Microbiological surveillance of flexible bronchoscopes after a high-level disinfection with peracetic acid: preliminary results from an Italian teaching hospital.

BACKGROUND: Flexible bronchoscopes are heat labile, complex and difficult to clean, and some nosocomial outbreaks related to bronchoscopy have been reported in literature. The aim of our study was to determine, through a systematic monitoring, whether bronchoscopes' cleaning and disinfection procedures have been correctly adopted by health operators. METHODS: We conducted a 19 months-long prospective study in the Unit of Pulmonology at Careggi Teaching Hospital (Florence, Italy), analyzing endoscopes that were reprocessed through a high-level disinfection procedure. Samples collection was performed weekly by two trained operators. Results were organized in a database and then exported for descriptive and inferential statistical analysis. RESULTS: From February 2016 to September 2017 we collected 218 samples from bronchoscopes' valves (N=109) and from their inner channels (N=109). Staphylococci were found in 34 samples (15.69% of all samples). Pseudomonas was found in 11 samples (5.04% of all samples). Pseudomonas aeruginosa wasn't found in any sample. CONCLUSIONS: Our results came out to be better than similar studies in literature and demonstrated that a correct endoscopes' hygiene should be part of a more complex strategy of surveillance and control of healthcare-associated infections. However, a continuous monitoring of endoscopes could provide a wider view about this problem, and more reliable results.

Mycotic and aflatoxin contamination in Myristica fragrans seeds (nutmeg) and Capsicum annum (chilli), packaged in Italy and commercialized worldwide.

Aflatoxins are secondary metabolites of moulds known to be carcinogenic for humans, and therefore should not be ingested in high doses. This study aimed to determine the level of mould and aflatoxin contamination in dehydrated chilli and nutmeg imported from India and Indonesia, respectively, packaged in Italy, and commercialized worldwide. We tested 63 samples of chilli (22 sanitized through heat treatment and 41 not heat-treated) and 52 samples of nutmeg (22 heat-treated and 30 not heat-treated) for aflatoxin, moulds and moisture content. Heat-treated samples were less contaminated than untreated samples. Spices in powder form (both chilli and nutmeg) were more contaminated than whole ones. In untreated spices, we observed a positive correlation between mould and moisture content. Of the powdered nutmeg and chilli samples, 72.5% and 50% tested positive for aflatoxin contamination, with a range of 0-17.2 µg kg(-1) and 0-10.3 µg kg(-1), respectively. The steam treatment of spices would be useful in reducing the initial amount of moulds. Although the risk from the consumption of spices contaminated with aflatoxins is minimal, owing to the small amount used in food, preventive screening of the whole food chain is very important, especially because the most frequently identified toxin was B1, which is the most dangerous of the four toxins (B1, B2, G1, G2).

In vitro activity of Aloe vera inner gel against microorganisms grown in planktonic and sessile phases.

The failure of traditional antimicrobial treatments is becoming a worldwide problem. The use of Aloe vera is of particular interest for its role as curative agent and its efficacy in complementary therapies for a variety of illnesses. This study evaluated the antimicrobial activity of A. vera inner gel against a panel of microorganisms, Gram-positive and -negative bacteria, and Candida albicans. In addition to A. vera inner gel being used in the treatment of peptic ulcers, in dermatological treatments, and wound healing, it was also tested on the sessile phase of clinical Helicobacter pylori strains (including multi-drug-resistant strains) and on planktonic and sessile phase of Staphylococcus aureus/Pseudomonas aeruginosa clinical isolates from venous leg ulcers.A. vera inner gel expresses its prevalent activity against Gram-negative bacteria and C. albicans in respect to Gram-positive bacteria. The results of the A. vera antibiofilm activity showed a decrease of the produced biomass in a concentration-dependent-way, in each analyzed microorganism. The data obtained show that A. vera inner gel has both an antimicrobial and antibiofilm activity suggesting its potential use for the treatment of microbial infections, in particular for H. pylori gastric infection, especially in case of multi-drug-resistance, as well as for an effective wound dressing.

Effect of temperature on the release of carvacrol and cinnamaldehyde incorporated into polymeric systems to control growth and biofilms of Escherichia coli and Staphylococcus aureus.

This study assessed the effect of temperature on the release of essential oil components incorporated by melt compounding into polymeric films. Specifically, polyethylene-co-vinylacetate (EVA) films containing carvacrol (CAR) and cinnamaldehyde (ALD), alone and in combination, were prepared and their surface and mechanical properties and antibacterial and anti-biofilm activity against Escherichia coli and Staphylococcus aureus were evaluated. The addition of ALD and CAR did not provoke variation in the surface morphology of EVA and allowed their delivery. At 37°C, films containing CAR, ALD or their combination (25+75%) were found to have the strongest bactericidal effect, whereas at lower temperatures a lower killing rate was observed. There was no clear evidence of the influence of temperature on the anti-biofilm activity of the essential oil component-based polymeric films. The biomass formed on EVA containing ALD, CAR or their combination (25+75) was significantly lower (60-80% reduction) than that formed on the EVA control at both 37° and 22°C.

Occurrence of Acrylamide in breakfast cereals and biscuits available in Italy.

INTRODUCTION: Acrylamide, produced during thermal processing of carbohydrate-rich foods, is classified as "probably carcinogenic to humans" by the International Agency for Research on Cancer. OBJECTIVE: The aim of this study was to determine acrylamide levels in biscuits and breakfast cereals considering the widespread use of these products for all ages. METHOD: Acrylamide determination was carried out in 56 samples by HPLC-UV technique. RESULTS AND DISCUSSION: The results showed a considerable variability in the contents of acrylamide in the samples analysed, most likely due to differences in industrial processing and ingredients. The percentages of contaminated samples tested were very high (95.5% of the biscuits and 75% of the breakfast cereals) with a wide range of contamination: from 30 µg/kg to 940 µg/kg. Our results showed that 22.7% of biscuits and 33% of breakfast cereals exceeded the indicative values recommended by EC 2013/647 set at 500 µg/kg and 200-400 µg/kg (according to the composition) respectively. CONCLUSIONS: Our findings suggest concern about the risk for human health.

Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready-to-eat salads, ham, and raw meat.

Food specimens were analyzed in order to research Enterococcus spp.: 636 samples of raw meat (227 beef, 238 poultry, and 171 pork), 278 samples of cheese (110 fresh soft cheese and 168 mozzarella cheese), 214 samples of ready-to-eat salads, and 187 samples of ham. 312 strains of Enterococcus spp samples were isolated, then identified and submitted to susceptibility tests against 11 antimicrobial agents. The predominant species were Enterococcus faecalis in raw meat and Enterococcus faecium in retail products. Low percentages of microorganisms were resistant to vancomycin (3.53%), teicoplanin (2.24%), linezolid (0.32%), and amoxicillin in combination with clavulanic acid (0.32%). A high percentage of resistance was noted in E. faecalis at high level gentamicin (21.9%) and tetracycline (60.6%). In general, strains of E. faecalis were more resistant than E. faecium. Enterococci should be considered not only potential pathogens, but also a reservoir of genes encoding antibiotic resistance which can be transferred to other microorganisms. Continuous monitoring of their incidence and emerging resistance is important in order to identify foods which potentially represent a real risk to the population, and to ensure effective treatment of human enterococcal infections.

Development and characterization of essential oil component-based polymer films: a potential approach to reduce bacterial biofilm.

The development of new polymeric materials aimed to control the bacterial biofilm appears to be an important practical approach. The goal of the present study was to prepare and characterize poly(ethylene-co-vinyl acetate) copolymer (EVA) films containing citronellol, eugenol, and linalool and evaluate their efficiency on growth and biofilm formation of Listeria monocytogenes, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, and Pseudomonas aeruginosa in monospecies and dual species. The results showed that the addition of oil components influenced the elastic modulus (15 % decrease), the tensile stress (30 % decrease), the elongation at break (10 % increase), and the contact angle values (10-20° decrease) while leaving the homogeneity of the surface unaltered. Among the polymeric films, EVA + citronellol and EVA + eugenol at 7 wt% had the best inhibitory effect. After 24-48 h of incubation, EVA + citronellol was more effective against the growth (30-60 % reduction) than EVA + eugenol (15-30 % inhibition). However, this inhibition decreased after 240 h of incubation. On the contrary, the biofilm evaluation revealed a strong inhibition trend also after prolonged incubation time: the amount of biomass per square centimeter formed on copolymer with oil components was significantly less (40-70 % decrease) than that on pure copolymer control for L. monocytogenes, S. aureus, and E. coli. When polymeric materials were simultaneously inoculated with combinations of S. aureus and E. coli, the biomass accumulated was higher for EVA + citronellol and lower for EVA + eugenol than that in monoculture biofilm. The findings were similar to the results obtained by 2,3-bis[2-methyloxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assay that measures the metabolic activity of viable cells.

Study on carvacrol and cinnamaldehyde polymeric films: mechanical properties, release kinetics and antibacterial and antibiofilm activities.

Polyethylene-co-vinylacetate (EVA) films with different concentrations (3.5 wt% and 7 wt%) of essential oil constituents, carvacrol or cinnamaldehyde, were prepared and characterized by mechanical, antibacterial and antibiofilm properties. The incorporation of the compounds into copolymer films affected their elastic modulus, tensile stress and elongation at break. Carvacrol and cinnamaldehyde act as plasticizers which reduce the intermolecular forces of polymer chains, thus improving the flexibility and extensibility of the film. The analysis of the surface characteristics demonstrated that essential oil constituents lowered the contact angle values without causing any remarkable variation of the surface roughness. The films allowed progressive diffusion of the bioactive molecules and the kinetic of release was correlated with the damaging effect on bacterial growth. The kill curves proved that the film with essential oil constituents (7 wt%) had a significant bactericidal effect (reduction of 4 and 2 log CFU) against Staphylococcus aureus and Escherichia coli and a bacteriostatic effect against Staphylococcus epidermidis and Listeria monocytogenes (reduction of about 1 log CFU). With regard to biofilm formation the biomass formed on polymeric films surface was significantly reduced if compared with the pure copolymer control. The results were confirmed by fluorescence microscopy images by Live/dead staining. The reduction in the surface tension coupled to an inherent bactericidal property of carvacrol and cinnamaldehyde could in turn affect the initial attachment phase of bacteria and compromise the normal biofilm development.

Design, synthesis and antibacterial evaluation of a polycationic calix[4]arene derivative alone and in combination with antibiotics.

The growing antibiotic resistance phenomenon continues to stimulate the search for new compounds and strategies to combat bacterial infections. In this study, we designed and synthesized a new polycationic macrocyclic compound (2) bearing four N-methyldiethanol ammonium groups clustered and circularly organized by a calix[4]arene scaffold. The in vitro activity of compound 2, alone and in combination with known antibiotics (ofloxacin, chloramphenicol or tetracycline), was assessed against strains of Staphylococcus aureus (ATCC 6538 and methicillin-resistant isolate 15), S. epidermidis (ATCC 35984 and methicillin-resistant isolate 57), and Pseudomonas aeruginosa (ATCC 9027 and antibiotic-resistant isolate 1). Calix[4]arene derivative 2 showed significant antibacterial activity against ATCC and methicillin-resistant Gram positive Staphylococci, improved the stability of tetracycline in water, and in combination with antibiotics enhanced the antibiotic efficacy against Gram negative P. aeruginosa by an additive effect.

Impact of routine infant and adolescent hepatitis B vaccination in Tuscany, Central Italy.

BACKGROUND: In Italy routine infant and adolescent immunization against hepatitis B was introduced in 1991. OBJECTIVE: Evaluation of (1) coverage with three doses of hepatitis B vaccine in infants and adolescents; (2) seroconversion to anti-hepatitis B surface antigen antibody (anti-HBs) in adolescents receiving three doses of vaccine; (3) concordance of coverage rates in infants with prevalence of neutralizing antibodies in sera from anonymous children; (4) trend of notified cases of acute hepatitis B. METHODS: A sample of infants and adolescents living in Tuscany was studied during 6 years (1992 through 1997) by matching birth records and immunization certificates. Sera from 139 adolescents who completed the vaccination course and from 159 anonymous children belonging to immunized cohorts (1 to 5 years) were tested with a quantitative anti-HBs assay. Incidence of acute hepatitis B by age was calculated from regional statistics on notified infectious diseases between 1992 and 1996. RESULTS: Overall 10,606/11,164 (95%) infants and 10,599/11,100 (95%) adolescents received 3 doses of vaccine. Seroconversion to anti-HBs was detected in 98% of adolescent vaccinees. Anti-HBs titers > or =10 IU/l were detected in 87% of children. A 49% decline of acute hepatitis B cases was registered between 1992 and 1996 in 15- to 24-year-olds living in Tuscany. No case occurred in vaccinated adolescents. CONCLUSIONS: Coverage against hepatitis B is excellent in cohorts subject to mandatory immunization. If efforts to vaccinate are maintained at these levels, elimination of hepatitis B virus transmission could occur within few decades in Italy.

In vitro antibacterial activity of some aliphatic aldehydes from Olea europaea L.

In the present paper we report the 'in vitro' activity of eight aliphatic long-chain aldehydes from olive flavor (hexanal, nonanal, (E)-2-hexenal, (E)-2-eptenal, (E)-2-octenal, (E)-2-nonenal, (E)-2-decenal and (E,E)-2,4-decadienal) against a number of standard and freshly isolated bacterial strains that may be causal agents of human intestinal and respiratory tract infections. The saturated aldehydes characterized in the present study do not exhibit significant antibacterial activity, while the alpha,beta-unsaturated aldehydes have a broad antimicrobial spectrum and show similar activity against Gram-positive and Gram-negative microorganisms. The effectiveness of the aldehydes under investigation seems to depend not only on the presence of the alpha,beta-double bond, but also on the chain length from the enal group and on the microorganism tested.

The effect of Nepeta cataria extract on adherence and enzyme production of Staphylococcus aureus.

Nepeta cataria L., commonly known as catnip, is a perennial herb with a considerable folkloric reputation. A diethyl ether extract of this plant has been shown to have antimicrobial activity against fungi and Gram-positive bacteria. The aim of this work was to study the activity of N. cataria extract on 44 Staphylococcus aureus strains, some resistant to methicillin, and S. aureus 6538P (American Type Culture Collection) by evaluating the effect of subminimum inhibitory concentrations on coagulase, DNAse, thermonuclease and lipase production, and on in-vitro adherence. DNAse, thermonuclease and lipase were inhibited by concentrations equal to 1/2 and 1/4 MIC. A reduction of adherence was also observed.

Effects of Helichrysum italicum extract on growth and enzymatic activity of Staphylococcus aureus.

Helichrysum italicum G. Don (Compositae) is a shrub commonly found in dry, sandy and stony areas of Mediterranean regions. This plant is known for its anti-inflammatory, anti-allergic and antimicrobial activity. The aim of this study was to evaluate the effect of the diethyl ether extract on growth of Staphylococcus aureus (ATCC 6538P, MRSA and MSSA isolates) and the influence of subminimum inhibitory concentrations (subMICs) on some enzymes which are considered virulence factors. The results indicate that the H. italicum extract had an inhibitory effect on S. aureus strains reducing both their growth and some of the enzymes such as coagulase, DNAse, thermonuclease and lipase. Helichrysum italicum extract could be a novel antimicrobial agent, less toxic to human skin and tissues, worthy of further studies.

Uptake and retention of Vibrio cholerae non-O1, Salmonella typhi, Escherichia coli and Vibrio harvey by mussels in seawater.

Vibrio cholerae non O1 is known to persist in estuarine and freshwater environments. Experiments evaluated the amount of microorganisms accumulated in mussels maintained in static seawater, contaminated with 10(4) to 10(6) cells/ml and the depuration time required in circulating water. Accumulation and retention times were compared with those for Escherichia coli, Salmonella typhi and Vibrio harvey. E. coli and S. typhi accumulated to a greater extent and were released from mussels more quickly than vibrios which became undetectable 2 to 3 days later than E. coli. Seasonal seawater temperatures (14 to 21 degrees C) had a limited influence on depuration but vibrios appear to be retained with more efficacy over 16 degrees C while E. coli and S. typhi were eliminated to a greater extent. When mussels were contaminated with mixed culture, vibrios appeared to predominate on E. coli, while no interference was observed between E. coli and S. typhi.

Fatty acid profiles in pigmented and non-pigmented strains of S. marcescens.

Wild, pigmented strains of Serratia marcescens and their non-pigmented mutants were compared on the basis of fatty acid profiles and lipid content. Classic biochemical tests show only minor differences, as well as fatty acid ratio C18:C16. The total amount of lipid synthesized and the saturated/unsaturated fatty acids ratio disclose a sharp total lipid reduction and a high percentage of unsaturated fatty acids in the pigmented strains, placing them in separated clusters compared with the nonpigmented mutants. It is hypothesized that the synthesis of the polyacetate required for the completion of the prodigiosin molecule may result in waste of methyl groups and thus affect the total amount of lipids.

Evaluation of antiherpesvirus-1 and genotoxic activities of Helichrysum italicum extract.

The antiherpes virus-1 and genotoxic activities of diethyl ether extract from flowering tops of Helichrysum italicum (Compositae) were investigated. The extract showed significant antiviral activity at concentrations ranging from 400 to 100 microg/ml. This activity was not due to cytotoxic effect of the extract since Vero cells exhibited altered morphology or growth characteristics indicative of cytotoxic effects at higher concentration (800 microg/ml). Moreover H. italicum extract showed no DNA-damaging activity at concentrations up to 2000 microg/disk.