Comparison of random amplified polymorphic DNA analysis and enterobacterial repetitive intergenic consensus-PCR for epidemiological studies of Salmonella.

Discrimination of 70 Salmonella strains previously studied by ribotyping was realized by RAPD and ERIC-PCR analysis. RAPD results on the 56 S. typhimurium isolates did not closely match those of ribotyping. With ERIC-PCR, two fingerprints only were obtained. For the 14 S. enteritidis strains, a helpful discrimination was obtained with RAPD analysis, while ERIC-PCR resulted in a single fingerprint.

Quantitative risk assessment of human salmonellosis from the consumption of a turkey product in collective catering establishments.

The quantitative risk assessment (QRA) approach recommended by the Codex Alimentarius Commission was used to assess the risk of human salmonellosis from the consumption of 'cordon bleu', a specific turkey product, in collective catering establishments (CCEs) of a French department. The complete process was modeled and simulated, from the initial storage in the CCE freezer to the consumption, using a Monte Carlo simulation software. Data concerning the prevalence of contaminated 'cordon bleu', the level of contamination of Salmonella, the cooking and storage process were collected from 21 CCEs and 8 retailers of 'cordon bleu' in the selected department. Thermal inactivation kinetics for Salmonella were established to estimate the effect of heat treatment on the concentration in the product and to calculate the dose that could be ingested by the consumer. The Beta-Poisson dose-response model of Rose and Gerba [Water Science and Technology 24 (1991) 29] with the specific parameters for Salmonella was used to estimate the probability of infection related to the ingestion of a particular dose and a factor was applied to estimate the probability of illness from ingestion. The individual risk of salmonellosis, the risk of outbreak and the number of cases were calculated using Monte Carlo simulation method. The risk of salmonellosis was close to zero when the 'cordons bleus' were cooked in the oven. Therefore, the risk was calculated for the fryer cooking since the insufficient cooking time observed was, sometimes, at the origin of low temperatures (37-89 degrees C). The influence of both the initial concentration of Salmonella in the product and the heat storage before consumption on the final risk was studied. For a high initial concentration of Salmonella in the product, when the 'cordons bleus' are fryer cooked, the average risk of salmonellosis was equal to 3.95 x 10(-3) without storage before consumption and 2.8 x 10(-4) if the product is consumed after storage. This paper presents the results of the QRA and discusses risk management options to minimize the risk of salmonellosis.

Evidence of transplacental transmission of bluetongue virus serotype 8 in goats.

During the incursion of bluetongue virus (BTV) serotype 8 in Europe, an increase in the number of abortions in ruminants was observed. Transplacental transmission of BTV-8 in cattle and sheep, with subsequent foetal infection, is a feature of this specific bluetongue serotype. In this study, BTV-8 ability to cross the placental barrier at the beginning of the second third of pregnancy and at the end of pregnancy was investigated in goats in two separate experiments. In the first experiment, nine goats were experimentally infected with BTV-8 at 61 days of pregnancy. Foetuses were collected 21 dpi. BTV-8 was evidenced by real time RT-PCR and by viral isolation using blood from the umbilical cord and the spleens of 3 out of the 13 foetuses. All dams were viraemic (viral isolation) at the moment of sampling of the foetuses. Significant macroscopic or histological lesions could not be observed in foetuses or in their infected dams (notably at the placenta level). In the second experiment, 10 goats were infected with BTV-8 at 135 days of pregnancy. Kids were born by caesarean section at the programmed day of birth (15 dpi). BTV-8 could not be detected by rt-RT-PCR in blood or spleen samples from the kids. This study showed for the first time that BTV-8 transplacental transmission can occur in goats that have been infected at 61 days of pregnancy, with infectious virus recovered from the caprine foetuses. The observed transmission rate was quite high (33%) at this stage of pregnancy. However, it was not possible to demonstrate the existence of BTV-8 transplacental transmission when infection occurred at the end of the goat pregnancy.

Evaluation of humoral response and protective efficacy of two inactivated vaccines against bluetongue virus after vaccination of goats.

Bluetongue serotype 8 has become a major animal health issue in the European Union and the European member States have agreed on a vaccination strategy, which involves only inactivated vaccines. In this study, the efficacy of two inactivated vaccines against bluetongue virus serotype 8 (BTV-8) used in Europe since 2008, BTVPUR ALSAP(®) 8 (MERIAL) and BOVILIS(®) BTV8 (Intervet/SP-AH), was evaluated in goats immunized and challenged with BTV-8 field isolates under experimental conditions. Serological, virological and clinical examinations were conducted before and after challenge. Three groups of 10 goats each (groups A, B and C) were randomly constituted and 2 groups (A and C) were subcutaneously vaccinated twice with one dose of the two commercial vaccines BTVPUR ALSAP 8 (group A) or BOVILIS BTV8 (group C) respectively. Animals of the groups A, C and B (B: controls) were challenged with a virulent inoculum containing BTV-8. During the experiment, it was found out that the BTV-8 challenge inoculum was contaminated with another BTV serotype. However, results demonstrated that vaccination of goats with two injections of BTVPUR ALSAP 8 or BOVILIS BTV8 provided a significant clinical protection against a BTV-8 challenge and completely prevented BTV-8 viraemia in all vaccinated animals. Qualitative data showed no difference in the kinetics and levels of the humoral response induced by these two inactivated vaccines.

Characteristics of Salmonella contamination of broilers and slaughterhouses in the region of Constantine (Algeria).

The present study provides the first data about the prevalence of Salmonella contamination of broilers and slaughterhouses in the region of Constantine, Algeria. The serotypes and anti-microbial resistance phenotypes of the isolates were determined, and risk factors contributing to the contamination were evaluated. A total of 2490 samples, 1800 originating from 30 broiler farms and 690 from 15 slaughterhouses, were taken during two periods: March 2005-June 2006 and September 2006-March 2007. Salmonella contamination concerned 37% of the broiler farms and 53% of the slaughterhouses. Among the 55 isolates recovered, 10 different serotypes were identified. The most frequently recovered serotypes in both slaughterhouses and breeder farms were S. Hadar (36%, n = 20), S. Virchow (16%, n = 9), S. Infantis (10.9%, n = 6), S. Albany (11%, n = 6) and S. Carnac (7%, n = 4). Isolates belonging to S. Heidelberg (2%, n = 1) and S. Rissen (2%, n = 1) were found only in farms, while those belonging to S. Typhimurium (9%, n = 5), S. Enteritidis (4%, n = 2) and S. Montevideo (2%, n = 1) were recovered only from slaughterhouses. Thirty-nine isolates (80%) were resistant to at least one anti-microbial and 51% were multi-resistant, i.e. resistant to two or more anti-microbial molecules. About 58% (n = 32) were resistant to streptomycin, 36% (n = 20) to tetracyclines, 27% (n = 15) to nalidixic acid, 13% (n = 7) to ofloxacin and one isolate to enrofloxacin. Finally, seven distinct anti-microbial resistance profiles were identified. In parallel, four risk factors were found to be significantly associated with Salmonella contamination. Together with the huge spread of Salmonella in the broiler production chain in Constantine, Algeria, these risk factors highlight the hazards of the broiler channels, particularly linked to poor technical and hygiene practices.

[Pathogenic power of Salmonellae: virulence factors and study models]. / Le pouvoir pathogène des salmonelles: facteurs de virulence et modèles d'étude.

Salmonellae are potentially pathogenic for humans as well as for numerous animal species. They possess numerous virulence factors, which allow them to adapt to various environmental conditions and to host response at each step of the pathogenic process. Key-steps such as the invasion of epithelial cells or survival within macrophages have been extensively studied. These studies have led to the discovery of an original protein secretion system and have demonstrated the existence of pathogenicity islands. This considerable progress is due to the development of numerous in vitro and in vivo models and of new identification strategies for the implicated genes. Recently, many original and elegant strategies have been recently proposed.

[Epidemiologic markers of Salmonella]. / Les marqueurs épidémiologiques des salmonelles.

Salmonellae are enterobacteria responsible for outbreaks of human and animal clinical diseases, with important hygienic and economic consequences. Accurate epidemiological studies require the use of efficient markers, which make it possible to trace the establishment and diffusion of different bacterial strains and also to evaluate the similarities between different isolates. Numerous phenotypic and genotypic markers applicable to Salmonella are available for these epidemiological studies. Nevertheless, the relative interest of those markers depends on the serotypes, and a different hierarchy can be achieved for the Typhimurium and Enteritidis serotypes.

Evaluation of IS200-PCR and comparison with other molecular markers To trace Salmonella enterica subsp. enterica serotype typhimurium bovine isolates from farm to meat.

A procedure that uses an original molecular marker (IS200-PCR) and that is based on the amplification of DNA with outward-facing primers complementary to each end of IS200 has been evaluated with a collection of 85 Salmonella enterica subsp. enterica serotype Typhimurium isolates. These strains were isolated from a group of 10 cows at different stages: during transportation between the farm and the slaughterhouse, on the slaughter line, from the environment, and from the final product (ground beef). The 85 isolates were characterized by their antibiotic resistance patterns and were compared by IS200-PCR and by use of four other genotypic markers. Those markers included restriction profiles for 16S and 23S rRNA (ribotypes) and amplification profiles obtained by different approaches: random amplified polymorphic DNA analysis, enterobacterial repetitive intergenic consensus PCR, and PCR ribotyping. The results of the IS200-PCR were in accordance with those of other molecular typing methods for this collection of isolates. Five different genotypes were found, which made it possible to refine the hypotheses on transmission obtained from phenotypic results. The genotyping results indicated the massive contamination of the whole group of animals and of the environment by one clonal strain originally recovered from one cow that excreted the strain. On the other hand, a few animals and their environment appeared to be simultaneously contaminated with genetically different strains.

Value of plasmid profiling, ribotyping, and detection of IS200 for tracing avian isolates of Salmonella typhimurium and S. enteritidis.

Seventy selected strains of Salmonella typhimurium and S. enteritidis isolated from related poultry flocks in three independent geographical areas were characterized by phenotypic and genotypic methods to compare the usefulness of the methods in epidemiological studies. The 56 S. typhimurium isolates were poorly discriminated by their biotypes, resistance patterns, and plasmid profiles. Nine different ribotypes were obtained after DNA digestion by BglII, PvuII, and SmaI. Seven IS200 types, characterized by six to nine copies of IS200 on the chromosome, were detected after digestion of genomic DNA by PstI. These studies resulted in the definition of 15 clonal lineages distributed in three clusters. The 14 S. enteritidis strains were not discriminated either by ribotyping or by detection of IS200 (IS200 typing), but were separated on the basis of antibiotic resistance and plasmid profiling. The stability of the insertion sequence type was confirmed by inoculation of an S. typhimurium strain to axenic chickens reared for 15 weeks in sterile isolators.