New Salmonella Serotype: Salmonella enteritidis ser. Ordonez

The antigenic formula of a new serotype, Salmonella enteritidis serotype Ordonez, was characterized as (1),13,23,37:y:l,w.     

Application of BAX system, Tecra Unique Salmonella test, and a conventional culture method for the detection of Salmonella in ready-to-eat and raw foods

AIMS: To compare the BAX system, the Tecra Unique Salmonella test, and a conventional culture method for the detection of Salmonella in various foods. METHODS AND RESULTS: Ready-to-eat and raw foods were inoculated with Salmonella serotype Typhimurium, Salmonella serotype Enteritidis, Salmonella serotype Typhi, or Salmonella serotype Derby. Incubated pre-enrichment cultures were examined using the BAX system, the Tecra Unique Salmonella test, and a conventional culture method. Salmonella could be detected in all ready-to-eat food samples inoculated with S. Typhimurium, S. Enteritidis, or S. Derby, with any of the three test methods. However, false negatives were obtained with the Tecra test and the culture method when samples with higher background flora were inoculated with S. Typhi. Sensitivity test results suggested the two rapid tests performed as well as the culture method in the detection of 10(1) CFU of S. Typhimurium in 25-g cooked or raw food. CONCLUSIONS: The BAX system and the Tecra Unique Salmonella test demonstrated results comparable with those of the culture method in the detection of Salmonella serotypes used except S. Typhi. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first evaluation of the BAX system, the Tecra Unique Salmonella test, and a culture method in the detection of Salmonella in a variety of western and oriental foods.     

Prevalence and serotypes of Salmonella associated with goats at two Australian abattoirs

Aims:  This study was carried out to determine the prevalence and serotype of Salmonella in goats presented for slaughter.
Methods and Results:  A total of 121 goats were examined for the presence of Salmonella in matching rumen, faecal and carcass samples. Samples were analysed for the presence of Salmonella following the Australian Standard AS 1766.2.5-1991. Salmonella was isolated from 56 (46·3%) faecal samples, 55 (45·5%) rumen samples and 35 (28·9%) carcass samples. The dominant serotypes isolated were Salmonella serotype Saintpaul (31%), Salmonella serotype Typhimurium (13%) and Salmonella serotype Chester (11%).
Conclusions:  Salmonella was isolated from at least one of the three sample sites in 68% of animals. Carcase contamination with faeces, compared with rumen liquor, is a greater hazard for Salmonella contamination of goat carcases. Goat meat is a potential source of Salmonella serovars associated with human disease.
Significance and Impact of the Study:  Goat carcases contaminated with Salmonella during slaughter could be a source of food-borne disease if consumed raw or inadequately cooked, or may be a source of cross-contamination to other foods.     

Characterization of the RpoS status of clinical isolates of Salmonella enterica

The stationary-phase-inducible sigma factor, sigma(S) (RpoS), is the master regulator of the general stress response in Salmonella and is required for virulence in mice. rpoS mutants can frequently be isolated from highly passaged laboratory strains of Salmonella: We examined the rpoS status of 116 human clinical isolates of Salmonella, including 41 Salmonella enterica serotype Typhi strains isolated from blood, 38 S. enterica serotype Typhimurium strains isolated from blood, and 37 Salmonella serotype Typhimurium strains isolated from feces. We examined the abilities of these strains to produce the sigma(S) protein, to express RpoS-dependent catalase activity, and to resist to oxidative stress in the stationary phase of growth. We also carried out complementation experiments with a cloned wild-type rpoS gene. Our results showed that 15 of the 41 Salmonella serotype Typhi isolates were defective in RpoS. We sequenced the rpoS allele of 12 strains. This led to identification of small insertions, deletions, and point mutations resulting in premature stop codons or affecting regions 1 and 2 of sigma(S), showing that the rpoS mutations are not clonal. Thus, mutant rpoS alleles can be found in freshly isolated clinical strains of Salmonella serotype Typhi, and they may affect virulence properties. Interestingly however, no rpoS mutants were found among the 75 Salmonella serotype Typhimurium isolates. Strains that differed in catalase activity and resistance to hydrogen peroxide were found, but the differences were not linked to the rpoS status. This suggests that Salmonella serotype Typhimurium rpoS mutants are counterselected because rpoS plays a role in the pathogenesis of Salmonella serotype Typhimurium in humans or in the transmission cycle of the disease.     

A new Salmonella serotype isolated in Canada:Salmonella sherbrooke (16:d:1,6)

A new Salmonella serotype classified in the Kauffman sub-genus I (Kauffman 1963) has been isolated in Canada from a stock of cacao beans from Nigeria. Salmonella sherbrooke shares the antigenic structure 16:d:1,6.     

Detection of Salmonella enterica Subpopulations by Phenotype Microarray Antibiotic Resistance Patterns

Three strains of Salmonella enterica serotype Enteritidis were compared to Salmonella enterica serotype Heidelberg, Salmonella enterica serotype Newport, and Salmonella enterica serovar Typhimurium for growth in the presence of 240 antibiotics arranged within a commercial high-throughput phenotype microarray. The results show that antibiotic resistances were different for subpopulations of serotype Enteritidis separated only by genetic drift.     

Multilocus sequence typing supports the hypothesis that cow- and human-associated Salmonella isolates represent distinct and overlapping populations

A collection of 179 human and 156 bovine clinical Salmonella isolates obtained from across New York state over the course of 1 year was characterized using serotyping and a multilocus sequence typing (MLST) scheme based on the sequencing of three genes (fimA, manB, and mdh). The 335 isolates were differentiated into 52 serotypes and 72 sequence types (STs). Analyses of bovine isolates collected on different farms over time indicated that specific subtypes can persist over time on a given farm; in particular, a number of farms showed evidence for the persistence of a specific Salmonella enterica serotype Newport sequence type. Serotypes and STs were not randomly distributed among human and bovine isolates, and selected serotypes and STs were associated exclusively with either human or bovine sources. A number of common STs were geographically widespread. For example, ST6, which includes isolates representing serotype Typhimurium as well as the emerging serotype 4,5,12:i:-, was found among human and bovine isolates in a number of counties in New York state. Phylogenetic analyses supported the possibility that serotype 4,5,12:i:- is closely related to Salmonella serotype Typhimurium. Salmonella serotype Newport was found to represent two distinct evolutionary lineages that differ in their frequencies among human and bovine isolates. A number of Salmonella isolates carried two copies of manB (33 isolates) or showed small deletion events in fimA (nine isolates); these duplication and deletion events may provide mechanisms for the rapid diversification of Salmonella surface molecules. We conclude that the combined use of an economical three-gene MLST scheme and serotyping can provide considerable new insights into the evolution and transmission of Salmonella.     

One-step triplex high-resolution melting analysis for rapid identification and simultaneous subtyping of frequently isolated Salmonella serovars

Salmonellosis is one of the most important food-borne diseases worldwide. For outbreak investigation and infection control, accurate and fast subtyping methods are essential. A triplex gene-scanning assay was developed and evaluated for serotype-specific subtyping of Salmonella enterica isolates based on specific single-nucleotide polymorphisms in fragments of fljB, gyrB, and ycfQ. Simultaneous gene scanning of fljB, gyrB, and ycfQ by high-resolution melting-curve analysis of 417 Salmonella isolates comprising 46 different serotypes allowed the unequivocal, simple, and fast identification of 37 serotypes. Identical melting-curve profiles were obtained in some cases from Salmonella enterica serotype Enteritidis and Salmonella enterica serotype Dublin, in all cases from Salmonella enterica serotype Ohio and Salmonella enterica serotype Rissen, from Salmonella enterica serotype Mbandaka and Salmonella enterica serotype Kentucky, and from Salmonella enterica serotype Bredeney, Salmonella enterica serotype Give, and Salmonella enterica serotype Schwarzengrund. To differentiate the most frequent Salmonella serotype, Enteritidis, from some S. Dublin isolates, an additional single PCR assay was developed for specific identification of S. Enteritidis. The closed-tube triplex high-resolution melting-curve assay developed, in combination with an S. Enteritidis-specific PCR, represents an improved protocol for accurate, cost-effective, simple, and fast subtyping of 39 Salmonella serotypes. These 39 serotypes represent more than 94% of all human and more than 85% of all nonhuman Salmonella isolates (including isolates from veterinary, food, and environmental samples) obtained in the years 2008 and 2009 in Austria.     

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.     

Enzyme-linked immunosorbent assay for Salmonella typhimurium in food: feasibility of 1-day Salmonella detection

A microtitration plate, antibody-capture, enzyme-linked immunosorbent assay was developed for detection of Salmonella typhimurium. The assay utilizes a monoclonal detector antibody which shows no cross-reactions with non-Salmonella species and only a slight cross-reaction with one other Salmonella serotype. By using only one cultural stage (in a nonselective, chemically defined medium) prior to the enzyme-linked immunosorbent assay, low numbers of cells in food (10 cells 25 g-1) were detected in 19 h. Non-Salmonella competing organisms did not interfere with detection of S. typhimurium even when present in the ratio of 10(6):1 (non-Salmonella/Salmonella spp.). The assay shows the feasibility of rapid, 1-day testing for Salmonella spp. with antibody technology.     

Clinical salmonellosis in a guinea pig colony caused by a new Salmonella serotype, Salmonella ochiogu

During a severe outbreak of clinical salmonellosis in an experimental guinea pig colony, a new strain of Salmonella was isolated and identified. The new serotype, with the antigenic structure 1,3,19 : Z38 : e, n, Z15, for which the name Salmonella ochiogu has been suggested, caused both enteric and systemic infection in the animal colony. During the outbreak a total of 127 animals died (26.9%). All ages of animal were affected. Treatment with oral tetracycline was successful when combined with strict hygienic measures.     

沙门氏菌Ⅲb的一个新血清型

An enterobacterium culture, S3188, providing with Salmonella biological characteristics, except exhibiting indole positive reaction, was isolated from the intestinal content of reptile a snake. It utilized malonate, did not ferment dulcitol, it attacked lactose promptly, and ONPG positive. H antigens appeared diphasic. By cross agglutination and absorption tests, it was demonstrated that the antigenic formula of this strain is 53:1, Z13:e, n,(Z15)... It was found to be a new serotype of Salmonella IIIb.     

Enzyme-linked immunosorbent assay for Salmonella typhimurium in food: feasibility of 1-day Salmonella detection.

A microtitration plate, antibody-capture, enzyme-linked immunosorbent assay was developed for detection of Salmonella typhimurium. The assay utilizes a monoclonal detector antibody which shows no cross-reactions with non-Salmonella species and only a slight cross-reaction with one other Salmonella serotype. By using only one cultural stage (in a nonselective, chemically defined medium) prior to the enzyme-linked immunosorbent assay, low numbers of cells in food (10 cells 25 g-1) were detected in 19 h. Non-Salmonella competing organisms did not interfere with detection of S. typhimurium even when present in the ratio of 10(6):1 (non-Salmonella/Salmonella spp.). The assay shows the feasibility of rapid, 1-day testing for Salmonella spp. with antibody technology.     

Microarray analysis of antimicrobial resistance genes in Salmonella enterica from preharvest poultry environment

Aims:  To detect antimicrobial resistance genes in Salmonella isolates from turkey flocks using the microarray technology.
Methods and Results:  A 775 gene probe oligonucleotide microarray was used to detect antimicrobial resistance genes in 34 isolates. All tetracycline-resistant Salmonella harboured tet(A) , tet(C) or tet(R) , with the exception of one Salmonella serotype Heidelberg isolate. The sul1 gene was detected in 11 of 16 sulfisoxazole-resistant isolates. The aadA , aadA1 , aadA2 , strA or strB genes were found in aminoglycoside-resistant isolates of Salm. Heidelberg, Salmonella serotype Senftenberg and untypeable Salmonella . The prevalence of mobile genetic elements, such as class I integron and transposon genes, in drug-resistant Salmonella isolates suggested that these elements may contribute to the dissemination of antimicrobial resistance genes in the preharvest poultry environment. Hierarchical clustering analysis demonstrated a close relationship between drug-resistant phenotypes and the corresponding antimicrobial resistance gene profiles.
Conclusions:  Salmonella serotypes isolated from the poultry environment carry multiple genes that can render them resistant to several antimicrobials used in poultry and humans.
Significance and Impact of the Study:  Multiple antimicrobial resistance genes in environmental Salmonella isolates could be identified efficiently by microarray analysis. Hierarchical clustering analysis of the data was also found to be a useful tool for analysing emerging patterns of drug resistance.     

Evaluation of methods for the identification of Salmonella enterica serotype Typhimurium DT104 from poultry environmental samples

An increase in the prevalence of Salmonella enterica serotype Typhimurium DT104 has been reported worldwide. This study examined the prevalence of this microorganism in poultry environmental samples from commercial layer flocks and pullet environments as well as the sensitivity and specificity of a PCR-based method, and multiple antibiotic resistance profile of Salmonella serogroup B isolates in relation to the serotype and phagetype reference method for the identification of Salmonella Typhimurium DT104. A total of 435 Salmonella isolates were obtained from poultry house environmental samples tested during a 20-month period representing a prevalence of 5.5%. Of these, 313 (72%) isolates were identified as Salmonella serogroup B isolates. These isolates were tested by a PCR-based assay, and for resistance to five antibiotics: ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT) for the rapid identification of Salmonella Typhimurium DT104. Upon comparing the antibiotic resistance and PCR results with serotype and phage type data, the sensitivity and specificity for the identification of Salmonella Typhimurium DT104 of both methods were found to be 100%, and 99.6%, respectively. Both methods can be completed within 24 h after obtaining an isolate, while serotyping and phagetyping required more than 5 days to complete.     

Salmonella in raccoons (Procyon lotor) in southern Ontario, Canada

Numerous serotypes of Salmonella have been detected in a variety of wild animals, including raccoons (Procyon lotor). Raccoons are common, mid-size omnivores that live in close association with people in urban and rural areas in Ontario. Although raccoons are known to shed Salmonella, little is known about their potential long-term role in maintaining Salmonella infections. We sampled feces from raccoons in three areas of Ontario: one primarily urban site around Niagara, one primarily rural site north of Guelph, and the grounds of the Toronto Zoo, in 2007 to identify which serotypes of Salmonella were commonly shed by raccoons in southern Ontario. In addition, we conducted a longitudinal study at the Toronto Zoo site to determine if raccoons remain persistently infected with Salmonella. Salmonella was found in 45% of samples. The prevalence of Salmonella in raccoon feces ranged from 27% at the rural site to 65% at the urban site. We detected 16 serotypes of Salmonella in 83 positive samples. The most common serotype detected in raccoons from the rural and zoo sites was Salmonella enterica serotype Typhimurium, whereas Salmonella Newport was detected most commonly in the urban site. Only one raccoon of 11 that were captured in four or more consecutive trapping sessions shed the same Salmonella serotype for two consecutive months, suggesting that raccoons regularly acquire new Salmonella serotypes from the environment.     

辽宁食源性沙门菌血清型、 耐药谱及PFGE分型特征

目的分析辽宁食源性沙门菌血清型、耐药谱及脉冲场凝胶电泳(PFGE)型别,探讨辽宁沙门菌污染的同源性,为食源性疾病溯源和预警提供基础。方法对辽宁省2015年食品中、食源性疾病中分离的41株沙门菌进行血清学分型、耐药试验、PFGE分子分型,采用Bio Numerics version 6.6软件分析,比较同源性。结果 41株菌分为15个血清型,居前三位的是15株肠炎沙门菌、5株德尔卑沙门菌、5株姆班达卡沙门菌(辽宁省内少见血清型);对41株菌进行15种抗生素的耐药试验,对单一一种抗生素的耐药率为100.0%,其中红霉素97.6%,萘啶酸61.0%,氨苄西林53.7%;41株菌共分为18种PFGE带型,带型分布分散,只有两种优势,一种带型包含20株菌,有14株肠炎沙门菌,6株其他沙门菌,相似度为92.7%~100%;另一种包含5株菌,4株姆班达卡沙门菌,1株鼠伤寒沙门菌,相似度为96.6%~100.0%。结论辽宁省食源性沙门菌的血清型以肠炎沙门菌为主,生肉制品是其主要污染来源;血清型与PFGE图谱带型分布广泛,相同血清型沙门菌的PFGE带型聚集成簇、菌株具有高度同源性;相同PFGE型别的菌株耐药谱一致或相似;沙门菌的耐药情况较严重。     

The prevalence of Salmonella spp. in bovine faecal,rumen and carcass samples at a commercial abattoir

AIMS: To determine the prevalence, serotype and antibiotic resistance profile of Salmonella isolates in cattle and on carcasses at a commercial Irish abattoir. METHODS AND RESULTS: Faecal, rumen and carcass samples were collected from a beef abattoir over a 12-month period and examined for the presence of Salmonella spp. Isolates were serotyped, phage typed (when serotype was found to be S. Typhimurium) and tested for susceptibility to a panel of antibiotics. Salmonella was isolated from 2% of faecal, 2% of rumen and 7.6% of carcass samples. Salmonella was most frequently isolated from samples taken during the period August to October. S. Dublin was isolated from 72% of positive samples. S. Agona and S. Typhimurium definitive type (DT)104 were each isolated from 14% of positive samples. All S. Typhimurium DT104 isolates were resistant to ampicillin, chloramphenicol, streptomycin, sulphafurazole and tetracycline (ACSSuT). On occasion, from a single animal, the same serotype was isolated from more than one sample (i.e. faeces and rumen; faeces and carcass; rumen and carcass; faeces, rumen and carcass). CONCLUSIONS: Salmonella is present in cattle at slaughter and on beef carcasses at an Irish abattoir, with a higher frequency of occurrence during the period August to October. Most isolates from the study are not commonly associated with human clinical infection, with the exception of S. Typhimurium DT104 (R-type ACSSuT). SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides epidemiological data that is necessary for the understanding of beef as a source of human Salmonella infection.     

Salmonella resistant to extended-spectrum cephalosporins: prevalence and epidemiology

Salmonella resistant to extended-spectrum cephalosporins (ESCs) have emerged worldwide since 1988. By 2004, 43 countries had reported this public health problem. Resistance was mediated by classical extended-spectrum beta-lactamases, plasmid-mediated cephalosporinases, and recently a class A carbapenemase. Of these, CMY-2 is the most widely disseminated enzyme. Salmonella enterica serotype Typhimurium and S. enterica serotype Enteritidis are the most common serovars associated with ESC resistance in human infections. Many outbreaks in humans have been reported, most often among children and neonates. ESC-resistant Salmonella is frequently recovered from animals and food, with poultry as primary food source, suggesting that humans are often infected by these routes.     

Fate of Salmonella in dry confectionery raw materials

Aims:  To study the behaviour of salmonellae inoculated in the dry, raw materials, related to chocolate confectionery manufacture, as affected by the strain, cell preparation method and storage temperature.
Methods and Results:  Outbreak and non outbreak-associated salmonellae were used for the inoculation of cocoa butter oil, crushed cocoa and hazelnut shells, cocoa beans and almond kernels. Dry matrices were inoculated with lawn-collected and broth-grown cells and were stored at 5 and 21°C for 21 days. Results demonstrated that all strains survived in all dry matrices. Lawn-collected cells survived considerably better than broth-collected cells, at either temperature, and outbreak-associated strains of Salmonella serotype Enteritidis PT30 and Salmonella serotype Oranienburg appeared to be among the best surviving strains.
Conclusions:  The work demonstrated that salmonellae can survive storage for 3–4 weeks in dry raw materials and that survival is dependent on the source of strains, cell preparation and inoculation methodology, and storage temperature.
Significance and Impact of the Study:  The data contributes to understanding the parameters to consider when assessing the risk of dry raw materials as the most likely source of salmonellae in chocolate processing plants. It also demonstrates the importance of implementing effective lethal processes and segregation procedures to prevent cross-contamination to ensure the safety of confectionery products regarding Salmonella .