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PubMed1979

Compatible results obtained from biotyping and serotyping in Serratia marcescens.

The correspondence between complete serotype and biotype (P.A.D. Grimont and F. Grimont, J. Clin. Microbiol. 8:73-83, 1978) of 474 Serratia marcescens strains was studied. Of 127 serotypes, 70 were represented by two or more strains of the same serotype belonged to one biotype. However, for 91% of serotypes, strains of the same serotype belonged to one biogroup--i.e., a group of closely related biotypes. Biogroups are A1 (A1a, A1b); A2/6 (A2a, A2b, A6a, A6b); A3 (A3a, A3b, A3c, A3d); A4 (A4a, A4b); A5/8 (A5, A8a, A8b, A8c); and TCT (TCT, TT). Only two serotypes were composed of a mixture of pigmented and nonpigmented biogroups. Pigmented biogroups (A1 and A2/6) were otherwise differentiated from nonpigmented biogroups (A3, A4, A5/8, and TCT) by serotyping. Some biogroups preferentially occurred in some O serogroups: A4 in 01; A2/6 in O6, O8, and O14; and A3 in O9, O12, and O15. Three H serogroups were found to be biochemically homogeneous: H1, H7, and H20 were respectively and uniquely composed of biogroups A4, TCT, and A3. A square matrix of O versus H serogroups, with the corresponding biogroup for each O X H combination, was used for comparisons between O groups and between H groups. Identical patterns of biogroups were shown by serogroups O6, O8, and O14. Taxonomical, ecological, and practical consequences of these findings are discussed.

Carbon↗
PubMed2001

Detection of salmonellae in chicken feces by a combination of tetrathionate broth enrichment, capillary PCR, and capillary gel electrophoresis.

This report describes a rapid detection procedure for salmonellae from chicken feces by the combination of tetrathionate primary enrichment (preenrichment [PE])-bacterial lysis-capillary PCR and capillary gel electrophoresis. Pure Salmonella enterica serovar Enteritidis 64K was reisolated and detected by capillary PCR after buffered peptone water and nutrient broth, tetrathionate broth base Hajna (TTBH), and tetrathionate broth (TTB) preenrichments. When the same culture was mixed with intestinal homogenate, bacteriological reisolation and capillary PCR detection was achieved only by TTBH and TTB preenrichments. Capillary gel electrophoresis revealed that a Salmonella genus-specific 281-bp PCR product was detected when Salmonella strains but not non-Salmonella strains were tested. The detection limit of capillary PCR with whole-cell DNA extracted from pure Salmonella enterica serovars Enteritidis 64K, Typhimurium LT2-CIP60-62, and Gallinarum 64K was 3, 3, and 9 CFU ml(-1), respectively. The detection limit of capillary PCR from whole-cell DNA extracted from intestinal homogenate artificially contaminated with the same three strains was 3, 3, and 7 CFU ml(-1), respectively. We compared the results of the capillary PCR and bacteriological examination from the natural samples. Thirty-five of 53 naturally contaminated samples produced a specific PCR product. In 9 of the 35 PCR-positive samples, Salmonella could not be detected bacteriologically either by PE or a primary and delayed secondary enrichment (DSE) combination. In the 18 PCR-negative samples, 4 samples were found to harbor Salmonella by both PE and DSE and 14 samples were positive after DSE. Fifty-three additional intestinal homogenate samples, which were negative by their PE and DSE in bacteriological examination, were found to be also negative by their PCRs. The total time required to detect Salmonella with the capillary PCR method we used was approximately 20 h. If samples are from clinically diseased birds, the total time for PCR and detection is reduced to 2 h since the 18-h PE is not required. These results indicate that TTB enrichment, bacterial lysis, and genus-specific capillary PCR combined with capillary gel electrophoresis constitute a sensitive and selective procedure which has the potential to rapidly identify Salmonella-infected flocks.

Animals↗
PubMed1975

Comparison of several methods for the isolation of salmonellae from egg products.

Lactose broth and buffered peptone water proved to be equally effective as preenrichment media in the isolation of salmonellae from egg products. With tetrathionate bile brilliant green broth, incubated at 43 degrees C as enrichment medium, more samples were found to be positive than with selenite cystine broth incubated at 37 degrees C or 43 degreesC and selenite brilliant green broth incubated at 37 degrees C. As selective agar, brilliant green agar proved to be more reliable than bismuth sulfite agar. These results indicate that the ISO method for the isolation of salmonellae from meat and meat products can be successfully used for the examination of egg products.

Agar↗
PubMed1981

Plasmid DNA in acidophilic, chemolithotrophic thiobacilli.

Plasmid patterns were determined in 15 strains of iron-oxidizing Thiobacillus ferrooxidans. In four of these strains plasmid DNA was not detected. In the other strains the molecular weights of plasmids ranged from 5 x 10(6) to 50 x 10(6) and each strain had a different plasmid composition. The change of growth substrate from ferrous iron to tetrathionate did not affect the plasmid pattern in T. ferrooxidans nor did it in T. acidophilus, which was adapted from glucose to grow on tetrathionate.

Ferrous Compounds↗
PubMed2001

Comparison of GN Hajna and tetrathionate as initial enrichment for salmonellae recovery from swine lymph nodes and cecal contents collected at slaughter.

An epidemiologic survey was conducted to determine the prevalence of salmonellae in swine from 5 farms of an integrated swine operation. The purpose of this study was to evaluate the recovery efficiencies for salmonellae from swine lymph nodes and cecal contents when GN Hajna and tetrathionate were compared as initial enrichments. Salmonellae were isolated from 61% of 645 pigs at slaughter; 324 positive cultures were from lymph nodes, and 224 were from cecal contents. Frequently, pigs had salmonellae isolated from both the lymph nodes and cecal contents. Total isolations, regardless of source, were similar for GN Hajna (247) and tetrathionate (301). There was no difference (P > 0.05) in the number of isolations from lymph nodes when GN Hajna enrichment was compared with tetrathionate enrichment (174 vs. 150). However, there was a significant (P < 0.05) advantage of utilizing tetrathionate when compared with GN Hajna for isolations from cecal contents (151 vs. 73).

Abattoirs↗
PubMed1991

A comprehensive study on the isolation and characterization of the HeLa S3 nuclear matrix.

Different agents have been employed to extract the histones and other soluble components from isolated HeLa S3 nuclei during nuclear matrix isolation. We report that 0.2M (NH4)2SO4 is a milder extracting agent than NaCl and LIS (lithium 3,5-diiodosalicylate), on the basis of the apparent preservation of the elaborate fibrogranular network and the residual nucleolus that resemble the in situ structures in whole cells and nuclei, minimal aggregation, and sufficient solubilization of DNA and histones. The importance of intermolecular disulfide bonds, RNA and 37 degrees C stabilization on the structural integrity of the nuclear matrix was examined in detail using sulfydryl alkylating, reducing and oxidizing agents, and RNase A. The data suggest that any disulfides formed during the isolation are not essential for maintaining the structural integrity of the in vitro matrix. However, structural integrity of the matrix is dependent upon RNA and to some degree on disulfides that presumably existed in situ. Sodium tetrathionate and 37 degrees C stabilization of isolated nuclei resulted in nuclear matrices containing an approximately twofold greater amount of protein, RNA and DNA than control preparations. The 37 degrees C incubation, unlike the sodium tetrathionate stabilization, does not appear to induce intermolecular disulfide bond formation. Neither stabilizations resulted in significant differences of the major matrix polypeptide pattern on two-dimensional (2-D) gels stained with Coomassie Blue as compared to that of unstabilized matrix. The major nuclear matrix proteins, other than the lamins, did not react to the Pruss murine monoclonal antibody (IFA) that recognizes all known intermediate filament proteins, suggesting that the internal matrix proteins are not related to the lamins in intermediate filament-like quality.

Blotting, Western↗
PubMed1981

Effect of sodium tetrathionate on the activities of some enzymes in kidney and urine.

The activities of lactate dehydrogenase, glutamate dehydrogenase, aspartate aminotransferase, beta-galactosidase, N-acetyl-beta-D-glucosaminidase, leucine aminopeptidase, gamma-glutamyltransferase and alkaline phosphatase in renal tissue and urine of rats treated with sodium tetrathionate were determined. A decrease of enzyme activities in renal tissue and an increase in urine were observed. The largest decrease in the glutamate dehydrogenase of renal tissue amounted to 0.7 times the control value, and was correlated with an appropriate increase in the urine. Increases in urinary enzyme activity were especially marked for beta-galactosidase and N-acetyl-beta-D-glucosaminidase (3 and 6 times the control values, respectively). The increase in enzyme activities was not accompanied by a corresponding change in the urinary protein. Characterization of urinary lactate dehydrogenase and N-acetyl-beta-D-glucosaminidase isoenzymes also indicates the renal origin of these enzymes. The abnormally high enzyme activities of the urine correlated with the nature and degree of renal damage shown by electron microscopy.

Acetylglucosaminidase↗
PubMed1988

Inactivation of type MM phosphoglycerate mutase by sulfhydryl group reagents during facial embryogenesis.

Type MM phosphoglycerate mutase from free dissected mandibular processes from embryonic rats was reversibly inactivated by tetrathionate, p-chloromercuribenzoate, and Hg2+. Titration with p-chloromercuribenzoate showed the existence of two sulfhydryl groups per enzyme subunit, the modification of which produced a progressive decline in enzyme activity. The apparent Km values for substrate and cofactor were not affected by tetrathionate treatment. Phosphoglycerate mutase inactivated by tetrathionate and by p-chloromercuribenzoate was unable to form the functionally active phosphorylenzyme when mixed with glycerate-2,3-P2. Glycerate-2,3-P2 protected against tetrathionate but failed to protect against Hg2+ and p-chloromercuribenzoate.

Animals↗
PubMed1993

Sulfane-activated reduction of cytochrome c by glutathione.

The inorganic sulfane tetrathionate (-O3SSSSO3-) resembles glutathione trisulfide (GSSSG) in that it remarkably activates the reduction of cytochrome c by GSH, both under aerobic and anaerobic conditions. These observations can be explained by the formation of the persulfide GSS-, due to nucleophilic displacements of sulfane sulfur. The GSS- species has previously been proposed to act as a chain carrier in the catalytic reduction of cytochrome c, and perthiyl radicals GSS., formed in the reduction step, were thought to recycle to sulfane via dimerization to GSSSSG.2 The present study provides some arguments in favour of a chain mechanism involving the GSS. + GS-<-->(GSSSG).- equilibrium and sulfane regeneration by a second electron transfer from (GSSSG).- to cytochrome c. Thiosulfate sulfurtransferase (rhodanese) is shown to act as a cytochrome c reductase in the presence of thiosulfate and GSH, and again the generation of GSS- can be envisaged to explain this result.

Animals↗
PubMed1999

Isolation of Salmonella organisms from the mesenteric lymph nodes of horses at necropsy.

OBJECTIVES: To determine the prevalence of Salmonella infections in horses at necropsy. DESIGN: Cross-sectional prevalence survey. ANIMALS: 102 horses. PROCEDURE: Mesenteric lymph nodes were collected from horses that were necropsied. Horses had died or were euthanatized because of severe disease or at the request of the owner. Twenty-eight of the horses were racehorses euthantized following acute catastrophic injuries on the racetrack. Mesenteric lymph nodes were submitted for Salmonella culture via direct plating of tissue specimens on MacConkey agar and by use of 4 enrichment culture techniques that used tetrathionate and selenite enrichment broth and brilliant green and Salmonella-Shigella selective plating media. RESULTS: Salmonella typhimurium was isolated from the mesenteric lymph nodes of 2 foals (2/102, 1.96% of the horses). Salmonella organisms were not isolated from the mesenteric lymph nodes of adult horses. CONCLUSIONS AND CLINICAL RELEVANCE: Prevalence of Salmonella infections in horses of our study (1.96%) suggests that the results of cross-sectional surveys, using bacteriologic culture to determine prevalence of Salmonella infection, should be interpreted with caution. Prevalence of Salmonella infections determined in a single facility may not reflect the prevalence of Salmonella-infected horses in the general population; furthermore, obtaining a Salmonella isolate from a horse does not establish that the horse is a chronic Salmonella carrier.

Agglutination Tests↗
PubMed2001

Mechanism of oxidation of inorganic sulfur compounds by thiosulfate-grown Thiobacillus thiooxidans.

Thiobacillus thiooxidans was grown at pH 5 on thiosulfate as an energy source, and the mechanism of oxidation of inorganic sulfur compounds was studied by the effect of inhibitors, stoichiometries of oxygen consumption and sulfur, sulfite, or tetrathionate accumulation, and cytochrome reduction by substrates. Both intact cells and cell-free extracts were used in the study. The results are consistent with the pathway with sulfur and sulfite as the key intermediates. Thiosulfate was oxidized after cleavage to sulfur and sulfite as intermediates at pH 5, the optimal growth pH on thiosulfate, but after initial condensation to tetrathionate at pH 2.3 where the organism failed to grow. N-Ethylmaleimide (NEM) inhibited sulfur oxidation directly and the oxidation of thiosulfate or tetrathionate indirectly. It did not inhibit the sulfite oxidation by cells, but inhibited any reduction of cell cytochromes by sulfur, thiosulfate, tetrathionate, and sulfite. NEM probably binds sulfhydryl groups, which are possibly essential in supplying electrons to initiate sulfur oxidation. 2-Heptyl-4-hydroxy-quinoline N-oxide (HQNO) inhibited the oxidation of sulfite directly and that of sulfur, thiosulfate, and tetrathionate indirectly. Uncouplers, carbonyl cyanide-m-chlorophenylhydrazone (CCCP) and 2,4-dinitrophenol (DNP), inhibited sulfite oxidation by cells, but not the oxidation by extracts, while HQNO inhibited both. It is proposed that HQNO inhibits the oxidation of sulfite at the cytochrome b site both in cells and extracts, but uncouplers inhibit the oxidation in cells only by collapsing the energized state of cells, delta muH+, required either for electron transfer from cytochrome c to b or for sulfite binding.

2,4-Dinitrophenol↗
PubMed2001

Determination of salmonellae from municipal wastewaters.

This study compared the efficiency of culture methods for salmonellae detection in wastewaters collected from three Finnish municipal treatment plants and from one laboratory-scale plant. The performance of one-step enrichment in Preuss tetrathionate broth was better than that of two-step enrichment (buffered peptone water pre-enrichment (BPW) and selective enrichment in Rappaport-Vassiliadis (RV) medium. The best combinations for Salmonella isolation were xylose-lysine-deoxycholate (XLD) and Rambach (RB) agars after Preuss enrichment and did not differ when brilliant green-magnesium chloride (BM) or brilliant green phenol red (BP) agars were used. The two-step enrichment inhibited the growth of both salmonellae and interfering accompanying flora. Salmonella-positive plates were generally easier to read when inoculated from RV than from Preuss medium because of less growth of competing flora. XLD and BM agars supported growth of salmonellae and inhibited growth of competing flora better than BP and Rambach agars. XLD and BM agars gave the highest numbers of salmonellae isolations but XLD and Rambach agars gave the best differentiation. Salmonella levels were < 3- > 1100 MPN/100 mL.

Agar↗