Lewis M. Graves

An Outbreak of Gastroenteritis and Fever Due to Listeria monocytogenes in Milk

BACKGROUND After an outbreak of gastroenteritis and fever among persons who attended a picnic in Illinois, chocolate milk served at the picnic was found to be contaminated with Listeria monocytogenes. METHODS In investigating this outbreak, we interviewed the people who attended the picnic about what they ate and their symptoms. Surveillance for invasive listeriosis was initiated in the states that receive milk from the implicated dairy. Stool and milk samples were cultured for L. monocytogenes. Serum samples were tested for IgG antibody to listeriolysin O. RESULTS Forty-five persons had symptoms that met the case definition for illness due to L. monocytogenes, and cultures of stool from 11 persons yielded the organism. Illness in the week after the picnic was associated with the consumption of chocolate milk. The most common symptoms were diarrhea (present in 79 percent of the cases) and fever (72 percent). Four persons were hospitalized. The median incubation period for infection was 20 hours (range, 9 to 32), and persons who became ill had elevated levels of antibody to listeriolysin O. Isolates from stool specimens from patients who became ill after the picnic, from sterile sites in three additional patients identified by surveillance, from the implicated chocolate milk, and from a tank drain at the dairy were all serotype 1/2b and were indistinguishable on multilocus enzyme electrophoresis, ribotyping, and DNA macrorestriction analysis. CONCLUSIONS L. monocytogenes is a cause of gastroenteritis with fever, and sporadic cases of invasive listeriosis may be due to unrecognized outbreaks caused by contaminated food.

Establishment of a Universal Size Standard Strain for Use with the PulseNet Standardized Pulsed-Field Gel Electrophoresis Protocols: Converting the National Databases to the New Size Standard

ABSTRACT The PulseNet National Database, established by the Centers for Disease Control and Prevention in 1996, consists of pulsed-field gel electrophoresis (PFGE) patterns obtained from isolates of food-borne pathogens (currently Escherichia coli O157:H7, Salmonella, Shigella, and Listeria) and textual information about the isolates. Electronic images and accompanying text are submitted from over 60 U.S. public health and food regulatory agency laboratories. The PFGE patterns are generated according to highly standardized PFGE protocols. Normalization and accurate comparison of gel images require the use of a well-characterized size standard in at least three lanes of each gel. Originally, a well-characterized strain of each organism was chosen as the reference standard for that particular database. The increasing number of databases, difficulty in identifying an organism-specific standard for each database, the increased range of band sizes generated by the use of additional restriction endonucleases, and the maintenance of many different organism-specific strains encouraged us to search for a more versatile and universal DNA size marker. A Salmonella serotype Braenderup strain (H9812) was chosen as the universal size standard. This strain was subjected to rigorous testing in our laboratories to ensure that it met the desired criteria, including coverage of a wide range of DNA fragment sizes, even distribution of bands, and stability of the PFGE pattern. The strategy used to convert and compare data generated by the new and old reference standards is described.

Multistate outbreak of Listeria monocytogenes infection linked to delicatessen turkey meat.

BACKGROUND Despite a decreasing incidence of listeriosis in the United States, molecular subtyping has increased the number of recognized outbreaks. In September 2000, the New York City Department of Health identified a cluster of infections caused by Listeria monocytogenes isolates with identical molecular subtypes by pulsed-field gel electrophoresis (PFGE) and ribotyping. METHODS To determine the magnitude of the outbreak and identify risk factors for infection, we notified state health departments and conducted a case-control study. A case was defined as a patient or mother-infant pair infected with Listeria monocytogenes whose isolate yielded the outbreak PFGE pattern. Controls were patients infected with Listeria monocytogenes whose isolate yielded a different PFGE pattern. Patients were asked about food and drink consumed during the 30 days before the onset of illness. RESULTS Between May and December 2000, there were 30 clinical isolates of Listeria monocytogenes with identical PFGE patterns identified in 11 US states. Cases of infection caused by these isolates were associated with 4 deaths and 3 miscarriages. A case-control study implicated sliced processed turkey from a delicatessen (Mantel-Haenszel odds ratio, 8.0; 95% confidence interval, 1.2-43.3). A traceback investigation identified a single processing plant as the likely source of the outbreak, and the company voluntarily recalled 16 million pounds of processed meat. The same plant had been identified in a Listeria contamination event that had occurred more than a decade previously. CONCLUSIONS Prevention of persistent L. monocytogenes contamination in food processing plants presents a critical challenge to food safety professionals.

Nationwide outbreak of listeriosis due to contaminated meat

We used molecular subtyping to investigate an outbreak of listeriosis involving residents of 24 US states. We defined a case as infection with Listeria monocytogenes serotype 4b yielding one of several closely related patterns when subtyped by pulsed-field gel electrophoresis. Patients infected with strains yielding different patterns were used as controls. A total of 108 cases were identified with 14 associated deaths and four miscarriages or stillbirths. A case-control study implicated meat frankfurters as the likely source of infection (OR 17.3, 95% CI 2.4-160). The outbreak ended abruptly following a manufacturer-issued recall, and the outbreak strain was later detected in low levels in the recalled product. A second strain was recovered at higher levels but was not associated with human illness. Our findings suggest that L. monocytogenes strains vary widely in virulence and confirm that large outbreaks can occur even when only low levels of contamination are detected in sampled food. Standardized molecular subtyping and coordinated, multi-jurisdiction investigations can greatly facilitate detection and control of listeriosis outbreaks.

Listeriosis Outbreaks and Associated Food Vehicles, United States, 1998–2008

Outbreak investigations can identify industrial gaps and regulatory measures to protect food.

Multistate outbreak of listeriosis linked to Turkey deli meat and subsequent changes in US regulatory policy

BACKGROUND Listeriosis, a life-threatening foodborne illness caused by Listeria monocytogenes, affects approximately 2500 Americans annually. Between July and October 2002, an uncommon strain of L. monocytogenes caused an outbreak of listeriosis in 9 states. METHODS We conducted case finding, a case-control study, and traceback and microbiological investigations to determine the extent and source of the outbreak and to propose control measures. Case patients were infected with the outbreak strain of L. monocytogenes between July and November 2002 in 9 states, and control patients were infected with different L. monocytogenes strains. Outcome measures included food exposure associated with outbreak strain infection and source of the implicated food. RESULTS Fifty-four case patients were identified; 8 died, and 3 pregnant women had fetal deaths. The case-control study included 38 case patients and 53 control patients. Case patients consumed turkey deli meat much more frequently than did control patients (P = .008, by Wilcoxon rank-sum test). In the 4 weeks before illness, 55% of case patients had eaten deli turkey breast more than 1-2 times, compared with 28% of control patients (odds ratio, 4.5; 95% confidence interval, 1.3-17.1). Investigation of turkey deli meat eaten by case patients led to several turkey processing plants. The outbreak strain was found in the environment of 1 processing plant and in turkey products from a second. Together, the processing plants recalled > 30 million pounds of products. Following the outbreak, the US Department of Agricultures Food Safety and Inspection Service issued new regulations outlining a L. monocytogenes control and testing program for ready-to-eat meat and poultry processing plants. CONCLUSIONS Turkey deli meat was the source of a large multistate outbreak of listeriosis. Investigation of this outbreak helped guide policy changes designed to prevent future L. monocytogenes contamination of ready-to-eat meat and poultry products.

Listeria marthii sp. nov., isolated from the natural environment, Finger Lakes National Forest.

Four isolates (FSL S4-120(T), FSL S4-696, FSL S4-710, and FSL S4-965) of Gram-positive, motile, facultatively anaerobic, non-spore-forming bacilli that were phenotypically similar to species of the genus Listeria were isolated from soil, standing water and flowing water samples obtained from the natural environment in the Finger Lakes National Forest, New York, USA. The four isolates were closely related to one another and were determined to be the same species by whole genome DNA-DNA hybridization studies (>82 % relatedness at 55 degrees C and >76 % relatedness at 70 degrees C with 0.0-0.5 % divergence). 16S rRNA gene sequence analysis confirmed their close phylogenetic relatedness to Listeria monocytogenes and Listeria innocua and more distant relatedness to Listeria welshimeri, L. seeligeri, L. ivanovii and L. grayi. Phylogenetic analysis of partial sequences for sigB, gap, and prs showed that these isolates form a well-supported sistergroup to L. monocytogenes. The four isolates were sufficiently different from L. monocytogenes and L. innocua by DNA-DNA hybridization to warrant their designation as a new species of the genus Listeria. The four isolates yielded positive reactions in the AccuProbe test that is purported to be specific for L. monocytogenes, did not ferment L-rhamnose, were non-haemolytic on blood agar media, and did not contain a homologue of the L. monocytogenes virulence gene island. On the basis of their phenotypic characteristics and their genotypic distinctiveness from L. monocytogenes and L. innocua, the four isolates should be classified as a new species within the genus Listeria, for which the name Listeria marthii sp. nov. is proposed. The type strain of L. marthii is FSL S4-120(T) (=ATCC BAA-1595(T) =BEIR NR 9579(T) =CCUG 56148(T)). L. marthii has not been associated with human or animal disease at this time.

Outbreak of Listeriosis among Mexican Immigrants as a Result of Consumption of Illicitly Produced Mexican-Style Cheese

BACKGROUND In 2000, an outbreak of listeriosis among Hispanic persons was identified in Winston-Salem, North Carolina. The objectives of the present study were to identify the source of, strains associated with, and risk factors for Listeria monocytogenes infection for patients affected by the outbreak. METHODS Microbiological, case-control, and environmental investigations were conducted. Participants in the case-control study were case patients who became infected with L. monocytogenes between 1 October 2000 and 31 January 2001 and control subjects who were matched with case patients on the basis of ethnicity, sex, age, and pregnancy status. All participants were residents of Winston-Salem. RESULTS We identified 13 patients, all of whom were Hispanic, including 12 females who were 18-38 years of age. Eleven case patients were pregnant; infection with L. monocytogenes resulted in 5 stillbirths, 3 premature deliveries, and 3 infected newborns. Case patients were more likely than control subjects to have eaten the following foods: fresh, unlabeled, Mexican-style cheese sold by door-to-door vendors (matched odds ratio [MOR], 17.5; 95% confidence interval [CI], 2.0-152.5); queso fresco, a Mexican-style soft cheese (MOR, 7.3; 95% CI, 1.4-37.5); and hot dogs (MOR, 4.6; 95% CI, 1.1-19.4). L. monocytogenes isolates recovered from 10 female case patients, from cheese bought from a door-to-door vendor, from unlabeled cheese from 2 Hispanic markets, and from raw milk from a local dairy had indistinguishable patterns on pulsed-field gel electrophoresis. CONCLUSIONS This outbreak of listeriosis was caused by noncommercial, fresh, Mexican-style cheese made from contaminated raw milk traced to 1 local dairy. We recommend educating Hispanic women about food safety while they are pregnant, enforcing laws that regulate the sale of raw milk and dairy products made by unlicensed manufacturers, making listeriosis a reportable disease in all states, routinely interviewing case patients, and routinely subtyping clinical L. monocytogenes isolates.

Invasive Listeriosis in the Foodborne Diseases Active Surveillance Network (FoodNet), 2004–2009: Further Targeted Prevention Needed for Higher-Risk Groups

BACKGROUND Listeriosis can cause severe disease, especially in fetuses, neonates, older adults, and persons with certain immunocompromising and chronic conditions. We summarize US population-based surveillance data for invasive listeriosis from 2004 through 2009. METHODS We analyzed Foodborne Diseases Active Surveillance Network (FoodNet) data for patients with Listeria monocytogenes isolated from normally sterile sites. We describe the epidemiology of listeriosis, estimate overall and specific incidence rates, and compare pregnancy-associated and nonpregnancy-associated listeriosis by age and ethnicity. RESULTS A total of 762 listeriosis cases were identified during the 6-year reporting period, including 126 pregnancy-associated cases (17%), 234 nonpregnancy-associated cases(31%) in patients aged <65 years, and 400 nonpregnancy-associated cases (53%) in patients aged ≥ 65 years. Eighteen percent of all cases were fatal. Meningitis was diagnosed in 44% of neonates. For 2004-2009, the overall annual incidence of listeriosis varied from 0.25 to 0.32 cases per 100,000 population. Among Hispanic women, the crude incidence of pregnancy-associated listeriosis increased from 5.09 to 12.37 cases per 100,000 for the periods of 2004-2006 and 2007-2009, respectively; among non-Hispanic women, pregnancy-associated listeriosis increased from 1.74 to 2.80 cases per 100,000 for the same periods. Incidence rates of nonpregnancy-associated listeriosis in patients aged ≥ 65 years were 4-5 times greater than overall rates annually. CONCLUSIONS Overall listeriosis incidence did not change significantly from 2004 through 2009. Further targeted prevention is needed, including food safety education and messaging (eg, avoiding Mexican-style cheese during pregnancy). Effective prevention among pregnant women, especially Hispanics, and older adults would substantially affect overall rates.

Listeria monocytogenes Infection from Foods Prepared in a Commercial Establishment: A Case-Control Study of Potential Sources of Sporadic Illness in the United States

BACKGROUND Listeria monocytogenes has been estimated to cause >2500 illnesses and 500 deaths annually in the United States. Efforts to reduce foodborne listeriosis have focused on foods frequently implicated in outbreaks. Potential sources for L. monocytogenes infection not associated with outbreaks remain poorly understood. METHODS The Foodborne Diseases Active Surveillance Network conducts surveillance for culture-confirmed listeriosis at clinical laboratories in 9 states. After excluding outbreak-associated cases, we attempted to enroll eligible case patients with L. monocytogenes infection in a case-control study from 2000 through 2003. Control subjects were recruited through health care providers and were matched to case patients by state, age, and immunosuppression status. Data were collected about exposures occurring in the 4 weeks before specimen collection from the case patients. RESULTS Of the 249 case patients with L. monocytogenes infection, only 12 (5%) had cases that were associated with outbreaks; 6 other patients were ineligible for other reasons. Of 231 eligible case patients, 169 (73%) were enrolled in the study. We classified 28 case patients as having pregnancy-associated cases. We enrolled 376 control subjects. In multivariable analysis, L. monocytogenes infection was associated with eating melons at a commercial establishment (odds ratio, 2.6; 95% confidence interval, 1.4-5.0) and eating hummus prepared in a commercial establishment (odds ratio, 5.7; 95% confidence interval, 1.7-19.1). CONCLUSIONS Most cases of L. monocytogenes infection were not associated with outbreaks. Reducing the burden of foodborne listeriosis may require interventions directed at retail environments and at foods, such as melons and hummus, that are not commonly recognized as high risk. Because of the severity of listeriosis, pregnant women and other persons at risk may wish to avoid eating these newly implicated foods.