Biomedical subjects

G Salvat

Publications and source records attributed to G Salvat.

At least 37 records · Page 2Linked to original sources

PubMed2003

Effect of two candidate genes on the Salmonella carrier state in fowl.

Selection for increased resistance to Salmonella carrier-state (defined as the persistency of the bacteria 4 wk after inoculation) could reduce the risk for the consumer of food toxi-infections. The effects of two genomic regions on chromosomes 7 and 17 harboring two genes, NRAMP1 (SLC11A1) and TLR4, known to be involved in the level of chicken infection 3 d after inoculation by Salmonella were thus tested on a total of 331 hens orally inoculated at the peak of lay with 10(9) bacteria. The animals and their parents were genotyped for a total of 10 microsatellite markers mapped on chromosomes 7 and 17. Using maximum likelihood analysis and interval mapping, it was found that the SLC11A1 region was significantly involved in the control of the probability of spleen contamination 4 wk after inoculation. Single nucleotide polymorphisms (SNP) within the SLC11A1 and TLR4 gene were tested on those animals as well as on a second batch of 279 hens whose resistance was assessed in the same conditions. As the former was significantly associated with the risk of spleen contamination and the number of contaminated organs, SLC11A1 appears to be involved in the control of resistance to Salmonella carrier state. The involvement of the TLR4 gene was also highly suspected as a significant association between SNP within the gene, and the number of contaminated organs was detected.

Animals↗

PubMed2001

Risk factors for Campylobacter spp. contamination in French broiler-chicken flocks at the end of the rearing period.

Our objectives were to identify risk factors for contamination of French broiler flocks by Campylobacter. We used 75 broiler farms in western France. A questionnaire was administered to the farmers and samples of fresh droppings were taken to assess the Campylobacter status of the broiler flocks. 42.7% of the flocks were positive for Campylobacter spp. The risk of contamination of the broiler flocks by Campylobacter was increased in summer/autumn, in houses with static air distribution, when two or more people took care of the flock, in poultry farms with three or more houses and when the drinking water for the chickens was acidified. The presence of litter-beetles in the change room also increased the risk of contamination. The administration of an antibiotic treatment following a disease decreased the risk of a flock being contaminated by Campylobacter.

Animals↗

PubMed2001

Tracing of Salmonella spp. in two pork slaughter and cutting plants using serotyping and macrorestriction genotyping.

AIMS: The origin of Salmonella contamination of pork products is not well established. In order to further this knowledge, the transmission of Salmonella spp. from live pigs to pork cuts was investigated in two pork slaughter and cutting plants. METHODS AND RESULTS: Salmonella spp. were isolated from both pork (pigs, carcasses, cuts) and the environment before and during slaughterhouse activities. Eight serotypes were identified. XbaI and SpeI macrorestriction distinguished 20 genotypes of Salmonella Typhimurium and 16 genotypes of Salmonella Derby. A major cluster of Salmonella Typhimurium genotypes was common to both plants and all pig-related genotypes, while a predominant pig-related Salmonella Derby genotype was common to both plants. CONCLUSION: None of the Salmonella strains persisted for long periods in the pork-processing environments. SIGNIFICANCE AND IMPACT OF THE STUDY: This work shows that contaminated live pigs, because of bacterial spread due to the process and ineffective cleaning procedures, are involved in Salmonella contamination.

Abattoirs↗

PubMed2001

Campylobacter contamination in French chicken production from farm to consumers. Use of a PCR assay for detection and identification of Campylobacter jejuni and Camp. coli.

AIMS: Campylobacter contamination in French chicken production from the farm to the consumer was determined using a PCR assay for bacteria detection and identification. METHODS AND RESULTS: Samples were bird droppings from poultry houses, neck skins, livers, hearts, gizzards, wings, legs and escalopes from slaughterhouses and gizzards, legs, drumstick, breast and escalopes from a supermarket. Bacterial DNA extraction was performed after an enrichment step in a broth and was followed by PCR. An internal control (IC) was used for both DNA extraction and PCR. Campylobacter were detected in 79.2% of poultry houses. Of the 303 samples, 201 were Campylobacter-positive (i.e. 66.3%) including 43.2% faecal samples, 5.6% slaughterhouse samples and 17.5% supermarket samples. There was no significant difference between the molecular method and the conventional culture technique for Campylobacter detection whatever the samples. The sensitivity was 5 UFC g(-1) of samples and 1.5 x 10(3) UFC ml(-1) of enrichment broth. The use of IC revealed PCR inhibition in 13 samples and problems in the DNA extraction in five samples. CONCLUSION: Significant Campylobacter contamination affects all stages of French chicken production. SIGNIFICANCE AND IMPACT OF THE STUDY: The understanding of Campylobacter contamination at different levels of chicken production and the determination of the best place(s) for intervention are important for significantly decreasing Campylobacteriosis. Our technique is rapid and can be used on different chicken samples for Campylobacter detection and identification.

Abattoirs↗

PubMed2001

Molecular epidemiology of Listeria monocytogenes isolates collected from the environment, raw meat and raw products in two poultry- and pork-processing plants.

AIMS: In order to study the transmission of Listeria monocytogenes in a poultry and a pork meat plant, we analysed the contamination by this pathogen over several months. METHODS AND RESULTS: Five hundred and two isolates of L. monocytogenes were collected and characterized by genotyping and serotyping. Thirty-seven genotypes were obtained by ApaI-restriction analysis-pulsed field gel electrophoresis (REA-PFGE) and 35 by SmaI-REA-PFGE and resulted in 50 combined genotypes. The tracing of the contamination in both plants showed that some clones were able to survive for several months. However, some other clones were found only during processing operations, were not detectable after cleaning and seemed to enter continuously into the plant. CONCLUSIONS: Some L. monocytogenes strains may persist for a long period in the plant environment. Different genotypes can be associated with poultry as well as pork meat. SIGNIFICANCE AND IMPACT OF THE STUDY: Listeria monocytogenes contamination can be due to contaminated raw materials, bacterial spread and also ineffective cleaning procedures.

Animals↗

PubMed2000

Some safety aspects of Salmonella vaccines for poultry: in vivo study of the genetic stability of three Salmonella typhimurium live vaccines.

Live vaccine strains of Salmonella should be avirulent, immunogenic and genetically stable. Some isolates of three commercially available live vaccine strains of Salmonella typhimurium, sampled during a study on their persistence in a vaccinated flock of chickens, were analyzed for genetic stability using macrorestriction analysis of their genome. Two out of the three vaccine strains showed genetic instabilities. Two of the 51 isolates of Zoosaloral vaccine strain and nine of the 32 analyzed isolates of chi(3985), a genetically modified organism, were variants and showed different macrorestriction profiles.

Animals↗

PubMed2000

Physicochemical surface properties of five Listeria monocytogenes strains from a pork-processing environment in relation to serotypes, genotypes and growth temperature.

Physicochemical surface properties, related to electrostatic, van der Waals and Lewis acid-base interactions, of five Listeria monocytogenes strains isolated from pork-processing environments were determined after two subcultures at 37 degrees C and a final culture at three temperatures: 37, 10 and 4 degrees C. Three strains (Lm1, Lm114 and Lm191) were genetically related while two were unrelated (Lm25 and Lm74) according to ApaI-macrorestriction and pulsed-field gel electrophoresis (PFGE) typing. Listeria monocytogenes cell surfaces were generally negatively charged regardless of pH and tended to be hydrophilic due to a basic character. However, variable physicochemical surface properties of the five Listeria monocytogenes isolates were observed after growth at 37 degrees C. After growth at 10 degrees C, the three genetically related isolates exhibited similar surface properties and were slightly more hydrophilic and basic than the others. After growth at 4 degrees C, the five isolates displayed the same weak affinity for all kinds of solvents and low electrophoretic mobility values. A sharp decrease of temperature and subsequent growth of various Listeria monocytogenes strains resulted in loss of the physicochemical surface property variability, which may suggest the role of common chill adaptation mechanisms affecting surface properties.

Abattoirs↗

PubMed1999

Listeria monocytogenes in pork slaughtering and cutting plants. Use of RAPD, PFGE and PCR-REA for tracing and molecular epidemiology.

In order to determine the origin of pork cuts contamination by Listeria monocytogenes, 287 isolates, collected from five French pork slaughtering and cutting plants, from live pigs to pork cuts, were characterised using three molecular typing methods: random amplification of polymorphic DNA (RAPD) carried out with five different primers, genomic macrorestriction using ApaI with pulsed-field gel electrophoresis (PFGE) and a PCR-restriction enzyme analysis (PCR-REA) based on the polymorphism existing within the inlA and inlB genes. Results obtained from RAPD and PFGE were closely related and distinguished respectively 17 RAPD types (r1-r17) and 17 PFGE types (a1-a17) among the 287 isolates, whereas the PCR-REA analysis only yielded two profiles (p1 and p2). Considering the combined results obtained with the three molecular typing methods, 19 Listeria monocytogenes genotypes (1-19) were distinguished. Serotyping led at least four serotypes being distinguished: 1/2a, 3a, 1/2c and 3c. The application of genotyping identified the predominance of a Listeria monocytogenes strain of type (1) and other very closely related ones (5, 9, 10, 12, 13, 14, 16 and 19) which were present on pork as well as in the environment within the five investigated plants. This study also pointed out the presence of these closely related Listeria monocytogenes strains over a 1-year period in the environments of two plants, even after cleaning and disinfection procedures. This highlights the possibility for some Listeria monocytogenes strains to persist in pork processing environments and raises the problem of the efficiency of cleaning and disinfection procedures used in pork slaughterhouses, chilling and cutting rooms.

Abattoirs↗

PubMed1999

An immunoconcentration-PCR assay to detect Salmonella in the environment of poultry houses.

An immunoconcentration-PCR assay was developed for the rapid and specific detection of Salmonella. This assay was evaluated against a conventional bacteriological method for the detection of Salmonella from environmental swabs of poultry houses. The 120 samples investigated were pre-enriched in phosphate buffered peptone water and Salmonella was separated by an immunoconcentration process using an automated system (VIDAS bioMérieux, Marcy l'Etoile, France) prior to PCR. The specificity of the assay was high as no false-positives were found. The sensitivity of the assay was 70%. The correlation between the ICS-PCR assay and the bacteriological method was 84%.

Animals↗

PubMed1999

Evaluation of a multiplex PCR assay for simultaneous identification of Salmonella sp., Salmonella enteritidis and Salmonella typhimurium from environmental swabs of poultry houses.

A Multiplex PCR-based assay (m-PCR) with three sets of primers was developed for the detection of all serotypes of Salmonella enterica and the identification of Salmonella Enteritidis and Salmonella Typhimurium. This method was evaluated against a bacteriological method for the analysis of environmental swabs of poultry houses. Samples were preenriched in phosphate-buffered peptone water for 24 h and subjected to three different protocols prior to PCR: (i) an immunomagnetic separation using Dynabeads anti-Salmonella (Dynal); (ii) a DNA extraction procedure using the Instagene matrix; (iii) an additional step of culture on an MSRV medium. With protocols 1 and 2, eight positive results were found by PCR and 20 with the bacteriological method. Protocol 3 combining MSRV and PCR gave similar results to those obtained from bacteriological methods and allowed Salmonella detection within 2 days.

Animals↗

PubMed1999

Identification by a multiplex PCR-based assay of Salmonella typhimurium and Salmonella enteritidis strains from environmental swabs of poultry houses.

A multiplex-PCR-based assay (m-PCR) was developed for the detection of Salmonella and for the identification of the two serotypes Enteritidis and Typhimurium. Three sets of primers selected from different genomic sequences amplified a 429 bp fragment specific for the genus Salmonella within a randomly cloned sequence, a 559 bp target specific for Salmonella Typhimurium within the fliC gene and a 312 bp fragment specific for Salmonella Enteritidis within the sefA gene. The m-PCR-based assay was used for detecting Salmonella from 1078 environmental swabs of poultry houses. Prior to PCR, these swabs were pre-enriched in phosphate-buffered peptone water for 18-20 h and then sub-cultured on a Modified Semi-solid Rappaport Vassiliadis medium (MSRV) for 18-20 h. The m-PCR combined with MSRV had a better sensitivity (95%) than the bacteriological method (92.5%). The MSRV-m-PCR assay and the bacteriological method had an agreement rate of 95.6%.

Animals↗

PubMed1999

Measurement of the residual contamination of post-weaning facilities for pigs and related risk factors.

A survey was designed in France to assess residual contamination in post-weaning rooms within the 2 h prior to restocking when all the pigs of the previous batch had been removed (all in-all out) and after application of the 'in use' cleaning procedures. The protocol was based on the results of a pilot study and was considered 'a good compromise'. Special agar plates (Rodac plates) with a prominent agar medium surface were prepared and used to collect the bacteria. They were applied by impression onto the floors and the partitions between the pens. In all, 24 plates were applied per room. The culture medium was VRBG which mainly permits the growth of enterobacteria. The plates were incubated for 24 h (37 degrees C) and the Colony-Forming Units (CFU) counted. After sampling, measurements were made and information collected from the farmer to obtain potential explanations for the resulting microbiological data. The same scientist sampled 129 post-weaning rooms with slatted floors. Of the 3045 plates, 18.4% were negative and 12.8% were heavily colonized (> 300 CFU). A score combining six criteria based on the counts was produced, and indicated a wide spread of the rooms. The profile of circumstances associated with the different levels of the score was extracted. The following points should be respected to achieve low residual contamination: removal of the slurry from the pit below the slatted floor; damping to be started soon after pig removal; prolonged damping; thorough washing; disinfecting soon after washing; and attention to the recommended dosage of disinfectant. Smooth surfaces were found to be less contaminated than rough ones. It is concluded that cleaning and disinfecting operations must not be considered as minor tasks and should clearly be integrated into the process of pig production.

Animals↗

PubMed1999

Molecular characterization of the diversity of Campylobacter spp. isolates collected from a poultry slaughterhouse: analysis of cross-contamination.

Investigations of a free-range broiler flock during the rearing period and at the slaughterhouse by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) of the flagellin (flaA) gene (flaA typing) have shown that poultry carcasses are contaminated by Campylobacter spp. strains which were previously present in the poultry faces. Moreover, the investigation of the previous and the following batches in the processing plant using flaA typing have shown that cross-contamination between batches coming from different flocks occurs and is also a risk factor for the presence of Campylobacter spp. on poultry carcasses.

Abattoirs↗

PubMed1999

Development of a m-PCR assay for simultaneous identification of Campylobacter jejuni and C. coli.

Multiplex PCR assay (m-PCR) with three sets of primers was developed for simultaneous identification of Campylobacter jejuni and C. coli. Poultry faecal samples were enriched in Preston broth for 24 h and streaking on selective media was performed before and after enrichment. m-PCR was applied on bacterial cultures harvested from media plates. The data showed a selective effect of Preston broth which favoured the growth of C. coli. Identification of the species by the hippurate hydrolysis test and by the m-PCR was performed on 294 isolates of Campylobacter. The efficiency of the identification by the biochemical test is only 34% in comparison to 100% efficiency with the PCR. The use of our m-PCR in combination with the culture method allowed reliable detection and identification of C. jejuni and C. coli within 3-4 d.

Animals↗

PubMed1998

Comparison of five typing methods for the epidemiological study of Listeria monocytogenes.

Five typing methods were compared in a study designed to adapt a strategy for epidemiologically typing large numbers of Listeria monocytogenes strains. The methods studied were serotyping, electrophoretic typing of esterases (zymotyping), restriction fragment length polymorphism of ribosomal DNA (ribotyping), random amplified polymorphic DNA (RAPD) and pulsed-field gel electrophoresis (PFGE). Data were analysed by computer-assisted statistical analysis. Included in the analysis were 35 strains of L. monocytogenes, including 14 epidemic strains isolated during outbreaks in France in 1992 and 1993, and 21 strains isolated from food and the environment. Five serotypes, eight zymotypes, ten ribotypes, 13 RAPD patterns and 12 PFGE patterns were identified among the 35 strains. The most discriminating combination of typing methods was ribotyping and PFGE typing [27 types, discriminatory index (D.I.) = 0.978]. A factorial analysis of correspondence for each method differentiated the epidemic strains from the environmental strains. This study shows that computer-assisted statistical treatment of the data, combined with the use of discriminating typing methods, is a powerful tool for the epidemiological analysis of Listeria monocytogenes.

Animals↗

PubMed1998

Development of a new culture medium for the rapid detection of Salmonella by indirect conductance measurements.

The main difficulties in conductance medium development are to allow Salmonella to grow and produce a conductance signal while impeding growth of related species such as Escherichia coli and Citrobacter freundii. Various selective agents were screened for these capacities and a new medium was derived, named KIMAN (Whitley Impedance Broth basal medium supplemented with three selective components: novobiocin, malachite green and potassium iodide). This medium supported the growth of Salmonella serotypes and inhibited non-salmonella strains in pure cultures.

Bacteriological Techniques↗

PubMed1998

Development of a multiplex PCR gene fingerprinting method using gyrA and pflA polymorphisms to identify genotypic relatedness within Campylobacter jejuni species.

The two genes gyrA and pflA, whose sequence variability had been previously described, were evaluated separately for their potential value in discriminating strains of Campylobacter jejuni. A single method was then developed by which the two loci were simultaneously amplified using a multiplex PCR procedure, and banding patterns were generated using a pre-selected set of restriction endonuclease enzymes. The method was applied to 18 strains of Camp. jejuni from different poultry sources varying in geographical origin and year of isolation. Results were combined and compared by means of numerical analysis with the classification obtained by flaA-typing and macrorestriction SmaI and KpnI. The usefulness of PCR fingerprinting of the gyrA/pflA genes for rapid ordering of strains by genotypic relatedness and providing additional information for estimating the degree of linkage between strains was demonstrated.

Animals↗

PubMed1998

Antimicrobial efficacy of AvGard carcase wash under industrial processing conditions.

1. The efficacy of the AvGard Trisodium Phosphate (TSP) immersion carcase wash process was evaluated during 5 industrial trials against Salmonella, Enterobacteriaceae, thermotolerant coliforms and total aerobic count. The effect against Pseudomonas was also studied in the first 3 trials. 2. Dramatic reductions in Salmonella incidence were seen using a whole carcase rinse method. In 4 of the 5 trial sites, only one positive sample was found after AvGard treatment (average 0.5% incidence), in spite of an average control incidence of 57.7%. In the 5th site, a water-chilled broiler plant, an average control incidence of 74.0% was reduced to 9.4% after AvGard treatment. 3. In the latter case, Most Probable Number (MPN) analyses were performed on some of the Salmonella positive samples taken from the control and post-treatment series; the average MPN count per carcase on controls was 115, whereas for AvGard treated birds the figure was only 0.6 per carcase, a greater than 2 log reduction. 4. In addition, AvGard treatment gave average log reductions for all trials of: Enterobacteriaceae; 2.5 log; Coliforms; 2.7 log, and Total Aerobic Count; 1.1 log, leading to carcases substantially free of Gram negative pathogens. 5. Pseudomonas was reduced by an average of 1.7 log in the first 3 trials, dramatically reducing the carcase loading of this important spoilage organism.

Animals↗