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Biomedical subjects

F Humbert

Publications and source records attributed to F Humbert.

At least 19 recordsLinked to original sources

Evaluation of a French ELISA for the detection of Salmonella Enteritidis and Salmonella Typhimurium in flocks of laying and breeding hens.

In France, the regular and compulsory detection of Salmonella Enteritidis (SE) and Salmonella Typhimurium (ST) in flocks of breeding and laying hens is based on bacteriological examination of environmental swabs and faeces samples. The aim of this study was to compare this bacteriological examination with a serological method (ELISA) developed in our laboratory. This ELISA was first evaluated by use of artificially infected hens. During these experimental infection studies, several groups of hens were inoculated with SE, ST, different vaccines and different Salmonella serovars to calculate the experimental parameters of our ELISA. Then, in a field study, 43 flocks were followed monthly using two bacteriological samples (environmental swab and pool of faeces) and 20 serological samples (sera or yolks). Twenty-seven flocks without SE or ST gave a negative serological response throughout their surveillance. Among the 10 various serovars different from SE and ST isolated in this study, S. Heidelberg, S. Agona and S. Hadar gave seropositive results in seven flocks. Consequently, this ELISA was not specific of SE and ST as it detected serovars sharing or not common antigens with SE and ST. Seropositive results were also obtained each month for two flocks where no Salmonella could be isolated. Finally, in seven flocks found infected with SE or ST, the positive ELISA results appeared later than the bacteriological detection. Therefore, for the detection of chicken flocks recently infected with SE or ST, bacteriological examination currently used in France seems to be more appropriate than this ELISA.

Animals↗

Use of avilamycin for growth promotion and avilamycin-resistance among Enterococcus faecium from broilers in a matched case-control study in France.

We assessed the putative link between avilamycin-resistant Enterococcus faecium carriage and avilamycin consumption in broilers. As part of the French programme of monitoring for antimicrobial resistance, broilers sampled at slaughterhouse in 1999 and 2000 and carrying avilamycin-resistant E. faecium were matched by slaughterhouse, slaughter month and production type (free-range, standard, light) with control broilers carrying avilamycin-susceptible strains. History of antibiotics consumption (either for growth promotion or therapeutic purpose) in the broiler flocks sampled was collected from the monitoring programme and consumption of each antibiotic class was screened as a potential risk factor. Avilamycin was a risk factor for avilamycin-resistant E. faecium carriage: OR=2.3.

Abattoirs↗

Characterization of fibrosurfin, an interfibrillar component of sea urchin catch connective tissues.

The Sea URchin Fibrillar (SURF) domain is a four-cysteine module present in the amino-propeptide of the sea urchin 2alpha fibrillar collagen chain. Despite numerous international genome and expressed sequence tag projects, computer searches have so far failed to identify similar domains in other species. Here, we have characterized a new sea urchin protein of 2656 amino acids made up of a series of epidermal growth factor-like and SURF modules. From its striking similarity to the modular organization of fibropellins, we called this new protein fibrosurfin. This protein is acidic with a calculated pI of 4.12. Eleven of the 17 epidermal growth factor-like domains correspond to the consensus sequence of calcium-binding type. By Western blot and immunofluorescence analyses, this protein is not detectable during embryogenesis. In adult tissues, fibrosurfin is co-localized with the amino-propeptide of the 2alpha fibrillar collagen chain in several collagenous ligaments, i.e., test sutures, spine ligaments, peristomial membrane, and to a lesser extent, tube feet. Finally, immunogold labeling indicates that fibrosurfin is an interfibrillar component of collagenous tissues. Taken together, the data suggest that proteins possessing SURF modules are localized in the vicinity of mineralized tissues and could be responsible for the unique properties of sea urchin mutable collagenous tissues.

Amino Acid Sequence↗

Potentials of radio-frequency field gradient NMR microscopy in environmental science.

An understanding of transport, flow, diffusivity and mass transfer processes is of central importance in many fields of environmental biotechnology such as biofilm, bioreactor and membrane engineering, soil and groundwater bioremediation, and wastewater treatment. Owing to its remarkable sensitivity to molecular displacements and to its noninvasive and nondestructive character, pulsed field gradient (PFG) nuclear magnetic resonance (NMR) can be a valuable tool for investigating such processes. In conventional NMR microscopy, spatial encoding is achieved by using static magnetic field gradients (B(0) gradients). However, an interesting alternative is to use radio-frequency magnetic field gradients (RF or B(1) gradients). Although the latter are less versatile than the former, RF field gradient microscopy is particularly suitable for dealing with heterogeneous systems such as porous media because of its quasi-immunity to background static magnetic field gradients arising from magnetic susceptibility inhomogeneities, unlike the B(0) gradients microscopy. Here, we present an overview of basic principles and the main features of this technique, which is still relatively unused. Different examples of diffusion imaging illustrate the potentialities of the method in both micro-imaging and the measurement of global or local diffusion coefficients within membranes and at liquid-solid interfaces. These examples suggest that a number of environmental problems could benefit from this technique. Different future prospects of application of B(1) gradient NMR microscopy in environmental biotechnology are considered.

Journal Article↗

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↗

Changes in culturability and virulence of Salmonella typhimurium during long-term starvation under desiccating conditions.

The survival of Salmonella typhimurium under desiccation and starvation conditions commonly associated with farm buildings was investigated in a desiccation model system: filtration onto polycarbonate membranes placed in a sealed desiccator with 0.0067 g/m3 absolute humidity. Heterogeneities within bacterial populations in relation to time of desiccation were investigated on a single-cell basis by epifluorescence microscopy coupled with an image analysis system in conjunction with fluorescent dyes Chemchrome V6 and DAPI. Changes in cellular states were compared to the results of plate counts (colony forming units, CFU) on selective (modified semi-solid Rappaport Vassiliadis (MSRV)) and non-selective (nutrient agar (NA) and R2A agar) media, and to the measurements of infectivity and virulence using two animal models (chicks and mice). During 9 weeks of experimental desiccation, total cell counts (DAPI) of starved S. typhimurium remained stable, as did esterase activity (Chemchrome V6), but DAPI fluorescence intensity decreased slowly. Bacterial cells entered gradually into non-culturable states (decrease of CFU counts on MSRV, NA and R2A agar media) and the total loss of culturability on NA (defined as probability of presence of 1 CFU on the membrane inferior to 10 (-6)) was obtained after 9 weeks. Loss of chick infectivity and mice virulence in animal models occurred more rapidly, within three weeks of experimental desiccation.

Animals↗

Nuclear longitudinal relaxation time images by radiofrequency field gradients.

Combination of the Super Fast Inversion Recovery (SUFIR) method (D. Canet, J. Brondeau, and K. Elbayed, J. Magn. Reson. 77, 483 (1988)) and imaging procedures by radiofrequency field gradients (P. Maffei, P. Mutzenhardt, A. Retournard, B. Diter, R. Raulet, J. Brondeau, and D. Canet, J. Magn. Reson. A 107, 40 (1994)) provides spatially resolved maps of longitudinal relaxation times (T1). In addition to accurate T1 values, enhanced spatial resolution is obtained.

Equipment Design↗

Maps of self-diffusion coefficients by radiofrequency field gradient NMR microscopy

The methods of measurement of spatially resolved diffusion coefficients using radiofrequency field gradient (E. Mischler et al., J. Magn. Reson. B 106, 32, 1995; R. Kimmich et al., J. Magn. Reson. A 112, 7, 1995) produce 1D profiles whose amplitude is not only a function of the local self-diffusion coefficient but also is modulated by cosine functions of spatial coordinates. Due to this modulation diffusion-weighted images cannot be obtained unless cumbersome data processing is used. Here, we present a new sequence which avoids this modulation and yields in a straightforward manner true self-diffusion coefficient maps; this is in contrast with conventional methods which use static field gradients and which are therefore altered by background gradients. The feasibility and the reliability of the method are demonstrated with phantoms; it is also applied to different systems of interest such as solvent swelled rubber, membranes, and plants. Copyright 1999 Academic Press.

Journal Article↗

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↗

Self-Diffusion Measurements by Carbon-13 NMR Using Radiofrequency Field Gradients

This paper demonstrates the feasibility of self-diffusion measurements by radiofrequency field gradients with equipment tuned to the carbon-13 resonance frequency (i.e., 50 MHz for a field of 4.7 T). For the first time, self-diffusion coefficients of carbon disulfide and carbon tetrachloride are measured by NMR. The agreement with results from tracer methods is excellent. Copyright 1998 Academic Press. Copyright 1998 Academic Press

Journal Article↗

Diffusion measurements using radiofrequency field gradient: artifacts, remedies, practical hints

The two major advantages of experiments carried out with radiofrequency (RF) field-gradient NMR are the instrumental simplicity and the insensitivity to background static magnetic field gradients. These features combined with large RF gradients, which became available only recently, should make this technique especially attractive for molecular translational diffusion studies. However, a critical evaluation of the method shows that under some circumstances (small and/or heterogeneous samples, weak diffusion coefficients, very short relaxation times) the quality of measurements may be affected by a number of artifacts. Their origin has been investigated and several remedies have been considered; in particular, a new improved sequence is presented. The success of various experimental tests demonstrates the efficiency of the proposed solutions which thus open the way to much wider application fields. Copyright 1998 Academic Press.

Journal Article↗

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↗

Bacteriological monitoring of Salmonella enteritidis carrier birds after decontamination using enrofloxacin, competitive exclusion and movement of birds.

Two hundred and forty, four-week-old laying birds naturally infected with Salmonella enteritidis PT33 (Pasteur Institute phage typing system) were randomly divided twice (before and during the treatments) to obtain four separately housed groups of 60 birds and to study the efficacy of three decontamination treatments: enrofloxacin either with or without the movement of birds to a clean area, and enrofloxacin combined with movement of birds and a competitive exclusion treatment. The control group remained untreated. In each group contamination with S enteritidis was checked bacteriologically, every week from two months before until two months after the treatments began. All the samples taken from all the birds before the treatments began were S enteritidis-positive. After the treatments it was not possible to isolate salmonella either from the environment or from the faeces of the three treated groups. All the birds were humanely sacrificed at 22 weeks of age and samples of liver, spleen, ovaries and caeca were analysed for the presence of salmonella. The results demonstrated that although antibiotic therapy, the movement of birds into a clean house and competitive exclusion, either combined or not, had some efficacy in reducing infection levels, it was not possible to decontaminate all the birds completely.

Animal Husbandry↗

Evaluation of a new enrichment broth for the isolation of Salmonella spp. from poultry products.

Four selective enrichment broths were compared for the detection of Salmonella spp. in naturally contaminated poultry products and the recovery of atypical Salmonella strains in suspensions of pure cultures. In analysis of 100 poultry samples, the sensitivities observed were 94.0% for Müller-Kauffmann Tetrathionate-Brilliant Green (MKTBG), 97.6% for Rappaport Vassiliadis (RV), 42.2% for Selenite Cystine (SC) and 97.6% for the new broth KIMAN (Whitley Impedance Broth supplemented with 20 mg/l of novobiocin sodium salt, 10 mg/l of malachite green oxalate and 40 g/l of potassium iodide). The two most efficient broths--RV and KIMAN for recovery of atypical Salmonella strains (gallinarum biotypes gallinarum and pullorum, typhi, paratyphi A) were less toxic than MKTBG but more toxic than s.c. broth. According to these results, the use of RV and KIMAN could be a good combination to assure maximal recovery of Salmonella strains.

Animals↗

A selective medium for the rapid detection by an impedance technique of Pseudomonas spp. associated with poultry meat.

A new medium for detecting and enumerating Pseudomonas spp. associated with poultry meat spoilage by a rapid impedance technique was developed, after testing potential growth promoters for eight Pseudomonas strains and inhibitors against eight competing strains (Enterobacteriaceae) able to grow on the medium of Mead and Adams (1977). Four basal media (brain heart infusion, brucella broth, Shaedler broth and Whitley impedance broth (WIB)) and a synthetic medium were evaluated. Whitley impedance broth was the best basal medium for detecting variations in impedance in relation to Pseudomonas growth. The efficiency of WIB was improved by adding compounds which enhanced the growth of Pseudomonas on the synthetic medium. Among the incubation temperatures tested, 22 degrees C proved to be the best compromise between growth of Pseudomonas associated with poultry meat spoilage and inhibition of competitors. Among the 15 inhibitory substances evaluated against Pseudomonas competitors, five were chosen for inclusion in the final medium: metronidazole, carbenicilline, cetrimide, cycloheximide and diamide (MCCCD medium). Preliminary results obtained from experiments with beef and pork meat showed that this medium could also be used without diamide and at an incubation temperature of 25 degrees C. The impedance technique using MCCCD medium was then compared with an official method which uses the medium of Mead and Adams (1977) on 106 samples of poultry neck skin. The linear regression coefficient between the two techniques was approximately r = 0.85. Impedance was able to detect 10(3) Pseudomonas g-1 within less than 19 h making it a promising technique for predicting poultry meat spoilage.

Animals↗

Effects of AvGard treatment on the microbiological flora of poultry carcases.

1. The efficiency of the AvGard (or Assur-Rince in the USA) trisodium phosphate poultry carcase decontamination process was evaluated during both manual and industrial trials against total aerobic mesophilic count (TAMC), thermotolerant coliforms, Pseudomonas, Enterobacteriaceae, Campylobacter, Listeria monocytogenes and Salmonella. 2. The TSP treatment proved to have significant effects on the bacterial decontamination of poultry neck skin, lowering the contamination by a factor of about 10 for TAMC and of 100 for Coliform and Pseudomonas. 3. Numeration of Salmonella with an innovative miniaturised most probable number method has proved that the effect upon these micro-organisms was also close to 2 log10 reduction. 4. The effect of TSP treatment on the ecological balance of psychrotrophic bacterial flora was also investigated to study the origin of the shelf-life flora of treated carcases (Pseudomonas being reduced to the limit of detection) and to ascertain whether L. monocytogenes might gain a competitive advantage. In fact AvGard reduced the number of L. monocytogenes on poultry carcases. 5. As a consequence of the virtual elimination of the Pseudomonas usually present, Brochothrix thermosphacta became the main species responsible for putrefaction. 6. Because the growth rate of Brochothrix thermosphacta was greater than that of L. monocytogenes at refrigeration temperature, it was considered that putrefaction would occur before the emergence of large numbers of L. monocytogenes.

Animals↗

[The risk of transmission of salmonellae in poultry farming: detection and prevention in Europe].

While salmonellas can cause disease problems among poultry, they remain essentially a concern for public health, as a cause of outbreaks of food poisoning. The principal site of multiplication of these bacteria is the digestive tract, particularly the caecum, which may result in widespread contamination of the environment. The pathogenicity of salmonellas depends on the invasive properties and the ability of the bacteria to survive and multiply within cells, particularly macrophages. These properties are the source of vertical transmission which, in the case of survival of the embryo, can result in contamination of a flock or, in the case of embryonic mortality, can result in an explosion of contaminated eggs. Salmonella infection can be diagnosed by isolating the bacteria and/or serological testing of the flock. European Union Directive 92/117/EC, modified by Directive 97/22/EC, stipulates either the destruction of infected flocks of breeding birds, or decontamination of the flock in an effective way, before normal trade in products can be resumed. Noteworthy examples of effective measures suitable for prophylaxis of Salmonella infection in poultry flocks include the slaughter of infected breeding stock, the creation of sanitary barriers at building entrances, heat treatment of feed, the use of competitive exclusion, selection of breeds genetically resistant to Salmonella, and occasional vaccination and antibiotic treatment. However, the most effective means of reducing food poisoning remains adequate cooking of food and maintenance of the cold chain.

Animals↗