a new epicardial dual chambered device with cardiopulmonary bypass: experience with seven cases.

Seven patients with infected transvenous dual chambered pacemakers have undergone removal of the device using cardiopulmonary bypass. There were four women and three men with a mean age of 58 years. Six patients had localized infection in the generator pocket (mean of 4.6 previous unsuccessful operations for surgical sterilization). Four infections were due to Staphylococcus epidermidis, two to Staphylococcus aureus, and one patient presented septicemia caused by Staphylococcus epidermidis and Pseudomonas aeruginosa. The atrial and ventricular transvenous electrodes were removed under direct vision using cardiopulmonary bypass. A new dual chambered epicardial pacemaker was implanted. The procedure was well-tolerated, and all patients are infection free with working pacemakers after a mean follow-up of 25.4 months.

Cardiopulmonary Bypass↗