Relationship between virulence gene profiles of atypical enteropathogenic Escherichia coli and Shiga toxin-producing E. coli isolates from cattle and sheep in New Zealand.

Virulence gene profiles of atypical enteropathogenic Escherichia coli (aEPEC) and Shiga toxin-producing E. coli (STEC) from cattle, sheep, and humans were examined to determine the relationship between pathotypes. Shared virulence factors (intimin, EHEC hemolysin, serine protease, and a type II secretion system) were identified, suggesting a dynamic evolutionary relationship between aEPEC and STEC.

Comparison of PCR binary typing (P-BIT), a new approach to epidemiological subtyping of Campylobacter jejuni, with serotyping, pulsed-field gel electrophoresis, and multilocus sequence typing methods.

To overcome some of the deficiencies with current molecular typing schema for Campylobacter spp., we developed a prototype PCR binary typing (P-BIT) approach. We investigated the distribution of 68 gene targets in 58 Campylobacter jejuni strains, one Campylobacter lari strain, and two Campylobacter coli strains for this purpose. Gene targets were selected on the basis of distribution in multiple genomes or plasmids, and known or putative status as an epidemicity factor. Strains were examined with Penner serotyping, pulsed-field gel electrophoresis (PFGE; using SmaI and KpnI enzymes), and multilocus sequence typing (MLST) approaches for comparison. P-BIT provided 100% typeability for strains and gave a diversity index of 98.5%, compared with 97.0% for SmaI PFGE, 99.4% for KpnI PFGE, 96.1% for MLST, and 92.8% for serotyping. Numerical analysis of the P-BIT data clearly distinguished strains of the three Campylobacter species examined and correlated somewhat with MLST clonal complex assignations and with previous classifications of "high" and "low" risk. We identified 18 gene targets that conferred the same level of discrimination as the 68 initially examined. We conclude that P-BIT is a useful approach for subtyping, offering advantages of speed, cost, and potential for strain risk ranking unavailable from current molecular typing schema for Campylobacter spp.

Multilocus sequence typing of Campylobacter jejuni, and the correlation between clonal complex and pulsed-field gel electrophoresis macrorestriction profile.

The characterization of Campylobacter jejuni has been significantly improved by the use of multilocus sequence typing (MLST), which allows the relationship between isolates to be determined. The sequence types (STs) of 261 isolates of C. jejuni from New Zealand were determined. Isolates were obtained from a range of sources including chicken meat, cattle, pigs, duck, sheep, water and human infections. Thirty-two new alleles and 44 new STs were identified. Comparison of the MLST data and pulsed-field gel electrophoresis macrorestriction profiles showed that the macrorestriction profiles were good predictors of the clonal complex (CC) but not ST. All the major CCs identified elsewhere in the world were found in New Zealand as well as the association of certain CCs with particular animal niches. The majority of new STs identified were from river water isolates.

Molecular subtyping and distribution of the serine protease from shiga toxin-producing Escherichia coli among atypical enteropathogenic E. coli strains.

Atypical enteropathogenic Escherichia coli (aEPEC) and Shiga toxin-producing E. coli (STEC) were examined to determine the prevalence and sequence of espP, which encodes a serine protease. These analyses indicated shared espP sequence types between the two E. coli pathotypes and thus provide further insights into the evolution of aEPEC and STEC.

Salmonella in uncooked retail meats in New Zealand.

A national quantitative survey of Salmonella in five types of uncooked retail meats in New Zealand was undertaken from August 2003 to May 2005 to establish baseline proportionality data. The overall prevalence of Salmonella in 1,108 meat samples was 1.1% (95% confidence interval, 0.6 to 1.9). Low prevalences of Salmonella in each meat type were observed, with 3% (1.2 to 6.1) in chicken, 1.3% (0.3 to 3.8) in lamb and mutton, 0.5% (0 to 3.0) in unweaned veal, 0.4% (0 to 2.4) in beef, and 0% (0 to 1.6) in pork. The Salmonella serotypes isolated were Salmonella Infantis from beef; Salmonella Typhimurium PT1 from unweaned veal and chicken; Salmonella sp. 6,7:k:-, Salmonella Enteritidis PT9a, Salmonella sp. 4,5,12:-:-, Salmonella sp. 4,12:-:-, and Salmonella Typhimurium PT160 from chicken; and Salmonella sp. 4:-:2 and Salmonella Brandenburg from lamb. Four of the isolates from chicken, Salmonella sp. 4,5,12:-:- (two isolates), Salmonella sp. 4,12:-:-, and Salmonella Typhimurium PT1, were very similar phenotypically and serologically to the attenuated Salmonella vaccine strain used in MeganVacl for poultry. One lamb sample yielded a count of Salmonella Brandenburg of 4.24 most probable number (MPN)/g, while all other positive samples were <1.0 MPN/g. The results provide baseline proportionality data for Salmonella in retail uncooked meats that will contribute invaluably toward future risk assessment in light of other information, such as consumption data that can be used for risk characterization.

Prevalence, numbers, and subtypes of Campylobacter jejuni and Campylobacter coli in uncooked retail meat samples.

A national quantitative survey of Campylobacter jejuni and Campylobacter coli in 1,011 uncooked retail meat samples (beef, unweaned veal, chicken, lamb and mutton, and pork) was undertaken from August 2003 to June 2004 to establish baseline proportionality data. The presence, number, and type of Campylobacter present in each sample was assessed. Prevalences of C. jejuni and C. coli were 89.1% in chicken, 9.1% in pork, 6.9% in lamb and mutton, 3.5% in beef, and 10% in unweaned veal. C. jejuni was identified in the majority of positive samples (246 of 259). In chicken samples positive for C. jejuni, 40.2% had counts of <0.3 most probable number (MPN)/g, 50.5% had 0.3 to 10.0 MPN/g, 8.8% had 10.1 to 50.0 MPN/g, and 0.5% had 110 MPN/g. In other meats (49 samples), Campylobacter counts were < or = 0.3 MPN/g, except for one unweaned veal sample at > 10.9 MPN/g. Penner serotyping and SmaI macrorestriction genotyping using pulsed-field gel electrophoresis with 247 isolates revealed 17 Penner serotypes and 56 electrophoresis profiles. Seven Penner serotypes (HS1 complex, 2, 4 complex, 6, 11, 27, and 42) were represented by 10 or more isolates from chicken. When data from both typing methods were combined, 62 sero-genotypes were generated. In a comparison of these sero-genotypes with historical data for isolates from human cases, 71% of the beef isolates, 50% of the lamb and mutton isolates, 50% of the pork isolates, 41% of the chicken isolates, and 25% of the unweaned veal isolates were common to both sources. These results provide baseline proportionality profiles of Campylobacter in these five meats and will facilitate exposure assessment in combination with other information such as consumption data and subsequent quantitative risk assessment.

Isolation of Vibrio cholerae O1 strains similar to pre-seventh pandemic El Tor strains during an outbreak of gastrointestinal disease in an island resort in Fiji.

Five strains of Vibrio cholerae O1, one each from an Australian and a New Zealand tourist with gastrointestinal illness returning from an island resort in Fiji and the remaining three from water sources located in the same resort, were extensively characterized. Conventional phenotypic traits that are used for biotyping of O1 V. cholerae categorized all five strains as belonging to the El Tor biotype. Genetic screening of 11 regions that are associated with virulence in V. cholerae showed variable results. The absence of genes comprising Vibrio seventh pandemic island-I (VSP-I) and VSP-II in all the strains indicated that these strains were very similar to the pre-seventh pandemic V. cholerae O1 El Tor strains. The ctxAB genes were absent in all strains whereas orfU and zot were present in four strains, indicating that the strains were non-toxigenic. Four strains carried a truncated CTX prophage. Although epidemiological and molecular studies suggested that these strains did not cause cholera amongst tourists at the resort, their similarity to pre-seventh pandemic strains, their prior association with gastrointestinal illness and their presence in the island resort setting warrant more attention.

Application of pulsed-field gel electrophoresis to identify potential outbreaks of campylobacteriosis in New Zealand.

Since 2002, New Zealand's incidence of campylobacteriosis has exceeded 300 cases per 100,000 people per annum. To evaluate genetic variation in human isolates, 183 Campylobacter isolates were collected from a single clinical laboratory in Christchurch: 77 during an 8-week period in spring, and the rest 3 months later over a second 8-week period in autumn. Isolates were identified to the species level and subtyped using Penner serotyping (Campylobacter jejuni only) and pulsed-field gel electrophoresis (PFGE) using both SmaI and KpnI. Approximately two-thirds of the isolates could be grouped into clusters of between 2 and 26 isolates with indistinguishable SmaI and KpnI patterns. Less than 10% of the isolates were of the same type between the two sampling periods. The epidemiological relevance of the PFGE clusters was supported by temporal clustering, some spatial clustering, and some statistically significant demographic similarities among cases in a cluster. Conversely, patient cases yielding isolates which did not cluster with isolates from other cases were more likely to report recent overseas travel and less likely to live within larger urban centers. To identify whether these clusters actually represent common-source outbreaks, however, would require the detailed, rapid, and reiterative epidemiological investigation of cases within a PFGE cluster. The combined and timely application of subtyping and epidemiological investigation would appear to be a promising strategy for understanding campylobacteriosis in New Zealand.

Outbreak of campylobacteriosis following pre-cooked sausage consumption.

OBJECTIVES: A small outbreak of campylobacteriosis involving three cases was investigated in terms of Campylobacter types present in the suspect food (pre-cooked sausages) and clinical samples from the cases. METHOD: Foods and faecal samples from people involved in the incident, which occurred in Christchurch, New Zealand, were tested for the presence of Campylobacter and identification of the species made. Isolates were typed by Penner serotyping and macrorestriction analysis using pulsed-field gel electrophoresis (PFGE). Investigations were conducted as to whether the contamination was on the surface or the interior of the sausages. RESULTS: All isolates from food and faecal samples were identified as C. jejuni and were indistinguishable from one another by the typing methods employed. Only the surfaces of the sausages were contaminated. Three other isolates of an indistinguishable subtype were isolated from campylobacteriosis cases in Christchurch occurring over approximately the same period. CONCLUSIONS AND IMPLICATIONS: Given the rarity of the subtype isolated from the three family members and the three other cases, it is possible that the outbreak was larger than the initial investigation revealed. It is likely that the sausages were contaminated after they had been cooked by the retailer and were not reheated prior to consumption. This report illustrates the role of cross-contamination in an outbreak with an unusual food vehicle for campylobacteriosis. Physical separation of cooked and raw product is necessary to prevent recurrences of outbreaks similar to the one described here.

First incursion of Salmonella enterica serotype typhimurium DT160 into New Zealand.

An outbreak of human Salmonella enterica serotype Typhimurium DT160 infection in New Zealand was investigated from May to August 2001. Handling of dead wild birds, contact with persons with diarrheal illness, and consumption of fast food were associated with infection. Contaminated roof-collected rainwater was also detected.