Investigation of multidrug-resistant Salmonella serotype typhimurium DT104 infections linked to raw-milk cheese in Washington State.

CONTEXT: Multidrug-resistant Salmonella Typhimurium DT104 has recently emerged as a cause of human and animal illness in Europe and North America. In early 1997, health officials in Yakima County, Washington, noted a 5-fold increase in salmonellosis among the county's Hispanic population. OBJECTIVES: To characterize bacterial strains and identify risk factors for infection with Salmonella Typhimurium in Yakima County. DESIGN: Laboratory, case-control, and environmental investigations. SETTING AND PARTICIPANTS: Patients with culture-confirmed Salmonella Typhimurium infection living in Yakima County and age- and neighborhood-matched control subjects. MAIN OUTCOME MEASURES: Food vehicle implication based on case-control study and outbreak control. RESULTS: Between January 1 and May 5, 1997, 54 culture-confirmed cases of Salmonella Typhimurium were reported. The median age of patients was 4 years and 91% were Hispanic. Patients reported diarrhea (100%), abdominal cramps (93%), fever (93%), bloody stools (72%), and vomiting (53%); 5 patients (9%) were hospitalized. Twenty-two patients and 61 control subjects were enrolled in the case-control study. Seventeen case patients (77%) reported eating unpasteurized Mexican-style soft cheese in the 7 days before onset of illness compared with 17 control subjects (28%) (matched odds ratio, 32.3; 95% confidence interval, 3.0-874.6). All case-patient isolates were phage definitive type 104 (DT104) (n = 10) or DT104b (n = 12), and 20 (91%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfamethoxazole, and tetracycline. The cheese produced and eaten by 2 unrelated patients was made with raw milk traced to the same local farm. Milk samples from nearby dairies yielded Salmonella Typhimurium DT104. The incidence of Salmonella Typhimurium infections in Yakima County returned to pre-1992 levels following interventions based on these findings. CONCLUSIONS: Multidrug-resistant Salmonella Typhimurium DT104 emerged as a cause of salmonellosis in Yakima County, and Mexican-style soft cheese made with unpasteurized milk is an important vehicle for Salmonella Typhimurium DT104 transmission. We postulate that recent increases in human salmonellosis reflect the emergence of Salmonella Typhimurium DT104 among dairy cows in the region. Continued efforts are needed to discourage consumption of raw milk products, promote healthier alternatives, and study the ecology of multidrug-resistant Salmonella Typhimurium.

An outbreak of Escherichia coli O157:H7 infections associated with leaf lettuce consumption.

In July 1995, 40 Montana residents were identified with laboratory-confirmed Escherichia coli O157:H7 infection; 52 residents had bloody diarrhea without laboratory confirmation. The median age of those with laboratory-confirmed cases was 42 years (range, 4- 86); 58% were female. Thirteen patients were hospitalized, and 1 developed hemolytic-uremic syndrome. A case-control study showed that 19 (70%) of 27 patients but only 8 (17%) of 46 controls reported eating purchased (not home-grown) leaf lettuce before illness (matched odds ratio, 25.3; 95% confidence interval, 3.9-1065.6). Pulsed-field gel electrophoresis identified a common strain among 22 of 23 isolates tested. Implicated lettuce was traced to two sources: a local Montana farm and six farms in Washington State that shipped under the same label. This outbreak highlights the increasing importance of fresh produce as a vehicle in foodborne illness. Sanitary growing and handling procedures are necessary to prevent these infections.

An international outbreak of Salmonella infections caused by alfalfa sprouts grown from contaminated seeds.

An outbreak of Salmonella serotype stanley infections occurred in the United States and Finland in 1995. The outbreak was investigated through case-control studies in Arizona, Michigan, and Finland; by isolate subtyping; and by tracing and culturing of the implicated food. Alfalfa sprout consumption was the only exposure associated with S. stanley infections in Arizona (matched odds ratio [MOR] = 11.1; 95% confidence interval [CI], 1.4-513), Michigan (MOR = 5.5; CI, 1.6-23), and Finland (MOR undefined; CI, 4.9-infinity). US and Finnish patient isolates were a unique outbreak strain distinct from S. stanley isolates not linked to the outbreak. Alfalfa sprouts eaten by patients in 6 US states and Finland were traced to seed shipped by a Dutch shipper. Thus, it was concluded that alfalfa sprouts grown from contaminated seed caused an international outbreak of > or =242 S. stanley infections in > or =17 US states and Finland. This outbreak illustrates a new mechanism through which contamination of fresh produce can cause large, widely dispersed outbreaks.

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.

Bartonella vinsonii subsp. berkhoffii subsp. nov., isolated from dogs; Bartonella vinsonii subsp. vinsonii; and emended description of Bartonella vinsonii.

Two bacterial strains, one isolated from the blood of a dog with valvular endocarditis and one isolated from the blood of a healthy dog, were similar to Bartonella species, as determined by a number of phenotypic criteria, including growth characteristics, biochemical reactions, and cell wall fatty acid composition. The results of 16S rRNA gene sequence similarity studies confirmed that these strains are closely related and belong in the genus Bartonella and that Bartonella vinsonii is their closest relative (the 16S rRNA of isolate 93-C01T [T = type strain] was 99.37% identical to the 16S rRNA of the type strain of B. vinsonii, the 16S rRNA of isolate G7464 was 99.61% identical to the 16S rRNA of the type strain, and the 16S rRNAs of the dog isolates were 99.77% identical to each other). The 16S rRNAs of both strains contained a 12-base insertion that was not present in the 16S rRNA of the type strain of any Bartonella species. DNA relatedness tests revealed that these strains were related at the species level to the type strain of B. vinsonii. They were, however, significantly more closely related to each other than to B. vinsonii. On the basis of their unique 16S rRNA sequence insertion, their preferentially high level of relatedness, and their similar origins (dogs), we believe that strains 93-C01(T) and G7464 should be placed in a separate subspecies of B. vinsonii, for which we propose the name B. vinsonii subsp. berkhoffii subsp. nov. The type strain of B. vinsonii subsp. berkhoffii is strain 93-C01 (= ATCC 51672). The description of B. vinsonii is emended to accommodate the new subspecies, and B. vinsonii subsp. vinsonii is described.

The role of heightened surveillance in an outbreak of Escherichia coli O157.H7.

After instituting laboratory screening for Escherichia coli O157.H7, a Connecticut hospital isolated the organism from four persons in September 1993. As a result, an outbreak of E. coli O157.H7 associated with a country club was detected. The club had served hamburger from the same shipment at two picnics. Attendees of two picnics were interviewed, stool cultures were obtained from symptomatic persons, and the remaining hamburger was cultured. Twenty (22%) of 89 persons who ate hamburger became ill, compared with 1 of 60 who did not eat hamburger (relative risk = 13.5, 95% confidence interval 3.2-56.3). Among persons who ate hamburgers, illness was strongly associated with eating hamburger that was not thoroughly cooked (P < 0.001). All 20 samples from 5 remaining boxes of patties yielded E. coli O157.H7. Isolates from hamburger and case-patients were indistinguishable by pulsed-field gel electrophoresis. Heightened surveillance can rapidly identify outbreaks and may mitigate their impact. However, continued review of food safety issues is necessary if E. coli O157.H7 outbreaks are to be prevented.

Isolation and characterization of a beta-D-glucuronidase-producing strain of Escherichia coli serotype O157:H7 in the United States.

A phenotypic variant of Escherichia coli serotype O157:H7 (G5101) was isolated from a patient with bloody diarrhea. Strain G5101 does not ferment sorbitol but is beta-D-glucuronidase and urease positive. Serotyping and colony hybridization using a serotype-specific DNA probe confirmed that the isolate was O157:H7. G5101 produces Shiga-like toxins I and II and contains an eae gene that is highly conserved in the O157:H7 serotype. This strain would have been missed by laboratories that screen for the sorbitol-negative, beta-D-glucuronidase-negative phenotype in isolating E. coli O157:H7 from clinical and food specimens.

Listeria monocytogenes infection and serotype distribution among HIV-infected persons in Los Angeles County, 1985-1992.

Persons infected with human immunodeficiency virus (HIV) are at greater risk of infection with Listeria monocytogenes (LM) than the general population. We quantify the risk of listeriosis in persons with acquired immune deficiency syndrome (AIDS) and HIV infection in Los Angeles County (LAC) and report the LM serotype distribution among HIV-infected patients with listeriosis. Active surveillance for listeriosis was performed in LAC during most of the period from 1985 through 1992. Thirty-four (10%) of 351 nonperinatal cases of listeriosis reported in LAC from 1985 through 1992 were in HIV-infected persons, 25 of whom met the 1987 AIDS case definition. The incidence of listeriosis was 95.8 and 8.8 cases per 100,000 person-years among persons with AIDS and all HIV-infected persons, respectively, but only 1.0 case per 100,000 person-years in the total population. Excluding cases from a 1985 listeriosis epidemic associated with consumption of contaminated Mexican-style cheese, 11 (65%) of 17 HIV-infected persons with available isolates were infected with LM serotype 1/2b, whereas only 64 (31%) of 208 other persons with listeriosis and available isolates were infected with LM serotype 1/2b (odds ratio = 4.1; 95% confidence interval = 1.3-14.1). LM serogroup 1/2b may have been more common among HIV-infected persons in LAC than among other persons with listeriosis because of differences in diet or sexual practices, or to chance alone. Persons with HIV-infection, especially those with AIDS, should be educated in avoiding foods at high risk of listerial contamination, such as soft cheeses, foods sold from delicatessen counters, and undercooked chicken.

Role of foods in sporadic listeriosis. II. Microbiologic and epidemiologic investigation. The Listeria Study Group.

OBJECTIVE: To evaluate the role of foods in sporadic listeriosis. DESIGN: Microbiologic survey of foods collected from refrigerators of patients with listeriosis identified through active laboratory-based surveillance. Patient and food Listeria monocytogenes isolates were subtyped to identify foods contaminated with the same strain of L monocytogenes that caused illness in the patient; samples of these foods were obtained from the retail source. SETTING: Multistate population-based study conducted between 1988 and 1990. RESULTS: Listeria monocytogenes grew from at least one food specimen in the refrigerators of 79 (64%) of 123 listeriosis patients; 11% of more than 2000 food specimens collected in the study contained L monocytogenes. Twenty-six (33%) of 79 refrigerators with foods that grew L monocytogenes contained at least one food isolate of the same strain as that in the corresponding patient, a frequency much higher than would be expected by chance (P less than .001). Multivariate analysis showed that of the food specimens that grew L monocytogenes, foods that were ready-to-eat, foods that grew L monocytogenes by a direct-plating method (a measure of the level of contamination), and foods that contained serotype 4b isolates were independently associated with an increased likelihood of containing the patient-matching strain. CONCLUSION: We identified specific food and L monocytogenes isolate characteristics--ready-to-eat foods, foods containing higher concentrations of L monocytogenes, and foods containing serotype 4b--which were associated with disease-causing strains. These results can provide guidance to industry and regulatory agencies in developing strategies to prevent listeriosis.

Comparison of cold enrichment and U.S. Department of Agriculture methods for isolating Listeria monocytogenes from naturally contaminated foods. The Listeria Study Group.

We compared the cold enrichment (CE) and U.S. Department of Agriculture (USDA) methods for isolating Listeria monocytogenes by examining 402 food samples. The food samples were collected from refrigerators of listeriosis patients as part of a multistate active surveillance project to determine the role of foods in sporadic listeriosis in the United States. L. monocytogenes was isolated from 51 food samples (13%). The USDA method was significantly better (P less than 0.001) than the CE method. The isolation efficiencies of the USDA and CE methods were 96 and 59%, respectively. Quantitation of L. monocytogenes in the food samples revealed that many food samples containing less than 0.3 CFU/g were negative as determined by the CE method but positive as determined by the USDA method.

Investigation of an outbreak of listeriosis: new hypotheses for the etiology of epidemic Listeria monocytogenes infections.

From December 1986 to March 1987 an outbreak of Listeria monocytogenes infection occurred in the Philadelphia metropolitan area. A patient-control study showed patients were more likely than controls to have had an ill family member and to have used antidiarrheal medication during the month before their illness. Diet histories showed patients were significantly more likely to have eaten ice cream or salami than were controls, and to have shopped at one grocery store chain. Subtyping of L. monocytogenes isolates of patients showed no predominant strain, and cultures of food products eaten by patients were negative except for Brie cheese eaten by one patient. With no predominant strain of L. monocytogenes in the patients, a common source for this outbreak is unlikely. Thus, the identified risk factors may have been associated with carriage of L. monocytogenes and a coinfecting organism may have precipitated disseminated disease. Possible cofactors should be considered in investigations of future outbreaks of listeriosis.

Evaluation of enrichment and plating media for isolating Listeria monocytogenes.

We compared selective enrichment broths used by the U.S. Food and Drug Administration (FDA) and the U.S. Department of Agriculture (USDA), Food Safety and Inspection Service, for their efficiency in the quantitative recovery of Listeria monocytogenes from a naturally contaminated Brie cheese that was obtained as part of an epidemic investigation. Quantitative recovery of Listeria in FDA broth (greater than 2.4 x 10(5) colony forming units/mL) was significantly better than recovery in USDA broth (9.3 x 10(3) colony forming units/mL). When USDA broth was supplemented with D-glucose and Phytone (papaic digest of soy protein), its recovery efficiency improved but did not equal that of FDA broth for isolating L. monocytogenes from Brie cheese. A comparison of 4 selective plating media [modified McBride's agar, gum base nalidixic acid agar, lithium chloride-phenylethanol-moxalactam agar (LPM), and acriflavine-ceftazidime agar (AC)] showed that 3 L. monocytogenes strains belonging to serotype 1/2a were partially or completely inhibited on LPM and AC agars. One strain of serotype 1/2a formed microcolonies on modified McBride's agar after 48 h of incubation.

Commercial latex agglutination tests for detection of Haemophilus influenzae type b and Streptococcus pneumoniae antigens in patients with bacteremic pneumonia.

The validity of commercial latex agglutination kits for detection of Haemophilus influenzae type b and Streptococcus pneumoniae antigens in serum and urine specimens was studied. We tested serum and urine specimens from 44 patients with bacteremic pneumonia (23 S. pneumoniae, 13 H. influenzae type b, 11 other) with commercial latex agglutination kits (Directigen, Bactigen) for S. pneumoniae and H. influenzae type b antigens. All specimen samples were randomized and read blindly by two readers. Interreader reproducibility was 100%. The sensitivity and specificity of both kits for H. influenzae type b antigens in serum and urine were greater than 90%. None of the 24 urine samples from S. pneumoniae bacteremic patients were positive by either kit, although 6 ng of type 3 polysaccharide could be detected in spiked urine. Sensitivity for S. pneumoniae antigens in serum was 27% for Directigen and 38% for Bactigen. Specificity for S. pneumoniae antigens in serum was 95% for Directigen and 74% for Bactigen. The results suggest that the kits are useful in diagnosing H. influenzae type b pneumonia. However, the commercially available S. pneumoniae reagents tested appear to have limited utility for diagnosing S. pneumoniae pneumonia because both kits lack sensitivity and Bactigen lacks specificity, as well.

Isolation of Listeria monocytogenes from raw milk.

During a recent outbreak of listeriosis, we examined 121 raw milk samples and 14 milk socks (filters). Listeria monocytogenes was recovered from 15 (12%) of 121 milk specimens and 2 (14%) of 14 milk socks. The optimal processing method consisted of cold enriching diluted milk for 1 month with culture to selective broth, followed by plating.

Infections with Mycobacterium chelonei in patients receiving dialysis and using processed hemodialyzers.

Between April and November 1982, 27 of 140 patients in a hemodialysis center in Louisiana were infected with rapidly growing mycobacteria; 14 had bacteremia alone, 3 had soft-tissue infections, 1 had an access-graft infection, and 9 had widely disseminated disease. Of 26 identified isolates, 25 were Mycobacterium chelonei ssp. abscessus, and one was an M. chelonei-like organism. One factor common to all patients was exposure to processed hemodialyzers (artificial kidneys). Environmental sampling of the water-treatment system showed widespread contamination with nontuberculous mycobacteria, which were also recovered from the patient's side (blood compartment) of five of 31 hemodialyzers that had been processed and were ready for use. The formaldehyde concentration was less than 2% in two of three such contaminated dialyzers tested. We hypothesize that patients became infected when their blood circulated through processed dialyzers that contained viable rapidly growing mycobacteria. This outbreak demonstrates that hemodialysis patients may be at risk for developing infections with rapidly growing mycobacteria and that such infections may go unrecognized when routine culture methods are used. It also emphasizes the importance of using effective procedures to disinfect dialyzers in hemodialysis centers.

Risk of group A meningococcal disease: bacterial interference and cross-reactive bacteria among mucosal flora.

During outbreaks of group A meningococcal disease in Seattle, Wash., and Portland, Oreg., we studied the mucosal flora of the affected population and comparison groups to identify possible determinants of susceptibility and resistance to disease. Antimeningococcal immunoglobulin A can block the bactericidal activity of specific antibodies of other classes and has been associated with susceptibility in adults. We used immunoprecipitation and fluorescent-antibody techniques to detect mucosal microorganisms cross-reactive with group A meningococci that might have stimulated such antibodies. Cross-reactive strains of Bacillus pumilus and Streptococcus faecalis were found. Bacterial interference on mucosal surfaces has been shown to reduce susceptibility to other pathogens. With an agar overlay technique, we sought nasopharyngeal microorganisms that inhibited the growth of group A meningococci. Forty-five percent of subjects carried inhibitory strains representing at least nine different species. Inhibitory strains were less common (32%) in residents from "skid row" areas (see D.J. Bogue, Skid Row in American Cities, University of Chicago Press, for a comprehensive definition of these areas) than in a comparison group that did not experience meningococcal disease (61%), suggesting that their presence may be associated with resistance to acquisition of meningococci or to meningococcal disease.

Staphylococcus aureus isolates from patients with nonmenstrual toxic shock syndrome. Evidence for additional toxins.

Although toxic shock syndrome toxin-1 (TSST-1) has been proposed as the toxin responsible for toxic shock syndrome, its role in this disease has not been proved. To study this question, we examined Staphylococcus aureus strains isolated from normally sterile sites in patients with nonmenstrual toxic shock syndrome for the presence of TSST-1 production. Only 20 (62.5%) of 32 produced TSST-1, compared with 41 (93%) of 44 vaginal isolates from patients with menstrual toxic shock syndrome. Of strains of S aureus from patients with nonmenstrual toxic shock syndrome, TSST-1-negative isolates were more likely to be associated with a fatal outcome and to not be phage group 1 than TSST-1-positive isolates. Seven of the TSST-1-negative strains were evaluated in a rabbit subcutaneous chamber model of toxic shock syndrome. Fifteen (60%) of 25 rabbits developed a toxic shock syndrome-like illness and nine died. Clinical signs and histopathologic findings in the rabbits were similar to those seen in rabbits inoculated with TSST-1-positive S aureus isolates. These results suggest that other, as yet unrecognized, toxins play a role in toxic shock syndrome, and that TSST-1 production may not be essential to the pathogenesis of toxic shock syndrome.

Pasteurized milk as a vehicle of infection in an outbreak of listeriosis.

Between June 30th and August 30th, 1983, 49 patients in Massachusetts acquired listeriosis. Seven cases occurred in fetuses or infants and 42 in immunosuppressed adults; 14 patients (29 per cent) died. Of 40 Listeria monocytogenes isolates available for testing, 32 were serotype 4b. Two case-control studies, one matching for neighborhood of residence and the other for underlying disease, revealed that the illness was strongly associated with drinking a specific brand of pasteurized whole or 2 per cent milk (odds ratio = 9, P less than 0.01 for the neighborhood-matched study; odds ratio = 11.5, P less than 0.001 for the illness-matched study). The association with milk was further substantiated by four additional analyses that suggested the presence of a dose-response effect, demonstrated a protective effect of skim milk, associated cases with the same product in an independent study in another state, and linked a specific phage type with the disease associated with milk. The milk associated with disease came from a group of farms on which listeriosis in dairy cows was known to have occurred at the time of the outbreak. Multiple serotypes of L. monocytogenes were isolated from raw milk obtained from these farms after the outbreak. At the plant where the milk was processed, inspections revealed no evidence of improper pasteurization. These results support the hypothesis that human listeriosis can be a foodborne disease and raise questions about the ability of pasteurization to eradicate a large inoculum of L. monocytogenes from contaminated raw milk.