Serosurvey for West Nile virus in horses in southern France.
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Animals↗
Biomedical subjects
G Dauphin
Publications and source records attributed to G Dauphin.
Identification of bluetongue virus serotype 2 (Corsican strain) by reverse-transcriptase PCR reaction analysis of segment 2 of the genome.
In October 2000, bluetongue virus was detected on the French island of Corsica. The disease was also reported in Sardinia, Calabria, Sicily and on the Spanish islands of Majorca and Minorca. This paper describes the use of molecular techniques for a rapid identification and serotype determination of serotype 2 of the virus. The nucleotide sequences of segments 2 and 7 of the genome of the Corsican strain were determined and its phylogenetic relationships are described.
Animals↗
Use of PFGE typing for tracing contamination with Listeria monocytogenes in three cold-smoked salmon processing plants.
The sites of Listeria monocytogenes contamination in three cold-smoked salmon (Salmo salar) processing plants were detected by sampling salmon and the plant's environment and equipment at different production stages. Of the 141 samples collected from three processing plants, 59 (42%) were contaminated with L. monocytogenes. The rates of contamination varied as to the plant and the sample source. L. monocytogenes isolates from 17 various contaminated seafood products (fresh, frozen and smoked fishes, cooked mussels) were also studied. A total of 155 isolates from the three plants and the various seafoods were characterized by genomic macrorestriction using ApaI and SmaI with pulsed-field gel electrophoresis (PFGE) and 82 isolates were serotyped. Macrorestriction yielded 20 pulsotypes and serotyping yielded four serovars: 1/2a, 1/2b, 1/2c, 4b (or e), with 77 (93%) belonging to serovar 1/2a. One clone of L. monocvtogenes predominated and persisted in plant I and was the only pulsotype detected in the final product although it was not isolated from raw salmon. No L. monocytogenes was detected in the smoked skinned salmon processed in plant II, even though 87% of the raw salmon was contaminated. All the smoked salmon samples collected in plant III were contaminated with a unique clone of L. monocytogenes, which may have occurred during slicing. In the three plants, the contamination of final products did not seem to originate from the L. monocytogenes present on raw salmon, but from the processing environment.
Animals↗
Evidence of Borna disease virus genome detection in French domestic animals and in foxes (Vulpes vulpes).
Borna disease virus (BDV) is an enveloped, non-segmented negative-stranded RNA virus which belongs to the Bornaviridae family. BDV is an aetiological agent of encephalitis in horses, sheep and several other vertebrate species. In order to extend our knowledge about the presence of BDV in France, a study based on BDV RNA detection by RT-nested-PCR was done with 196 animal tissues: 171 brain samples collected from different animal species (75 horses, 59 foxes, 31 cattle, 4 dogs, 1 sheep, 1 roe deer) and 25 horse blood samples. An RNA internal standard molecule was constructed and was co-amplified with the test template. This study reports the first detection of BDV RNA in France in 10 brain samples collected from horses, foxes and cattle, and from 14 horse blood samples. Detection of the BDV genome in the brains of six red foxes is the first evidence of BDV infection in this species.
Animals↗
Generalised linear mixed models analysis of risk factors for contamination of Danish broiler flocks with Salmonella typhimurium.
We present a retrospective observational study of risk factors associated with the occurrence of Salmonella typhimurium (ST) in Danish broiler flocks. The study is based on recordings from 1994 in the ante-mortem database maintained by the Danish Poultry Council. The epidemiological units are the broiler flocks (about 4000 flocks) which are clustered within producers. Broiler flocks with ST-infected parent stocks show increased risk of salmonella infection, and also the hatchery affects the salmonella status significantly. Among the rearing factors, only the use of medicine as well as the time of rearing, and the sampling method are significant. Epidemiological control would seem most efficient on starting at the top levels of the production hierarchy from which a major part of the ST contamination is derived. A secondary purpose of the study is to evaluate different statistical approaches and software for the analysis of a moderately-sized data set of veterinary origin. We compare the results from five analyses of the generalised linear mixed model (GLMM) type. The first observation is that the results agree reasonably well and lead to similar conclusions. A closer look reveals certain patterns of bias and estimation accuracy that correspond well with theoretical findings and practical experience reported in the statistical literature.
Animal Husbandry↗
Ionophore properties of monensin derivatives studied on human erythrocytes by 23Na NMR and K+ and H+ potentiometry: relationship with antimicrobial and antimalarial activities.
Eight derivatives of monensin with a modified C25-C26 moiety were synthesized. Their ionophore properties were studied on human erythrocytes by measuring Na+ influx with 23Na NMR and concomitant K+ and H+ efflux by potentiometry. Modification of OH-26 led to inversion of selectivity of transport in favor of K+/Na+ in comparison with monensin. This selectivity disappeared by suppression of the C26-OH moiety. Finally the ionophore ability was lost if the head-to-tail chelation of the monensin skeleton was prevented by blocking the terminal OH-25 and -26 functions. All the compounds were inactive on Gram-negative bacteria and fungi. MIC measured on Bacillus cereus showed that derivatives with increased K+/Na+ selectivity were clearly the most active against Bacillus growth. Most of the compounds showed potential antimalarial properties in the nanomolar range when tested in vitro against Plasmodium falciparum. The IC50S measured were correlated with the whole Na+ and K+ transport efficiency rather than with the ionic selectivity. In both cases determination of initial fluxes of transport for both cations (Na+ and K+) was necessary to investigate the relationship between biological and ionophore properties.
Adult↗
Na+ and K+ transport by 4-chlorophenylurethane-monensin in Enterococcus hirae de-energized and energized cells studied by 23Na-NMR and K+ atomic absorption.
Na+ and K+ movements induced by 4-chlorophenylurethane-monensin, which presents an inverted ion selectivity (K+ > Na+) in model systems compared with monensin, were followed on Enterococcus hirae cells by 23Na-NMR and K+ atomic absorption. For de-energized cells, the urethane derivative is much more selective for K+ than monensin, but only at low concentrations (10(-3)-10(-4) mM). For higher concentrations, as previously shown for monensin, the sodium and potassium movements are driven by the ion gradients present. On energized cells, both K+ and Na+ gradients were highly perturbed, and this can be related to the higher toxicity in mice and bacteria for this derivative.
Biological Transport↗
Conditions modulating the ionic selectivity of transport by monensin examined on Enterococcus hirae (Streptococcus faecalis) by 23Na-NMR and K+ atomic absorption.
Factors likely to modulate the ionic selectivity of monensin were examined on Enterococcus hirae (Streptococcus faecalis) in two states previously characterized: the resting (de-energized) cell and the active (energized) cell. Internal and external Na+ were followed by corresponding 23Na-NMR resonances K+ concentrations were measured by atomic absorption. For a given cellular population of de-energized cells, the apparent transport rates and the final cationic concentrations reached at the steady state were decreasing with the ionophore dose. Monensin was selective for sodium only at low concentrations, in the range 1 mM-10(-4) mM the transport was depending on the effective cationic gradients. Comparison of the activity curves for two cell populations (7.10(9) and 7.10(10) cells/ml) showed the importance of the ratios of monensin/mg phospholipid and also of the ratios of external/internal volumes. On energized cells, except for low monensin concentrations, the main effect was a K(+)-induced efflux and not a Na+ influx. Two factors were modulating the resulting selectivity of this ionophore: the response of the intrinsic bacterial carriers and the generation of the gradients (mainly the external pH) which were favourable to a K+/Na+ transport. Once again the results obtained for two cell populations could be compared, the determining factors were the ratio external/internal volume and the generation of the pH gradient.
Biological Transport↗
Futile cycling of glycogen in Fibrobacter succinogenes as shown by in situ 1H-NMR and 13C-NMR investigation.
Glycogen was synthesized during all the growth phases in the rumen anaerobic cellulolytic bacterium Fibrobacter succinogenes. Glycogen synthesis and degradation were monitored using in situ 13C and 1H-NMR spectroscopy in resting cells of F. succinogenes. The cells were incubated at 37 degrees C under anaerobic conditions with [1-13C]glucose and [2-13C]glucose. 1H-NMR spectra were used to quantify enrichment by 13C of metabolism products. Glucose was utilized for energy requirements of the bacterium, essentially via the Embden-Meyerhof pathway, leading to the synthesis of succinate and acetate, while glycogen was stored. From [1-13C]glucose, labeling occurred on C2 of succinate and acetate, and on both C1 and C6 of glycogen, the labeling on C1 being predominant. The C6-labeling of glycogen may be explained by scrambling and reversal of the glycolytic pathway at the triose-phosphate and fructose 1,6-bisphosphate level. When the bacteria were incubated first with [1-13C]glucose, then washed and incubated with [2-13C]glucose, the pattern of 13C labeling in the products of the metabolism, as shown by 13C and 1H-NMR spectra, indicated that glycogen was degraded at the same time as it was being stored, suggesting futile cycling of glycogen. The hydrolysis of previously stored glycogen can provide, in the presence of glucose, up to 30% of the carbon source for the bacteria.
Bacterial Proteins↗
Phosphate-dependent sodium transport in S. faecalis investigated by 23Na and 31P NMR.
Na+ movements in S. faecalis were studied by 23Na NMR. They proved to be dependent on phosphate concentration in the buffer during the de-energization step. K+ and H+ were also studied respectively by potentiometry and 31P NMR and were shown not to be implicated. For de-energized cells the internal phosphate concentration, on the contrary, was directly linked to the external phosphate contained in the buffer. The experiments showed a Na+/Pi dependence in this prokaryote so far known only in eukaryotes.
Biological Transport↗
Bioconversion of monensin by a soil bacterium, Sebekia benihana.
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Bacteria↗
Study by NMR of the mode of action of monensin on Streptococcus faecalis de-energized and energized cells.
Streptococcus faecalis was used as a bacterial model for studying the mode of action of monensin by NMR investigations. Experiments were carried out in two states, characterized by several complementary methods: (i) the resting (de-energized) cell which was considered as an inert biological membrane, on which cationic transport induced by the ionophore alone can be investigated; (ii) the active (energized) cell where the ionophore-sensitive response of the living organism, particularly the cation pumps and the glycolysis, is probed. Studies of resting cells were performed, with changing external ionic concentrations, in the presence of monensin, which is preferentially a sodium carrier. Internal and external Na+ and H+ were followed by corresponding 23Na and 31P (inorganic phosphate) NMR resonances, K+ fluxes were measured by atomic absorption. It was shown that the induced cationic movements were linked to the existing ionic gradients for K+ and Na+. 31P and 13C NMR spectra for the intermediary metabolites detected in active cells showed that glycolysis is dramatically modified in the presence of monensin.
Adenosine Triphosphate↗
Study of a delta-hydroxy ketone-hemiketal equilibrium by 2D-NMR exchange spectroscopy for the antibiotic grisorixin: involvement in cationic transport through membranes.
Study of a delta-hydroxyketone-hemiketal equilibrium in the polyether antibiotic grisorixin was performed with 2D-NMR spectroscopy. The efficiency of 13C chemical exchange spectroscopy for the assignment of 1H and 13C resonances, in the 2 forms, was shown, making possible a conformational investigation of both forms. This equilibrium was observed for grisorixin in solvents of varying polarity, such as CD2Cl2, CDCl3, CD3CN, or CD3OD, but not in C6D12 or C6D6. Other related antibiotics with the same terminal heterocycle were described only in the closed hemiacetalic structure. The low ionic fluxes measured in a bulk chloroformic membrane for grisorixin were explained by this equilibrium, which competed unfavorably with the cation capture process at the water-chloroform interface. This equilibrium would not be present in a phospholipidic bilayer membrane containing the ionophore, published experimental results are taken into account. The peculiar tautomeric equilibrium observed for grisorixin could be linked to the specific axial stereochemistry of the C7-C8 bond, which creates tension in the globular conformation.
Anti-Bacterial Agents↗
Microbial conversion of nigericin in three successive steps, by Sebekia benihana.
A strain of Sebekia benihana NRRL 11111 was found to transform nigericin in three successive steps, giving three compounds which were isolated. Their structure were determined by IR, 1H and 13C NMR, and fast atom bombardment mass spectra. The first compound resulted from the reduction of the terminal hemiketal ring it was then transformed into the two other compounds as a result of the oxidation of methyl (C-33) into a CH2OH and COOH group respectively. All these products had lost the ionophoric and antibiotic properties of nigericin and thus were products of a detoxification process.
Anti-Bacterial Agents↗
Semi-synthesis of A23187 (calcimycin) analogs. III. Modification of benzoxazole ring substituents, ionophorous properties in an organic phase.
Ten semi-synthetic analogs of A23187 (calcimycin), with only the benzoxazole ring substituents modified together with the ionophore X14885A were studied with regard to their calcium and magnesium carrier properties through an organic phase (toluene - butanol, 70:30). The results indicate that the carboxylic group and the oxazolic nitrogen, maintained in the ortho position are essential for the ionophorous properties. Further, the introduction of a substituent in place of the NHCH3 group, producing steric hindrance of the carboxylic group leads to a destabilization of the 2:1 associations with cations.
Benzoxazoles↗
Semisynthesis of A23187 (calcimycin) analogs. II. Introduction of a methyl group on the benzoxazole ring.
Semisynthesis of two demethylamino A23187 with a methyl group in the 4- or 5-position on the benzene ring were carried out via the cleavage of A23187 oxazole ring and rebuilding of modified benzoxazoles. These compounds were shown to release Ca++ and MG++ from mitochondria and to keep part the antibacterial activity of the natural metabolite.
Calcimycin↗
Microbial conversion of grisorixin, a monovalent cation ionophorous antibiotic.
The isolation and identification of a bioconversion product of grisorixin from a strain of Streptomyces rimosus is reported. The structure of this product was elucidated from physicochemical data, in particular 13C NMR spectra. Its ionophorous and antibiotic properties are markedly different from those of grisorixin.
Anti-Bacterial Agents↗