[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.

[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.

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.

Enumeration of Escherichia coli and coliforms in surface water by multiple tube fermentation and membrane filter methods.

The current investigation was carried out in order to compare directly the multiple tube fermentation method (MTF), using standard procedures (lactose broth, LB) and the Colilert reagent, with the membrane filter method (MF) using Les Endo agar (LEA), m-faecal coliform agar (mFCA) and chromogenic coliform agar (CCA), for recovery of coliforms and Escherichia coli in 80 surface water samples. Total coliforms were isolated from 100% of samples by all methodologies. Faecal coliforms/E. coli were detected in 100% of samples by MTF methods, but only in 75.5% by MF-mFCA and in 86.2% by MF-CCA. Even if MTF-LB counts were consistently higher, the Colilert reagent accurately determined total coliforms and E. coli levels within 24 h with no additional confirmatory tests. Therefore, it could be a powerful tool for rapidly assessing possible faecal contamination and a suitable alternative to the traditional MTF and MF techniques utilized for coliform detection.

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.

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.

Routine preoperative investigation. Results of a multicenter survey in Italy. Collaborator Group.

We conducted a study to acquire information on the current behavior of a sample of Italian surgeons and anesthesiologists about prescribing, interpreting, and using routine preoperative investigations. Consultants in surgery and anesthesiology in 60 hospitals in northern, central, and southern Italy were interviewed. Prescription of these procedures by doctors were driven more by personal experience than by updated scientific knowledge. This practice often led to ineffective and inefficient clinical practice, with healthy patients undergoing useless, time-consuming, costly, and sometimes harmful procedures.

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.

Prevalence of antibodies against enterically transmitted viral hepatitis (HAV and HEV) among adolescents in an inland territory of central Italy.

A study on the seroprevalence of HAV and HEV infections among adolescents in an inland territory of central Italy (Molise region) was carried out. The prevalence of antibodies was respectively 3.3% and 0.4%. The results indicated that (1) anti-HAV prevalence in children is low but HAV infection is always present, and (2) HEV infection could be endemic and not necessarily imported.

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.

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.

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.

Study of an endemic goiter area in northern Campania.

The purpose of this paper was the determination, in the endemic goiter area of Teano (Caserta, Italy), of: i) The goiter prevalence in a group of 920 patients who attended the Outpatient Endocrinology Department; ii) The urinary iodine excretion in 150 adults (20-73-year-old) and 502 children (10-16-year-old; iii) The thyroid size in the 502 children; iv) The environmental iodine levels. Out of 920 patients a total of 750 (81.5%) goiters were detected. Out of these 750 cases, 415 (55.3%) were of grades 1b and 2, 335 (44.7%) of grades 3 and 4. A statistically significant association between goiter size and age was found (p less than 0.05). Laboratory data were entirely available for 506 goitrous patients. Serum TG levels was increased with goiter size and age, whereas there was a progressive decrease in mean serum TSH levels with increasing goiter size and age. The screening program performed on 502 schoolchildren aged 10 to 16 yr found a 68.3% prevalence of grade 1 goiter and a mean urinary ratio iodine/creatinine of 52 +/- 32 (SD) micrograms/g. A mean urinary ratio iodine/creatinine of 60 +/- 27 (SD) micrograms/g was reported in a sample of 150 adult inhabitants. Iodine measurements in water supplies showed levels equal to or less than 1 microgram/l. The area investigated can be identified as a moderate iodine deficient area and classified as grade 1-2 according to the Pan American Health Organization criteria.