Outbreak of vancomycin-resistant Enterococcus spp. in an Italian general intensive care unit.

Following the identification of two clinical isolates of vancomycin-resistant enterococci (VRE) from intensive care unit (ICU) patients, a surveillance programme detected that six of eight ICU patients were colonised by VRE. Standard epidemic control measures were instituted in the ICU. During a 16-month period, 13 (2.5%) of 509 ICU patients had VRE-positive swabs upon admission, and 43 (8.7%) of 496 VRE-negative patients were colonised by VRE in the ICU. Patients who acquired VRE in the ICU had a longer ICU stay (p < 0.0001). No other statistically significant differences were demonstrated. Two patients had documented infection (infection/colonisation index, 3.6%; overall VRE infection frequency, 0.4%), but both recovered and were discharged. VRE colonisation did not increase the mortality rate. Automated ribotyping identified three clusters containing, respectively, the first 52 Enterococcus faecium isolates, two Enterococcus faecalis isolates, and two further isolates of E. faecium. Multilocus sequence typing demonstrated that two E. faecium isolates representative of the two ribotypes belonged to sequence types 78 and 18, and that these two isolates belonged to the epidemic lineage C1, which includes isolates with a wide circulation in northern Italy. The outbreak was controlled by continuous implementation of the infection control programme, and by the opening of a new unit with an improved structural design and hand-washing facilities.

Acid tolerance and fecal recovery following oral administration of Saccharomyces cerevisiae.

Probiotic microorganisms to be used as biotherapeutic agents have to resist the rigors of the upper human gastrointestinal tract. In this study we evaluated the acid tolerance in vitro and the fecal recovery in vivo after oral administration of a Saccharomyces cerevisiae strain to healthy volunteers. At the lowest pH value (pH 1.0) the yeast load in tablets decreased slightly. From pH 1.0 to pH 7.0 the release of S. cerevisiae in buffer solutions increased. The selected yeast strain showed good tolerance to low pH which mimic the gastric environment. After one month of treatment at a dose of 100 million cells per day, S. cerevisiae grew from the feces of 6 (37.5%) of the 16 healthy, treated volunteers. Based on the results of the present experiments the yeast studied can be considered a strain that tolerates adverse conditions comparable to those of the human gastrointestinal tract, and when administered orally may colonize the bowel of healthy volunteers and even replace resident Candida species.

Bactericidal activity of Pistacia lentiscus mastic gum against Helicobacter pylori.

In this study we evaluated the antibacterial activity of mastic gum, a resin obtained from the Pistacia lentiscus tree, against clinical isolates of Helicobacter pylori. The minimal bactericidal concentrations (MBCs) were obtained by a microdilution assay. Mastic gum killed 50% of the strains tested at a concentration of 125 microg/ml and 90% at a concentration of 500 microg/ml. The influence of sub-MBCs of mastic gum on the morphologies of H. pylori was evaluated by transmission electron microscopy. The lentiscus resin induced blebbing, morphological abnormalities and cellular fragmentation in H. pylori cells.

Vancomycin-resistant Enterococcus faecium infection in three children given allogeneic hematopoietic stem cell transplantation: clinical and microbiologic features.

BACKGROUND AND OBJECTIVES: The emergence of vancomycin-resistant enterococci (VRE) as nosocomial pathogens is a major problem in the US; in Europe, VRE nosocomial infections are uncommon and only rarely have been reported in Pediatric or Neonatal Units. The aim of this study is to report on the clinical and microbiological features of VRE infections in 3 children given hematopoietic stem cell transplantation (HSCT). PATIENTS AND METHODS: Five episodes of vancomycin-resistant Enterococcus faecium (VREF) infection were diagnosed in 3 children given an allogeneic HSCT. Molecular methods, such as random amplification of polymorphic DNA (RAPD) fingerprinting and automated ribotyping, were used in order to define the circulation of strains. RESULTS: All the isolates were resistant to all commercially available agents and showed the VanA genotypic profile. All children were successfully treated with the combination of quinupristin/dalfopristin (QD) plus teicoplanin (TEC), although treatment was not sufficient to eradicate the micro-organism promptly from the gastrointestinal tract. All our children are still alive. After the first isolation of VRE, a surveillance protocol was started and we documented that the rate of colonization in children and their mothers was less than 1.5%. The RAPD method demonstrated the possible nosocomial transmission of one strain. INTERPRETATION AND CONCLUSIONS: Our experience demonstrates that VRE infection is a life-threatening complication in children given HSCT. Prompt diagnosis of this infection and its treatment with the combination of QD and TEC can successfully manage this severe infection in profoundly immunocompromised patients.

Regulation of inflammatory responses to Bordetella pertussis by N(G)-monomethyl-L-arginine in mice intranasally infected.

To investigate effect of MMLA, an inhibitor of nitric oxide (NO) production, on regulation of inflammatory responses to Bordetella pertussis infection, mice were infected intranasally, and treated with various concentrations of MMLA. Ten days after infection, mice treated with MMLA at dosage of 100 mg/kg, given intraperitoneally in a single dose or for 5 consecutive days, showed at histopathologic examination, a significant decrease of intensity of inflammation (scores, 0.6 +/- 0.2 and 0.9 +/- 0.5 respectively). A decrease of cellular accumulation of neutrophils and lymphocytes in the bronchoalveolar lavage (BAL) fluid was observed in infected mice treated with MMLA, especially at dosage of 10 mg/kg, given in a single dose intraperitoneally. In addition, BP-infected mice treated with MMLA (100 mg/kg, intraperitoneally) for 5 consecutive days showed higher mortality rate than untreated mice infected with B. pertussis, and the number of B. pertussis in lungs of mice treated with MMLA was significantly increased. However, MMLA treatment of infected mice had some effect on levels of IFN-gamma and nitrite/nitrate (end-stable products of NO) in the BAL fluid. This study indicates that NO may play a role either as microbiocidal agent or as a modulator of immune regulation, inasmuch as it may upregulate tissue inflammatory response to B. pertussis.

Comparative in vitro activity of silver sulfadiazine, alone and in combination with cerium nitrate, against staphylococci and gram-negative bacteria.

Silver sulfadiazine (SSD), a topical antimicrobial agent, has been widely used for the prophylaxis and treatment of burn infections during the past 30 years. We determined the antimicrobial activity of SSD, alone and in combination with cerium nitrate (CN), gentamicin and amikacin against 130 recent clinical isolates, including multiresistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) or Pseudomonas aeruginosa. The overall activity of SSD was good against all the tested strains and it was particularly high against MRSA (MIC90 100 microg/ml). CN showed no inhibitory effect, even up to 800 microg/ml, on bacterial strains tested. The combination of SSD and CN was as active as SSD alone. In conclusion, SSD has a broad spectrum of activity at concentrations lower than those commonly used in clinical preparations. All strains were inhibited by less then one-fiftieth of the SSD "in use" concentration (10 mg/ml). Our data confirm the efficacy of this topical agent in the prevention and treatment of infections in burns or other surgical wounds and suggest its possible use in clearing staphylococcal carriage as an alternative to mupirocin.

Effects of recombinant murine (rm) interleukin-12 and rm interferon-gamma in mice infected with Bordetella pertussis.

The recombinant cytokines interferon (IFN)-gamma and interleukin (IL)-12 stimulate several macrophage-mediated functions that are important in host defense. An experimental pertussis model showed that intraperitoneal (i.p.) administration of 10,000 U of recombinant murine (rm) IFN-gamma to mice at the time of Bordetella pertussis infection caused a marked and significant reduction in the number of colony-forming units of bacteria in the lungs. Administration i.p. of 1 microgram of rmIL-12 or 1 microgram of rmIL-12 at the time of and for 5 consecutive days after B. pertussis challenge also induced a significant reduction in the number. However, i.p. administration of 1 microgram of rmIL-12 with 10,000 U of IFN-gamma at the time of B. pertussis challenge did not provide protection. These findings indicate that exogenous administration of rmIL-12 and rmIFN-gamma enhances resistance of mice to B. pertussis infection.

In vitro and in vivo induction of nitric oxide by murine macrophages stimulated with Bordetella pertussis.

Nitric oxide (NO) exhibits potent antimicrobial activity in vitro. The function of NO in host defenses in vivo, however, is presently unclear. Experiments were undertaken to determine the production of NO in vitro from murine peritoneal and alveolar macrophages, and murine macrophage cell line (J774A.1) stimulated with Bordetella pertussis or pertussis toxin (PT). In addition, we determined circulating levels of NO in the sera and bronchoalveolar lavage (BAL) fluids of mice infected intranasally with B. pertussis. The results of this study showed that in vitro murine peritoneal macrophages induce production of NO in response to B. pertussis and PT. In addition, murine macrophage cell line, J774A.1 also induces NO production after stimulation with B. pertussis. NO production was also detected in alveolar macrophages from mice infected intranasally with B. pertussis. Finally, a significant increment of circulating levels of NO was noted, in the sera but not in the BAL fluids, of mice infected intranasally with B. pertussis.

Ocular infections: antibiotics and bacterial adhesion on biomaterials used in ocular surgery.

The in vitro antibacterial activity of ofloxacin, sagamycin and other antibiotics was evaluated against 85 bacterial isolates [coagulase-negative staphylococci (CNS), n = 37, Staphylococcus aureus, n = 28, and Pseudomonas aeruginosa, n = 20] obtained from patients with ocular infections. The antistaphylococcal activity of ofloxacin was quite elevated with a 90% minimal inhibitory concentration (MIC90) of 1.56 mg/l against CNS and S. aureus. Rokitamycin and erythromycin showed a good activity against methicillin-sensitive staphylococci, but were less active than ofloxacin and sagamycin against methicillin-resistant strains (MIC90 > 100 mg/l). Sagamycin was highly effective against staphylococci (MIC90 0.78 mg/l) and appeared to be the most active compound against P. aeruginosa (MIC90 6.25 mg/l), followed by ofloxacin, tobramycin and gentamicin. In a successive part of the study, the adhesive properties of slime-producing staphylococci were tested on biomaterials used in ocular surgery. Intraocular lenses, Silastic sheetings, circling bands and grooved strips showed a high affinity for slime-producing strains, while round silicone sponges were not covered by bacterial biofilm. In the last part of our study, we demonstrated how subMIC levels of ofloxacin increased the adhesion of slime-producing staphylococci. Our data confirmed the excellent activity of ofloxacin and sagamycin against ocular pathogens and the key role of adhesion in promoting colonization and infections of biomaterials.

Effects of recombinant human gamma interferon on intracellular survival of Bordetella pertussis in human phagocytic cells.

Several studies have demonstrated that Bordetella pertussis has the ability to enter and survive intracellularly within human polymorphonuclear leukocytes (PMNL) and human monocytes/macrophages. The effects of human recombinant gamma interferon (IFN-gamma) on the survival of B. pertussis in PMNL and human monocytes, and on the oxidative burst activity of PMNL and human monocytes in response to B. pertussis were assessed in this study. IFN-gamma partially increased intracellular killing of phagocytosed B. pertussis in human monocytes, as determined by an orange acridine-crystal violet assay. In contrast, IFN-gamma did not enhance intracellular killing of B. pertussis in PMNL. No significant increase of superoxide production was noted in human monocytes in response to B. pertussis when stimulated with various concentrations of IFN-gamma. The partial increase of B. pertussis killing by IFN-gamma within monocytes, together with poor production of superoxide may explain how B. pertussis can survive within human phagocytic cells, and thus cause a more prolonged course of the disease.

Activity of daptomycin against enterococci and coagulase-negative staphylococci (CNS): relationship between CNS susceptibility and slime production.

We compared the in vitro activity of daptomycin, a new lipopeptide antibiotic, with that of vancomycin and other selected agents against 95 coagulase negative staphylococci (CNS) isolates causing septicemia or foreign-body infections in immunocompromised patients. These strains were classified as follows: 51 methicillin-susceptible CNS (23 slime producers); 44 methicillin-resistant CNS (23 slime producers). We also investigated the activity of daptomycin against 50 Enterococcus faecalis isolates. Minimal inhibitory concentrations (MICs) were determined by the broth microdilution method. Daptomycin at a concentration of 2 mg/L was inhibitory for all the evaluated strains. Vancomycin and ciprofloxacin showed good activity: 90% of the strains were inhibited by these agents at 8 mg/L. The activity of netilmicin, rifampin and trimethoprim-sulfamethoxazole was instead limited. Resistance to the antimicrobial agents tested was seen with increased frequency among slime producing strains. Daptomycin and teicoplanin were the most active agents tested against E. faecalis (MIC90 0.25 mg/L and 0.12 mg/L).

Interferon-gamma levels in serum and bronchoalveolar lavage fluid of mice infected with Bordetella pertussis.

The adherence of Bordetella pertussis to respiratory cilia and its survival in neutrophils and macrophages is crucial to the pathogenesis of whooping cough. To investigate the role of endogenous interferon (IFN)-gamma in acute infection, levels of IFN-gamma in bronchoalveolar lavage (BAL) fluid of mice infected intranasally with B. pertussis were determined. Since pertussis toxin is released during infection by B. pertussis either locally or systemically, serum levels of IFN-gamma in mice injected intravenously with pertussis toxin were also determined. A persistent and significant increase of IFN-gamma levels with concomitant peripheral blood lymphocytosis was observed after 5 and 10 days. The results of this study showed an early but transitory production of endogenous IFN-gamma in BAL fluid of mice infected with B. pertussis.

Effect of teicoplanin and vancomycin on Staphylococcus aureus ultrastructure.

Morphological changes induced by teicoplanin and vancomycin on Staphylococcus aureus have been comparatively evaluated. The most pronounced structural damages were found on the cell wall which was uniformly thickened on bacteria exposed to teicoplanin while it was of irregular thickness and often absent around the cells in those exposed to vancomycin. The inhibitory action of teicoplanin on proteoglycan polymerization but not on nucleic acid and protein synthesis indicates that the effect in the wall is due to the accumulation at this level of proteoglycan soluble precursors continuously produced by the cell. The different effects induced by vancomycin on cell wall morphology can also be related to the compatibility of this antibiotic in altering cytoplasmatic membrane function and ribonucleic acid synthesis.

[Lactitol and neomycin: monotherapy or combined therapy in the prevention and treatment of hepatic encephalopathy?]. / Lattitolo e neomicina: monoterapia o terapia combinata nella prevenzione e nel trattamento dell'encefalopatia epatica?

The beneficial effect of disaccharides, lactulose and lactitol, in prevention and treatment of hepatic encephalopathy is well established but their use in combination with neomycin is still controversial. We studied in vitro the fecal bacterial growth, acid and gas formation in presence of lactitol (beta-galactoside-sorbitol) and neomycin alone or in combination. The results indicate that neomycin only inhibits the growth of susceptible bacteria (E. coli, Staph. aureus) which, conversely, are poor lactitol fermenters. The resistant organisms (Lactobacillus acidophilus, Clostridium perfringens) that are efficient disaccharide fermenters continue to metabolize lactitol still when antibiotic is added. Addition of lactitol 10% increased the inhibitory effect of neomycin on bacterial growth by 25-50% within 60-70 min. These preliminary data suggest that lactitol and neomycin may have additional or synergistic effects in vivo when used together in presence of favourable intestinal microbial environment.

Prophylactic co-trimoxazole versus norfloxacin in neutropenic children--perspective randomized study.

Co-trimoxazole or norfloxacin were randomly administered to 44 granulocytopenic children with malignancies in order to prevent bacterial infections. Although more patients in the co-trimoxazole group had febrile episodes (p less than 0.01), the mean of febrile days and the mean of days with systemic antibiotics did not differ significantly in the two groups. Five patients in the co-trimoxazole group had a microbiologically documented infection (four with septicemia) due to Escherichia coli (n = 2), Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus sp. There were four septicemic episodes in the norfloxacin group due to P. aeruginosa, Streptococcus pneumoniae, Streptococcus mitis and Streptococcus faecalis. Compliance was good during administration of both drugs. No signs or symptoms of arthropathy were seen in the norfloxacin group. The number of gram-negative bacilli resistant to co-trimoxazole isolated from stools significantly increased during prophylaxis with co-trimoxazole (p less than 0.001). Norfloxacin did not select resistant strains and was very active in eradicating gram-negative bacilli from stools (27.5% of positive cultures).

Clinical safety and efficacy of ofloxacin.

In this study we report about the efficacy and tolerability of ofloxacin in the treatment of 15 patients with severe and moderately severe infections including osteomyelitis (5), soft tissue infections (5), salmonellosis in AIDS patients (2), acute or chronic pulmonary infections (2) and mediastinitis (1). The following organisms were isolated in culture specimens: Staphylococcus aureus (4), Pseudomonas aeruginosa (4), Staphylococcus epidermidis (3), Serratia marcescens (1), Escherichia coli (1), Aeromonas hydrophila (1), Klebsiella oxytoca (1), Klebsiella pneumoniae (1), Salmonella cholerae-suis (1), Salmonella sp. (1), Enterobacter cloacae (1). All isolates were sensitive to the drug. Of 5 cases with osteomyelitis, 2 were cured and 3 improved clinically (with bacteriological eradication of the pathogens). The best results were obtained in patients with soft tissue infections: 4 patients were cured and 1 improved. Two patients with salmonella bacteremia and AIDS experienced a recurrence 1 month and 2 months respectively after stopping therapy. The patient with mediastinitis was successfully treated. Improvement was recorded for 2 patients with bronchiectasis and exacerbation of chronic bronchitis. The drug was well tolerated, only one episode of mild nausea and vomiting was reported and did not require discontinuation of the therapy. The study indicates that ofloxacin is a safe and effective agent in the treatment of various infections.

Antistaphylococcal activity of teicoplanin in association with other antibiotics.

The in vitro activity of antibiotic combination of teicoplanin and gentamicin (or netilmicin) and teicoplanin and cephalotin on Staphylococcus aureus was evaluated using the checkerboard method and time-kill curves. With few exceptions neither antagonism nor synergism was seen for the combination of teicoplanin and aminoglycosides using the checkerboard method. Using time-kill curves synergism was often found for the combination of teicoplanin and netilmicin (even at sub-minimal inhibitory concentrations of netilmicin) and for the combination of teicoplanin and cephalotin. No antagonistic interactions occurred.

[Epidemiologic study on the prevalence of Enterobacter, Serratia and Pseudomonas strains, producers of cefoxitin-inducible beta-lactamases]. / Studio epidemiologico sulla prevalenza di stipiti di Enterobacter, Serratia e Pseudomonas produttori di betalattamasi cefoxitina-inducibili.

Resistance of Enterobacter, Serratia and pseudomonas strains to newer cephalosporins is often associated with stable derepression of synthesis of the chromosomal betalactamases. Similar resistance is developed by enzyme inducible strains in response to betalactamases inducers. This finding poses many clinical problems including emergence of resistance during therapy with the drugs. In this study we evaluated the MICs of several new betalactam compounds against 76 Enterobacter, Serratia and Pseudomonas strains before and after cefoxitin-induction of betalactamases. The MICs against several Enterobacter strains (45%) after cefoxitin induction were elevated four fold or more. Serratia strains showed no significant variations of the MICs after cefoxitin induction. The MICs of piperacillin against many Pseudomonas strains (78%) after cefoxitin induction were elevated four fold or more. These data were confirmed using cefoxitin disk approximation test. Outbreaks of nosocominal infection with these multiresistant bacteria and spread of the strains throughout the hospital are already being seen. Control of these problems can only be achieved through the judicious and restricted use of these new antibiotics.