[Vibrio spp. infections of clinical significance and implication for public health]. / Infezioni da Vibrio spp. di interesse clinico e problematiche di Sanitå Pubblica.

Vibrio spp. infections still are a Public Health concern. Vibrio spp. can be found in marine, estuarine, and freshwater environments, and can be able to cause diseases in fish, shellfish, mammals, as well as in humans. Since '80 to date, the number of species within the genus increased from 21 to more than 100. The most important is Vibrio cholerae, the etiological agent of the cholera, responsible of seven pandemics; serotypes O1 and O139 can produce cholera toxin, while serotypes non-O1/non-O139 are generally associated with sporadic cholera cases and extraintestinal infections. Vibrio parahaemolyticus is an important cause of gastroenteritis associated with contaminated seafood consumption, whereas Vibrio vulnificus and V. alginolyticus can be related to wound infections or seafoodborne primary septicemia in immunocompromised patients. Disease prevention is mainly based on the application of proper individual or collective preventive measures.

Typing of Listeria monocytogenes strains isolated in Italy by inlA gene characterization and evaluation of a new cost-effective approach to antisera selection for serotyping.

AIMS: In this study, 105 Listeria monocytogenes strains isolated from humans, foods and environmental samples were characterized using several typing methods. Moreover, serotyping procedure was evaluated, and a cost-effective methodological approach based on preliminary PCRs screening was proposed. METHODS AND RESULTS: The isolates were analysed by conventional serotyping, multiplex-PCRs for serogroup and lineage identification and PCR-RFLP of inlA gene to identify potentially noninvasive L. monocytogenes. Among the strains, only the serotypes 1/2a, 1/2c, 1/2b, 4b and 3a were identified. The isolates were classified into serogroups I (58.10%), II (22.85%), III (12.38%) and IV (6.67%). Among clinical strains, lineage I was more represented (68.75%) than lineage II; whereas, lineage II was more associated with food (90.24%) and environmental (85.72%) isolates. Most of food (89.02%) and environmental (85.71%) isolates were classified into truncated InlA profiles, whereas the 93.75% of clinical strains were associated with a complete form of the protein. CONCLUSION: Molecular techniques were sensitive and specific for classifying strains into serogroup and lineage and in agreement with the serotyping. Moreover, a preliminary PCRs-based screening was proposed to select only the necessary antisera by a flow chart; this methodological approach allows cost saving up to 42%. Our results further suggest the role of InlA protein in human listeriosis, particularly in immunocompetent individuals, and a correlation between truncated protein and serotype. SIGNIFICANCE AND IMPACT OF THE STUDY: This study further validates molecular methods for L. monocytogenes analysis and proposed a new cost-effective approach for serotyping. It could help to improve a national surveillance network for L. monocytogenes infections in Italy.

[Alcohol consumption, pregnancy and fetal alcohol syndrome: implications in public health and preventive strategies]. / Alcolismo, gravidanza e fetopatia alcolica: implicazioni in Sanità Pubblica e strategie preventive.

Alcohol consumption during pregnancy is a significant public health problem and is an established cause of serious birth defects and developmental delay collectively described as fetal alcohol syndrome (FAS). FAS is caused by congenital alcohol-induced damages and is a cause of mental retardation. It is characterised by facial abnormalities and growth deficiency. Infants affected by the syndrome show intellectual impairment, and difficulties in learning, memory, problem-solving, and attention as well as experiencing additional problems with mental health and social interactions. However, an absence of the characteristic facial defects and growth deficiency may result in a failure to identify children with prenatal alcohol exposure, which can further present as alcohol-related neurodevelopment disorder (ARND) or alcohol-related birth defects (ARBD). Estimates of prevalence of FAS in U.S.A. range between 0.3 to 2.2 per 1,000 live births, but much higher rates occurring in some communities. Harmonisation of the methodology used for epidemiological studies, with research activities that establishes real baseline prevalence of FAS and identification of women who are at highest risk of bearing a FAS-affected child, are an essential prerequisite to prevention. In addition, it is essential to assess different FAS preventive approaches through carefully controlled studies. Universal, selected, and indicated preventive strategies have been identified, targeting different kind of populations. Since FAS and other adverse effects of drinking during pregnancy are theoretically completely preventable, it is vital to make more efforts to improve the application of the most appropriate interventions. Although in Italy alcohol consumption has constantly increased, mainly amongst the young (including women of childbearing age), knowledge on FAS and alcohol-related effects is completely lacking. Because of the high cost for care of individuals with this syndrome, it is essential to apply appropriate interventions to prevent this problem.

[Prevalence of Listeria spp. in dairy farm and evaluation of antibiotic-resistance of isolates]. / Prevalenza di Listeria spp. in aziende produttrici di latte e valutazione dell'antibiotico-resistenza dei ceppi isolati.

The study was performed to estimate the prevalence and antimicrobial sensitivities of Listeria spp. in raw milk, feaces end environmental samples isolated from 10 dairy in Molise Region. A total of 454 samples were collected, which comprised 40 raw milk, 40 animal faeces and 374 environmental samples. Listeria monocytogenes was never isolated from raw milk specimens; one was isolated from faeces speciments and two were isolated from environmental samples. All isolates were resistant to two or more of antimicrobial agents tested (cephalotin, ampicillin, tetracycline, co-trimoxazole, erytromicin, clindamycin, gentamicin, oxacillin). One isolate of L. monocytogenes was susceptible to all antimicrobial agents tested except oxacillin. This study indicates that faeces, equipments and environment are important reservoirs of Listeria spp. in dairy farm, and can represent potential source of contamination of raw milk. However, the contamination of milk, and the risk of infection, can be effectively eliminated by pasteurisation process.

Occurrence of Vibrio and other pathogenic bacteria in Mytilus galloprovincialis (mussels) harvested from Adriatic Sea, Italy.

Sixty-two samples of Mytilus galloprovincialis (mussels) harvested from approved shellfish waters in the Adriatic Sea were examined for the presence of Vibrio, Salmonella, Campylobacter, and verocytotoxin producing Escherichia coli. Vibrio spp. were isolated from 48.4% of samples; the species most frequently found were V. alginolyticus (32.2%) and V. vulnificus (17.7%), followed by V. cincinnatiensis (3.2%), V. parahaemolyticus (1.6%), V. fluvialis (1.6%) and V. cholerae non-O1 (1.6%). V. parahaemolyticus resulted negative to Kanagawa-phenomenon and to PCR amplification of tdh gene. V. cholerae resulted negative to PCR amplification of sto gene. No Salmonella, Campylobacter, or E. coli verocytotoxin-producing strains were isolated. The results of this study suggest the potential risk of ingesting raw or undercooked mussels due to the frequent presence of potentially pathogenic Vibrio species.

Prevalence of Helicobacter-like organisms in porcine gastric mucosa: a study of swine slaughtered in Italy.

Recent reports described some cases of gastritis in man caused by an uncultured gram-negative spiral bacterium morphologically identical to organisms observed in the stomachs of mammalians (e.g. cats, dogs, pigs). The aim of the present study is to confirm the presence of these bacteria in Italian swine. Tightly spiralled organisms (Gastrospirillum suis) were found in the stomach of eight (9.4%) out of 85 pigs examined. The bacteria were always associated with macroscopic lesions indicative of gastritis. Attempts to culture H. pylori or Helicobacter-like organisms were unsuccessful. The possibility that Gastrospirillum may be a zoonotic pathogen, with transmission occurring from pigs to humans, is discussed.

Evaluation of immunomagnetic separation and plating media for recovery of Salmonella from meat.

Immunomagnetic separation (IMS) was compared with selective enrichment in selenite cystine (SC) broth for isolation of Salmonella from 86 artificially contaminated ground beef samples. Both Rambach agar (RA) and Hektoen enteric (HE) agar were used as selective plating media. The highest count of Salmonella colonies per plate was obtained after enrichment in SC broth and plating on RA (mean value: 111.1+/-58.1 CFU per plate); the lowest count was obtained after IMS and plating on HE agar (mean value: 65.4+/-36.6 CFU per plate). Salmonella in preenrichment was concentrated 1.7-fold by IMS and represented 31% of the microbial population captured by the beads, but nonspecific binding was high. As a result of the large numbers of competing bacteria, isolations on both RA and HE agar following IMS were quite difficult (mean value for Salmonella colonies: 79.9+/-42.7 CFU per plate; mean value for non-Salmonella colonies: 144.1+/-75.7 CFU per plate; ratio of Salmonella to non-Salmonella colonies: 0.8). This study indicates that SC broth is superior to IMS in the isolation of Salmonella from raw ground meat.

Dental health in a group of drug addicts in Italy.

The caries prevalence, periodontal health, and oral hygiene status were assessed in 124 drug addicts aged 18-34 yr in Mercato San Severino (Salerno), Italy. The years of drug addiction ranged from 1 to 15 yr. The majority used heroin (96%) but all the subjects indicated that they had used more than one drug. The percentage of drug addicts caries free was 6.5%, ranging from no caries free subjects in the group over 29 yr of age, to 11.4% in the group aged 26-29 yr old. Mean DMFT and DMFS scores for all age groups were 12.9 and 36.2, respectively; mean Periodontal Index (PI) and mean Simplified Oral Hygiene Index (OHI-S) scores were 1.37 and 1.71, respectively. The poor dental health in this target group indicates the need for more extensive collaboration between the social and dental health care sectors.

Dentists and AIDS: a survey of knowledge, attitudes, and behavior in Italy.

OBJECTIVES: Italy, together with Spain, is second only to France in the total number of AIDS cases in Europe, with over 16,800 as of March 1993. The purpose of this study was to evaluate knowledge, attitudes, and behavior concerning AIDS and infection control among Italian dentists. METHODS: A questionnaire was mailed to 1,000 dentists randomly selected from the Italian Dental and Maxillo-Facial Association's register of dentists. RESULTS: Of the 715 dentists responding, 70.7 percent of dentists knew all the main risk groups able to transmit the infection and that semen is a biologic fluid potentially contaminated by HIV virus. This knowledge was greater if the number of patients per week was not higher than 55 and if the dentist had had a previous contact with an HIV-seropositive patient. Only a few (21.1%) knew all the oral manifestations of AIDS. Over 65 percent of the dentists indicated that they would treat HIV-seropositive patients (71.9%) or those with AIDS (66.8%). Dentists were more willing to care for an HIV-seropositive patient if they were involved in specialties with high blood contact, if they had a previous contact with an HIV patient, as the average number of patients per week increased, and if they did not consider saliva as a possible route of transmission of HIV. A small percentage of dentists who had the opportunity to treat patients at risk for AIDS (12.1%) or HIV seropositive (9.4%) refused to treat them. Willingness to treat was the most significant predictor of actual treatment of an infected patient. Only 24.4 percent routinely used all barrier techniques (gloves, masks, and protective eyewear). Predictors of routine use of all barrier techniques were specialties with high blood contact, considering saliva a possible route of transmission of HIV infection, average number of patients per week fewer than 40, and number of years of practice. CONCLUSIONS: Educational efforts for improving knowledge and finding and implementing ways to motivate dentists to the correct and routine use of infection control procedures are needed.

[Detection of Salmonella, Listeria spp., Vibrio spp., and Yersinia enterocolitica in frozen seafood and comparison with enumeration for faecal indicators: implication for public health]. / Ricerca di Salmonella, Listeria spp., Vibrio spp. e Yersinia enterocolitica in prodotti ittici congelati e surgelati del commercio e correlazione con gli indici di contaminazione fecale: implicazioni in Sanità Pubblica.

Infections transmitted through consumption of contaminated seafood is a significant source of human morbidity. The aim of this study was to compare the detection of Salmonella, Listeria, Vibrio, and Yersinia enterocolitica in frozen seafood with results from enumeration of conventional faecal indicators. A total of 213 crustaceans or molluscs were purchased from local vendors in Italy: 74% were harvested in Italy, 25% from other European countries and 1% from outside Europe. Listeria spp. was isolated from 20% of samples, Vibrio spp. from 11%, Salmonella from 3% and Y. enterocolitica from 1%. Listeria species isolated were L. monocytogenes, L. innocua, L. welshimeri, L. ivanovii and L. seeligeri. Vibrio species isolated were V. alginolyticus and V. fluvialis. The most contaminated shellfish for both faecal indicator microrganism and pathogens were hen clams (6% contained Salmonella, 27% Listeria spp. and 3% Y. enterocolitica), while from 27% of shrimps Vibrio spp. was recovered. Higher levels of faecal indicators were recovered from samples harvested outside Europe, and 66% of samples harvested in Thailand were contaminated from Salmonella. Significant differences were found in the levels of contamination of seafoods depending upon the freezing regime, but there was a limited association between presence of potential pathogens (particularly Vibrio spp.) and conventional faecal indicators. Hence, we suggest reconsideration of current legal parameters to evaluate microbiological quality of seafood.

[Levels of anti-Legionella antibodies in a healthy Neapolitan population]. / Livelli di anticorpi anti-legionella in popolazione sana Napoletana.

Legionella pneumophila was firstly recognized as the causative agent of a serious pneumonia about a decade ago. Since then a number of studies was carried out in order to define the epidemiological and clinical features of the microorganism. So far a high variability of the geographic distribution of the infection has been reported, with prevalences up to 50% of healthy populations. Therefore, a seroepidemiological survey was carried out on prevalence of Legionella pneumophila infections among healthy people in Naples. 562 serum specimens were randomly collected and examined by the indirect fluorescent antibody technique for antibodies against serogroups 1 to 6 of Legionella pneumophila, using 1:256 as the cut off titer. An overall prevalence of 23% was observed among the 562 persons tested. 24.5% among males and 21.9% among females had antibodies against Legionella pneumophila. There were no significant differences associated with sex and age, even though an apparent increase with age was observed. The highest prevalence was found for antibodies against serogroup 3 (9.4%). This finding was confirmed in all the age-sex groups, but among males aging between 21 and 40 years and among elderly people, 12.1% of persons tested had antibodies against only one serogroup of Legionella pneumophila, whilst 5.3%, 3.4% and 2.1% of subjects had antibodies against 2, 3 and 4 serogroups respectively. No serologic cross reactivity of Legionella pneumophila groups was found among seropositivities to different serogroups.

[Evaluation of the effectiveness of disinfectants used in several Neapolitan hospitals]. / Valutazione in campo dell'efficacia di disinfettanti utilizzati in alcuni ospedali Napoletani.

Among the various and complex aspects concerning the etiology of nosocomial infections, a relevant role is played by the indiscriminate and incorrect use of disinfectants: they may change paradoxically from devices used for nosocomial infection prevention into carriers of infections. Critical situations are: a) inadequate cleaning of the container; b) inaccurate concentrations set up; c) use of unsterilized water as a diluent; d) introduction of gauze or cotton flocks into the container; e) disinfectant contact with inactivating substances such as soaps, detergents, and organic substances; f) solution stability; etc.. Therefore, in the light of the experience we acquired in the field of nosocomial infections, we believed interesting to carry out a study in order to detect contamination of disinfectants used in various hospitals in Naples as well as their usage. In the period from January to May 1988, 243 disinfecting solutions routinely used in surgical departments of the 2nd School of Medicine and of three large neapolitan hospitals were tested for microbiological contamination. The analysis was carried out according to Kelsey and Maurer. 13 disinfectants were contaminated. The bacterial count exceeded the value of 5000 col/ml in 6 cases, while for he other disinfectants there were levels between 2500 and 4500 col/ml. 64% of the microorganisms isolated belonged to Pseudomonas spp. (cepacia, aeruginosa, fluorescens, maltophilia), while Acinetobacter calcoaceticus and Staphylococcus epidermidis were found in 21% and 15% of specimens respectively. A major consideration arising out from our study is the little attention played to health problems related to disinfection by hospital staff, together with a lack of knowledge about the proper way of using disinfectants.(ABSTRACT TRUNCATED AT 250 WORDS)

Detection of Cryptosporidium parvum oocysts in experimentally contaminated lettuce using filtration, immunomagnetic separation, light microscopy, and PCR.

Recovery of Cryptosporidium parvum oocysts in a fecal suspension that experimentally contaminated onto lettuce leaves was investigated. Material recovered from the lettuce samples by washing in detergent solutions were concentrated by filtration using the Envirochek Sampling Capsule. Oocysts were concentrated by immunomagnetic separation (IMS) and detected by microscopy following modified Ziehl-Neelsen (MZN) staining. Cryptosporidal DNA was detected using a nested-PCR assay for amplification of a fragment of the Cryptosporidium oocyst wall protein (COWP) gene, which was applied to DNA extracted from both filtrates, and material recovered from MZN stained smears on glass slides after microscopy. No Cryptosporidium were detected by microscopy or by PCR of un-inoculated lettuce leaves. After IMS, means of 0-6.5% of the total numbers of oocysts inoculated were recovered and detected by microscopy. Detection by PCR was less sensitive than microscopy. There was a strong association between successful PCR amplification, the numbers of oocysts detected by microscopy and the numbers of oocysts in the inoculum. This study confirms that C. parvum oocysts can be recovered from contaminated lettuce using filtration and IMS, and detected by microscopy and PCR. However, further developments are required to improve recovery of this parasite.

In situ mapping of urease-positive areas in porcine gastric mucosa.

Urease activity is a feature of gastric helicobacters, and its abundant production provides an indirect means of detecting their colonization. A method for mapping urease-positive areas directly on the gastric mucosa was developed, and 57.8% of pigs had evidence of Helicobacter colonization based on urease assay. Moreover, 89.2% of urease-positive pigs had gastritis, confirming that the known association found in man between Helicobacter and gastritis was found also in pigs. The proposed urease assay allowed detection of all urease-positive areas on the gastric mucosa, thus overcoming the biopsy sampling problems derived from the patchy distribution of helicobacters. In this way, gastric mucosa specimens from urease-positive areas could be usefully utilized for culture attempts and for microscopic examination.

Immunomagnetic separation and conventional culture procedure for detection of naturally occurring Salmonella in raw pork sausages and chicken meat.

The aim of the study was to compare immunomagnetic separation (IMS) and conventional selective enrichment procedures using selenite cystine broth (SC) and Rappaport-Vassiliadis broth (RV) in 137 naturally contaminated food samples (69 raw pork sausages and 68 chicken meat). The utilization of SC or IMS appeared to be the most appropriate enrichment procedure: 15 out of 18 Salmonella-positive samples (83.3%) were detected by SC and 12 (66.7%) by IMS; RV yielded only seven positive isolations (38.9%). However, RV yielded the highest count of Salmonella colonies per plate and the lowest interference by competing organisms. IMS could become a reliable alternative to standard enrichment procedures and a combined IMS and selective enrichment broth could increase the chance of Salmonella recovery.