Serotype Typhimurium Infection Research Articles

Inhibition of multiple staphylococcal growth states by a small molecule that disrupts membrane fluidity and voltage.

New molecular approaches to disrupting bacterial infections are needed. The bacterial cell membrane is an essential structure with diverse potential lipid and protein targets for antimicrobials. While rapid lysis of the bacterial cell membrane kills bacteria, lytic compounds are generally toxic to whole animals. In contrast, compounds that subtly damage the bacterial cell membrane could disable a microbe, facilitating pathogen clearance by the immune system with limited compound toxicity. A previously described small molecule, D66, terminates Salmonella enterica serotype Typhimurium (S. Typhimurium) infection of macrophages and reduces tissue colonization in mice. The compound dissipates bacterial inner membrane voltage without rapid cell lysis under broth conditions that permeabilize the outer membrane or disable efflux pumps. In standard media, the cell envelope protects Gram-negative bacteria from D66. We evaluated the activity of D66 in Gram-positive bacteria because their distinct envelope structure, specifically the absence of an outer membrane, could facilitate mechanism of action studies. We observed that D66 inhibited Gram-positive bacterial cell growth, rapidly increased Staphylococcus aureus membrane fluidity, and disrupted membrane voltage while barrier function remained intact. The compound also prevented planktonic staphylococcus from forming biofilms and a disturbed three-dimensional structure in 1-day-old biofilms. D66 furthermore reduced the survival of staphylococcal persister cells and of intracellular S. aureus. These data indicate that staphylococcal cells in multiple growth states germane to infection are susceptible to changes in lipid packing and membrane conductivity. Thus, agents that subtly damage bacterial cell membranes could have utility in preventing or treating disease.IMPORTANCEAn underutilized potential antibacterial target is the cell membrane, which supports or associates with approximately half of bacterial proteins and has a phospholipid makeup distinct from mammalian cell membranes. Previously, an experimental small molecule, D66, was shown to subtly damage Gram-negative bacterial cell membranes and to disrupt infection of mammalian cells. Here, we show that D66 increases the fluidity of Gram-positive bacterial cell membranes, dissipates membrane voltage, and inhibits the human pathogen Staphylococcus aureus in several infection-relevant growth states. Thus, compounds that cause membrane damage without lysing cells could be useful for mitigating infections caused by S. aureus.

A Dose-Response Investigation of a Micronized Porous Ceramic Particle to Improve the Health and Performance of Post-weaned Pigs Infected With Salmonella enterica Serotype Typhimurium

The objective was to investigate the effects of supplementing increasing concentrations of PowerGuard (PG), a micronized ceramic particle, to weaned pigs on health and performance following a Salmonella enterica serotype Typhimurium infection. Forty barrows were transported to the USDA facility in Liberty, TX, USA. Pigs were randomly assigned to one of five treatments (n = 8): (1) uninfected control (CON), no Salmonella typhimurium (ST) and no PG treatment; (2) infected control (ST), infected with ST but no PG treatment; (3) PG0.05, infected with ST and supplemented with 0.05% PG; (4) PG0.25, infected with ST and supplemented with 0.25% PG; and (5) PG0.50, infected with ST and supplemented with 0.5% PG. All pigs were enrolled at 21.5 ± 1.33 days of age and did not differ in initial BW (1.98 ± 0.09 kg). Pigs were anesthetized to insert temperature recording devices into the abdominal cavity. Pigs were offered feed and water ad libitum. Pigs in ST, PG0.05, PG0.25, and PG0.50 were infected orally with 1.75 × 107 colony-forming units of Salmonella typhimurium on day 7. Pig body weights and peripheral blood samples were collected on days 0, 7, 10, 14, and 21. Pigs were harvested on day 21 and ileum and liver samples were collected for histopathological analyses. There was no treatment difference for final BW (P ≥ 0.201). There was a tendency (P = 0.087) for a treatment difference in the fecal score; ST and PG0.50 had more loose fecal scores than CON and PG0.25. There was a treatment × time interaction for intraperitoneal temperature (P < 0.0001); PG0.05, PG0.25, and PG0.50 had attenuated febrile responses during the acute post-infection period compared with ST. There was a treatment × time interaction for total leukocyte counts (P = 0.007); PG treatments reduced leukocytosis post-infection compared with ST. Supplementing PG0.25 improved many health and performance variables when pigs were infected with Salmonella Typhimurium. Furthermore, supplementing PG0.05 attenuated the febrile response and many hematological variables. However, supplementing PG0.5 did not improve many aspects of health or performance. Therefore, supplementing PowerGuard between 0.05 and 0.25% of the diet may play a role in protecting weaned pigs from disease caused by Salmonella.

Pathology of Salmonella enterica Subspecies enterica Serotype Typhimurium Infection in Chinchillas (Chinchilla lanigera)

Septicaemia is the main pathological manifestation of Salmonella infection in chinchillas (Chinchilla lanigera), although information on its pathology is limited. We now describe the gross, histological and immunohistochemical features of Salmonella enterica subspecies enterica serotype Typhimurium (S. Typhimurium) infection in ranched chinchillas. Eighty-five adult pregnant chinchillas had anorexia, prostration, weight loss, hyperthermia and abortion. Necropsy of 13 animals revealed splenomegaly, hepatomegaly, mesenteric lymphadenomegaly, prominent Peyer's patches, pulmonary oedema, white pinpoint lesions in multiple organs and petechiae in the urinary bladder. In all cases, histological lesions were consistent with septicaemia, characterized by acute necrotizing hepatitis, splenitis, lymphadenitis, cystitis, pneumonia, enterocolitis, gastritis and/or nephritis. Immunohistochemistry for Salmonella spp on seven cases revealed intralesional immunolabelling in all affected organs. Salmonella sp was isolated from liver, spleen, intestinal contents and blood of 10 chinchillas. Isolates of four animals were identified as S. Typhimurium.

Toll-like Receptor 11 (TLR11) Prevents Salmonella Penetration into the Murine Peyer Patches

Toll-like receptors (TLRs) are key molecular sensors used by the mammalian innate immune system to detect microorganisms. Although TLR functions in colonic immune homeostasis and tolerance to commensal bacteria have been intensively researched, the precise roles of different TLRs in response to pathogen infection in the gut remain elusive. Peyer patches are the major entrance of Salmonella infection and antigen transportation in intestine. Here, we report that, in contrast to TLR5 as a "carrier of Salmonella," TLR11 works as a "blocker of Salmonella" to prevent highly invasive Salmonella from penetrating into the murine Peyer patches and spreading systemically. TLR11 plays an important role in mediating TNF-α induction and systemic inflammation in response to Salmonella infection. Remarkably, in mice lacking TLR11, apparent hemorrhages at Peyer patches are induced by highly invasive Salmonella, a phenotype resembling human Salmonella infection. Therefore, our results indicate a potentially important role for TLR11 in preventing murine intestinal infection and modulating antigen transportation in the gut and imply an important role for various TLRs in cooperation with tight control of pathogens penetrating into Peyer patches. The TLR11 knock-out mouse can serve as a good animal model to study Salmonella infection.

Ecto-5'-nucleotidase (CD73) regulates host inflammatory responses and bacterial persistence during murine Salmonellosis (169.10)

Abstract Salmonella spp. is responsible for the vast majority of foodborne illnesses in the US. Adenosine is an antiinflammatory mediator that limits tissue damage during inflammation. Surface enzymes CD39 & CD73 mediate the synthesis of extracellular adenosine & can regulate immune responses. We studied the expression of CD39 & CD73 in liver & spleen after infection of C57BL/6 mice with Salmonella enterica serotype Typhimurium (ST) & evaluated the role of CD73 after oral infection of wildtype & CD73-KO mice, in regulating immune responses & bacterial persistence. Liver & spleen cytokine mRNA expression was tested by RT-PCR. Cytokines were assayed by ELISA from cultured supernatant of activated splenocytes. Splenocyte-derived Th cells were tested for intracellular cytokines by FACS. Inflammatory responses in infected CD73-/- mouse livers were tested in H&E sections. Bacterial count was done from liver tissues. After ST infection of wildtype mice, both CD39 & CD73 transcript levels declined in spleen & liver. Splenocytes from ST-infected CD73-/- mice produced significantly more IFN-γ & IL17a. Infected CD73-/- mice had higher expressions of IFN-γ & TNF-α mRNA in liver. Histological examinations of liver sections showed significantly greater number of “inflammatory-foci”. Liver bacterial load was significantly lower in CD73-/-mice. CD73 expression contributes to adenosine accumulation that attenuates inflammation during Salmonellosis & may impair immunity to favor bacterial persistence.

An Outbreak ofSalmonellaSerotype Typhimurium Infections with an Unusually Long Incubation Period

A 1998 investigation of an outbreak of Salmonella serotype Typhimurium infections among children tasting unpasteurized milk during tours of a dairy farm demonstrated a distribution of unusually long incubation periods (median, 8 days; interquartile range [IQR], 6-14 days). Bacterial isolates were highly acid tolerant and contained genes associated with protection against destructive phagocytic reactive oxygen intermediates. We hypothesize that exposure to low-dose oral inoculum of a pathogen with these properties could have contributed to cases of non-typhoidal salmonellosis with the longest incubation period reported to the Centers for Disease Control and Prevention (CDC).

Multistate outbreak ofSalmonellaserotype Typhimurium infections associated with consumption of restaurant tomatoes, USA, 2006: hypothesis generation through case exposures in multiple restaurant clusters

Multiple salmonellosis outbreaks have been linked to contaminated tomatoes. We investigated a multistate outbreak of Salmonella Typhimurium infections among 190 cases. For hypothesis generation, review of patients' food histories from four restaurant-associated clusters in four states revealed that large tomatoes were the only common food consumed by patients. Two case-control studies were conducted to identify food exposures associated with infections. In a study conducted in nine states illness was significantly associated with eating raw, large, round tomatoes in a restaurant [matched odds ratio (mOR) 3·1, 95% confidence interval (CI) 1·3-7·3]. In a Minnesota study, illness was associated with tomatoes eaten at a restaurant (OR 6·3, mid-P 95% CI 1·05-50·4, P=0·046). State, local and federal regulatory officials traced the source of tomatoes to Ohio tomato fields, a growing area not previously identified in past tomato-associated outbreaks. Because tomatoes are commonly eaten raw, prevention of tomato contamination should include interventions on the farm, during packing, and at restaurants.

Will the Food Safety Modernization Act Help Prevent Outbreaks of Foodborne Illness?

Large-scale outbreaks of foodborne illness have recently focused attention on the ability of the U.S. food-safety system to protect the public health. The nationwide outbreak of Salmonella enterica serotype Typhimurium infection associated with peanut products that is described by Cavallero et al. is one example.1 This contamination, which was ultimately traced to the Peanut Corporation of America (PCA), took a high toll — 714 people were affected, about 200 were hospitalized, and 9 died. Investigators found multiple potential routes of contamination at PCA facilities, such as rain leakage and cross-contamination between raw and roasted peanuts. Although the outbreak was eventually . . .

Adenosine synthesized by CD73 and CD39 regulates inflammation during murine Salmonella infection (162.10)

Abstract Salmonella spp. is a leading cause of hospitalization & death in the US associated with foodborne illness. Salmonella causes acute gut inflammation. Adenosine is an antiinflammatory mediator that limits tissue damage during inflammation. Surface enzymes CD39 & CD73 mediate the synthesis of adenosine. We studied the expression of CD39 & CD73 in liver, spleen, & CD4+ Th cells after infection of C57BL/6 mice with Salmonella enterica serotype Typhimurium (ST) & evaluated the role of CD73 in regulating ST-induced inflammation. Proinflammatory cytokine mRNA expression was assayed by RT-PCR arrays & cytokine proteins were tested by FACS & ELISA after ST infection or ST antigen (Ag) treatment. Function of CD73 was tested by examining proinflammatory cytokine production with or without CD73-inhibitor treatment of activated splenocytes exposed to ST Ag. Inflammatory responses to ST infection in CD73-/- mice were evaluated in liver & spleen tissue in H&E sections & in activated CD4+ Th cells stained for intracellular IL17a & IFN-γ. ST Ag-treated & CD73-inhibited splenocytes produced more proinflammatory cytokines (IL17A). Splenocytes from ST-infected CD73-/- mice produced more IL17a cytokine & Th17 cells & infected CD73-/- mice also had greater infiltration of inflammatory cells in the liver. Extracellular adenosine synthesized by CD73 may regulate host inflammation during murine Salmonellosis.

Enteric Pathology and Salmonella-Induced Cell Death in Healthy and SIV-Infected Rhesus Macaques

The goal of this study was to morphologically characterize a ligated ileal loop model of Salmonella enterica serotype Typhimurium infection in rhesus macaques (Macaca mulatta) and to verify the occurrence of Salmonella-induced cell death in vivo. Eight adult healthy male rhesus macaques were used for ligated ileal loop surgery. Four macaques had been intravenously inoculated with simian immunodeficiency virus (SIV) mac251. Ileal ligated loops were inoculated with wild-type (WT) S. Typhimurium strain IR715 (ATCC14028 nal (r)), an isogenic noninvasive mutant strain (ATCC14028 nal (r) ΔsipAΔsopABDE2), or sterile Luria Bertani broth. Loops were surgically removed at 2, 5, and 8 hours post-inoculation (hpi). Intestinal samples were processed for histopathology, immunohistochemistry for detecting Salmonella, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL), and transmission electron microscopy. Combined histopathology scores were similar between SIV-infected and control macaques. As expected, the invasion-deficient mutant was less pathogenic than WT S. Typhimurium. Neutrophil infiltrate in the intestinal mucosa correlated with bacterial loads (r = 0.7148; P < .0001) and fluid accumulation (r = 0.6019; P < .0001) in the lumen of the intestinal loops. Immunolabeled WT S. Typhimurium was observed in the epithelium and lamina propria at the tip of the villi at 2 hpi, progressing toward deeper lamina propria at 5-8 hpi. Most TUNEL-positive cells localized to the lamina propria, and some had morphological features of macrophages. Ultrastructurally, bacteria were observed intracellularly in the lamina propria as well as within apoptotic bodies. This study provides morphological evidence of Salmonella-induced cell death in vivo in a relevant nonhuman primate model.

DNA microarray analysis of Salmonella serotype Typhimurium strains causing different symptoms of disease

BackgroundSalmonella enterica subsp. enterica is one of the leading food-borne pathogens in the USA and European countries. Outcome of human Salmonella serotype Typhimurium infections ranges from mild self-limiting diarrhoea to severe diarrhoea that requires hospitalization. Increased knowledge of the mechanisms that are responsible for causing infection and especially the severity of infection is of high interest.ResultsStrains were selected from patients with mild infections (n = 9) and patients with severe infections (n = 9) and clinical data allowed us to correct for known underlying diseases. Additionally, outbreak isolates (n = 3) were selected. Strains were analyzed on a DNA-DNA microarray for presence or absence of 281 genes covering marker groups of genes related to pathogenicity, phages, antimicrobial resistance, fimbriae, mobility, serotype and metabolism. Strains showed highly similar profiles when comparing virulence associated genes, but differences between strains were detected in the prophage marker group. The Salmonella virulence plasmid was present in 72% of the strains, but presence or absence of the virulence plasmid did not correspond to disease symptoms. A dendrogram clustered strains into four groups. Clustering confirmed DT104 as being a clonal phagetype. Clustering of the remaining strains was mainly correlated to presence or absence of the virulence plasmid and mobile elements such as transposons. Each of the four clusters in the tree represented an almost equal amount of strains causing severe or mild symptoms of infection.ConclusionsWe investigated clinical significance of known virulence factors of Salmonella serotype Typhimurium strains causing different disease symptoms, and conclude that the few detected differences in Salmonella serotype Typhimurium do not affect outcome of human disease.

Contribution of Flagellin Pattern Recognition to Intestinal Inflammation duringSalmonella entericaSerotype Typhimurium Infection

Salmonella enterica serotype Typhimurium causes acute inflammatory diarrhea in humans. Flagella contribute to intestinal inflammation, but the mechanism remains unclear since most mutations abrogating pattern recognition of flagellin also prevent motility and reduce bacterial invasion. To determine the contribution of flagellin pattern recognition to the generation of innate immune responses, we compared in two animal models a nonmotile, but flagellin-expressing and -secreting serotype Typhimurium strain (flgK mutant) to a nonmotile, non-flagellin-expressing strain (flgK fliC fljB mutant). In vitro, caspase-1 can be activated by cytosolic delivery of flagellin, resulting in release of the interferon gamma inducing factor interleukin-18 (IL-18). Experiments with streptomycin-pretreated caspase-1-deficient mice suggested that induction of gamma interferon expression in the murine cecum early (12 h) after serotype Typhimurium infection was caspase-1 dependent but independent of flagellin pattern recognition. In addition, mRNA levels of the CXC chemokines macrophage inflammatory protein 2 and keratinocyte-derived chemokine were markedly increased early after serotype Typhimurium infection of streptomycin-pretreated wild-type mice regardless of flagellin expression. In contrast, in bovine ligated ileal loops, flagellin pattern recognition contributed to increased mRNA levels of macrophage inflammatory protein 3alpha and more fluid accumulation at 2 h after infection. Collectively, our data suggest that pattern recognition of flagellin contributes to early innate host responses in the bovine ileal mucosa but not in the murine cecal mucosa.

Interleukin-23 Orchestrates Mucosal Responses to Salmonella enterica Serotype Typhimurium in the Intestine

Salmonella enterica serotype Typhimurium causes an acute inflammatory reaction in the ceca of streptomycin-pretreated mice that involves T-cell-dependent induction of gamma interferon (IFN-gamma), interleukin-22 (IL-22), and IL-17 expression (genes Ifn-gamma, Il-22, and Il-17, respectively). We investigated here the role of IL-23 in initiating these inflammatory responses using the streptomycin-pretreated mouse model. Compared to wild-type mice, the expression of IL-17 was abrogated, IL-22 expression was markedly reduced, but IFN-gamma expression was normal in the ceca of IL-23p19-deficient mice during serotype Typhimurium infection. IL-23p19-deficient mice also exhibited a markedly reduced expression of regenerating islet-derived 3 gamma, keratinocyte-derived cytokine, and reduced neutrophil recruitment into the cecal mucosa during infection. Analysis of CD3(+) lymphocytes in the intestinal mucosa by flow cytometry revealed that alphabeta T cells were the predominant cell type expressing the IL-23 receptor in naive mice. However, a marked increase in the number of IL-23 receptor-expressing gammadelta T cells was observed in the lamina propria during serotype Typhimurium infection. Compared to wild-type mice, gammadelta T-cell-receptor-deficient mice exhibited blunted expression of IL-17 during serotype Typhimurium infection, while IFN-gamma expression was normal. These data suggested that gammadelta T cells are a significant source, but not the sole source, of IL-17 in the acutely inflamed cecal mucosa of mice. Collectively, our results point to IL-23 as an important player in initiating a T-cell-dependent amplification of inflammatory responses in the intestinal mucosa during serotype Typhimurium infection.

A comparison of cecal colonization of Salmonella enterica serotype Typhimurium in white leghorn chicks and Salmonella-resistant mice

BackgroundSalmonellosis is one of the most important bacterial food borne illnesses worldwide. A major source of infection for humans is consumption of chicken or egg products that have been contaminated with Salmonella enterica serotype Typhimurium, however our knowledge regarding colonization and persistence factors in the chicken is small.ResultsWe compared intestinal and systemic colonization of 1-week-old White Leghorn chicks and Salmonella-resistant CBA/J mice during infection with Salmonella enterica serotype Typhimurium ATCC14028, one of the most commonly studied isolates. We also studied the distribution of wild type serotype Typhimurium ATCC14028 and an isogenic invA mutant during competitive infection in the cecum of 1-week-old White Leghorn chicks and 8-week-old CBA/J mice. We found that although the systemic levels of serotype Typhimurium in both infected animal models are low, infected mice have significant splenomegaly beginning at 15 days post infection. In the intestinal tract itself, the cecal contents are the major site for recovery of serotype Typhimurium in the cecum of 1-week-old chicks and Salmonella-resistant mice. Additionally we show that only a small minority of Salmonellae are intracellular in the cecal epithelium of both infected animal models, and while SPI-1 is important for successful infection in the murine model, it is important for association with the cecal epithelium of 1-week-old chicks. Finally, we show that in chicks infected with serotype Typhimurium at 1 week of age, the level of fecal shedding of this organism does not reflect the level of cecal colonization as it does in murine models.ConclusionIn our study, we highlight important differences in systemic and intestinal colonization levels between chick and murine serotype Typhimurium infections, and provide evidence that suggests that the role of SPI-1 may not be the same during colonization of both animal models.

ERRATUM

In an article in the 1 February 2004 issue of the Journal (Martin LJ, Fyfe M, Doré K, et al. Increased burden of illness associated with antimicrobial-resistant Salmonella enterica serotype Typhimurium infections. J Infect Dis 2004; 189:377–384), there is an error in line 3 of the right-hand column on page 378. Not all isolates included in our study were obtained from stool samples: 4 isolates were obtained from unknown sources, 2 from urine samples, and 1 from a wound specimen. In addition, in the righthand column on page 381, in lines 6–9 of the first paragraph of the “Multivariate regression” subsection of the “Results,” the sentence should read as follows: “After eliminating case subjects treated with ineffective antimicrobial treatments and antimicrobial treatments of unknown effectiveness, these associations remained positive and significant.” The authors regret these errors.

A Multistate Outbreak of Salmonella enterica Serotype Typhimurium Infection Linked to Raw Milk Consumption—Ohio, 2003

In December 2002, the Ohio Department of Health was notified of two children with Salmonella infection. Both had a history of drinking raw milk from a combination dairy-restaurant-petting zoo (dairy). The dairy was the only establishment in Ohio licensed to sell raw milk and reported 1.35 million visitors annually. We investigated to determine the extent of the outbreak and identify illness risk factors. A case patient was any person with pulsed-field gel electrophoresis–matched Salmonella enterica serotype Typhimurium from 30 November 2002 to 18 February 2003. Sixty-two met the confirmed case definition. Forty dairy case patient patrons were included in a case-control study; 56 controls were their well meal companions. Consumption of raw milk was found to be associated with illness (odds ratio, 45.1; 95% confidence interval, 8.8 to 311.9). The dairy discontinued selling raw milk. Because 27 other states still allow the sale of raw milk, awareness of the hazards of its consumption should be raised and relevant regulations carefully reviewed.

Multidrug-resistant Salmonella Typhimurium infection from milk contaminated after pasteurization.

An outbreak of multidrug-resistant Salmonella enterica serotype Typhimurium infections occurred in Pennsylvania and New Jersey. A case-control study implicated pasteurized milk from a dairy, and an inspection indicated the potential for contamination after pasteurization. Dairy cattle are the likely reservoir, and milk may be an important vehicle of Salmonella transmission to humans.

Salmonella serotype Typhimurium infection of bovine Peyer's patches down-regulates plasma membrane calcium-transporting ATPase expression.

To identify host genes differentially expressed during Salmonella enterica serotype Typhimurium infection, an RNA differential display was made with total RNA extracted from ileal loops that were infected with Salmonella Typhimurium 2.5 h after infection. Down-regulated cDNA was identified in bovine Peyer's patches after infection that was highly homologous to a human plasma membrane calcium-transporting ATPase (PMCA). Differential expression of PMCA, evaluated by Northern analysis, was found to have more than a 4.6-fold decrease in expression of mRNA (size, approximately 5.1 kb). PMCA mRNA was detected by in situ hybridization exclusively within epithelial cells in the Peyer's patches. cDNA (4.4 kb) was amplified by rapid amplification of cDNA ends, cloned, and sequenced and showed a high homology to hPMCA. Bovine PMCA is down-regulated in epithelial cells of Peyer's patches after infection with Salmonella Typhimurium and, subsequently, may influence cellular calcium levels that contribute to the inflammatory processes in the pathogenesis of diarrhea.

Salmonella Typhimurium infections transmitted by chlorine-pretreated clover sprout seeds.

Raw seed sprouts have caused numerous outbreaks of enteric infections. Presoaking seeds in a 20,000 mg/liter (ppm) calcium hypochlorite solution before sprouting is recommended to reduce bacterial contamination and infection risk. In 1999, the authors investigated an outbreak of Salmonella serotype Typhimurium infections in Colorado. In a case-control study, they matched 20 cases with 58 controls by age, sex, and telephone prefix; 10 (52%) of 19 cases and no controls recalled eating raw alfalfa-style sprouts in the 5 days before the patient's illness (p < 0.00001). Traceback implicated clover sprouts grown from seeds shared by two sprouters. The time period and region over which these sprouts were sold matched the occurrences of 112 culture-confirmed illnesses. Only one of the sprouters presoaked seeds as recommended, and fewer infections were attributable to this sprouter (0.29 vs. 1.13 culture-confirmed infections/50-pound (110.1-kg) bag of seed). After recall of the implicated sprouts and seed, S. Typhimurium illnesses declined. Contaminated raw clover sprouts can cause outbreaks of enteric illness. Presoaking contaminated seeds in a 20,000 mg/liter calcium hypochlorite solution reduces, but does not eliminate, the risk of infection. Until safer production methods are developed, persons eating raw sprouts continue to risk developing potentially serious gastrointestinal illness.

Outbreaks of Multidrug-ResistantSalmonellaTyphimurium Associated With Veterinary Facilities—Idaho, Minnesota, and Washington, 1999

CDC received reports in 1999 from three state health departments of outbreaks of multidrug-resistant Salmonella serotype Typhimurium infections in employees and clients of small animal veterinary clinics and an animal shelter. Salmonella infections usually are acquired by eating contaminated food; however, direct contact with infected animals, including dogs and cats, also can result in exposure and infection. This report summarizes clinical and epidemiologic data about these outbreaks and reviews methods of reducing the likelihood of Salmonella transmission in veterinary settings by avoiding fecal-oral contact.