Common Cutaneous Bacteria Isolated from Snakes Inhibit Growth of Ophidiomyces ophiodiicola.

There is increasing concern regarding potential impacts of snake fungal disease (SFD), caused by Ophidiomyces ophiodiicola (Oo), on free-ranging snake populations in the eastern USA. The snake cutaneous microbiome likely serves as the first line of defense against Oo and other pathogens; however, little is known about microbial associations in snakes. The objective of this study was to better define the composition and immune function of the snake cutaneous microbiome. Eight timber rattlesnakes (Crotalus horridus) and four black racers (Coluber constrictor) were captured in Arkansas and Tennessee, with some snakes exhibiting signs of SFD. Oo was detected through real-time qPCR in five snakes. Additional histopathological techniques confirmed a diagnosis of SFD in one racer, the species' first confirmed case of SFD in Tennessee. Fifty-eight bacterial and five fungal strains were isolated from skin swabs and identified with Sanger sequencing. Non-metric multidimensional scaling and PERMANOVA analyses indicated that the culturable microbiome does not differ between snake species. Fifteen bacterial strains isolated from rattlesnakes and a single strain isolated from a racer inhibited growth of Oo in vitro. Results shed light on the culturable cutaneous microbiome of snakes and probiotic members that may play a role in fighting an emergent disease.

Tolerance of high-dose (3,000 mg/day) coenzyme Q10 in ALS.

An open-label dose-escalation trial was performed to assess the safety and tolerability of high doses of coenzyme Q10 (CoQ10) in ALS. CoQ10, a cofactor in mitochondrial electron transfer, may improve the mitochondrial dysfunction in ALS. In this study, CoQ10 was safe and well tolerated in 31 subjects treated with doses as high as 3,000 mg/day for 8 months.

Results from the first computerised Italian surveillance of human Salmonella isolates. The Italian SALM-NET Working Group.

We report all the first computerised data collected in Italy for the surveillance of Salmonella isolates. Primarily, within the wide framework of the European Community Human Salmonella Surveillance Project (SALM-NET), we report data on the most commonly isolated serotypes in Italy from January 1994 to December 1996. In addition, we report all computerised data historically collected by some Italian regions regarding the period 1980-1993. Total data included in the Italian SALM-NET data base account for 59,336 Salmonella isolates. In the list of the most frequent isolates starting from 1989, S. Enteritidis always ranked first, followed by S. Typhimurium.

Results of the three-year surveillance by the Italian SALM-NET System: human isolates of Salmonella serotypes.

Within the wide framework of the European Community Human Salmonella Surveillance Project (SALM-NET), the data on the most commonly isolated serotypes in Italy from January 1994 to December 1996 are reported. The total data included in the Italian SALM-NET data base account for 34,412 Salmonella isolates. In the list of the most frequent isolates, S. enteritidis ranked always first in the years 1994, 1995 and 1996 with 5435 (43.4%), 4589 (37.1%) and 4044 (42.4%), respectively, over the total number of Salmonella isolates. This serotype is followed by S. typhimurium in the list of the top ten isolates, with 2236 (17.9%), 2831 (22.9%) and 2239 (23.5%). The other serotypes included in this list accounted for a much lower number and percentages ranging from 505 isolates of S. derby (4.0%) to 99 isolates of S. brandenburg (0.8%).

The antibiotic resistance patterns of Salmonella Typhi isolates in Italy, 1980-96. The Italian SALM-NET Working Group. Salmonella Network.

In this paper we report the distribution of Salmonella Typhi isolates in Italy and their resistance patterns to antibiotics. The data were collected by the Italian SALM-NET surveillance system in a pilot retrospective study of the period 1980-96. Data on drug-resistance were available for 82 isolates out of 176 S. Typhi isolated in Italy. Of these 82 isolates, 32 (39%) were resistant or intermediate to 1 or more antibiotics. Eight isolates were resistant and 7 intermediate to streptomycin; 4 isolates were resistant to ampicillin alone or in association with other antibiotics; only 2 strains (1 isolated in Lombardia in 1993 and the other 1 in Lazio in 1994) were resistant to chloramphenicol, and 2 (isolated in Sardegna and Piemonte in 1995 and 1996, respectively) showed intermediate resistance to chloramphenicol. The strains showing resistance to 3 or more antibiotics were very scarce: 1 (with 5 complete resistances) was isolated in Lazio in 1994, and another 1 (with complete resistance to 10 antibiotics and intermediate resistance to 2 antibiotics) was isolated in Molise in 1988. In conclusion, besides the routine activities to control typhoid fever, an accurate and continuous surveillance is necessary in order to quickly identify multidrug-resistant (MDR) S. Typhi strains and prevent their spread, even though their level, in our country, is still quite low.

Serotype and phage type distribution of salmonellas from human and non-human sources in Italy in the period 1973-1995.

According to the data collected at the Rome-based National Reference Centre for Enterobacteria, 266,547 Salmonella strains isolated from human sources (226,513) and from non human sources (40,034) were characterised in Italy during the period 1973-1995. The serotype of all isolates, and the antibiotype and phage type of selected isolates were determined. Human-source isolations grew from 4372 in 1973 to 12,310 in 1995: non-human source isolations, from 339 in 1973 to 3459 in 1995. Salm. Typhimurium ranked first in the list of the most common serotypes isolated from both sources in the period 1973-1988 except in the years 1975 and 1976 when it was overtaken by Salm. Wien. Since 1983 Salm. Enteritidis has been among the top ten isolates from animals, and ranked first in the list of isolates from humans in 1988 and from non human sources in 1991. During the last years the number of multidrug-resistant strains, mostly belonging to phage types 104 and 193 of Salm. Typhimurium has been rising. Salmonella strains have also been isolated from numerous extraintestinal infections, almost exclusively caused by Salm. Enteritidis and Typhimurium.

An outbreak of Salmonella hadar associated with food consumption at a building site canteen.

A biphasic outbreak of gastroenteritis caused by Salmonella hadar affected canteen employees and workers at a construction site in central Italy in September 1994. There were 448 symptomatic cases, from 61 of whom group C Salmonella was isolated. Six cases were canteen employees. Twenty-two other individuals were asymptomatic excreters. There were 10 secondary cases. Working as a food handler at the canteen constituted an increased risk of infection, independently of ingestion of the food (odds ratio: 62.1; 95% confidence interval (CI): 9.5-406.6). Having eaten at the canteen on the 19th and 20th September was identified as risk factor for subjects symptomatic within 72 hours (relative risk (RR): 17.0, 95% CI: 2.3-124.3), and cooled meat salad was identified as the vehicle of infection (RR: 36.6, 95% CI: 14.3-93.8). The use of portable toilets was another possible route of transmission of infection for all cases (RR: 1.3, 95% CI: 1.0-1.6). The index case was a cook who had symptoms five days before the peak of the outbreak. From 27 individuals both symptomatic and asymptomatic excreters group B, group D and not-typed Salmonellas were isolated. This study underlines the problem of improper food handling in salmonellosis outbreaks and emphasizes the role of several vehicles in the transmission of salmonellosis in a community.

Phenotypic and genotypic biotyping of environmental strains of Vibrio cholerae non-O1 isolated in Italy.

The purpose of this study was to characterize strains of Vibrio cholerae non-O1 isolated in Italy from different sources by biochemical and serological assays, antibiotic susceptibility testing, and molecular biotyping. Serotyping and genomic analysis by pulsed-field gel electrophoresis proved to be useful in discriminating the isolates. The data obtained show a wide heterogeneity at the genomic level, and in keeping with this, the serogrouping classification provided evidence of a high variability of the investigated strains. In addition, none of the strains tested produced cholera-like toxins.

Epidemiological analysis of strains of Salmonella enterica serotype Enteritidis from foodborne outbreaks occurring in Italy, 1980-1994.

Polymerase chain reaction (PCR-) ribotyping was performed on 243 strains of Salmonella enterica serotype Enteritidis isolated during the years 1980-1994 from 58 foodborne outbreaks occurring in different regions of Italy. The majority (37) of the outbreaks were attributed to phage type (PT) 4, followed by PT1 (seven outbreaks); the latter was identified in 1993 in Italy in epidemic strains of Enteritidis. In eight cases more than one phage type was recognised from a single event. Nine PCR-ribotypes (PCR-RTs) were detected, with a strong prevalence of PCR-RTs f7 and e5 (23 and 21 outbreaks, respectively). In two instances two distinct PCR-RTs were identified within strains from a single outbreak. All but one of the PT1 outbreaks were caused by PCR-RT f7, whereas PT4 outbreaks could be subdivided into six subsets. Clustering of isolates was consistent with data obtained from epidemiological investigations. PCR-ribotyping proved to be an effective and reliable tool for subtyping isolates of Enteritidis belonging to the most frequent phage types. Nevertheless, in terms of laboratory expertise and lack of inter-laboratory standardisation, this typing technique is best suited for reference laboratories.

Conventional and molecular approaches to isolates of Salmonella hadar from sporadic and epidemic cases.

In September 1994 an outbreak of gastroenteritis occurred in 437 people who had consumed lunch in the canteen of a factory in Central Italy. Salmonella sp. was isolated from stools of 99 patients and in 73 of them Salmonella hadar was identified. This is the first outbreak caused by this serotype described in Italy. In order to examine the genotypic basis of the epidemic strains, molecular typing was applied to sporadic strains isolated before and after the outbreak episode. For this purpose phage type, resistance to antibiotics, DNA plasmid profile and sites of insertion of the mobile element of IS200 were determined. The epidemic strains were genetically distinct from the non-epidemic isolates; they were shown to be phage type 26, harbouring four small plasmids, were resistant to nalidixic acid and showed a unique characteristic IS200 fingerprint. The typing methods used in this study allowed the identification and discrimination of the outbreak strains from related isolates. They can thus be considered as a tool for epidemiological purposes. In addition we should point out the emerging resistance to nalidixic acid, largely used in veterinary medicine, in Salm. hadar.

Foodborne outbreaks caused by Salmonella in Italy, 1991-4.

This report summarizes studies on 1699 foodborne outbreaks, in Italy, reported to the Istituto Superior di Sanità (ISS) (the National Institute of Health of Italy, Rome) during the period 1991-4. The most frequently reported foodborne outbreaks were caused by salmonellae (81%), in particular by Salmonella enteritidis and non-serotyped group D salmonella (34% and 33% of the total salmonella outbreaks, respectively). A vehicle was implicated in 69% of the salmonella outbreaks; eggs were implicated in 77% of the outbreaks for which a vehicle was identified or suspected. Salmonella strains isolated in 54 outbreaks were studied for phenotypic and genotypic characteristics. The isolates belonged to S. enteritidis (50 outbreaks), S. typhimurium (three outbreaks) and S. hadar (one outbreak). In the S. enteritidis outbreaks, phage type 4 was most frequently isolated (64.8%), followed by phage type 1 (14.8%). The virulence plasmid of 38 megadaltons was found in many different phage types of S. enteritidis.

Phage types and ribotypes of Salmonella enteritidis in southern Italy.

Differently from other European countries, Southern Italy was affected by a considerable increase in human infections due to Salmonella enterica subsp. enterica serovar Enteritidis (S. Enteritidis) only after 1990. On the present investigation, two groups of S. Enteritidis strains isolated during the low-incidence period 1980-1984 and the epidemic period 1990-1993, respectively, have been submitted to phage-typing and ribotyping in order to ascertain whether the epidemic increase was determined by the spread of a foreign bacterial clone or not. Among the 150 isolates relative to the aforesaid two periods, 12 different phage types (PTs) were observed. PT4 was the most common phage type among the strains isolated in 1980-1984 (61%) as well as in those of the epidemic period 1990-1993 (72%). PT8 was the second most frequent (33%) phage type in 1980-1984. It was substituted by PT1 (19%) in the 1990-1993 period. Analysis of rDNA patterns obtained after Hinc II digestions and Escherichia coli rRNA hybridizations showed 8 different patterns, A to H. The great majority of the strains studied (140 isolates, 93%) belonged to the ribotype A, showing a similar frequency both in 1980-1984 (36 of 39, 92%) and in 19901993 (104 of 111, 94%). The predominance of PT4 and ribotype A among both preepidemic and epidemic strains is in agreement with the hypothesis that host genetic diversity decline and modern farming practices in the poultry industry have facilitated a widespread dissemination of preexisting endemic strains. This hypothesis urges to plan new strategies in preventing S. Enteritidis infections.

Salmonella enteritidis in Italy.

During the period 1982-1992 the percentages of Salmonella enteritidis isolations in Italy have increased from 2.4 to 57.1% from human beings and from 0.5 to 22.8% from food. Seven hundred and fifty-seven isolates, 702 from man and 55 from food, were characterized. Phage type 4 accounted for the 76.8% of these isolates. The majority of strains were sensitive to the antibiotics tested; only 18 (2.4%) showed resistance to three or more antibiotics by nine different patterns.

Simultaneous infection with three different S. enteritidis strains in a nursing home resident.

A culture taken from a nursing home resident as part of a S. enteritidis outbreak was found to have a mixed infection due to three different strains of S. enteritidis. One of the three strains belonged to phage type (PT) 4, one to PT6 and one reacted with phages but did not conform to any typing scheme (RDNC). All three strains had the 38.9 megadaltons (MDa) plasmid found in the isolates from the outbreak-related cases, in addition the PT6 and RDNC strains harboured a 69.9 MDa plasmid. The importance of phage typing and plasmid analysis for S. enteritidis strain characterization and their epidemiologic and bacterial significance are discussed.

Characterisation of Yersinia species isolated from a kennel and from cattle and pig farms.

Between April 1984 and November 1986, 126 faeces samples were collected from puppies and bitches in a kennel and examined for Yersinia species; 45 (35.7 per cent) of them were positive. Thirty-eight isolates were Y enterocolitica, five were Y frederiksenii and two were Y intermedia. Twenty-one of the Y enterocolitica isolates belonged to serogroup 0:3 and 17 of these were L-sorbose negative; all these isolates were from puppies. One strain of Y frederiksenii and all the L-sorbose-negative Y enterocolitica serogroup 0:3 isolates were resistant to antimicrobial agents and showed four different patterns of resistance (ampicillin, cephalothin and tetracycline; ampicillin, cephalothin, streptomycin and tetracycline; ampicillin, chloramphenicol, cephalothin, streptomycin and sulfathiazole; and ampicillin, chloramphenicol, cephalothin, streptomycin, sulfathiazole and tetracycline. In January 1986, investigations were conducted on a cattle farm and a pig farm close to the kennel. Of 19 bovine faeces samples 11 (57.8 per cent) were positive for Yersinia species; eight yielded Y enterocolitica and four yielded Y frederiksenii. None of the Y enterocolitica isolates belonged to serogroup 0:3. Of 20 porcine faeces samples eight (40 per cent) were positive for Yersinia species; all eight yielded Y enterocolitica and four also yielded Y pseudotuberculosis. Two of the isolates of Y frederiksenii and two of the isolates of Y enterocolitica from the farms had the following resistance patterns: ampicillin, cephalothin and streptomycin; ampicillin, streptomycin and tetracycline; ampicillin, chloramphenicol, cephalothin, streptomycin, trimethoprim-sulphamethoxazole and tetracycline; and ampicillin, chloramphenicol, cephalothin, streptomycin, sulphatiazole, trimethoprim-sulphamethoxazole and tetracycline.(ABSTRACT TRUNCATED AT 250 WORDS)

Epidemiological markers of Salmonella typhimurium isolates in Rome.

The phage type, antimicrobial resistance pattern, colicinogenic activity and DNA plasmid content of 172 strains of Salmonella typhimurium isolated in Rome from 1984 to 1986 were determined; 142 isolates were from patients with enteritis, 30 were from asymptomatic subjects. Most of the phage types identified were isolated during 2 or 3 of the study years; others, e.g., PT141, PT 204c and PT 194 were isolated during 1 year only, and only the last of these was isolated in large numbers. Phage typing proved most valuable in identifying epidemiologically related strains; DNA plasmid analysis was most useful in characterising further cultures of the same phage type, especially those isolated during suspected epidemics.

Italian experience in Salmonella enteritidis 1978-1988: characterization of isolates from food and man.

Salmonella enteritidis accounted for 5.45% of the 118.685 Salmonella isolates from man and for 2.65% of the 3.315 Salmonella isolates from food in Italy in the eleven year period 1978 to 1988. In the years 1978-1982 no S. enteritidis strain was isolated from eggs and poultry; in the years 1983-1988 the 53% of S. enteritidis isolates from food were from eggs and poultry. In 1989 S. enteritidis accounted for 744 isolates from man and 22 from food of which 80% were from eggs and poultry (partial data). In that year 18 outbreaks caused by S. enteritidis were reported to the National Centre of Enteric Pathogens in Rome. Characteristics of 81 S. enteritidis isolates were examined of which 27 were from sporadic cases involving humans and 40 from outbreaks in humans; 14 isolates were from food, all but one connected with the outbreaks. All the isolates studied were sensitive to the antibiotics tested; plasmid profile analysis showed a predominant profile pattern in both epidemic and non-epidemic strains; lysine decarboxylase was present in all the strains tested. Although in at least three epidemics a common supplier of eggs was proved, the source was not identified. Unfortunately it was not possible to determine the phage type of isolates because of the unavailability of specific phages.

Siderophore production by Salmonella species isolated from different sources.

A total of 230 Salmonella strains were screened for enterobactin and aerobactin production, sensitivity to bacteriocins and resistance to antibiotics. All the isolates produced the phenolate siderophore enterobactin. Amongst these, 74 strains, most belonging to S. enteritidis, were sensitive to colicin B. Only 26 isolates, all belonging to S. wien, produced an additional iron chelator, i.e. the siderophore aerobactin, and 22 out of these were sensitive to cloacin DF13. Analysis of iron repressible outer membrane proteins and plasmid profiles in S. wien strains showed that the expression of a 74-kDa iron-repressible outer membrane protein and the presence of large plasmids were associated with multiple antibiotic resistance, aerobactin production and sensitivity to cloacin DF13. The incidence of aerobactin-producing strains among S. wien isolates was higher during years 1974-1985; the epidemiological implications of these results are discussed.

Phage type and DNA plasmid profile of Salmonella typhimurium isolates in the area of Isernia, Italy.

Thirty-eight Salmonella typhimurium strains isolated from December 1987 to March 1988 in Isernia, Central Italy, were characterized on the basis of their phage type, resistance to antimicrobials and plasmid profiles. According to their phage types, the isolates could be assigned to one of six groups, the prevalent one being PT 195 which accounted for 73.6% of isolates. On the basis of their plasmid content, the isolates could be assigned to one of ten groups. The prevalent plasmid profile (60.0; 6.0; 4.3; 4.0; 3.2 megadaltons) was found in 60.4% of isolates. All the isolates from a particular food (salsicce), and as most of isolates from humans who had consumed this food belonged to phage type 195 and were of the same plasmid profile. The combined use of phage typing and DNA plasmid analysis proved to be a useful tool in identifying epidemiologically related isolates in this investigation.