J. McLauchlin

Listeria monocytogenes and listeriosis: a review of hazard characterisation for use in microbiological risk assessment of foods

Considerable effort has been put into the application of quantitative microbiological risk assessment for Listeria monocytogenes, and data are available for England and Wales (probably more so than most other countries) on the adverse health effects, together with incidence data on different age and risk groups for human L. monocytogenes infections. This paper reviews aspects of Listeria and human listeriosis, especially from a public health perspective and provide hazard characterisation data, i.e. the qualitative and/or quantitative evaluation of the adverse health effect associated with the hazard, which is the relationship between exposure levels (dose) and frequency of illness. The majority of cases of human listeriosis are food-borne; however, the disease process is complex with multiple routes of infection. The dose-response relationship is poorly understood, and data from human volunteer studies are not available and would be unethical to produce. Data are available from a range of different animal and in vitro models, although these poorly mimic the natural disease process in route of infection, end point, host and history of prior exposure to the bacterium. Epidemiological data provide some information on infective doses and dose responses, but because of the characteristics of the disease (the hugely variable and potentially very long incubation periods, the low attack rates and the rarity of identification of specific food vehicles), this also provides limited data for calculation of dose responses. There is some, albeit limited, evidence for strain variation, but this is an area of considerable uncertainty despite great advances in the genetic basis of the virulence of this bacterium, and almost all strains seem capable of causing serious disease. A variety of mathematical approaches have been used to model dose responses. The review is written to provide a clinical and epidemiological background to the mathematically oriented, as well as to outline the mathematical approaches to those interested in food-borne infection.

Detection by PCR of eight groups of enteric pathogens in 4,627 faecal samples: re-examination of the English case-control Infectious Intestinal Disease Study (1993–1996)

The English case-control Infectious Intestinal Disease Study (1993–1996) failed to detect an enteric pathogen or toxin in 49% of cases of gastroenteritis. In the present study, polymerase chain reaction (PCR) assays were applied to DNA and cDNA generated from 4,627 faecal samples from cases and controls archived during the original study for the detection of norovirus, rotavirus, sapovirus, Campylobacter spp., Salmonella spp., enteroaggregative Escherichia coli, Cryptosporidium spp., and Giardia spp. The percentage of archived samples from cases and from controls in which at least one agent (or toxin) was detected increased from 53% in the original study to 75% and from 19 to 42%, respectively, after the application of PCR assays. Among cases, the following percentages of enteric pathogens were detected: norovirus 36%, rotavirus A 31%, sapovirus 4%, Salmonella spp. 6%, Campylobacter jejuni 13%, Campylobacter coli 2%, other Campylobacter spp. 8%, enteroaggregative E. coli 6%, Giardia spp. 2%, and Cryptosporidium spp. 2%. The present study provides additional insight into the aetiology of infectious intestinal disease in England and highlights the occurrence of viral infections in cases as well as in asymptomatic individuals. Other notable findings include the frequent presence of Campylobacter spp. other than C. jejuni or C. coli, the high frequency of multiple agents in 41% of cases and in 13% of controls, and the variation in the aetiology and rate of infection found for different age groups. The results demonstrate the greater sensitivity of PCR-based methods compared to current conventional methods.

Changing Pattern of Human Listeriosis, England and Wales, 2001–2004

Disease has reemerged, mainly in patients ≥60 years of age with bacteremia.

Biological risks associated with consumption of reptile products.

The consumption of a wide variety of species of reptiles caught from the wild has been an important source of protein for humans world-wide for millennia. Terrapins, snakes, lizards, crocodiles and iguanas are now farmed and the consumption and trade of their meat and other edible products have recently increased in some areas of the world. Biological risks associated with the consumption of products from both farmed and wild reptile meat and eggs include infections caused by bacteria (Salmonella spp., Vibrio spp.), parasites (Spirometra, Trichinella, Gnathostoma, pentastomids), as well as intoxications by biotoxins. For crocodiles, Salmonella spp. constitute a significant public health risk due to the high intestinal carrier rate which is reflected in an equally high contamination rate in their fresh and frozen meat. There is a lack of information about the presence of Salmonella spp. in meat from other edible reptilians, though captive reptiles used as pets (lizards or turtles) are frequently carriers of these bacteria in Europe. Parasitic protozoa in reptiles represent a negligible risk for public health compared to parasitic metazoans, of which trichinellosis, pentastomiasis, gnathostomiasis and sparganosis can be acquired through consumption of contaminated crocodile, monitor lizard, turtle and snake meat, respectively. Other reptiles, although found to harbour the above parasites, have not been implicated with their transmission to humans. Freezing treatment inactivates Spirometra and Trichinella in crocodile meat, while the effectiveness of freezing of other reptilian meat is unknown. Biotoxins that accumulate in the flesh of sea turtles may cause chelonitoxism, a type of food poisoning with a high mortality rate in humans. Infections by fungi, including yeasts, and viruses widely occur in reptiles but have not been linked to a human health risk through the contamination of their meat. Currently there are no indications that natural transmissible spongiform encephalopathies (TSEs) occur in reptilians. The feeding of farmed reptiles with non-processed and recycled animal products is likely to increase the occurrence of biological hazards in reptile meat. Application of GHP, GMP and HACCP procedures, respectively at farm and slaughterhouse level, is crucial for controlling the hazards.

Prevalence and Level of Listeria monocytogenes and Other Listeria Species in Selected Retail Ready-to-Eat Foods in the United Kingdom

Although listeriosis is a rare cause of human disease in the United Kingdom, an increase in the number of cases has been observed since 2001, almost exclusively in persons older than 60 years. This increase prompted this study on the microbiological safety of ready-to-eat (RTE) foods, which included those types potentially linked to cases of listeriosis. Between May 2006 and April 2007, 6,984 RTE foods were sampled (2,168 sliced meats, 1,242 hard cheese, 1,088 sandwiches, 878 butter, 725 spreadable cheese, 515 confectionery products containing cream, and 368 probiotic drinks). The food types with the highest prevalence of Listeria monocytogenes were sandwiches (7.0%) and sliced meats (3.7% within shelf life, 4.2% end of shelf life). L. monocytogenes at > 100 CFU/g (exceeding the European Commissions food safety criteria limit) only occurred in sandwiches (0.4%) and sliced meats (0.7% within shelf life, 1.0% end of shelf life). Contamination with L. monocytogenes at >100 CFU/g was more frequent in meats that were prepacked and/or of pack size > or = 300 g and in sandwiches that were supplied prepacked that contained salad vegetables as an ingredient. Satisfactory microbiological quality was associated with premises on which the management was trained in food hygiene and those that complied with hazard analysis and critical control point principles. This study provides important information about the microbiological safety of RTE foods and demonstrates that the control of L. monocytogenes in such foods, and in particular sandwiches and sliced meats, is essential in order to minimize the risk of this bacterium being present at levels hazardous to health at the point of consumption.

Blinded application of microscopy, bacteriological culture, immunoassays and PCR to detect gastrointestinal pathogens from faecal samples of patients with community-acquired diarrhoea

A blinded trial in two different laboratories was performed to compare the detection of selected enteric pathogens in 92 unselected faecal samples collected from patients with community-acquired diarrhoea by conventional and PCR-based techniques. Conventional techniques detected a single potential etiological agent in 15% of the samples, whereas results of PCR detected evidence of at least one agent in 41% of the samples. Overall, the detection rates for the different pathogens were as follows: adenovirus serogroup F, 1%; Campylobacter spp., 7.6%; Salmonella spp., 4%; enteroaggregative Escherichia coli, 9.8%; enteropathogenic E. coli, 6.5%; enterotoxigenic Clostridium perfringens, 3%; Cryptosporidium spp., 13%; and Giardia spp., 11%. Results for the detection of Salmonella spp., Campylobacter spp. and C. perfringens were similar by both techniques, whereas Cryptosporidium and Giardia spp. were detected 22 times more often by PCR than by conventional microscopy. It was not possible to compare the results for detection of enteroaggregative E. coli and enteropathogenic E. coli since these were only investigated by PCR. The results of this small study clearly demonstrate the advantages of PCR-based methods compared to conventional techniques for the detection of gastrointestinal pathogens.

The microbiological safety of ready-to-eat specialty meats from markets and specialty food shops: a UK wide study with a focus on Salmonella and Listeria monocytogenes.

From 2359 specialty meats (continental sausages, cured/fermented, dried meats) sampled from markets and specialty food shops, 98.9% of samples were of satisfactory or acceptable microbiological quality. However, 16 (0.7%) were unsatisfactory as a result of Escherichia coli, Staphylococcus aureus or Listeria spp. contamination (>or=10(2) CFU/g), and nine (0.4%) were unacceptable due to presence of Salmonella spp. or Listeria monocytogenes (>10(2) CFU/g). Meats with unacceptable levels of L. monocytogenes were within shelf life (range: 8-143 days remaining). Nine different subtypes of L. monocytogenes were detected with sero/AFLP type 1/2c VII predominating (37%), although this subtype was not overrepresented in any particular meat type (P > 0.05). Ninety-six percent of continental sausages and cured/fermented products were stored at <8 degrees C at premises, including seven of the nine unacceptable samples. These nine meats were all pre-packed prior to supply to retail premises (OR = 0.1 P = 0.003) indicating that contamination with bacterial pathogens occurred earlier in the production chain. Most samples (72.7%, 8/11) with unsatisfactory levels of E. coli were sliced on request, suggesting cross-contamination at point of sale. This study highlights the importance of ensuring that products do not become contaminated before final packaging, that storage conditions are controlled, and that durability dates are an accurate indication of the shelf life of the product so as to minimise the potential for L. monocytogenes to be present at levels hazardous to health at the point of sale.

Multilocus Analysis of Cryptosporidium hominis and Cryptosporidium parvum Isolates from Sporadic and Outbreak-Related Human Cases and C. parvum Isolates from Sporadic Livestock Cases in the United Kingdom

ABSTRACT Cryptosporidium parvum and Cryptosporidium hominis isolates from sporadic, drinking water-associated, and intrafamilial human cases together with C. parvum isolates from sporadic cases in livestock were collected in the United Kingdom between 1995 and 1999. The isolates were characterized by analysis of three microsatellite markers (ML1, GP15, and MS5) using PCR amplification. Within C. hominis, four alleles were detected within the GP15 and MS5 loci, and a single type was detected with ML1. C. parvum was more polymorphic; 12 alleles were detected with GP15, 6 were detected with MS5, and 3 were detected with ML1. Multilocus analysis of polymorphisms within the three microsatellite loci was combined with those reported previously for an extrachromosomal small double-stranded RNA. Forty multilocus types were detected within these two species: 9 were detected in C. hominis, and 31 were detected in C. parvum. In C. hominis, heterogeneity was almost exclusively found in samples from sporadic cases. Similarity analysis identified three main groups within C. parvum, and the group that predominated in human infection was also found in livestock. Multilocus types of C. parvum previously identified only in humans were not detected in livestock. Isolates of both C. hominis and C. parvum from separate waterborne outbreaks were genetically homogeneous, suggesting preferential or point source transmission of certain types of these two species of parasites.

Methods for determining disease burden and calibrating national surveillance data in the United Kingdom: the second study of infectious intestinal disease in the community (IID2 study)

BackgroundInfectious intestinal disease (IID), usually presenting as diarrhoea and vomiting, is frequently preventable. Though often mild and self-limiting, its commonness makes IID an important public health problem. In the mid 1990s around 1 in 5 people in England suffered from IID a year, costing around £0.75 billion. No routine information source describes the UKs current community burden of IID. We present here the methods for a study to determine rates and aetiology of IID in the community, presenting to primary care and recorded in national surveillance statistics. We will also outline methods to determine whether or not incidence has declined since the mid-1990s.Methods/designThe Second Study of Infectious Intestinal Disease in the Community (IID2 Study) comprises several separate but related studies. We use two methods to describe IID burden in the community - a retrospective telephone survey of self-reported illness and a prospective, all-age, population-based cohort study with weekly follow-up over a calendar year. Results from the two methods will be compared. To determine IID burden presenting to primary care we perform a prospective study of people presenting to their General Practitioner with symptoms of IID, in which we intervene in clinical and laboratory practice, and an audit of routine clinical and laboratory practice in primary care. We determine aetiology of IID using molecular methods for a wide range of gastrointestinal pathogens, in addition to conventional diagnostic microbiological techniques, and characterise isolates further through reference typing. Finally, we combine all our results to calibrate national surveillance data.DiscussionResearchers disagree about the best method(s) to ascertain disease burden. Our study will allow an evaluation of methods to determine the community burden of IID by comparing the different approaches to estimate IID incidence in its linked components.

Survey of Salmonella contamination of edible nut kernels on retail sale in the UK

Consumption of nut kernels has shown an upward trend due to peoples increasing tendency to eat healthy snacks. The purpose of this survey was to establish the microbiological safety of retail edible nut kernel samples of different varieties. Overall Salmonella spp. and Escherichia coli were detected from 0.1% and 0.8% of 2886 edible nut kernels, respectively. S. Senftenberg and S. Tennessee were detected from two pre-packed samples of Brazil nuts (0.4%) and S. Anatum from a pre-packed mixed nuts sample (0.9%; mix: almonds, Brazils, cashews, peanuts, walnuts) indicating a risk to health. The levels of Salmonella ranged from <0.01 to 0.23/g. E. coli at unsatisfactory levels (150/g) was present in another pre-packed Brazils nuts sample (0.2%). E. coli was additionally found at lower levels (range: 3.6-43/g) in Brazils (1.9%), macadamia (1.5%), pistachios (1.1%), walnuts (0.7%), peanuts (0.7%), hazels (0.5%), cashews (0.4%), and almonds (0.3%). Levels of E. coli did not correlate with the presence of Salmonella. The batches contaminated with Salmonella were recalled and Food Standards Agency food alerts were issued to advise against the consumption of the affected products. The presence of Salmonella is unacceptable in ready-to-eat foods and follows that the need for applying good agricultural and hygiene practices and effective decontamination procedures during the production of edible kernels cannot be overemphasized.