Splenic gene expression profiling in White Leghorn layer inoculated with the Salmonella enterica serovar Enteritidis

Salmonella enterica serovar Enteritidis (SE) is a foodborne pathogen that can threaten human health through contaminated poultry products. Live poultry, chicken eggs and meat are primary sources of human salmonellosis. To understand the genetic resistance of egg‐type chickens in response to SE inoculation, global gene expression in the spleen of 20‐week‐old White Leghorn was measured using the Agilent 4 × 44 K chicken microarray at 7 and 14 days following SE inoculation (dpi). Results showed that there were 1363 genes significantly differentially expressed between inoculated and non‐inoculated groups at 7 dpi (I7/N7), of which 682 were up‐regulated and 681 were down‐regulated genes. By contrast, 688 differentially expressed genes were observed at 14 dpi (I14/N14), of which 371 were up‐regulated genes and 317 were down‐regulated genes. There were 33 and 28 immune‐related genes significantly differentially expressed in the comparisons of I7/N7 and I14/N14 respectively. Functional annotation revealed that several Gene Ontology (GO) terms related to immunity were significantly enriched between the inoculated and non‐inoculated groups at 14 dpi but not at 7 dpi, despite a similar number of immune‐related genes identified between I7/N7 and I14/N14. The immune response to SE inoculation changes with different time points following SE inoculation. The complicated interaction between the immune system and metabolism contributes to the immune responses to SE inoculation of egg‐type chickens at 14 dpi at the onset of lay. GC, TNFSF8, CD86, CD274, BLB1 and BLB2 play important roles in response to SE inoculation. The results from this study will deepen the current understanding of the genetic response of the egg‐type chicken to SE inoculation at the onset of egg laying.     

T cell receptor-transgenic mouse models for studying cellular immune responses to Salmonella in vivo

Cellular immune responses are crucial both for protective immunity against salmonellosis, and for the immunogenicity of oral vaccines based on avirulent live Salmonella as antigen carriers. The crucial early steps of T cell induction are difficult to investigate in conventional animals, but recently developed T cell receptor (TCR)-transgenic models allow visualization of antigen-specific T cells in vivo while they become induced. In this review, the results obtained with four different TCR-transgenic Salmonella infection models are described, and advantages and potential limits of each of the different models are compared.     

Pathogen specific carbohydrate antigen microarrays: a chip for detection of Salmonella O-antigen specific antibodies

A Salmonella O-antigen microarray was developed by covalent coupling of oligosaccharide antigens specific for serogroups Salmonella enterica sv. Paratyphi (group A), Typhimurium (group B) and Enteritidis (group D). Antibodies were correctly detected in sera from patients with culture verified salmonellosis. High serogroup-specificity was seen with the disaccharide antigens. With the larger antigens, containing the backbone sequence Manα1–2Rhaα1–2Gal (MRG), common backbone-specific antibodies (O-antigen 12) were also detected. This is “proof of principle” that pathogen-specific carbohydrate antigen microarrays constitute a novel technology for rapid and specific serological diagnosis in either individual patients or larger sero-epidemiological and vaccine studies.     

Climate variations and salmonellosis transmission in Adelaide,South Australia: a comparison between regression models

This is the first study to identify appropriate regression models for the association between climate variation and salmonellosis transmission. A comparison between different regression models was conducted using surveillance data in Adelaide, South Australia. By using notified salmonellosis cases and climatic variables from the Adelaide metropolitan area over the period 1990–2003, four regression methods were examined: standard Poisson regression, autoregressive adjusted Poisson regression, multiple linear regression, and a seasonal autoregressive integrated moving average (SARIMA) model. Notified salmonellosis cases in 2004 were used to test the forecasting ability of the four models. Parameter estimation, goodness-of-fit and forecasting ability of the four regression models were compared. Temperatures occurring 2 weeks prior to cases were positively associated with cases of salmonellosis. Rainfall was also inversely related to the number of cases. The comparison of the goodness-of-fit and forecasting ability suggest that the SARIMA model is better than the other three regression models. Temperature and rainfall may be used as climatic predictors of salmonellosis cases in regions with climatic characteristics similar to those of Adelaide. The SARIMA model could, thus, be adopted to quantify the relationship between climate variations and salmonellosis transmission.     

Public health investigations of Salmonella Enteritidis in catering raw shell eggs, 2002-2004

AIMS: In response to a dramatic change in the epidemiology of Salmonella Enteritidis in England and Wales thought to be associated with raw shell eggs, the Health Protection Agency initiated public health investigations to establish the incidence of Salmonella contamination and origin of eggs used by catering premises implicated in outbreaks of Salm. Enteritidis. METHODS AND RESULTS: Between October 2002 and November 2004, 16 971 eggs were sampled and Salmonella were recovered from 3.4%. Salmonella was isolated from 5.5% and 6.3% of Spanish and eggs of unknown origin, respectively, used in catering premises linked to outbreaks, a level significantly higher than that (1.1%) found in nonLion Quality UK eggs sampled. The small sample of UK Lion Quality eggs tested (reflecting their lack of use in premises visited) did not contain Salmonella. Several phage types of Salm. Enteritidis other than phage type 4 (PT 4) were identified with nonUK eggs. CONCLUSIONS: Eggs from Spain were implicated as a major source of infection. Eggs were contaminated more frequently with Salmonella when shells were dirty and/or cracked, and stored at above 8 degrees C. SIGNIFICANCE AND IMPACT OF THE STUDY: The use of Spanish eggs by the catering sector has been identified as a consistent significant factor in many of the outbreaks caused by Salm. Enteritidis nonPT4 in England and Wales during 2002-2004. Advice to caterers and hospitals that raw shell eggs should not be used in food that will either not be cooked or only lightly cooked should be reinforced.     

SEF17 fimbriae are essential for the convoluted colonial morphology of Salmonella enteritidis

Salmonella enteritidis isolated from poultry infections generated a convoluted colonial morphology after 48 h growth on colonisation factor antigen (CFA) agar at 25°C. A mutant S. enteritidis defective for the elaboration of the SEF17 fimbrial antigen, in which the agf gene cluster was inactivated by insertion of an ampicillin resistance gene cassette, and other wild-type S. enteritidis transduced to this genotype failed to produce convoluted colonies. However, growth of SEF17⁻ mutants at 25°C on CFA agar supplemented with 0.001% Congo red resulted in partial recovery of the phenotype. Immunoelectron microscopy demonstrated that copious amounts of the SEF17 fimbrial antigen were present in the extracellular matrix of convoluted colonies of wild-type virulent S. enteritidis isolates. Bacteria were often hyperflagellated also. Immunoelectron microscopy of SEF17⁻ mutants grown on CFA agar+0.001% Congo red demonstrated the elaboration of an as yet undefined fimbrial structure. Isolates of S. enteritidis which were described previously as avirulent and sensitive to environmental stress failed to express SEF17 or produce convoluted colonies. These data indicate an essential role for SEF17, and possibly for another fimbria and flagella, in the generation of the convoluted colonial phenotype. The relationship between virulence and colonial phenotype is discussed.     

Quantitative microbiological risk assessment: principles applied to determining the comparative risk of salmonellosis from chicken products

Since causal links have been established between food-borne illness and particular microorganisms, it has been possible to assess the public health risks of their presence in foods and propose measures to ensure the safety of customers. Effective control measures for food safety have been based on knowledge of the resistance of pathogenic microorganisms to the treatments used for preservation (e.g. acidification or reduced water activity) or decontamination (e.g. pasteurisation or sterilisation). Using this principle, food safety has been managed informally and successfully within the food industry for many years. Recently, formal risk assessment schemes, for example from Codex Alimentarius, have developed and placed the elements of decision-making on suitable control measures into a formal framework with clearly identifiable stages. The output of the four stages of a formal risk assessment (hazard identification, hazard characterisation, exposure assessment and risk characterisation) provides the basis for decisions on actions needed to control the identified hazard. There are many difficulties in ensuring that risk assessments are realistic and accessible to potential users, in many cases their value is limited by the data available. The study reported here on control of Salmonella in poultry products illustrates that it is possible to produce a comparative risk assessment based on published data. This study is not a full quantitative risk assessment, but provide useful pointers for a risk manager. Differences are discussed between the exposure assessment data needed to propose controls for infectious or toxigenic pathogens. For infectious pathogens, the presence of viable and infectious microorganisms is itself the hazard, but for toxigenic microorganisms, absence or destruction of viable cells at ingestion does not in itself ensure the absence of toxin. For toxin hazards, exposure assessment needs to consider previous conditions that may have led to toxin formation and persistence, rather than just the level of microbes at ingestion.     

Monitoring wild greenfinch (Carduelis chloris) for Salmonella enterica typhimurium

The identification and monitoring of emerging infectious diseases in free living wild birds is a challenge to wildlife biologists. In this study, a non-invasive methodology for identifying salmonellosis in wild garden birds was developed. We focussed on greenfinch, Carduelis chloris, which were found to have a seasonal pattern in the occurrence of Salmonella Typhimurium DT 56(v). Principal components analysis of biometric data indicated that low fat and low weight could be useful indicators of Salmonella positive greenfinch. A combination of biometrics taken from live birds, faecal analysis, and behavioural observations provide an effective and efficient system for identifying the presence of salmonellosis within greenfinch.