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[Prevalence of Salmonella enteritidis and Salmonella typhimurium in laying hen flocks battery and on floor housing. Comparative studies using bacteriological and serological demonstration methods].

The prevalence of Salmonella (S.) enteritidis and Salmonella typhimurium is described in 34 flocks of laying chickens. In an attempt to detect the rate of infection comparative bacteriological and serological methods were employed. By cultural methods 47.0% of the floor flocks and 35.3% of the battery flocks were S.e. or S. tm. infected. The same tendency was established with the Bommeli ELISA. Flocks on floor were positive in 64.7% and flocks in batteries were positive in 47.0%. Both methods--bacteriological isolation and serology--yielded more S.e. positive flocks kept on the floor than in battery systems. Long-term studies of four flocks demonstrate a long persistence of antibodies against S.e. in serum and yolk even after one year. Almost all S.e. infected flocks prove a correlation between increase of positive cultures and antibody positive sera. Also, a correlation between levels of absorption values of sera tested in the Bommeli ELISA and positive fecal samples is evident. This is particularly true for eight of 10 flocks. If S.e. infected laying flocks need to be detected, then high absorption values in sera or yolk and a high rate of antibody positive hens provide reasonable good conditions to attempt subsequent bacteriological examinations.

Animals↗

Molecular analysis of ampicillin-resistant sporadic Salmonella typhi and Salmonella paratyphi B clinical isolates.

OBJECTIVE: To study the mechanisms of antibiotic resistance in Salmonella typhi and Salmonella paratyphi B clinical isolates, and the clonality of resistant strains. METHODS: Antibiotic susceptibility was tested by disk-agar diffusion. Conjugation experiments and plasmid analysis by agarose gel electrophoresis after EcoRI digestion were followed by hybridization to a digoxigenin-labeled TEM-type beta-lactamase probe. DNA fingerprints were obtained by pulsed-field gel electrophoresis of Xbal-digested chromosomal DNA. RESULTS: Three S. typhi isolates (7% of the isolates studied), of which one was ampicillin resistant and the other two multiresistant (ampicillin, chloramphenicol, tetracycline, sulfamethoxazole/trimethoprim and streptomycin), and two ampicillin-resistant S. paratyphi B isolates (25% of the isolates studied) were further evaluated. A 34-MDa conjugative plasmid, previously isolated from Salmonella enteritidis, conferred ampicillin resistance. A 100-MDa conjugative plasmid encoded resistance to chloramphenicol, tetracycline and sulfamethoxazole/trimethoprim, as well as ampicillin. Chromosomal fingerprinting revealed two distinct resistant strains for each serovar which were different from a matched set of sensitive S. typhi strains. CONCLUSIONS: Two conjugative, TEM-type beta-lactamase-encoding plasmids conferred ampicillin resistance to S. typhi and S. paratyphi B. The 34-MDa plasmid was identical to that previously characterized from S. enteritidis, while the 100-MDa plasmid also encoded resistance to chloramphenicol, tetracycline and sulfamethoxazole/trimethoprim. Resistant isolates did not belong to a single clone but rather represented distinct strains.

Journal Article↗

Adaptive acid tolerance response in Salmonella enterica serovar Typhimurium and Salmonella enterica serovar Typhi.

The survival of bacteria in various environments depends on a number of protective responses including acid tolerance response (ATR). In this study, ATR phenomenon was compared in Salmonella enterica serovar Typhi 6 and Salmonella enterica serovar Typhimurium 98 under different culture conditions. Survival of the adapted culture (pre-acid shocked to pH 5.5) was significantly better (p < 0.05) as compared to control, unadapted culture after acid shock at pH 3.3. However, the ATR varied with the serovar, incubation temperature and the growth medium used (all p-values < 0.05). S. Typhi 6 failed to grow in pH 3.3 at 45 degrees C. The addition of tetracycline or chloramphenicol (1.0 microg ml(-1)) to adapted cultures during or after acid shock (pH 3.3) had no effect on ATR expression. In S. Typhimurium 98, growth was increased by 10% or greater in adapted culture (when grown at pH 3.3) as compared to growth observed with an unadapted culture (when grown at pH 7.3) on transfer to fresh growth medium at pH 7.3. A poor ATR observed in non-growing S. Typhimurium 98 suspensions clearly showed that ATR is an energy-consuming process. Storage of S. Typhimurium 98 cultures in pH 4.5 nutrient broth at 4 degrees C demonstrated that prolonged exposure to acidic conditions is more detrimental in comparison to the cultures stored at pH 7.3 at this temperature.

Acids↗

Restriction map of the Salmonella enteritidis virulence plasmid and its homology with the plasmid of Salmonella typhimurium.

A restriction map of the 61 kb virulence plasmid of Salmonella enteritidis was constructed and compared with the Salmonella typhimurium virulence plasmid map. Polymerase chain reaction was used to detect a region homologous to rck gene in the S. enteritidis plasmid which was localized in the map by hybridization. Regions homologous to traT and vagD/vagC loci were not detected in this plasmid by this method.

Base Sequence↗

High efficiency transformation of Salmonella typhimurium and Salmonella typhi by electroporation.

Salmonella typhimurium and S. typhi were transformed with high efficiency by electroporation. Transformation efficiencies of up to 10(10) transformants per microgram of pBR322 were obtained. In contrast to chemical transformation methods, neither the smooth lipopolysaccharide of S. typhimurium nor the Vi capsular polysaccharide of S. typhi greatly affected transformation efficiency. The introduction of a ga1E mutation slightly improved transformation efficiency in S. typhimurium (less than tenfold) while the Vi antigen of S. typhi had no detectable effect. The transformation efficiency of S. typhimurium with DNA derived from Escherichia coli was increased greatly by the removal of the hsd restriction system (100-fold). Under these conditions electroporation can be used for the routine and direct transformation of Salmonella strains with partially purified (alkaline lysis) plasmid DNA from E. coli.

Antigens, Bacterial↗

Association with HeLa cells of LPS mutants of Salmonella typhimurium and Salmonella minnesota in relation to their physicochemical surface properties.

Different LPS mutants of Salmonella typhimurium and Salmonella minnesota have been investigated with respect to (1) their tendency to associate with HeLa cell monolayers, and (2) their physicochemical surface properties. Aqueous biphasic partitioning, hydrophobic interaction chromatography, and ion exchange chromatography have been used to characterize the bacterial cell surface properties with respect to charge and hydrophobicity. Liability to hydrophobic interaction was defined either by the change of partition in a dextran-polyethylene-glycol (PEG) system by the addition of PEG-palmitate (P-PEG), or by the elution pattern from Octyl-Sepharose. Accordingly, charge was asssessed by the effect of positively charged trimethylamino-PEG (TMA-PEG) on the partition, and by the elution from DEAE-Sephacel. Bacterial being negatively charged and liable to hydrophobic interaction had the highest tendency to associate with HeLa cells. In some cases the methods for surface analysis gave conflicting results on charge and/or liability to hydrophobic interaction of the same LPS mutant. Possible reasons for these differences and the role of bacterial cell surface structures contributing to physicochemical character are discussed.

Chromatography↗

Modulation of arachidonic acid metabolism in experimental arthritis induced by Salmonella enteritidis. I. Effect of ibuprofen and flurbiprofen on experimental arthritis induced by Salmonella enteritidis.

Oral administration of two nonsteroidal anti-inflammatory drugs, ibuprofen and flurbiprofen, can suppress the Salmonella-induced arthritis in rats. The joint swelling index of arthritic paws showed suppression of arthritis in animals treated with the drugs, this effect being greater with flurbiprofen. Measurement of 5 eicosanoids in femoral vein plasma showed increase of arachidonic acid products in Salmonella-treated rats. Inhibition of joint inflammation resulting from treatment with ibuprofen and flurbiprofen is reflected by a decrease in concentration of all 5 eicosanoids which were found in the order: PGE2 greater than TXB2 greater than 6-keto-PGF1 alpha greater than PGF1 greater than PGF2 alpha. These studies indicate that flurbiprofen is a more powerful anti-inflammatory agent than ibuprofen. However, since the joint disease was not completely cured, optimal intervention is quite likely to require modulation of the lipoxygenase pathway.

Animals↗

Effect of dietary fatty acids on experimental manifestation of Salmonella-associated arthritis in rats. II. Effect of dietary fatty acids on experimental manifestation of Salmonella-associated arthritis in rats.

Dietary supplementation with a marine lipid concentrate rich in n-3 fatty acids and pure ethyl ester of dihomo-gamma-linolenic acid (DHLA) resulted in inhibition of the chronic phase of inflammation in Salmonella-associated arthritis. The anti-inflammatory effect of DHLA was much stronger than that of two n-3 fatty acids (eicosapentaenoic acid and docosahexaenoic acid) present in marine oil. Fatty acid profiles in phosphoglyceride fractions of red blood cells showed incorporation of the respective supplemented fatty acids. Concentrations of 4 cyclooxygenase products in femoral vein plasma were smaller in the fatty acid supplemented rats. These studies suggest that DHLA and marine n-3 fatty acids may have useful anti-inflammatory effects in Salmonella-associated arthritis.

Animals↗

Prophylaxis of Salmonella abortus ovis-induced abortion of sheep by a Salmonella typhimurium live vaccine.

A Salmonella typhimurium live vaccine with optimal level of attenuation for sheep, constructed by means of 'metabolic drift' mutations, was tested for its efficacy in preventing Salmonella abortus ovis-induced abortions. In two field trials in Kirgiziya, 78,000 to 100,000 first delivery sheep received a fully tolerated single dose of 10(9) c.f.u. live vaccine 2 months before to 4 months after insemination. Alternatively, they were immunized twice with commercial inactivated S. abortus ovis vaccine, or they served as non-immunized controls. The S. abortus ovis-induced abortion frequency in the controls was greater than or equal to 30%, in sheep immunized with inactivated vaccine greater than or equal to 11%. In flocks immunized with live vaccine, the S. abortus ovis-induced abortion frequency did not exceed 0.1%. Thus, use of 'metabolic drift' mutations for construction of stable vaccine strains optimally attenuated for the particular host species proved to be relevant to practice.

Abortion, Veterinary↗

Occurrence of Salmonella typhimurium virulence plasmid-specific sequences in different serovars of Salmonella.

We have subcloned the 96-kilobasepair (kb) virulence plasmid, pLT2, of Salmonella typhimurium line LT2 into 7 subfragments. Using these subclones as probes, 35 independent Salmonella isolates were tested for complementary DNA sequences Sequences homologous to pLT2 were present in 15 of the isolates. All of these contained sequences homologous to at least one specific probe representing 15 kb of pLT2. The traT gene from pLT2 was absent in 6 of these 15 isolates. Three strains reported to be cured of the plasmid were shown to harbour some sequences with homology to the pLT2 plasmid. Seven isolates were shown to contain homologous sequences with pBR322 but not with the pLT2 plasmid.

Cloning, Molecular↗

Adenylates and adenylate-energy charge in submerged and planktonic cultures of Salmonella enteritidis and Salmonella typhimurium.

Adenine nucleotide values and adenylate energy charge (AEC) were measured during the growth of Salmonella enteritidis and Salmonella typhimurium as submerged colonies in agarose gel and gelatin gel, and as planktonic cells in broth. Growth in all three systems showed similar trends with a ten-fold decrease in total adenylate pool during exponential growth, before attaining a fairly stable value throughout stationary phase. AEC values were generally low, (approximately 0.66), but did rise slightly during stationary phase. The large proportion of dead cells during early exponential phase may have contributed to the adenosine diphosphate and adenosine monophosphate pools, through cell lysis or excretion, and it is suggested that this was likely to account for the low values of AEC. In agarose and gelatin gelled cultures the percentage of adenosine triphosphate (ATP) in relation to the total adenylates showed random fluctuations. This was contrary to the broth culture where percentage ATP was highest after 12 h and the data formed a smooth curve. These data demonstrated that considerable physiological heterogeneity exists within a colony of bacteria growing in a gel matrix and by analogy a food material also, and that AEC is a poor indicator of cell viability in such systems.

Adenosine Triphosphate↗

Salmonella typhi and S typhimurium derivatives harbouring deletions in aromatic biosynthesis and Salmonella Pathogenicity Island-2 (SPI-2) genes as vaccines and vectors.

The S. typhimurium strain (TML deltaaroC deltassaV) WT05, harbouring defined deletions in genes involved in both the aromatic biosynthesis pathway (aroC) and the Salmonella Pathogenicity Island-2 (SPI-2) (ssaV) was shown to be significantly attenuated in C57 BL/6 interferon gamma knockout mice following oral inoculation. Similarly, the S. typhi strain (Ty2 deltaaroC deltassaV) ZH9 harbouring the aroC and ssaV mutations propagated less efficiently than wild type in human macrophages. These studies demonstrated the attractive safety profile of the aroC ssaV mutant combination. Strains S. typhimurium (TML deltaaroC deltassaV ) WT05 and S. typhi (Ty2 deltaaroC deltassaV) ZH9 were subsequently tested as vaccine vectors to deliver E. coli heat-labile toxin (LT-B) mucosally to mice. Mice inoculated orally with S. typhimurium (TML deltaaroC deltassaV) WT05 expressing LT-B (WT05/LT-B) elicited high titres of both LT-specific serum IgG and intestinal IgA, although no specific IgA was detected in the vagina. Similarly, intranasal inoculation of mice with S. typhi (Ty2 deltaaroC deltassaV) ZH9 expressing LT-B (ZH9/LT-B) elicited even higher titres of LT-specific serum antibody as well as LT-specific Ig in the vagina. We conclude that deltaaroC deltassaV strains of Salmonella are highly attenuated and are promising candidates both as human typhoid vaccines and as vaccine vectors for the delivery of heterologous antigens.

Animals↗

Formation of hybrids between coliphage lambda and Salmonella phage P22 with a Salmonella typhimurium hybrid sensitive to these phages.

An unusual Salmonella typhimurium hybrid with sensitivity to coliphage lambda and salmonella phage P22 has been recovered from matings between an Escherichia coli K-12 Hfr donor and an S. typhimurium recipient. The hybrid is an excellent host for achieving genetic recombination between lambda and P22. Two broad classes of hybrid phages were isolated. The lambda-P22 hybrid class, which has the protein coat of lambda, contains at least the c region of P22. The P22-lambda hybrid class has the protein coat of P22 and has inherited at least the c marker of lambda.

Animals↗

Barriers to recombination between closely related bacteria: MutS and RecBCD inhibit recombination between Salmonella typhimurium and Salmonella typhi.

Previous studies have shown that inactivation of the MutS or MutL mismatch repair enzymes increases the efficiency of homeologous recombination between Escherichia coli and Salmonella typhimurium and between S. typhimurium and Salmonella typhi. However, even in mutants defective for mismatch repair the recombination frequencies are 10(2)- to 10(3)-fold less than observed during homologous recombination between a donor and recipient of the same species. In addition, the length of DNA exchanged during transduction between S. typhimurium and S. typhi is less than in transductions between strains of S. typhimurium. In homeologous transductions, mutations in the recD gene increased the frequency of transduction and the length of DNA exchanged. Furthermore, in mutS recD double mutants the frequency of homeologous recombination was nearly as high as that seen during homologous recombination. The phenotypes of the mutants indicate that the gene products of mutS and recD act independently. Because S. typhimurium and S. typhi are approximately 98-99% identical at the DNA sequence level, the inhibition of recombination is probably not due to a failure of RecA to initiate strand exchange. Instead, these results suggest that mismatches act at a subsequent step, possibly by slowing the rate of branch migration. Slowing the rate of branch migration may stimulate helicase proteins to unwind rather than extend the heteroduplex and leave uncomplexed donor DNA susceptible to further degradation by RecBCD exonuclease.

Adenosine Triphosphatases↗

Choleriform syndrome and production of labile enterotoxin (CT/LT1)-like antigen by species of Salmonella infantis and Salmonella haardt isolated from the same patient.

Isolates of Salmonella infantis from the stool and blood and an isolate of Salmonella haardt from the stool of a patient with choleriform diarrhea produced labile enterotoxin (CT/LT1)-like antigen. Genetic and molecular experiments indicated that the production of CT/LT1-like antigen was chromosomally encoded.

Antigens, Bacterial↗

Mutations of the gyrA gene of clinical isolates of Salmonella typhimurium and three other Salmonella species leading to decreased susceptibilities to 4-quinolone drugs.

The region of the gyrA gene encoding nucleotides 72 to 557 in ten Salmonella typhimurium clinical isolates received from Vellore, India and six British clinical isolates of a number of Salmonella species has been sequenced. Those exhibiting a decreased susceptibility to 4-quinolone drugs (MICs of ciprofloxacin ranging from 0.128 mg/L to 1.0 mg/L) have been shown to possess a number of different mutations in this region of the gene (Asp-87 to Gly, Asp-87 to Asn and Ser-83 to Phe). The mutations Asp-87 to Gly and Ser-83 to Phe/Asp-87 to Asn have yet to be reported occurring in S. typhimurium.

Amino Acid Sequence↗

In vitro study of Salmonella enteritidis and Salmonella typhimurium definitive type 104: survival in egg albumen and penetration through the vitelline membrane.

Salmonella enteritidis and Salmonella typhimurium definitive type 104 (DT104) have been detected in the chicken oviduct, and their survival in egg albumen at the chicken body temperature of 42 degrees C may be important in oviductal and transovarian contamination of intact shell eggs. Eight S. enteritidis and 24 S. typhimurium DT104 strains were tested for their in vitro survival in egg albumen. The concentration of the organisms declined more rapidly when incubated at 42 degrees C than at 37 degrees C and dropped to nondetectable levels within 96 h at the higher, but not at the lower, temperature. In another experiment, 3 S. enteritidis and 3 S. typhimurium DT104 strains were randomly selected, and dosages of 20 and 200 cells of each strain were inoculated onto the vitelline membranes of egg yolks, which were then submerged in the original albumen and incubated for 24 h at 42 degrees C. Under these conditions, the organisms survived in albumen but did not penetrate the vitelline membrane. However, in a similar experiment, penetration did occur when the specimens were incubated at 30 degrees C for 72 h. The results suggest that low numbers of S. enteritidis and S. typhimurium DT104 can be expected to survive in egg albumen during the 24-h period of egg formation in the oviduct but would be unlikely to invade the yolk.before oviposition. However, depending on storage conditions following oviposition, S. enteritidis, as well as S. typhimurium DT104, could survive longer and may eventually invade the egg yolks.

Albumins↗