Salmonella in dairy operations in the United States: prevalence and antimicrobial drug susceptibility.

Salmonella serotypes are important foodborne pathogens of humans that can be acquired through consumption of contaminated meat and dairy products. Salmonella infection also can be a significant animal health issue. As part of a national study of U.S. dairy operations conducted between March and September 2002, fecal samples were collected from representative cows in 97 dairy herds in 21 states and were cultured to determine the prevalence of Salmonella shedding. Salmonella was recovered from the feces of at least one cow in 30.9% of the herds. Overall, 7.3% of fecal samples were culture positive for Salmonella. The three most frequently recovered serotypes were Salmonella Meleagridis (24.1%), Salmonella Montevideo (11.9%), and Salmonella Typhimurium (9.9%). The susceptibilities of Salmonella isolates recovered were determined using a panel of 16 antimicrobial drugs. Salmonella isolates recovered from dairy cows had relatively little resistance to these antimicrobial agents; 83.0% of the isolates were susceptible to all antimicrobials tested. This study provides updated information on the prevalence and susceptibility patterns of Salmonella in dairy herds and on cow and herd characteristics. These data contribute to our understanding of the ecology of Salmonella in the dairy farm environment.

Prevalence and antimicrobial susceptibility of Salmonella spp. isolates from US cattle in feedlots in 1999 and 2000.

AIMS: Faecal samples from cattle in US feedlots were evaluated for the presence of Salmonella. When Salmonella isolates were recovered the antimicrobial resistance patterns were determined. METHODS AND RESULTS: Faecal samples were collected from pen floors in 73 feedlots in 12 states during the period from October 1999 to September 2000. Pens of cattle selected for sampling were those that had been in the feedlot for the shortest period of time, the longest period of time and a randomly selected pen from the remaining pens. Faecal samples were cultured for Salmonella spp. and all Salmonella isolates were categorized by serotype. The susceptibilities of all isolates were determined using a panel of 17 antimicrobials. Overall, 6.3% (654/10,417) of the samples cultured positive for Salmonella spp. and 22.2% (94/422) of pens and 50.7% (37/73) of feedlots had one or more positive samples. There was little difference in the proportion of positive samples from short-fed (6.1%, 212/3482), random (6.4%, 217/3400) and long-fed (6.4%, 224/3485) pens of cattle. One of two pens of cattle that could not be attributed to a pen type had a single positive sample (2.0%, 1/50). Samples collected during the period of April to June (6.8%, 209/3054) and July to September (11.4%, 286/2500) were more likely to be positive than those collected during October to December (4.0%, 73/1838) and January to March (2.8%, 86/3025). The most common serotypes of Salmonella were dissimilar from those that are typically seen in human illness and cattle illness. The majority of isolates (62.8%, 441/702) were sensitive to all of the antimicrobials tested. Resistance was most frequently observed to tetracycline (35.9%, 252/702) followed by streptomycin (11.1%, 78/702), ampicillin (10.4%, 73/702) and chloramphenicol (10.4%, 73/702). Multiple resistance (resistance to > or =2 antimicrobials) was observed for 11.7% (82/702) of the isolates. CONCLUSIONS: Salmonella was isolated at low frequency from faeces of feedlot cattle and the serotypes were not those commonly associated with human illness. In addition most of the Salmonella isolates were sensitive to all the antimicrobials tested. SIGNIFICANCE AND IMPACT OF THE STUDY: This study contributes to understanding the ecology of Salmonella in cattle feedlots and the prevalence of resistance among potential food-borne pathogens.

Factors associated with fecal-shedding of Salmonella spp by horses on US operations

In a cross-sectional national study that included 972 operations with > 3 horses on 1/1/98 in 28 states in the USA, 8,417 fecal specimens were collected from horses and cultured to test for the presence of Salmonella spp. Operations were characterized as Salmonella spp-positive if at least one fecal specimen tested positive for Salmonella spp. Percentages of Salmonella spp-positive operations were computed by management and other factors (collected from operation-level questionnaires) that were hypothesized to be related to fecal shedding of Salmonella spp. A logistic-regression model was constructed to identify factors associated with horsesÆ shedding Salmonella spp in feces on an operation. The odds of an operation being Salmonella spp positive increased as the number of resident horses increased. In addition, the following factors were found to be associated with increased odds of an operation being Salmonella spp positive: horses were used primarily for breeding; operation cleanliness was characterized as poor by the data collector; and new resident equids had been added to the operation without routine quarantine

Fecal shedding of Salmonella spp by horses in the United States during 1998 and 1999 and detection of Salmonella spp in grain and concentrate sources on equine operations.

OBJECTIVE: To estimate prevalence of fecal shedding of Salmonella spp among horses in the US horse population and prevalence of Salmonella spp in grain or other concentrate used as horse feed on equine operations in the United States. DESIGN: Cross-sectional survey. SAMPLE POPULATION: Horses on 972 operations in 28 states. PROCEDURE: Fecal samples were collected from horses resident at each operation. Only a single sample was collected from any individual horse; number of horses from which samples were collected on each operation was determined on the basis of number of horses on the operation. A single sample of grain or concentrate was also collected from each operation. All samples were tested for Salmonella spp by means of bacterial culture. RESULTS: Overall, 0.8% (SE, 0.5) of resident horses shed Salmonella spp in their feces. The overall prevalence of operations positive for fecal shedding of Salmonella spp (i.e., operations with > or = 1 horse shedding Salmonella spp in its feces) was 1.8% (SE, 0.7). Prevalence of grain or other concentrate samples positive for Salmonella spp was 0.4%. Serotypes of Salmonella spp that were identified in grain or other concentrate were not those typically associated with clinical disease in horses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the national prevalence of fecal shedding of Salmonella spp by horses in the United States was 0.8%, and that prevalence of Salmonella spp in grain or other concentrate used for horse feed was 0.4%.

Evaluation of invasion-conferring genotypes and antibiotic-induced hyperinvasive phenotypes in multiple antibiotic resistant Salmonella typhimurium DT104.

Antibiotic resistance in pathogenic bacteria is a problem in both industrialized and developing countries. This is especially evident in Salmonella typhimurium, a foodborne pathogen that causes gastrointestinal and systemic disease throughout the world. S. typhimurium DT104 further poses a major health concern due to its apparent enhanced ability to acquire multiple antibiotic resistance genes and its putative hypervirulent phenotype. Recently, we demonstrated that multiresistant S. typhimurium do not appear to be more invasive than non-resistant cohorts. In the present study, we evaluated the presence of Salmonella pathogenicity island 1 (SPI1) flanking and internal sequences in over 400 isolates of multiresistant S. typhimurium. With these same isolates, we also used a tissue culture invasion assay to evaluate a potential relationship between antibiotic exposure and a hyperinvasive phenotype. Our studies revealed that SPI1 flanking sequences are similar in multiresistant and non-resistant S. typhimurium. Furthermore, we failed to identify any isolates that were hyperinvasive in the presence of any of the 14 antibiotics evaluated. These results further indicate that the putative hypervirulence of multiresistant S. typhimurium is not likely to occur at the level of invasion.

Multidrug-resistant Salmonella Typhimurium DT104 in poultry.

Salmonella Typhimurium isolates from feed ingredients or poultry sources isolated during 1995 to 1997 from different geographical locations within Minnesota were examined for the presence of Salmonella Typhimurium definitive type 104 (DT104). Antibiotic susceptibility studies indicated that 15 of 50 isolates of Salmonella Typhimurium had an antibiotic resistance pattern (ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline) that is usually observed with multidrug-resistant Salmonella Typhimurium DT104. Of the 15 isolates showing the antibiotic resistance pattern, 8 isolates were phage type 104, 3 isolates were typed as phage type 104 complex, and the remaining 4 isolates belonged to phage types 193, 81, and 126. DT104 was recovered from both feed ingredients and poultry samples. Of the seven feed ingredient-associated Salmonella Typhimurium isolates, four were DT104, whereas only 7 of 43 poultry-associated Salmonella Typhimurium isolates were DT104. A repetitive sequence-based polymerase chain reaction (rep-PCR) of 50 isolates of Salmonella Typhimurium representing 13 phage types identified seven distinct fingerprint profiles. No correlation between phage type and rep-PCR type was noticed. Eleven Salmonella Typhimurium isolates belonging to DT104 and its complex were grouped into two closely related rep-PCR types.

Augmentation of antibiotic resistance in Salmonella typhimurium DT104 following exposure to penicillin derivatives.

Antibiotic resistance in pathogenic bacteria has been a problem in both developed and developing countries. This problem is especially evident in Salmonella typhimurium, one of the most prevalent foodborne pathogens. While performing in vitro gentamicin protection-based invasion assays, we found that certain isolates of multiresistant S. typhimurium can be 'induced' to exhibit new resistance profiles. That is, bacteria become resistant to a wider range of antibiotics and they also exhibit quantitative increases in MIC values for antibiotics that were part of their pre-induction antibiograms. This 'induction' process involves growing the bacteria to stationary phase in the presence of antibiotics such as ampicillin, amoxicillin or ticarcillin. Since the isolates studied exhibited resistance to ampicillin, amoxicillin and ticarcillin prior to exposing the bacteria to these antibiotics, the observed phenomenon suggests that resistant Salmonella not only have a selective advantage over non-resistant Salmonella but their resistance phenotypes can be accentuated when an inappropriate antibiotic is used therapeutically.

Improved plating media for the detection of Salmonella species with typical and atypical hydrogen sulfide production.

The Salmonella detection ability of 2 surfactant-supplemental media, xylose-lysine-tergitol (Nia-proof) 4 (XLT4) and Miller-Mallinson (MM) agar, was compared against that of several commonly used plating media. XLT4 and MM appeared to be the most efficient in detecting Salmonella in meat products and food animal environments. MM was superior to XLT4 in detecting those increasingly more prevalent strains of Salmonella possessing weak to ultraweak H2S production characteristics.

Survey of Salmonella serotypes shed in feces of beef cows and their antimicrobial susceptibility patterns.

Salmonella prevalence on cow-calf operations was studied as a part of a national study of health and management of the U.S. beef cow-calf industry and was conducted as part of the National Animal Health Monitoring System. Within this study, the prevalence of Salmonella spp. shed in feces was determined. A total of 5,049 fecal samples were collected from 187 beef cow-calf operations each visited on a single occasion. The number of fecal samples collected per operation was predetermined based on herd size. Salmonellae were recovered from 1 or more fecal samples collected on 11.2% (21 of 187) of the operations. Overall 78 salmonellae representing 22 serotypes were recovered from 1.4% (70 of 5,049) of samples. Multiple serotypes were recovered from eight samples from a single operation. The five most common serotypes were Salmonella Oranienburg (21.8% of isolates), and Salmonella Cerro (21.8%), followed by Salmonella Anatum (10.3%), Salmonella Bredeney (9.0%), and Salmonella Mbandaka (5.1%). The most common serogroups identified were C1 (33.3%), K (21.8%), B (16.7%), and E (15.4%). Even though the recovery rate of salmonellae from fecal samples was very low, 43.6% (34 of 78) and 38.5% (30 of 78) of the isolates were among the 10 most common serotypes from cattle with clinical signs of disease or isolated from humans, respectively. The majority of the isolates (50 of 78; 64.1%) were recovered from fecal samples from two operations. All isolates were screened for resistance to a panel of 17 antimicrobics, and 87.2% (68 of 78) were susceptible to all of the antimicrobics. The resistant isolates were most commonly resistant to streptomycin (n = 9) and/or sulfamethoxazole (n = 9). Nine isolates showed multiple (> or =2 antimicrobics) resistance most commonly to streptomycin and sulfamethoxazole (n = 6).

Identification of diminished tissue culture invasiveness among multiple antibiotic resistant Salmonella typhimurium DT104.

Salmonella infections continue to cause gastrointestinal and systemic disease throughout the world. Salmonella typhimurium further poses a major health concern due to its apparent enhanced ability to acquire multiple antibiotic resistance genes. Currently it is unclear if multiresistant S. typhimurium are more or less pathogenic than non-resistant counterparts. Using an in vitro invasion assay, we evaluated the relative pathogenicity of over 400 multiresistant S. typhimurium isolates. Our studies failed to identify any <<<>>> isolates. However, we identified 12 isolates exhibiting invasive phenotypes that were constrained relative to controls. These <<<>>> strains were found in a variety of phagetypes all possessing at least a hexaresistant profile. Further studies revealed that the alterations in invasion were not due to changes in adherence. Limited studies exploring in vivo virulence revealed a mildly decreased ability to cause murine lethality for the hypoinvasive strain examined. These results indicate that the ability to cause disease is not increased but is rather mildly attenuated for certain isolates of multiresistant S. typhimurium.

Prevalence of fecal shedding of Salmonella organisms among captive green iguanas and potential public health implications.

OBJECTIVE: To determine prevalence of fecal shedding of Salmonella organisms among captive green iguanas (Iguana iguana). DESIGN: Cohort study. ANIMALS: 12 captive green iguanas. PROCEDURE: Iguanas were isolated in an environmental chamber, and fecal samples were collected weekly for 10 consecutive weeks. Samples were incubated aerobically in tetrathionate broth for 18 to 24 hours. Aliquots were then transferred to Hektoen and Salmonella-Shigella agar plates and incubated for an additional 18 to 24 hours. Isolated colonies were subcultured on nutrient agar slants, and Salmonella isolates were serogrouped and serotyped. RESULTS: All 12 iguanas were found to be shedding Salmonella organisms at least once during the study, and multiple serotypes were isolated from 7 of the 12. Salmonella organisms were isolated from 88 of 106 (83%) fecal samples; 21 samples contained multiple Salmonella serotypes. Overall, 11 Salmonella serotypes were identified. In 74 of 100 instances, when a particular Salmonella serotype was isolated from an individual iguana, the same serotype was also isolated from a subsequent fecal sample from that iguana. CLINICAL IMPLICATIONS: Results suggested that most iguanas have a stable mixture of Salmonella serotypes in their intestinal tracts and intermittently or continuously shed Salmonella organisms in their feces. Veterinarians should advise their clients on precautions for reducing the risk of acquiring these organisms from their pets. Public health officials trying to determine whether an iguana is the source of a specific Salmonella serotype that caused infection in human patients should submit at least 3 fecal samples collected from the iguana 1 week apart for bacterial culture.

Occurrence of Salmonella enteritidis in unpasteurized liquid egg in the United States.

In order to gain a greater understanding of the occurrence and distribution of Salmonella enteritidis in the United States, a survey of unpasteurized liquid egg (collected at 20 egg-breaking plants across the United States) was conducted over a 52-week period. Weekly liquid egg samples were submitted for Salmonella culturing at the National Veterinary Services Laboratories. Group D positive salmonellae were serotyped and phage-typed. On a regional basis, the Northern Region of the United States had the highest S. enteritidis recovery, with 20% of the samples submitted from plants in that region culture-positive for this serotype. Salmonella enteritidis positives from the Southeast, Central, and Western regions were 10%, 15%, and 6% of the samples submitted, respectively.

Plasmid profile analysis, phage typing, and antibiotic sensitivity of Salmonella dublin from clinical isolates in the United States.

One hundred clinical isolates of Salmonella choleraesuis subsp. choleraesuis serovar dublin (Salmonella dublin) were examined for phage sensitivity, antibiotic resistance patterns, and plasmid content. Computer analysis of the lysis patterns observed by using 27 typing phages divided the S. dublin isolates into 26 groups. One lytic pattern (Designated pattern 16) contained 52% of the isolates examined whereas 16 isolates had unique patterns, and nine patterns had fewer than ten members. Although 14 antibiotic resistance patterns were observed among the 100 isolates, 79% of the isolates grouped in three major patterns. Seven plasmid groups were identified and designated A-G based on the large plasmids found in the isolates. Of the 100 isolates, 28 contained the plasmid profile of Group A, 28 were Group B, 7 were Group C, 34 were Group D, and 1 isolate each was observed in Groups E, F, and G. The strong association between antibiotic resistance pattern and plasmid type suggest that the drug resistance genes are plasmid borne.

Molecular evolutionary genetics of the cattle-adapted serovar Salmonella dublin.

An electrophoretic analysis of allelic variation at 24 enzyme loci among 170 isolates of the serovar Salmonella dublin (serotype 1,9,12[Vi]:g,p:-) identified three electrophoretic types (Du 1, Du 3, and Du 4), marking three closely related clones, one of which (Du 1) is globally distributed and was represented by 95% of the randomly selected isolates. All but 1 of 114 nonmotile isolates of serotype 1,9,12:-:- recovered from cattle and swine in the United States were genotypically Du 1. The virulence capsular polysaccharide (Vi antigen) is confined to clone Du 3, which apparently is limited in distribution to France and Great Britain. For all 29 isolates of Du 3, positive signals were detected when genomic DNA was hybridized with a probe specific for the ViaB region, which contains the structurally determinant genes for the Vi antigen; and 23 of these isolates had been serologically typed as Vi positive. In contrast, all 30 isolates of Du 1 tested with the ViaB probe were negative. These findings strongly suggest that the ViaB genes were recently acquired by S. dublin via horizontal transfer and additive recombination. The clones of S. dublin are closely similar to the globally predominant clone (En 1) of Salmonella enteritidis (serotype 1,9,12:g,m:-) in both multilocus enzyme genotype and nucleotide sequence of the fliC gene encoding phase 1 flagellin. Comparative sequencing of fliC has revealed the molecular genetic basis for expression of the p and m flagellar epitopes by which these serovars are distinguished in the Kauffmann-White serological scheme of classification.

Experimental establishment of persistent infection in swine with a zoonotic strain of Salmonella newport.

An experiment was conducted to determine whether a persistent Salmonella newport infection could be established in swine, to determine duration of shedding and distribution of the organism in internal organs, and to determine whether changes occurred in antimicrobial susceptibility or plasmid profile of the organism during the course of long-term infection. Naturally farrowed Salmonella-free pigs (n = 22) were orally exposed to a multiply antimicrobial-resistant zoonotic strain of S newport when they were 7 weeks old. Tonsillar and rectal swab specimens were examined bacteriologically for S newport during the first week after exposure, then weekly for 7 weeks. Fecal samples were likewise examined weekly or every 2 weeks for 28 weeks after exposure. Necropsies of 2 or 3 randomly selected pigs were conducted at 2, 4, 8, 12, 16, 20, 24, and 28 weeks after exposure. A total of 45 specimens/pig representing the following internal organs or tissues were examined bacteriologically for S newport: liver, spleen, kidney, gallbladder, heart, heart blood, lung, stomach, and tonsils; segments of the intestinal tract with corresponding lymph nodes; and lymph nodes from lymphocenters of the head and neck, thoracic cavity, thoracic limbs, abdominal viscera, and abdominal wall. Exposure to S newport induced a mild and transient clinical response. The organism was recovered from 97% of tonsillar swab specimens and 89% of rectal swab specimens collected during 7 weeks after exposure and from 98% of fecal samples collected during 28 weeks after exposure.(ABSTRACT TRUNCATED AT 250 WORDS)