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PulseNet USA is the national molecular subtyping network system for foodborne disease surveillance. Sixty-four public health and food regulatory laboratories participate in PulseNet USA and routinely perform pulsed-field gel electrophoresis of Shiga toxigenic Escherichia coli isolated from humans, food, water, and the environment on a real-time basis. Clusters of infection are detected in three ways within this system: through rapidly alerting the participants in the electronic communication forum, the PulseNet Web conference; through cluster analysis by the database administrators at the coordinating center at the Centers for Disease Control and Prevention of the patterns uploaded to the central server by the participants; and by matching profiles of strains from nonhuman sources with recent human uploads to the national server. The strengths, limitations, and scope for future improvements of PulseNet are discussed with examples from 2002. In that year, notices of 30 clusters of Shiga toxigenic E. coli O157 infections were posted on the Web conference, 26 of which represented local outbreaks, whereas four were multistate outbreaks. Another 27 clusters were detected by central cluster detection performed at the Centers for Disease Control and Prevention, of which five represented common source outbreaks confirmed after finding an isolate with the outbreak pattern in the implicated food. Ten food isolates submitted without suspicion of an association to human disease matched human isolates in the database, and an epidemiologic link to human cases was established for six of them.

Twenty-one isolates of Listeria monocytogenes from food animal clinical cases that involved meningitis or meningoencephalitis, encephalitis, mastitis and abortion were characterized by serotyping and pulsed-field gel electrophoresis (PFGE) in order to improve our understanding of the genetic links between individual strains and strains recovered from human listeriosis cases. Results showed that five of the isolates were serotype 1/2a, six were 1/2b, nine were 4b, and one was untypeable. A caprine, two bovine and an ovine brain isolate shared identical PFGE patterns indicating that strains of L. monocytogenes are not host specific. Other isolates exhibited distinct patterns that were not shared, indicating a genetic diversity. Dendrogram analysis revealed that PFGE patterns of the isolates clustered primarily according to serotype. We compared the PFGE types obtained for these isolates with PFGE types for human clinical isolates present in the CDC national PulseNet database. Six (29%) of the twenty-one strains had patterns that were indistinguishable from pathogenic human isolates in the database. Our observations offer preliminary evidence that food animals could be significant reservoirs of L. monocytogenes that lead to human infections and support the inclusion of PFGE patterns of veterinary clinical isolates in the national PulseNet database for increased surveillance.

At the turn of the 20th century, typhoid fever was common in Wisconsin, and was a major impetus for the establishment of the Wisconsin State Laboratory of Hygiene (WSLH) in 1903. By the 1940s, typhoid was virtually eliminated in the United States due to public health measures such as disinfection of drinking water, sewage treatment, pasteurization, and shellfish bed sanitation. However, new food and waterborne pathogens have emerged to take the place of Salmonella Typhi. Infections with non-typhoidal Salmonella strains in the United States have increased almost 10-fold since the 1950s. In the last 20 years, the emergence of foodborne pathogens, such as Escherichia coli O157:H7, Cyclospora cayetanensis, Noroviruses (Norwalk-like viruses), Cryptosporidium parvum, Campylobacter jejuni, Yersinia enterocolitica, and multi-drug-resistant Salmonella, has identified a need for accurate laboratory diagnosis of enteric disease and outbreaks.

This report describes the investigation of a ground-beef-associated outbreak that involved five genetically distinct patient strains of Escherichia coli O157:H7. Human and product isolates were evaluated by pulsed-field gel electrophoresis (PFGE) with two endonucleases. The multiple-strain etiology of this outbreak underscores the importance of isolating and evaluating multiple colonies from outbreak-related products and comparing two endonuclease PFGE patterns of all product and human isolates identified during outbreak periods. This investigation emphasizes the importance of interviewing all confirmed and suspected case patients during the outbreak period, regardless of the PFGE pattern of their isolate, to confirm or rule out an epidemiologic link to the outbreak.

Objective: To investigate and control a nosocomial outbreak of Burkholderia cepacia lower respiratory tract infection., Design: Outbreak investigation and case-control study., Setting: A 260-bed community hospital., Patients: Participants were mechanically ventilated intensive care patients without cystic fibrosis. A case was defined as a hospitalized patient with a sputum culture positive for B. cepacia between January 1 and November 6, 1998., Methods: Respiratory therapy infection control policies and practices were reviewed; laboratory and environmental studies and a retrospective case-control study were conducted. Case-patients were matched with control-patients on age, gender, diagnosis, and type of intensive care unit., Results: Nine case-patients were identified; B. cepacia likely caused pneumonia in seven and colonization in two. Two respiratory therapy practices probably contributed to the transmission of B. cepacia: multidose albuterol vials were used among several patients, and nebulizer assemblies often were not dried between uses. B. cepacia was grown from cultures of three previously opened multidose vials; pulsed-field gel electrophoresis patterns of B. cepacia from seven case-patients and two multidose vials were indistinguishable. Case-patients had longer durations of heated humidified mechanical ventilation (mean, 9.8 days vs 4.4 days; P=.03) and were more likely to have exposure to one particular respiratory therapist than controls (odds ratio, undefined; 95% confidence interval, 4.7-infinity; P=.001). The association with the respiratory therapist, a temporary employee, persisted after controlling for duration of heated humidified ventilation. No new B. cepacia infections were identified after control measures were implemented., Conclusions: B. cepacia probably was transmitted among patients through use of extrinsically contaminated multidose albuterol vials. Respiratory therapy departments must pay close attention to infection control practices, particularly among new or temporary staff.

The toxin B assay was used to evaluate C. diff.-CUBE, a new dot-immunobinding assay (DIA) for the laboratory diagnosis of Clostridium difficile-associated diarrhea. The widely used latex agglutination test was also included for comparison. Stools from 100 patients suspected of having C. difficile-associated diarrhea were tested. The toxin B assay, latex agglutination, and DIA tests were positive for 12%, 9%, and 22% of the specimens, respectively. The sensitivity, specificity, and positive and negative predictive values of the DIA test were 67%, 84%, 36%, and 95%, respectively, compared with the toxin B assay. The specificity (98%) and positive predictive value (78%) for the latex agglutination test were significantly higher than those of the DIA test. Of 13 specimens solely positive by the DIA test, 11 were cultured and none were positive. Clinical assessment supported only two of the 13 positive DIA results. When clinical assessment was included in the analysis, the DIA positive predictive value rose to 45%. Although the sensitivity and negative predictive values of the DIA test are comparable to the latex agglutination test, the low specificity and positive predictive values of the DIA test make it an inappropriate method to use for screening in a population with a low prevalence of true positives.

Two CIE procedures (CIE-1, CIE-2) for the detection of Clostridium difficile in diarrheal stools were evaluated by comparison to cytotoxin assay and culture results and by comparison to a clinical likelihood of C. difficile-induced diarrhea. Using a combination of toxin assay and culture results for reference, the CIE-1 and CIE-2 procedures had sensitivities of 33% and 47%, specificities of 89% and 91%, and positive predictive values of 42% and 54%, respectively. Using clinical likelihood for reference, the best results were obtained by the CIE-2 method, which yielded a positive predictive value of 77%. Neither CIE procedure provided an acceptable sensitivity for the detection of C. difficile in stools.

The in vitro susceptibility of 20 isolates of Clostridium difficile to cefotaxime, moxalactam, and cefoperazone was determined by a standard agar dilution method. The median minimal inhibitory concentrations were 64, 32, and 32 mug/ml for cefotaxime, moxalactam, and cefoperazone, respectively.

We compared the selective blood agar medium of Gunn et al. (J. Clin. Microbiol. 5:650-655, 1977) which contains sulfamethoxazole plus trimethoprim (SXT-BA) to the conventional blood agar surface plate (SBA) and a modified blood agar pour plate plus broth method for the recovery of group A streptococci from throat swabs. The influence of CO(2) and ambient air incubation of the SXT-BA and SBA plates was also evaluated. A total of 696 throat swabs from symptomatic children were cultured simultaneously by the five methods and observed after overnight incubation; 204 positive cultures were detected overall. Recovery rates of each individual method were: SXT-BA (CO(2)), 90.7%; SXT-BA (air), 87.7%; pour plate plus broth, 83.3%; SBA (CO(2)), 79.4%; and SBA (air) 77%. Approximately one-half of the false-negative cultures in the SXT-BA (CO(2)) and SXT-BA (air) methods had colony counts of >/=10 to 100 colonies per plate. In contrast, for the SBA (CO(2)), SBA (air), and pour plate plus broth methods, approximately 70% of the false-negative cultures had colony counts of >/=10 to 100/plate. False-positive cultures obtained by the SXT-BA (CO(2)) and SXT-BA (air) methods were 11 and 12.7%, respectively-one-half as high as the rates obtained by the remaining methods. Beta-hemolytic streptococci, groups C, F, and G, are inhibited on the SXT-BA plates and were the primary cause of the higher false-positive rates on SBA and pour plate plus broth methods. An additional 3% positive cultures were obtained by incubating SXT-BA (CO(2)) plates up to 48 h before discarding as negative. We recommend either the SXT-BA (CO(2)) or the SXT-BA (air) method with up to 48 h of incubation for routine use in throat cultures.

The anaerobic broth disk (AnBD) method of Wilkins and Thiel and a new modification, designated the thioglycolate broth disk method, were compared with an agar dilution technique. The thioglycolate broth disk method was incubated aerobically (AeTBD) or anaerobically (AnTBD). One hundred anaerobic bacteria representing 15 species were tested with clindamycin, chloramphenicol, erythromycin, penicillin, and tetracycline. Agreement of results by the two methods with minimal inhibitory concentration determinations were: AnBD, 95.2%; AnTBD, 91.5%; AeTBD, 94.5%. With clindamycin, chloramphenicol, and penicillin, the agreement of the AeTBD and agar dilution results was 100%, 100%, and 95%, respectively. Using the AeTBD method, only 1.1% of all tests gave false susceptible readings, whereas 4.4% gave false resistant readings. All susceptibility testing errors occurred with tetracycline, erythromycin, and, to a lesser extent, penicillin. For each method, the changes in designation of bacteria as being susceptible or resistant to an antibiotic between trials primarily involved strains with minimal inhibitory concentrations which were +/- one dilution of the respective breakpoint value. The same situation was true for most bacteria that yielded false resistant readings within each trial. False resistant readings with tetracycline were determined to be unrelated to excess cation content of test media. These results reaffirm the reliability of the AnBD method and indicate that the AeTBD modification is equally reliable. The greater convenience and lower cost of the AeTBD method should make possible more widespread performance of susceptibility testing for anaerobic bacteria in hospital laboratories.

The antimicrobial activity of cefmetazole was compared with those of cefmenoxime, ceftizoxime, cefamandole, cefoperazone, cefotaxime, cephalothin, and latamoxef. In general, the activity of cefmetazole was less than those of the other cephalosporins. The in vitro activity of cefmetazole suggests that it will not prove useful as a broad-spectrum antimicrobial against gram-positive and gram-negative pathogens.