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Using a portable real-time PCR assay to detect Salmonella in raw milk.

The purpose of this study was to determine the efficacy of a portable real-time PCR system in detecting Salmonella spp. in raw milk. The 200 bulk milk samples chosen for this study constituted a subset of the samples for a larger study; this subset contained 24 samples that were culture positive for Salmonella and 176 that were culture negative. Milk was both plated directly on selective agar and plated after enrichment in selective media. Presumptive Salmonella colonies were isolated by direct culturing of five samples, while Salmonella was isolated from the remaining 19 positive samples only after enrichment. Presumptive Salmonella isolates were serotyped, and isolates from 22 samples were confirmed to be Salmonella isolates. PCR assays of culture-positive milk prior to enrichment yielded no evidence of Salmonella. DNA extracts of bacterial pellets from the enriched samples were analyzed for Salmonella by real-time PCR with the Ruggedized Advanced Pathogen Identification Device (RAPID). Fifty-four samples from the enrichment pellets tested positive for Salmonella by real-time PCR. Two samples that tested positive for Salmonella by culture and serotyping tested Salmonella negative by real-time PCR. Serotyping identified isolates from these samples as Salmonella Montevideo. All DNA extracts of Salmonella Montevideo isolates tested positive for Salmonella by real-time PCR. Thirty-three samples tested negative by culture and positive by real-time PCR. These results indicate that the portable real-time PCR system appears to be a useful tool for detecting Salmonella in raw milk. Additionally, the combination of enrichment and real-time PCR techniques used in this study can yield results in 24 h, compared with the 48 to 72 h required for traditional culture.

Animals↗

Fate of salmonellas and competing flora in meat sample enrichments in buffered peptone water and in Muller-Kauffmann's tetrathionate medium.

Studies have been carried out in which growth patterns of a Salmonella sp. and competing micro-organisms, especially other Enterobacteriaceae, were followed during pre-enrichment in buffered peptone water (BPw) and subsequent selective enrichment in tetrathionate broth (TBB). Pre-enrichment cultures were inoculated with minced meat and three reference samples containing nalidixic acid-resistant salmonellas. Irrespective of their initial numbers in BPw, Enterobacteriaceae increased to 10(8)/ml or more. During incubation in TBB at 43 degrees C, numbers of lactose-positive Enterobacteriaceae decreased in most enrichments which resulted in a positive salmonella isolation, but remained constant in the majority of those that did not. Levels of lactose-negative Enterobacteriaceae did not decrease in most salmonella-positive tests, but did so in half of the salmonella-negative ones. In the salmonella-positive tests the numbers of salmonellas had increased to 10(3)-10(7)/ml in BPw and after transfer to TBB slowly reached 10(4)/ml or more. In all cases the numbers of salmonellas exceeded those of the competing flora on brilliant green agar (BGA). In the salmonella-negative tests the numbers of salmonellas had increased less in BPw and decreased in most of the TBB enrichments. In none of these negative tests did the numbers of salmonellas exceed those of the competing flora on BGA. Escherichia coli dominated in most of the salmonella-negative tests. The results suggest more influence of lactose-positive than lactose-negative Enterobacteriaceae on the detection of salmonellas. The effect of competing microorganisms seems to depend not only upon their initial numbers, but also upon the types that can interact with salmonellas during selective enrichment.

Culture Media↗

Prevalence of Salmonella spp on conventional and organic dairy farms.

OBJECTIVE: To describe the occurrence of fecal shedding, persistence of shedding over time, and serogroup classification of Salmonella spp on a large number of dairy farms of various sizes. DESIGN: Longitudinal study. SAMPLE POPULATION: 22,417 fecal samples from cattle and 4,570 samples from the farm environment on 110 organic and conventional dairy farms in Minnesota, Wisconsin, Michigan, and NewYork. PROCEDURE: 5 visits were made to each farm at 2-month intervals from August 2000 to October 2001. Fecal samples from healthy cows, calves, and other targeted cattle groups and samples from bulk tank milk, milk line filters, water, feed sources, and pen floors were collected at each visit. Bacterial culture was performed at 1 laboratory. RESULTS: Salmonella spp were isolated from 4.8% of fecal samples and 5.9% of environmental samples; 92.7% of farms had at least 1 Salmonella-positive sample. The 75th percentile for median within-herd prevalence of Salmonella spp in cattle for 5 sampling visits to a given farm was 2.0% and for maximum within-herd prevalence of Salmonella spp was 13.6%. Farms with a median within-herd prevalence of Salmonella spp of > or = 2.0% accounted for 76.3% of Salmonella-positive samples. There was no significant difference in the prevalence of Salmonella spp between conventional and organic farms. Seasonal differences in Salmonella shedding were observed. More farms had at least 1 serogroup B isolate than any other serogroup, whereas serogroup E1 was the most common among all Salmonella-positive samples. More than 1 serogroup was isolated on 76.4% of Salmonella-positive farms. CONCLUSIONS AND CLINICAL RELEVANCE: Salmonella spp were isolated from > 90% of dairy farms; however, 25% of farms accounted for > 75% of Salmonella-positive samples. This information is critical for the direction of intervention strategies to decrease the prevalence of Salmonella spp on dairy farms.

Animals↗

Colonization of broiler chicks by Salmonella typhimurium definitive phage type 104.

The prevalence of an antibiotic-resistant strain of Salmonella Typhimurium definitive phage type 104 (DT104) has increased dramatically in recent years resulting in increased morbidity and mortality in both animals and humans. Colonization and shedding of Salmonella Typhimurium DT104 was studied in broiler chickens in two trials. In trial 1, 180 day-of-hatch chicks (n = 60 per group, n = 30 per replicate) were challenged with 10(6) CFU DT104 (wild-type isolate from poultry) or were commingled with a seeder chick challenged with 10(6) CFU DT104. In trial 2, 360 day-of-hatch chicks (n = 120 per treatment, n = 30 per rep) were divided into three groups. Chicks in the susceptible group were commingled with two seeder chicks that were orally challenged with 10(7) CFU/bird of a pan-sensitive strain of Salmonella Typhimurium DT104. Chicks in the resistant group were commingled with two seeder chicks that were orally challenged with 10(7) CFU/bird DT104 used in trial 1. For both trials, a control group was not exposed to DT104, composite fecal samples were evaluated twice weekly for levels of Salmonella shedding and 20 chicks per group were necropsied weekly and their cecal contents were cultured. At hatch all groups were colonized with naturally occurring Salmonella Senftenberg and Salmonella Mbandaka (trial 1) or Salmonella Senftenberg and Salmonella Ohio (trial 2) prior to exposure to DT104. Throughout the study, the level of Salmonella spp. shedding in feces (trial 1 means 3.1, 2.9, and 3.0 log10 CFU per g feces for challenged, seeder, and control groups, respectively) or ceca (trial 2 means 2.9. 2.9. and 2.5 log10 CFU per g ceca for resistant, susceptible, and control groups, respectively) did not differ among groups. In trial 1, colonization of DT104 remained constant at higher levels in the challenged group (mean 87%, P < 0.01), increased over time in the seeder group (10 to 50%, P < 0.02) and was not recovered from the control chicks. Salmonella Mbandaka colonization remained steady within each group with challenge and seeder groups maintaining higher levels of colonization than the control group. Salmonella Senftenberg colonization levels tended to decline (P = .058) over time in the challenged group (20 to 0%) and significantly decreased (P < 0.01) over time for both the seeder (80 to 0%) and control chicks (85 to 10%). In trial 2, the percentage of chicks colonized with susceptible DT104 declined (r = 0.90, P < 0.05) over the course of the trial from 45 to 0%, while recovery of the resistant DT104 persisted at a mean percentage of 27%. DT104 was not recovered from the control chicks. Salmonella Ohio colonization levels tended to decline (r = 0.79, P > 0.05) over time in the control group (75 to 20%) and significantly decreased (P < 0.05) over time in both susceptible and resistant groups (40 to 10%, r = 0.82 and 55 to 5%, r = 0.85, respectively). Salmonella Senftenberg was recovered from the control group at low frequency throughout the trial and was not recovered from the other groups. For either trial, no apparent affect on morbidity or mortality was observed. Introduction of DT104 by commingling may induce colonization resulting in persistent high levels of shedding in flocks simultaneously with other Salmonella species.

Animals↗

Salmonella enterica serotypes isolated from imported frozen chicken meat in the Canary islands.

To determine the prevalence of Salmonella enterica serotypes in imported frozen chicken meat, 406 samples (whole chicken, legs, and breast meat) were analyzed for Salmonella according to ISO6579 rules, serotypes were assigned, and phage typing was conducted for Salmonella serotypes Enteritidis, Typhimurium, and Heidelberg. The overall frequency of Salmonella isolation was 16.5%. By country of origin, the highest percentage of cases was found among the samples from France followed by samples from Brazil. The differences between legs and breast meat were significant. The most frequently isolated serotype of Salmonella was Enteritidis, followed by Salmonella Heidelberg, Salmonella Typhimurium, and Salmonella Virchow. By country of origin, we identified a large percentage of serotype Salmonella Enteritidis in the samples imported from Brazil. There was a greater diversity of serotypes isolated from the French samples, and Salmonella Enteritidis was not the dominant strain. In the samples from the United States, the only serotype isolated was Salmonella Kentucky, although a smaller number of samples was analyzed. The Salmonella Enteritidis phage type that prevailed in both France and Brazil was 4. Phage types 204c and 204 were identified for Salmonella Typhimurium, and phage types 8, 31, and 37 were identified for Salmonella Virchow.

Animals↗

[Salmonella bacteremia].

INTRODUCTION: Non-typhi salmonellae are invasive bacteria, which can, in certain conditions, get into the blood stream, and course bacteriemia or localized infections in different organ systems. Incidence of salmonella bacteriemia varied in different studies between 5.2-13.7%. The incidence is higher in patients younger than 1 year and the elderly. Among different serotypes, the most invasive are Salmonella typhimurium, Salmonella choleraesuis and Salmonella Virchow. The aim of our study was to determine the incidence of salmonella bacteriemia among patients hospitalized at the Clinic of Infectious Diseases Novi Sad during 1991-1998, and to point to certain risk factors for salmonella bacteriemia. RESULTS: During 1991-1998, 1309 patients were hospitalized with the diagnosis of Salmonella gastroenteritis. 12 patients (0.99%) had positive blood culture. The average age of salmonella bacteriemia patients was 17 (1-54 year), but 50% of them were younger than 10. 4/12 patients (33.33%) had positive stool and blood culture, but in 8/12 (66.66%) only positive blood culture had been established. The main serotype was Salmonella enteritidis (83.33%) and after that Salmonella Virchow (16.66%). In more than 50% of patients there was at least one risk factor responsible for dissemination: sideropenic anemia (25%), cerebral palsy (8.33%) and chronic cardiac disease (8.33%). Average duration of fever was 13.16 days and diarrhea 8.83. All patients presented with hepatosplenomegaly. One patient had a localized salmonella infection (periappendicular abscess) and she had undergone successful surgery. All our patients had been under cephalosporins of the third generation for 2 weeks and they recovered completely without sequelae. CONCLUSIONS: A relatively low incidence of salmonella bacteriemia (0.99%) in our patients can be explained with small number of children younger than 1 year and patients older than 70 years of age in whom bacteriemia is most common. Positive blood culture doesn't mean obligatory positive stool culture. Routine laboratory analyses are relatively nonspecific for the diagnosis of generalized infection. Cephalosporins of the third generation are considered to be the drug of choice in treatment of salmonella bacteriemia.

Adolescent↗

The incidence of antimicrobial-resistant Salmonella spp on freshly processed poultry from US Midwestern processing plants.

AIMS: To determine the incidence of antimicrobial-resistant Salmonella spp. on processed poultry (turkey) at Midwestern poultry plants. METHODS AND RESULTS: Two participating plants were visited at monthly intervals for a period of 1 year. Surface swabs were obtained from carcasses at two selected points on the production line, pre- and post-chill. In addition, samples of the chill water from chill tanks were also examined. Isolation and detection of Salmonella spp. from carcass swabs and chill water was carried out using standard enrichment techniques. Immunomagnetic separation was used to enhance the recovery of the pathogen. Salmonella isolates recovered were identified, serotyped and their antimicrobial resistance profiles determined using the National Antimicrobial Resistance Monitoring System. Results from the study indicated that the overall incidence of Salmonella was approx. 16.7%, with a greater incidence of the pathogen observed on pre-chill than post-chill carcasses. Salmonella isolates recovered displayed resistance to an average of four different antimicrobials. Approximately 15 different serotypes of Salmonella spp. were recovered, with Salmonella serotype Agona, Salmonella serotype Hadar, Salmonella serotype Heidelberg and Salmonella serotype Senftenberg being the most common. CONCLUSIONS: The incidence of Salmonella spp. was relatively low and isolates recovered showed significant degrees of antimicrobial resistance. Factors such as the processing plant examined, the season and farms that were presenting animals for processing influenced the incidence of the pathogen. SIGNIFICANCE AND IMPACT OF THE STUDY: Differences were observed in the serotypes of Salmonella recovered and the types of antimicrobial resistance found at the two plants. The study suggests that the use of antimicrobials at the farm level influences the creation of an environment that promotes the selection of antimicrobial-resistant Salmonella spp. The incidence, isolation and detection of Salmonella spp. on processed poultry are discussed.

Animals↗

Accessory replicons of species of Salmonella and Shigella.

Shigella and Salmonella strains isolated from clinical samples were examined. Out of 42 Shigella strains tested, 17 (40%) were found to be colicinogenic and another 3 were lysogenic. All three lysogens yielded a phage antigenically homologous to coliphage P2. Out of 30 strains tested, only 1 was found to be resistant to both neomycin and sulfamethoxazole. Out of 48 strains of Salmonella tested for drug resistance, only 2 showed multiple drug resistance. In contrast to Shigella isolates, the Salmonella isolates were infrequently (approximately 5%) bacteriocinogenic. The frequency of lysogeny in Salmonella strains was found to be 6% when tested on Salmonella typhimurium LT2, but by using a set of five indicators belonging to species Salmonella potsdam, Salmonella mbadanka, Salmonella dublin, Salmonella london, and Salmonella wandsworth, 50% of the strains were shown to be lysogenic. Salmonella phages related to P22 were recoverable from Salmonella saintpaul, Salmonella indiana, and Salmonella heidelberg. Some isolates of S. typhimurium yielded a temperature-sensitive and P22-heterologous phage which was found to be a more efficient transducer of bacterial genetic markers than P22. EcoRI-generated fragments of the DNA of some phages permitted the establishment of a clonal descent for some of the wild-type lysogenic bacterial strains. This last observation points out the potential usefulness of prophages as epidemiological markers.

Bacteriocins↗

Identification of Salmonellae with the 4-methylumbelliferyl caprilate fluorescence test.

We have tested 750 Salmonella strains and 130 strains of other species of the family Enterbacteriaceae with the 4-methylumbelliferyl caprilate reagent (MUCAP) test. The MUCAP test is a fluorescence test for rapid identification of Salmonella strains. The non-Salmonella strains were strains sent for identification as suspected Salmonella strains and thus have phenotypes similar to those of Salmonella strains. All 748 tested Salmonella strains of subgroups I, II, III, and IV were positive in the MUCAP test. Of the two tested rare Salmonella subgroup V strains, one was positive and the other was negative. In the selected material containing strains with phenotypes similar to those of Salmonella strains, only one Hafnia alvei strain of 130 Enterbacteriaceae bacteria tested was positive. The fluorescence of the H. alvei strain, the six tested Salmonella dublin strains, and the positive Salmonella subgroup V strain was weaker than that of the other salmonellae. The MUCAP assay is simple and is performed within 5 min. With an almost 100% sensitivity for Salmonella strains, apart from a single Salmonella subgroup V strain, we found the MUCAP test to be a convenient complement to traditional biochemical identification methods, especially for atypical and unusual Salmonella strains.

Bacteriological Techniques↗

Prevalence of Salmonella in two Botswana abattoir environments.

A 1-year study was carried out to investigate the prevalence of Salmonella in two abattoir environments coded "A" and "B" in Gaborone, Botswana. The total number of environmental samples collected from abattoirs A and B was 250 and 300, respectively. The samples were taken from soils in the corrals, knife blades, saw blades, cattle-drinking water, cattle feces, and feed. Preenrichment, enrichment, and selective/differential media, which enabled the favorable growth of Salmonella, were used in the study. Salmonellae were present in all sampled environments. The most common serotypes found in the environment at abattoir A were E1, C1, C2, and B. Serotypes B, C1, C2, C3, and E1 were common in abattoir B. Antigenic characterization of the salmonellae isolates showed that Salmonella Anatum, Salmonella Azteca, Salmonella Saintpaul, Salmonella Cerro, and Salmonella Westhampton were predominant in abattoir A, whereas Salmonella Anatum, Salmonella Mbandaka, Salmonella Molade, Salmonella Reading, and Salmonella Oranienburg were dominant in abattoir B. Implementing hazard analysis critical control point principles in work procedures would definitely reduce the gross contamination taking place in abattoirs.

Abattoirs↗

Relative effectiveness of the Bacteriological Analytical Manual method for the recovery of Salmonella from whole cantaloupes and cantaloupe rinses with selected preenrichment media and rapid methods.

Soak and rinse methods were compared for the recovery of Salmonella from whole cantaloupes. Cantaloupes were surface inoculated with Salmonella cell suspensions and stored for 4 days at 2 to 6 degrees C. Cantaloupes were placed in sterile plastic bags with a nonselective preenrichment broth at a 1:1.5 cantaloupe weight-to-broth volume ratio. The cantaloupe broths were shaken for 5 min at 100 rpm after which 25-ml aliquots (rinse) were removed from the bags. The 25-ml rinses were preenriched in 225-ml portions of the same uninoculated broth type at 35 degrees C for 24 h (rinse method). The remaining cantaloupe broths were incubated at 35 degrees C for 24 h (soak method). The preenrichment broths used were buffered peptone water (BPW), modified BPW, lactose (LAC) broth, and Universal Preenrichment (UP) broth. The Bacteriological Analytical Manual Salmonella culture method was compared with the following rapid methods: the TECRA Unique Salmonella method, the VIDAS ICS/SLM method, and the VIDAS SLM method. The soak method detected significantly more Salmonella-positive cantaloupes (P < 0.05) than did the rinse method: 367 Salmonella-positive cantaloupes of 540 test cantaloupes by the soak method and 24 Salmonella-positive cantaloupes of 540 test cantaloupes by the rinse method. Overall, BPW, LAC, and UP broths were equivalent for the recovery of Salmonella from cantaloupes. Both the VIDAS ICS/SLM and TECRA Unique Salmonella methods detected significantly fewer Salmonella-positive cantaloupes than did the culture method: the VIDAS ICS/SLM method detected 23 of 50 Salmonella-positive cantaloupes (60 tested) and the TECRA Unique Salmonella method detected 16 of 29 Salmonella-positive cantaloupes (60 tested). The VIDAS SLM and culture methods were equivalent: both methods detected 37 of 37 Salmonella-positive cantaloupes (60 tested).

Bacteriological Techniques↗

Characterization of antimicrobial-resistant Salmonella isolated from imported foods.

Two-hundred eight Salmonella isolates recovered from over 5,000 imported foods entering the United States in 2001 were tested for antimicrobial susceptibilities and further characterized for quinolone resistance mechanisms, integron carriage, and genetic relatedness. Salmonella Weltevreden (20%), Salmonella Newport (6%), Salmonella Lexington (5%), and Salmonella Thompson (4%) were the four most common serotypes recovered. Twenty-three (11%) isolates were resistant to at least one antimicrobial, and seven (3.4%) to three or more antimicrobials. Resistance was most often observed to tetracycline (9%), followed by sulfamethoxazole (5%), streptomycin (4%), nalidixic acid (3%), and trimethoprim/sulfamethoxazole (2%). One Salmonella Schwarzengrund isolate recovered from squid imported from Taiwan exhibited resistance to eight antimicrobials, including ampicillin, chloramphenicol, gentamicin, kanamycin, nalidixic acid, sulfamethoxazole, tetracycline, and trimethoprim/sulfamethoxazole. Six isolates (Salmonella Bareilly, Salmonella Derby, Salmonella Ohio and three Salmonella Schwarzengrund) contained class 1 integrons, which carried several resistance genes including dhfrI/dhfrXII, aadA, pse-1, and sat1, conferring resistance to trimethoprim/sulfamethoxazole, streptomycin, ampicillin, and streptothricin, respectively. Five of six nalidixic acid-resistant isolates possessed DNA point mutations at either Ser83 or Asp87 in DNA gyrase. One ciprofloxacin-resistant isolate possessed double mutations in DNA gyrase at positions Ser83 and Asp87 as well as a single mutation at Ser80 in parC. The top three serotypes identified, Salmonella Weltevreden (n = 41), Salmonella Newport (n = 13), and Salmonella Lexington (n = 11), were further characterized for genetic relatedness by pulsed-field gel electrophoresis. Fifty-five distinct pulsed-field gel electrophoresis patterns were observed among the 65 isolates, indicating extensive genetic diversity among these Salmonella serotypes contaminating imported foods.

Anti-Bacterial Agents↗

Distribution of Salmonella in swine herds in Québec.

Five porcine finishing units, previously identified as contaminated by Salmonella, were sampled to identify possible sources of contamination and to study the distribution of Salmonella within the herds. A total of 208 environmental samples were taken and 87 samples (42%) were found contaminated by Salmonella spp. Salmonella was recovered from several types of samples. Among these, fecal material from pens, building environment such as doors, floors, ventilation units, dust and farm accessories were most often found positive. Some of the flies and rodents were also positive. Two of the finishing units were part of an integrated production system and the prevalence and distribution of Salmonella spp. at different production steps of the integrated facilities were studied. Forty-one farms were sampled and a total of 1923 faecal samples in randomly selected pens were analysed. One hundred and fifty-one samples (7.9%) were positive for Salmonella spp. Among the farms sampled, 70.7% (29/41) were positive for isolation of Salmonella. The different levels in the integrated production were unevenly contaminated. Replacement sow (15.9%) and finishing unit for gilts (21.9%) were the most contaminated levels. Ten serotypes of Salmonella (n = 132) were identified in the production pyramid with a predominance of Salmonella Derby (37.1%) and Salmonella Typhimurium (34.1%). Pulse Field Gel Electrophoresis analysis of the various isolates from serotypes and Salmonella Typhimurium, Salmonella Derby and Salmonella Anatum showed no variation in the genetic profiles, within each serotype, suggesting a vertical contamination throughout the different production steps.

Agglutination Tests↗

[Occurence of salmonellae in laying hens in different housing systems and inferences for control].

As eggs represent now as ever the most important source for Salmonella infection in human beings and because of the currently occurring shift in housing conditions for laying hens from conventional cages to alternative systems it was studied whether the Salmonella prevalence in layers is influenced by the housing system. Following systems were considered: organic farming with free range management systems, floor management systems with free range, floor management systems without free range, conventional cages. 453 pooled faecal samples as single or double examination per herd from 329 flocks in different housing systems for table egg production from three Federal Lander were examined bacteriologically. The share of layer flocks which were Salmonella positive at least once independently of the housing system amounted to 32.2%. Analysis of the Salmonella findings in the single housing systems revealed that the share of Salmonella positive flocks was higher in conventional cage systems (46.3%) than in alternative housing systems (32.996% in organic farming with free range management systems, 21.9% in floor management systems with free range, 23.4% in floor management systems without free range). The results of the study clearly show that Salmonella Enteritidis (mostly phage type 4, other phage types rarely) presents with a share of 78% the dominant serovar in laying hens. The total number of all other serovars (apart from Salmonella Enteritidis and subspecies I rough) reached a share of ca. 14%, however, no other single serovar was dominant. As Salmonella Enteritidis is the predominant serovar in laying hens it is strongly recommended to use Salmonella Enteritidis vaccines for immunisation programmes of chickens during the rearing period. Because of the high prevalence of Salmonella organisms in the different housing systems, detailed information on the epidemiology of Salmonella in laying hens are needed to introduce effective control measures. Of particular interest is the question whether the Salmonella findings in laying flocks are the result of multiplication of already existing Salmonella organisms in the animals or whether the bacteria are introduced only during the laying period.

Animal Husbandry↗

The role of host cell death in Salmonella infections.

Salmonella enterica is an important enteric pathogen of humans and a variety of domestic and wild animals. Infection is initiated in the intestinal tract, and severe disease produces widespread destruction of the intestinal mucosa. Salmonella strains can also disseminate from the intestine and produce serious, sometimes fatal infections with considerable cytopathology in a number of systemic organs. A combination of bacterial genetic and cell biology studies have shown that Salmonella uses specific virulence mechanisms to induce host cell death during infection. Salmonella produces one set of virulence proteins to promote invasion of the intestine and a different set to mediate systemic disease. Significantly, each set of virulence factors mediates a distinct mechanism of host cell death. The Salmonella pathogenicity island-1 (SPI-1) locus encodes a type III protein secretion system (TTSS) that delivers effector proteins required for intestinal invasion and the production of enteritis. The SPI-1 effector SipB activates caspase-1 in macrophages, releasing IL-1beta and IL-18 and inducing rapid cell death by a mechanism that has features of both apoptosis and necrosis. Caspase-1 is required for Salmonella to infect Peyer's patches and disseminate to systemic tissues in mice. Progressive Salmonella infection in mice requires the SPI-2 TTSS and associated effector proteins as well as the SpvB cytotoxin. Apoptosis of macrophages in the liver is found during systemic infection. In cell culture, Salmonella strains induce delayed apoptosis dependent on SPI-2 function in macrophages from a variety of sources. This delayed apoptosis also requires activation of TLR4 on macrophages by the bacterial LPS. Downstream activation of kinase pathways leads to balanced pro- and antiapoptotic regulatory factors in the cell. NF-kappaB and p38 mitogen-activated protein kinase (MAPK) are particularly important for the induction of antiapoptotic factors, whereas the kinase PKR is required for bacterial-induced apoptosis. The Salmonella SPI-2 TTSS is essential for altering the balance in favor of apoptosis during intracellular infection, but the effectors involved remain poorly characterized. The SpvB cytotoxin has been shown to play a role in apoptosis in human macrophages by depolymerizing the actin cytoskeleton. A model for the role of bacteria-induced host cell death in Salmonella pathogenesis is proposed. In the intestine, the Salmonella SPI-1 TTSS and SipB mediate macrophage death by caspase-1 activation, which also releases IL-1beta and IL-18, promoting inflammation and subsequent phagocytosis by incoming macrophages and leading to dissemination to systemic tissues. Intracellular secretion of virulence effector proteins by the SPI-2 TTSS facilitates growth of Salmonella in these macrophages and the delayed onset of apoptosis in extraintestinal tissues. These infected, apoptotic cells are targeted for engulfment by incoming macrophages, thus perpetuating the cycle of cell-to-cell spread that is the hallmark of systemic Salmonella infection.

Animals↗

Salmonella in slaughter pigs: prevalence, serotypes and critical control points during slaughter in two slaughterhouses.

The purpose of this study was to show the distribution of Salmonella in slaughtered pigs and the environment of the slaughterhouse. 1,114 samples of slaughtered pigs (six different samples for Salmonella isolation and one serum sample for ELISA on antibodies per pig) and 477 samples of the slaughterhouse environment were collected in two slaughterhouses on two sampling days per slaughterhouse. Salmonella was isolated from one or more samples of 47% of the pigs. The highest prevalence of Salmonella was observed in rectal content samples (25.6%), whereas the lowest prevalence of Salmonella was observed on the carcasses (1.4%). The prevalence of Salmonella in other samples was: 19.6% in tonsils, 9.3% on livers, 9.3% on tongues, and 9.3% in mesenterial lymphnodes. The prevalence of Salmonella in environmental samples was high in the drain water samples in both slaughterhouses (61%) and on the carcass splitter in one slaughterhouse (33%). Salmonella typhimurium was the most frequently isolated serotype in pig samples and environmental samples in both slaughterhouses: 43% of the Salmonella isolates from pigs and 33% of the Salmonella isolates from the environment was S. typhimurium. The results of this study show that Salmonella prevalences in pigs differ a lot, depending on which part of the pig is sampled. Not all different samples of the pig will become available for human consumption, but collecting more than one sample per pig showed that Salmonella can be found in almost the whole pig. The result of surface samples of carcass and liver gives information about hygiene during the slaughter process; the result of tonsils, lymphnodes and rectal contents, combined with the serological result, gives information about infection of the pig before the slaughter process (on the farm, during transport or in lairage). It can be concluded that results of Salmonella isolation of slaughter pigs should always be carefully interpreted, depending on the type of sample that has been collected.

Abattoirs↗

Effects of in vitro growth phase on the pathogenesis of Salmonella typhimurium in mice.

The growth phase of a bacterial (Salmonella typhimurium) culture was shown to have pronounced effects on the pathogenic properties of the harvested bacteria. Salmonellae obtained from a culture in primary (exponential) growth phase (PP) were more readily cleared from the blood and more readily killed by phagocytes than were salmonellae obtained from a more slowly growing secondary growth phase (SP) culture. PP salmonellae were observed to cause death of mice sooner than SP salmonellae. This appeared to be because the more rapid growth of PP, as compared to SP, salmonellae continued in the liver and spleen for several hours following intravenous injection, and more than compensated for their high in vivo death rate. As a result, within 4 h there were approximately 10-fold more live salmonellae in the spleens and livers of mice that had received PP, as compared to SP, salmonellae. This 10-fold difference was maintained until the death of the mice, indicating that after the first 4 h post-inoculation, the net in vivo growth of the salmonellae was the same regardless of their growth phase in the inoculating culture. This transition between PP and SP salmonellae occurred long before a dense stationary phase culture was obtained. Salmonellae grown in minimal media exhibited the biological properties of SP salmonellae and never entered as rapid a growth phase as did salmonellae in complete media.

Animals↗

Prevalence of Salmonella spp. in poultry broilers and shell eggs in Korea.

This study was conducted to determine the presence of Salmonella spp. in raw broilers and shell eggs in Korea. In total, 135 dozen shell eggs and 27 raw broilers were tested. None of the egg yolks were found to contain Salmonella organisms but Escherichia coli, Escherichia hermanii, and Citrobacter freundii were isolated from egg shells. Salmonella spp. were detected in 25.9% of raw broilers, and Salmonella serotypes isolated from raw broilers were Salmonella Enteritidis, Salmonella Virchow, and Salmonella Virginia. D-values and antibiotic resistance of Salmonella isolates were also investigated. D-values of Salmonella enteritidis, Salmonella Virginia, and Salmonella Virchow in tryptic soy broth at 55 degrees C were 2.36, 2.13, and 0.70 min and 0.53, 0.37, and 0.20 min at 60 degrees C, respectively. All Salmonella isolates showed multiple antibiotic resistance patterns and were resistant to penicillin and vancomycin. One strain of Salmonella Enteritidis showed resistance to 12 antibiotics used in this study.

Animals↗