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Acarological risk was calculated as the probability of encountering at least one host-seeking Ixodes ricinus tick infected by the pathogen Borrelia burgdorferi sensu lato, in 100 m transects in the province of Genoa, Italy. The seasonal pattern of I. ricinus was studied using generalized estimating equations (GEE) with negative binomial error, to consider overdispersion of tick counts and repeated sampling of the same dragging sites from April 1998 to March 1999. Prevalence of infection by B. burgdorferi s.l. was evaluated by PCR and hybridization with genospecies-specific probes. Acarological risk (R) peaked in April (R=0·2, 95% CI 0·13–0·26) and November (R=0·29, 95% CI 0·10–0·46). Borrelia garinii and B. valaisiana were the most common genospecies at our study site suggesting a major role of birds as reservoirs. DNA from Anaplasma phagocytophilum, the agent of granulocytic ehrlichiosis in humans and animals, was amplified from an adult I. ricinus.

The sulfur K-edge x-ray absorption spectra for the amino acids cysteine and methionine and their corresponding oxidized forms cystine and methionine sulfoxide are presented. Distinct differences in the shape of the edge and the inflection point energy for cysteine and cystine are observed. For methionine sulfoxide the inflection point energy is 2.8 eV higher compared with methionine. Glutathione, the most abundant thiol in animal cells, also has been investigated. The x-ray absorption near-edge structure spectrum of reduced glutathione resembles that of cysteine, whereas the spectrum of oxidized glutathione resembles that of cystine. The characteristic differences between the thiol and disulfide spectra enable one to determine the redox status (thiol to disulfide ratio) in intact biological systems, such as unbroken cells, where glutathione and cyst(e)ine are the two major sulfur-containing components. The sulfur K-edge spectra for whole human blood, plasma, and erythrocytes are shown. The erythrocyte sulfur K-edge spectrum is similar to that of fully reduced glutathione. Simulation of the plasma spectrum indicated 32% thiol and 68% disulfide sulfur. The whole blood spectrum can be simulated by a combination of 46% disulfide and 54% thiol sulfur.

In August 2002, an exceptional flood devastated a suburban area in the surroundings of Vicenza, North-east Italy. A fatal case of haemorrhagic pneumonia, which was presumptively diagnosed as leptospirosis, was observed as a consequence of the inundation. A local seroepidemiological survey was activated thereafter, with the principal aim of evaluating the risk of infection with Leptospirae in the population exposed to the flood. A 6.8% seroconversion rate was found in the population studied; however, the case previously observed remained unique, since an overt outbreak of leptospirosis did not occur.

We have previously demonstrated that the alphaMbeta2 integrin (known as CR3 or Mac-1) expressed on neutrophils (PMNs) and/or on CHO Mac-1 transfected cells,in the presence of serum complement binds B. burgdorferi and promotes an increased non -opsonic adhesion, in the presence of serum complement. In this study we demonstrate that: 1) living motile B. burgdorferiand recombinant lipidated OspA and OspC, up-regulate CR3 expression on PMNs; 2) in the absence of serum, B. burgdorferi induces increased adhesion of CHO cells expressing CR3 to fibronectin, an extracellular matrix protein. Both the I-domain and the lectin-like domain of CR3 are involved in the binding recognition and activation because mAb anti I-domain and N-acetyl-glucosamine inhibit cell adhesion to fibronectin. These data indicate that B. burgdorferi whole cells, but not Osps, activate CR3 integrin; since this receptor plays a key role in priming neutrophils to important inflammatory events, the interaction of B. burgdorferi with neutrophils via the CR3 may enhance their role both in defence and in disease.

In the three-year period 1994 1996, 222 reports on human cases of leptospirosis were received by the Italian Ministry of Health. The average annual number of reports was 29.2% lower than in the preceding eight years. In all cases but two the infections were thought to have been acquired in Italy. As in previous years, the majority of cases was observed in the northern regions of the country (83.8%), mostly in males (88.9%). Cases occurred in all age groups, but were more common in the working-age population (15-64 years). There was no common-source outbreaks. The typical leptospiral seasonal course, with a peak in August, was observed. During 1994, leptospirosis was the reported cause of death in 19 patients. Mortality was higher among males than females. The overall fatality rate was 22.6%. During the study period, a total of 126 cases of leptospirosis were confirmed by the National Centre for Leptospirosis or one of the 12 Regional Leptospira Laboratories. Of the 103 patients for whom information on place of residence, contact with animals, occupational and recreational activities was available, 98 (95.1%) were people who live in rural areas or devote themselves to occupational or recreational activities at risk. The likely source of infection and the mode of exposure were known for 55 patients. Forty-five patients (81.8%) were likely infected by contaminating water (43 cases) or soil (2 cases), ten (18.2%) by direct contact with animals or animal urine. Both running (51.2%) and stagnant water (27.9%) have been reported as a source of infection. Rodents were implicated in 50.0% of the 10 cases involving animals. In comparison with the preceding eight-year period, the risk of contracting leptospirosis was found to have increased for recreational activities (from 34.7 to 38.2%) and decreased for occupational activities (from 45.8 to 32.7%). A large number of infections, however, was ascribed to accidental events (25.5%). As in the previous period, besides fever, the involvement of the liver was the most frequent clinical manifestation (70.8%). Influenza-like symptoms were the only signs of illness in 15.1% of cases. Infections by 9 different serogroups were detected. The most frequent antibodies were those against serovars icterohaemorrhagiae, poi, copenhageni and brattislava. The presence of co-agglutinins against serovars belonging to different serogroups prevented the identification of the presumptive infecting serogroup in 19.8% of subjects.

Little is known about the distribution of Human Granulocytic Ehrlichiosis (HGE) in Europe and even less is known in Italy, where no case of clinically documented HGE has been reported. In a previous study we reported the presence of Ehrlichia DNA in Ixodes ricinus ticks from Central Italy. By the use of an Ehrlichia-specific PCR we found that 24% of the ticks were positive. Furthermore, we demonstrated a simultaneous coinfection of the same tick by both, Borrelia burgdorferi and Ehrlichia phagocytophila. Since the Friuli-Venezia Giulia region (North-east of Italy) is endemic for Lyme borreliosis (LB) and the geographic distribution of HGE usually overlaps that of LB, we decided to carry out a survey concerning the presence of Ehrlichia spp. in a recreational area near Trieste where the presence of Lyme borreliosis and Borrelia burgdorferi sensu lato in ticks is well demonstrated. Ticks were analyzed in pools (because a low infection rate was expected): eleven samples out of 93 were found positive by Ehrlichia-PCR. Subsequent sequence analysis of some of the positive PCR products revealed a high homology with the HGE agent Ehrlichia (only one base substitution in almost 450 bp sequenced). These findings add new and interesting data on the Ehrlichia epidemiology in Italy. By now we have demonstrated the presence of two distinct granulocytic ehrlichiae in Italiyn ticks by the aid of a PCR-based analysis: Ehrlichia phagocytophila in Central Italy and an HGE-like Ehrlichia in the north-eastern Italy, in a region close to Slovenia where the first reported case of HGE in Europe occurred.

This paper describes the antigenic characteristics of a strain of Borrelia burgdorferi isolated from a patient seronegative for Lyme borreliosis, in the early stage of the illness. The strain was not recognized by a late serum sample from the patient; the isolate reacted in immunoblotting with some of the monoclonal antibodies directed against the immunodominant antigens of Borrelia burgdorferi. In addition to the OspA antigen this strain carries also the epitopes of OspB proteins, unlike the majority of European Borrelia burgdorferi strains.

After preliminary in vitro screening of 17 newly synthesized compounds belonging to the chemical class of N1-(aryliden)-2-pyridinecarboxyamidrazones, active against Mycobacterium tuberculosis H37Rv, the minimum inhibitory concentrations (MICs) of the ten most promising agents against three clinical isolates were determined by agar dilution. Compounds 12 and 14 were the most active, each inhibiting strain H37Rv at concentrations of 8 microg/ml and having a MIC of 16 microg/ml against the human isolates. The results obtained in this preliminary study confirmed the interesting antitubercular properties of these newly synthesized compounds and allowed us to carry out our investigations over a large number of isolated clinical strains.

The effects of sub-inhibitory concentrations (MIC 50 and MIC 25) of clindamycin, erythromycin and chloramphenicol on the adherence of Campylobacter jejuni to INT 407 cells were studied. The results indicated that the adhesion was inhibited at various extents, mostly by MIC 50, and that the inhibition increased with time. The interpretation of our data is that the antibiotics can interfere with the adhesin(s) synthesis.

Thirteen R plasmids derived from strains of E Enterobacteriaceae isolated from clinical material have been characterized. They belonged to many incompatibility groups and differ widely respecting other phenotypic characteristics, even if they come from bacteria isolated from a small geographic area.

The macroagglutination test, according Mailloux, was investigated for its feasibility in the rapid diagnosis of human and animal leptospirosis. Suspected sera examinated by Mailloux test, were also examinated by Complement Fixation and Microagglutination; the results suggest that: Mailloux macroagglutination is the serological test of choice, for screening of animal and human sera, mostly if it is not needed to know the infecting serovar.

The activity of Leptospira Laboratory in our Institute is briefly explained. The laboratory acts as International Reference for Leptospira typing, for microbiological diagnosis of leptospirosis and for basic research on pathogenesis and new trends in the serodiagnosis.