F. J. Bolton

Milkborne general outbreaks of infectious intestinal disease, England and Wales, 1992-2000.

From 1 January 1992 to 31 December 2000, 27 milkborne general outbreaks of infectious intestinal disease (IID) were reported to the Public Health Laboratory Service (PHLS) Communicable Disease Surveillance Centre (CDSC). These outbreaks represented a fraction (2%) of all outbreaks of foodborne origin (N = 1774) reported to CDSC, but were characterized by significant morbidity. Unpasteurized milk (52%) was the most commonly reported vehicle of infection in milkborne outbreaks, with milk sold as pasteurized accounting for the majority of the rest (37%). Salmonellas (37%), Vero cytotoxin-producing Escherichia coli (VTEC) O157 (33%) and campylobacters (26%) were the most commonly detected pathogens, and most outbreaks were linked to farms (67%). This report highlights the importance of VTEC O157 as a milkborne pathogen and the continued role of unpasteurized milk in human disease.

Isolation of Escherichia coli O157 from raw meat products.

This study has evaluated an enrichment and four subculture procedures for detection of Escherichia coli 0157 in raw meat products. The combination of enrichment in modified tryptone broth incubated at 42d̀C for 6 h, followed by immunomagnetic separation and subculture on to cefixime, tellurite sorbitol MacConkey agar was the most sensitive and selective procedure. Traditional subculture using 10 μl and 100 μl inocula and culture of centrifuged deposits were less satisfactory. A most probable number method was used to enumerate E. coli 0157 in naturally contaminated samples associated with human cases. The results indicated that the samples contained <0.3 to 2300 cfu g‐1 of E. coli 0157 which confirms that the infective dose for this organism is low.

Application of a new phagetyping scheme to campylobacters isolated during outbreaks

A new scheme for phagetyping campylobacters has been evaluated using strains isolated from five outbreaks. The phagetyping results have been compared with the results of Penner serotyping, Lior serotyping and Preston biotyping. Phagetyping recognized the causative strains in all of the incidents and also differentiated these strains from animal and environmental strains isolated during these investigations. In some outbreaks phagetyping proved to be more discriminatory than serotyping or biotyping, e.g. strains of Penner serotype 2, and serogroup 4, 13, 16, 50 were subdivided by this method. Phagetyping is to be recommended for typing strains from outbreaks and although the results indicate that it may be used alone we advocate that it should be used in conjunction with one of the established typing methods.

Presence of campylobacter and salmonella in sand from bathing beaches

The purpose of this study was to determine the presence of thermophilic Campylobacter spp. and Salmonella spp. in sand from non-EEC standard and EEC standard designated beaches in different locations in the UK and to assess if potentially pathogenic strains were present. Campylobacter spp. were detected in 82/182 (45%) of sand samples and Salmonella spp. in 10/182 (6%). Campylobacter spp. were isolated from 46/92 (50%) of samples from non-EEC standard beaches and 36/90 (40%) from EEC standard beaches. The prevalence of Campylobacter spp. was greater in wet sand from both types of beaches but, surprisingly, more than 30% of samples from dry sand also contained these organisms. The major pathogenic species C. jejuni and C. coli were more prevalent in sand from non-EEC standard beaches. In contrast, C. lari and urease positive thermophilic campylobacters, which are associated with seagulls and other migratory birds, were more prevalent in sand from EEC standard beaches. Campylobacter isolates were further characterized by biotyping and serotyping, which confirmed that strains known to be of types associated with human infections were frequently found in sand on bathing beaches.

E. coli O157 phage type 21/28 outbreak in North Cumbria associated with pasteurized milk.

In March 1999, a large community outbreak of Escherichia coli O157 infection occurred in North Cumbria. A total of 114 individuals were reported to the Outbreak Control Team (OCT); 88 had laboratory confirmed E. coli O157. Twenty-eight (32%) of the confirmed cases were admitted to hospital, including three children (3.4%) with haemolytic uraemic syndrome. There were no deaths. A case-control study found that illness was strongly associated with drinking pasteurized milk from a local farm (P = <0.0001) on single variable analysis. Microbiological investigations at the farm revealed E. coli O157 phage type (PT) 21/28 VT 2 which was indistinguishable from the human isolates by pulsed field gel electrophoresis. At the time of occurrence this was the largest E. coli O157 outbreak in England and Wales and the first E. coli O157 PT 21/28 VT 2 outbreak associated with pasteurized milk. This outbreak highlights lessons to be learnt regarding on-farm pasteurization.

Improved method for the isolation of Campylobacter jejuni and Campylobacter coli bacteriophages

A centrifugation and filtration method of isolating Campylobacter phages has been developed. Forty‐nine Campylobacter phages were isolated from 272 effluent samples of which 42 produced lysis with Campylobacter jejuni strains and seven with C. coli strains. Phages were recovered from pig manure, abattoir effluents, human faeces, sewage and poultry manure. Phages were not isolated from water samples, cattle and sheep faeces or farm pasture soil.

Phenotypic diversity of Campylobacter isolates from sporadic cases of human enteritis in the UK

Aims: The aim of this study was to identify and subtype a large collection of isolates of Campylobacter spp. to quantify diversity among strains causing human disease from geographically diverse sources in the United Kingdom.

Reference isolates for the clonal complexes of Campylobacter jejuni

Aims: To identify and make available through the National Collection of Type Cultures (NCTC) a set of reference isolates for the clonal complexes of Campylobacter jejuni.

New media for detection and counting of Clostridia in foods

The growth of pure cultures of Clostridium perfringens (ATCC 12922) and Cl. sporogenes (PA 3679) in five non‐selective media, fluid thioglycollate medium (FTM), rapid perfringens medium (RPM), Columbia broth Malthus (CBM), reinforced clostridial medium (RCM) and lactose sulphite (LS), was monitored using conductance measurements with a Malthus analyser. Only FTM and CBM gave useful results. The correlation of log10 plate counts on blood agar of the pure strain of Cl. sporogenes with detection times in FTM was highly significant (ru200a=u200a0·96, nu200a=u200a73), and with detection times in CBM less so (ru200a=u200a−0·909, nu200a=u200a33). The correlation of log10 counts on tryptose sulphite neomycin medium (TSN) of wild strains of Cl. sporogenes and Cl. perfringens with detection times with FTM in meat was also highly significant (ru200a=u200a0·933, nu200a=u200a54).

Evaluation of methods for the enumeration of Campylobacter jejuni and Campylobacter coli bacteriophages

The Spiral Plater System used for enumeration of bacteria was evaluated for the titration of Carnpylobacter phages. Twelve phages of C. jejuni were titrated using the conventional surface droplet technique, soft agar overlay‐pour plate technique, and the Spiral Plater System. Phage counts obtained by the three methods were similar but the Spiral Plater System showed greater precision than the other methods.