Maria Rita Gismondo

Review of probiotics available to modify gastrointestinal flora

There is evidence that the oral consumption of micro-organisms produces a protective effect on the gut flora. A significant number of studies suggests that probiotics might have beneficial effects on several microbial disorders of the gut, but it is very difficult to define the clinical efficacy of such products. In this review, we report the results of studies on travellers diarrhoea (TD), antibiotic associated diarrhoea (AAD) and acute diarrhoea (AD) in which various probiotic preparations have been used in controlled trials and been shown to have a beneficial therapeutic or prophylactic effect.

The use of probiotics in medical practice

Probiotics are defined as living organisms, beneficial to health when ingested. Different species of microorganisms such as lactic acid bacteria or yeasts have been proposed for human use. These microorganisms differ from each other and it is, therefore, unlikely that they will act in the same way. Probiotics could be used for several conditions such as diarrhoea, candidal vaginitis, urinary tract infections, immune disorders, lactose intolerance, hypercholesterolaemia and food allergy. The effects of probiotics in some of these conditions have been directly observed, in others it has been only suggested on the basis of in vitro studies and from experimental animal models. Controlled trials are needed to determine the scientific basis for their use, the correct formulation and ways of administration in different clinical situations.

In Vitro Antimicrobial Activity of Propolis Dry Extract

Abstract In this study the antibacterial and antifungal properties of propolis, a natural product of bees, have been investigated against different pathogens. Minimum inhibitory concentrations (MICs) and minimum bactericidal concentrations (MBCs) were determined according to NCCLS standards on 320 strains including Staphylococcus aureus, Group A beta-hemolytic streptococci, Streptococcus pneumoniae, Moraxella catarrhalis, Haemophilus influenzae, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa and Candida albicans. Time-kill curves were assessed for susceptible microorganisms, testing 0, 0.5, 1, 2, 4 × MIC for propolis, by counting viable bacteria after 0, 3, 6, 24 hours and viable yeasts after 0, 3, 6, 24 and 48 hours. Propolis showed good antimicrobial activity against most of the isolates, particularly S. pneumoniae, H. influenzae and M. catarrhalis, but not against Enterobacteriaceae. Time-kill curves demonstrated bacteriostatic rather than bactericidal activity of propolis, the latter being evident only at high concentrations.

In vitro evaluation of antibiotics' combinations for empirical therapy of suspected methicillin resistant Staphylococcus aureus severe respiratory infections

BackgroundMethicillin resistant Staphylococcus aureus (MRSA) is an increasingly common cause of nosocomial infections, causing severe morbidity and mortality worldwide, and accounting in some hospitals for more than 50% of all S. aureus diseases. Treatment of infections caused by resistant bacterial pathogens mainly relies on two therapeutic modalities: development of new antimicrobials and use of combinations of available antibiotics.Combinations of antibiotics used in the empiric treatment of infections with suspected methicillin resistant Staphylococcus aureus etiology were investigated.MethodsDouble (vancomycin or teicoplanin with either levofloxacin or cefotaxime) and triple (vancomycin or teicoplanin + levofloxacin + one among amikacin, ceftazidime, cefepime, imipenem, piperacillin/tazobactam) combinations were evaluated by means of checkerboard assay and time kill curves. Mutational rates of single and combined drugs at antimicrobial concentrations equal to the resistance breakpoints were also calculated.ResultsVancomycin or teicoplanin + levofloxacin showed synergy in 16/50 and in 9/50 strains respectively, while vancomycin or teicoplanin + cefotaxime resulted synergic for 43/50 and 23/50 strains, respectively. Triple combinations, involving teicoplanin, levofloxacin and ceftazidime or piperacillin/tazobactam gave synergy in 20/25 strains. Teicoplanin + levofloxacin gave synergy in triple combinations more frequently than vancomycin + levofloxacin.For single antibiotics, mutational frequencies ranged between 10-5 and <10-9 for levofloxacin, cefotaxime, amikacin and imipenem, and <10-9 for vancomycin and teicoplanin. When tested in combinations, mutational frequencies fell below 10-9 for all the combinations.ConclusionIn vitro evidence of synergy between glycopeptides, fluoroquinolones (levofloxacin) and β-lactams and of reduction of mutational frequencies by combinations are suggestive for a potential role in empirical therapy of severe pneumonia with suspected MRSA etiology.

In vitro antimicrobial activity of a novel propolis formulation (Actichelated propolis)

Aims: This study compared in vitro activities of Actichelated® propolis (a multicomposite material obtained with mechano‐chemichal activation) and of a hydroalcoholic extract of propolis.

Candida albicans cellular internalization: a new pathogenic factor?

The preliminary results of a study to show the possibility that Candida albicans can internalize into epithelial cells are reported. The study was performed on buccal, vaginal and HeLa cells. Buccal and vaginal cells, at a concentration of 5 x 10(4) cells/ml and HeLa monolayers were incubated for 2, 3 and 4 h with 10(5) colony forming units of a Candida albicans isolate. After incubation, non-internalised yeasts were eliminated and samples were processed for examination by Scanning Electron Microscopy. Results suggest that receptor-mediated endocytosis could be involved in the pathogenesis of recurrent oral and vaginal infections. This phenomenon could represent an interesting experimental model to testing drug interference in the development of therapeutic strategies against C. albicans infections.

Comparison of nested PCR and real time PCR of Herpesvirus infections of central nervous system in HIV patients

BackgroundMolecular detection of herpesviruses DNA is considered as the reference standard assay for diagnosis of central nervous system infections. In this study nested PCR and real time PCR techniques for detection of Herpes simplex virus type 1 (HSV-1), Cytomegalovirus (CMV) and Epstein-Barr virus (EBV) in cerebrospinal fluid of HIV patients were compared.MethodsForty-six, 85 and 145 samples previously resulted positive for HSV-1, CMV and EBV by nested PCR and 150 randomly chosen negative samples among 1181 collected in the period 1996–2003 were retrospectively reassessed in duplicate by real time PCR and nested PCR.ResultsSamples giving positive results for CMV, HSV-1 and EBV with nested PCR were positive also with real time PCR. One of the negative samples resulted positive for HSV and one for EBV. Real time PCR showed comparable sensitivity and specificity vs nested PCR.ConclusionReal time PCR proved to be a suitable method for diagnosis of herpesvirus infections in CNS, showing comparable sensitivity and being less time consuming than nested PCR.

Microbiological Evaluation of Commercial Probiotic products available in Italy

Abstract Scientific evidence of the prevention and therapy of some intestinal diseases is accumulating in regard to probiotic products. However, sufficient information on the use of probiotics in specific therapies is not yet available and, above all, there is no clear legislation about these products in Europe. In this study, we evaluated five different probiotic products commercially available in Italy for their qualitative and quantitative microbial content after about 12 and 22 months of storage. We also evaluated the stability of lactobacilli to 0.3% bile salts and to pH of 3.58 and 7.98. There were discrepancies between the declared content and our results found after storage for 4 of the tested products. Bile salts and basic pH did not affect the growth of the lactobacilli tested, while for 2 tested products 6 hours at acid pH produced a complete inhibition of bacterial growth. Our results suggest the need for clear legislation and adequate control of the manufacturing of probiotic products.

Activity of levofloxacin and ciprofloxacin in combination with cefepime, ceftazidime, imipenem, piperacillin-tazobactam and amikacin against different Pseudomonas aeruginosa phenotypes and Acinetobacter spp.

Background: Combination therapy is used to widen the antimicrobial spectrum, minimize toxicity and prevent the emergence of resistant mutants. Methods: Synergy between levofloxacin or ciprofloxacin and ceftazidime, cefepime, imipenem, piperacillin-tazobactam and amikacin was evaluated by checkerboard assay with 55 strains and by time-kill curves with 8 strains of Pseudomonasaeruginosa and Acinetobacter spp. Results: In the checkerboard assay, synergy and additivity were the most frequent effects observed among all the combinations against P. aeruginosa and Acinetobacter spp., with no significant differences between the two fluoroquinolones. No antagonism was observed. In the time-kill curves, synergy was evidenced against all the tested strains, at least for one combination at one of the time points considered. Levofloxacin and ciprofloxacin combined with ceftazidime, as well as levofloxacin plus amikacin, were synergistic for all the strains tested. Conclusion: Combinations of fluoroquinolones with β-lactams or amikacin show an enhanced activity against P. aeruginosa and Acinetobacter spp.

Escherichia coli: Effect of fosfomycin trometamol on some urovirulence factors

The aim of our study was to evaluate the interference of fosfomycin trometanol (F.T.), at subinhibitory concentrations (1/4 and 1/8 MICs), on some urovirulence factors of Escherichia coli (12 strains). We tested fimbriae production, adhesion to uroepithelial cells, hydrophobicity, motility and hemolysin production of E. coli grown in the presence or absence of F.T. The strains tested, grown in the presence of F.T. (1/8 MIC), were less capable of adhering to uroepithelial cells, had less hemagglutination and reduced motility. This behavior was enhanced at 1/4 MIC of F.T. The hemolysin production and hydrophobicity properties present in some of our tested strains also were significantly decreased when the E. coli were grown in the presence of sub-MIC concentrations of F.T. These results suggest that F.T. may be of clinical use as treatment for acute urinary tract infection and in pyelonephritis prophylaxis.