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Dechloromonas agitata gen. nov., sp. nov. and Dechlorosoma suillum gen. nov., sp. nov., two novel environmentally dominant (per)chlorate-reducing bacteria and their phylogenetic position.

Previous studies on the ubiquity and diversity of microbial (per)chlorate reduction resulted in the isolation of 20 new strains of dissimilatory (per)chlorate-reducing bacteria. Phylogenetic analysis revealed that all of the isolates were members of the Proteobacteria with representatives in the alpha-, beta- and gamma-subclasses. The majority of the new isolates were located in the beta-subclass and were closely related to each other and to the phototrophic Rhodocyclus species. Here an in-depth analysis of these organisms which form two distinct monophyletic groups within the Rhodocyclus assemblage is presented. Two new genera, Dechloromonas and Dechlorosoma, are proposed for these beta-subclass lineages which represent the predominant (per)chlorate-reducing bacteria in the environment. The type species and strains for these new genera are Dechloromonas agitata strain CKBT and Dechlorosoma suillum strain PST, respectively.

Betaproteobacteria↗

Improved method to measure erythrocyte filtration times increased extremely by chlorate.

A method is presented that allows the measurement of erythrocyte filtration times that are extremely prolonged. Filtration times through polycarbonate sieves are increased by a factor of 40 after 2 h incubation with 30 mM sodium chlorate. This increase in red cell rigidity offers an explanation for the haemolysis observed in chlorate poisoning in vivo.

Chlorates↗

Identification in various chlorate-resistant mutants of a protein involved in the activation of nitrate reductase in the soluble fraction of a chlA mutant of Escherichia coli K-12.

We report some properties of Protein PA which has been isolated from the soluble fraction of a chlB mutant after anaerobic growth in the presence of KNO3. This protein has been identified by its capacity to reactivate nitrate reductase present in the soluble fraction of a chlA mutant by the complementation process. The presence of active Protein PA in the chlB mutant is independent of the presence of oxygen or of nitrate during growth. In contrast, the addition of sodium tungstate to the growth medium leads to the formation of inactive Protein PA which is not able to activate nitrate reductase in the chlA-soluble extract by complementation. Inactive Protein PA has been quantitated immunologically. The partial purification of Protein PA has been achieved from various chlorate-resistant mutants (chlA-chlG). The establishment of particular complementation systems comprising the soluble extracts of chlA or chlB mutants and partially purified Protein PA from soluble fractions of different chlorate-resistant mutants, has allowed the quantitative estimation of this protein. The analysis by 'rocket immunoelectrophoresis' using an antiserum specific for Protein PA has shown that inactive Protein PA is present in approximately equivalent amounts in the chlA, chlE, chlG and chlD mutants.

Bacterial Proteins↗

Colletotrichum sublineolum genetic instability assessed by mutants resistant to chlorate.

The fungus Colletotrichum sublineolum, causal agent of sorghum anthracnose, presents high variability, genetic instability and host specialization. The aims of the present work were to investigate the mechanisms involved in the genetic instability in this species. Mutants resistant to chlorate and unable to use nitrate (Nit mutants), were obtained spontaneously, isolated and characterized for complementation pattern, reversion frequency and RAPD profile. The results showed that chlorate-resistant mutants could be divided into six phenotypic classes that probably represented mutations in the structural nitrate reductase locus (nit1), in the structural nitrite reductase locus (nit6 and niiA of Neurospora and Aspergillus, respectively), in the specific regulator locus (nit3), in the main regulator locus (nit2), in loci that codified the cofactor containing molybdenum necessary for nitrate reductase activity (NitM), and one or more genes responsible for nitrate intake (crn). In addition, the genetic control of this metabolism in C. sublineolum seems to be similar to other fungi species such as Aspergillus, Neurospora and Fusarium. The high reversion frequency (10(-4) to 10(-5)) presented by nit1 mutants suggests that the instability in evaluated strains could be a result of transposable elements activity. The RAPD analysis enabled confirmation that the Nit mutants have a similar genetic background to original strain, and that polymorphism exists among wild-type strains, nit1 mutants and revertants of C. sublineolum. These are important aspects for the later direction of molecular analysis, where these mutants will be used as a tool to isolate the active transposable elements in the C. sublineolum genome.

Chlorates↗

Determination of trace selenium by solid substrate-room temperature phosphorescence enhancing method based on potassium chlorate oxidizing phenyl hydrazine-1,2-dihydroxynaphthalene-3,6-disulfonic acid system.

A new method for the determination of trace selenium based on solid substrate-room temperature phosphorimetry (SS-RTP) has been established. This method was based on the fact that in HCl-KCl buffer solution, potassium chlorate could oxidize phenyl hydrazine to form chloridize diazo-ion after being heated at 100 degrees C for 20 min, and then the diazo-ion reacted with 1,2-dihydroxynaphthalene-3,6-disulfonic acid to form red azo-compound which could emit strong room temperature phosphorescence (RTP) signal on filter paper. Selenium could catalyze potassium chlorate oxidizing the reaction between phenyl hydrazine and 1,2-dihydroxynaphthalene-3,6-disulfonic acid, which caused the sharp enhancement of SS-RTP. Under the optimum condition, the relationship between the phosphorescence emission intensity (DeltaIp) and the content of selenium obeyed Beer's law when the concentration of selenium is within the range of 1.60-320 fg spot-1 (or 0.0040-0.80 ng ml-1 with a sample volume of 0.4 microl). The regression equation of working curve can be expressed as DeltaIp=13.12+0.4839CSe(IV) (fg spot-1) (n=6), with correlation coefficient r=0.9991 and a detection limit of 0.28 fg spot-1 (corresponding to a concentration range of 7.0x10(-13) g ml-1 Se(IV), n=11). After 11-fold measurement, R.S.D. were 2.8 and 3.5% for the samples containing 0.0040 and 0.80 ng ml-1 of Se(IV), respectively. This accurate and sensitive method with good repeatability has been successfully applied to the determination of trace selenium in Chinese wolfberry and egg yolk with satisfactory results. The mechanism of the enhancement of phosphorescence was also discussed.

Animals↗

Potential for chlorate interference in ion chromatographic determination of total nitrogen in natural waters following alkaline persulfate digestion.

Determination of total nitrogen in aqueous samples after thermal potassium peroxydisulfate (persulfate) digestion is a commonly used alternative to the tedious Kjeldahl procedure. When ion chromatography is used to quantify the nitrate formed during digestion, there is a potential for interference from a chlorate peak if the digested sample initially contained chloride in concentrations close to or greater than the concentration of nitrogen. It was determined that this interference can be avoided either by using chromatographic conditions which cleanly resolve the nitrate and chlorate peaks (e.g., the Dionex AG9-HG column) or by using digestion reagent concentrations chosen to maintain a high pH throughout the digestion. The second alternative is not a viable option for investigators using a single digestion for both total nitrogen (TN) and total phosphorus (TP) analysis.

Chlorates↗

Effect of experimental chlorate product administration in the drinking water on Salmonella typhimurium contamination of broilers.

The crop is a known source of Salmonella and Campylobacter contamination. Previously, we evaluated lactic acid in the drinking water during a simulated pretransport feed withdrawal (FW) and reported 0.44% lactic acid significantly (P < 0.05) reduced the number of Salmonella recovered in market-age broiler crops. However, total consumption of the organic acid-treated drinking water was reduced. Presently, we evaluated the effect of experimental chlorate product (ECP; 1x ECP is equivalent to a 15 mM chlorate ion concentration) during a 10-h pretransport FW. Market-age broilers were obtained from a commercial processing plant and randomly assigned to ECP-treated or control (nontreated) groups. Broilers were challenged by crop gavage with 10(8) Salmonella Typhimurium (ST) immediately upon arrival and 1 d prior to termination of the experiment. One day later, broilers were killed for ST enumeration (cfu) in the crop and ceca. Broilers provided ECP 24 h prior to slaughter consumed slightly more ECP water than broilers provided distilled water. Treatment with ECP caused a significant decrease (P < 0.05) in the incidence of ST in crop contents (2%) as compared to the controls (36.7%). Similarly, ECP treatment caused a significant decrease (P < 0.05) in number of ST (0.96 log10 ST/g cecal content) detected in the ceca when compared to controls (2.52 log10 ST). This study suggested that incorporation of ECP in the drinking water 24 to 48 h prior to slaughter could reduce Salmonella contamination in broilers.

Animals↗

Evaluation of Salmonella enteritidis in molting hens after administration of an experimental chlorate product (for nine days) in the drinking water and feeding an alfalfa molt diet.

The method most commonly used to induce molting and stimulate multiple egg-laying cycles in laying hens for commercial egg production is to fast the hens. Unfortunately, increased risk of Salmonella enteritidis (SE) infection may result from the use of this method. Methods to stimulate multiple egg-laying cycles without increasing the risk of SE infection are needed. Hens over 50 wk of age were divided into 12 groups of 11 hens each and placed in individual laying cages. One week prior to dietary changes, hens were placed on an 8-h light and 16-h dark photoperiod that continued for the 9-d molt. All hens were challenged orally with 10(6) cfu of SE on the fourth day of the molt. Treatments were nonfed hens with distilled water (NFD), nonfed hens with the experimental chlorate product (ECP, which provided 15 mM chlorate ion) water (NFECP), alfalfa diets with distilled water (ALD), and alfalfa diets with ECP water (ALECP). In the NFD hens, 67% (log10 2.74) of the crops and 94% (log10 5.62) of the ceca were colonized, whereas for the NFECP hens significant reductions to 22% (log10 1.05) of the crops and 61% (log10 2.44) of the ceca were observed. In the ALD hens, 61% (log10 2.52) of the crops and 94% (log10 4.06) of the ceca were colonized. In the ALECP hens, highly significant reductions to 11% (log10 1.26) of the crops and 39% (log10 1.12) of the ceca were observed. When compared with the NFD hens, significant reductions in SE invasion of the ovary, liver, and spleen occurred in all other treatments, except the ovary in the ALD hens. The low alfalfa intake is probably a factor in our lowered protection against SE when compared with previous results. For several parameters, these results suggest that ECP or the combination of ECP and alfalfa may be a useful tool to reduce the risk of SE during an induced molt.

Animal Feed↗

The effect of an experimental chlorate product on Salmonella recovery of turkeys when administered prior to feed and water withdrawal.

Previously, an experimental chlorate product (ECP) has been observed to reduce Escherichia coli and Salmonella infections in swine, cattle, and broilers. The following studies were performed to investigate the effects of different concentrations and durations of administering ECP on crop and ceca Salmonella typhimurium (ST) colonization of turkeys. In 2 separate trials, each conducted with 2 replicates, 15-wk-old turkey toms were challenged with 10(7) to 10(9) cfu of ST. In Experiment 1, toms were administered 0, 0.5, 1.0, 2.0, or 4.0x of ECP (a 1.0x concentration is equivalent to a 15 mM chlorate ion concentration) in the drinking water for 38 h. In Experiment 2, toms were administered a 2x concentration of ECP in the drinking water for 0, 14, 26, or 38 h prior to water withdrawal. All treatments were followed by a 10-h water withdrawal and an 8-h feed withdrawal prior to organ sampling. In Experiment 1, turkeys provided ECP had significantly (P < 0.05) lower populations and incidences of crop (>1.4 log reduction) and ceca (>0.6 log reduction) ST as compared with control birds (2.1 and 0.94 log ST average for all trials, respectively), with little or no additional benefit from administration of higher ECP concentrations. In Experiment 2, toms provided ECP had lower populations of crop (>2.2 log reduction) and ceca (>1.5 log reduction) ST when compared with controls (3.1 and 1.8 log ST, respectively). Again, there appeared to be little benefit in longer administration intervals on quantitative reduction of ST. These experiments suggest that the ECP significantly reduces Salmonella colonization in commercial turkeys when administered prior to feed and water withdrawal.

Animal Feed↗

Structure of tetraaquacopper(II) chlorate at 296 and 223 K.

[Cu(H2O)4](ClO3)2, Mr = 302.51, orthorhombic, Pbca, lambda (Mo K alpha) = 0.71069 A, Z = 4, F(000) = 604; T = 296 K, a = 12.924 (3), b = 9.502 (2), c = 7.233 (1) A, V = 888.3 (3) A3, D chi = 2.26 g cm-3, mu = 31.04 cm-1, R = 0.041 for 1174 unique reflections with I greater than sigma (I); T = 223 K, a = 12.853 (2), b = 9.492 (2), c = 7.216 (2) A, V = 880.4 (3) A3, D chi = 2.28 g cm-3, mu = 31.28 cm-1, R = 0.033 for 1279 unique reflections with I greater than sigma (I). The single type of copper ion is octahedrally coordinated by four water oxygens and two chlorate ion oxygens; the complex is characterized by three distinct pairs of Cu-O distances (which we have previously described as '2 + 2 + 2' coordination), one of which is substantially larger than the remaining two. The nominal 90 degrees O-Cu-O angles of the complex differ from that value by at most +/- 1.8 degrees. The observed structure of the complex is typical for copper(II) and is consistent with the Jahn-Teller effect. The single type of chlorate ion shows the expected trigonal pyramidal geometry, with average Cl-O bond lengths 1.485 (13) and 1.490 (11) A and average O-Cl-O bond angles 106.8 (13) and 106.8 (13) degrees for 296 and 223 K, respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Chemical Phenomena↗

Wound isolate of Salmonella typhimurium that became chlorate resistant after exposure to Dakin's solution: concomitant loss of hydrogen sulfide production, gas production, and nitrate reduction.

A strain of Salmonella typhimurium isolated from a decubitus ulcer that was being treated topically with half-strength Dakin's solution became H2S negative, nitrate negative, and unable to produce gas from glucose. Experimental data suggested that these effects were associated with the development of chlorate resistance. Thirty-five other strains of Salmonella spp. that were made chlorate resistant also became negative for these three tests.

Chlorates↗

Nitrates, chlorates and trihalomethanes in swimming pool water.

Water from swimming pools in the Miami area was analyzed for nitrates, chlorates and trihalomethanes. The average concentrations of nitrate and chlorate found in freshwater pools were 8.6 mg/liter and 16 mg/liter respectively, with the highest concentrations being 54.9 mg/liter and 124 mg/liter, respectively. The average concentration of total trihalomethanes found in freshwater pools was 125 micrograms/liter (mainly chloroform) and in saline pools was 657 micrograms/liter (mainly bromoform); the highest concentration was 430 micrograms/liter (freshwater) and 1287 micrograms/liter (saltwater). The possible public health significance of these results is briefly discussed.

Chlorates↗

Reduction of Salmonella Typhimurium in experimentally challenged broilers by nitrate adaptation and chlorate supplementation in drinking water.

The effects of two feed supplements on Salmonella Typhimurium in the ceca of market-age broilers were determined. Broilers orally challenged 6 days before slaughter with a novobiocin- and nalidixic acid-resistant strain of Salmonella Typhimurium were divided into one of four groups (20 birds each). The first group (the control group) received no treatment, the second group received sodium nitrate (SN) treatment (574 mg of NaNO3 per kg of feed), the third group received experimental chlorate product (ECP) treatment (15 mM NaClO3 equivalents), and the fourth group received ECP treatment in combination with SN treatment. The SN treatment was administered via feed for 5 days immediately before slaughter, and ECP was provided via ad libitum access to drinking water for the last 2 days before slaughter. Cecal contents were subjected to bacterial analysis. Significant (P < 0.05) Salmonella Typhimurium reductions (ca. 2 log units) relative to levels for untreated control broilers were observed for broilers receiving ECP in combination with SN. The ECP-only treatment resulted in significant (P < 0.05) reductions (ca. 0.8 log) of Salmonella Typhimurium in trial 2. We hypothesize that increasing Salmonella Typhimurium nitrate reductase activity resulted in increased enzymatic reduction of chlorate to chlorite, with a concomitant decrease in cecal Salmonella Typhimurium levels. On the basis of these results, preadaptation with SN followed by ECP supplementation immediately preharvest could be a potential strategy for the reduction of Salmonella Typhimurium in broilers.

Animals↗

[Adrenal cortex response to prolonged administration of low doses of magnesium chlorate in rats].

Rat adrenals were studied using histological and histochemical methods during prolonged intoxication with pesticide magnesium chlorate which was administered in a dose equal to 1/100 of LD50 (41 mg/kg of body weight). Animals that received distilled water and were kept in similar conditions were used as control. It was demonstrated that intoxication of rats for 3-7 days results in increased secretory activity of all the zones of adrenal cortex. Later (after 14-90 days) the zonal response to pesticide administration was variable. Magnesium chlorate treatment results in the disturbances of hormonal synthesis in adrenocorticocytes. Compensatory-adaptive capacities of zona fasciculata were found to be greater than those in zona glomerulosa and zona reticularis.

Administration, Oral↗

Determination of trace aluminum by fluorescence quenching method based on catalysis of potassium chlorate oxidizing alizarin red.

A new method for the determination of trace aluminum has been proposed. It is based on the fact that alizarin red can emit strong and stable fluorescence at 80 degrees C for 30 min and Al(3+) can effectively catalyze potassium chlorate oxidizing alizarin red to form non-fluorescence complex which cause the fluorescence quenching. The linear dynamic range of this method is 0.040-4.00 ngl(-1) with a detection limit of 5.3 pgl(-1). The regression equation can be expressed as DeltaI(f)=8.731+21.73c(Al(3+)) (ngl(-1)), with the correlation coefficient r=0.9992 (n=6). This sensitive, rapid and accurate method has been applied to the determination of trace aluminum(III) in human hair and tea samples successfully. What is more, the mechanism of catalyzing potassium chlorate oxidizing alizarin red by the fluorescence quenching method is also discussed.

Aluminum↗

Survey of bottled drinking waters sold in Canada for chlorate, bromide, bromate, lead, cadmium and other trace elements.

Mineral, spring and other bottled drinking waters sold in Canada in the winter of 1995-96 were surveyed for chlorate, bromide, bromate, Cr(VI), Li, B, Al, Mn, Cu, Zn, Sr, Ba, Be, V, Cr, Co, Ni, As, Se, Mo, Ag, Cd, Sb, Tl, Pb, Na, K, Ca and Mg. Chlorate and bromide were determined by ion chromatography (IC) with conductivity detection, Cr(VI) by IC with colorimetric detection, bromate by solvent extraction and gas chromatography (GC), trace elements by inductively coupled plasma mass spectrometry (ICPMS), and Na, K, Ca and Mg by flame atomic absorption spectrometry (FAA). Most chemicals in the 199 samples analysed were well within national and international drinking water guidelines. World Health Organization and/or Canadian drinking water guidelines were exceeded for B (22 samples), Al (9), Cr (1), Mn (5), Ni (1), As (10), Se (24) and Pb (1). Bromate levels are reported for information purposes and are considered as the maximum concentrations in the samples. In three distilled water products, unexpectedly high concentrations of Cu (88-147 micro g l(-1)) and Ni (16-35 micro g l(-1)) were found, and a comparison of distilled and non-distilled waters from two of the brands suggested the likely cause to be contamination during the distillation process. Li concentration in one sample was at a therapeutic dose and could pose an overdose risk to individuals on Li medication.

Canada↗

Molybdenum cofactor (chlorate-resistant) mutants of Klebsiella pneumoniae M5al can use hypoxanthine as the sole nitrogen source.

Selection for chlorate resistance yields mol (formerly chl) mutants with defects in molybdenum cofactor synthesis. Complementation and genetic mapping analyses indicated that the Klebsiella pneumoniae mol genes are functionally homologous to those of Escherichia coli and occupy analogous genetic map positions. Hypoxanthine utilization in other organisms requires molybdenum cofactor as a component of xanthine dehydrogenase, and thus most chlorate-resistant mutants cannot use hypoxanthine as a sole source of nitrogen. Surprisingly, the K. pneumoniae mol mutants and the mol+ parent grew equally well with hypoxanthine as the sole nitrogen source, suggesting that K. pneumoniae has a molybdenum cofactor-independent pathway for hypoxanthine utilization.

Chromosome Mapping↗

The complex structures of ATP sulfurylase with thiosulfate, ADP and chlorate reveal new insights in inhibitory effects and the catalytic cycle.

The ubiquitous enzyme ATP sulfurylase (ATPS) catalyzes the primary step of intracellular sulfate activation, the formation of adenosine 5'-phosphosulfate (APS). It has been shown that the enzyme catalyzes the generation of APS from ATP and inorganic sulfate in vitro and in vivo, and that this reaction can be inhibited by a number of simple molecules. Here, we present the crystal structures of ATPS from the yeast Saccharomyces cerevisiae complexed with compounds that have inhibitory effects on the catalytic reaction of ATPS. Thiosulfate and ADP mimic the substrates sulfate and ATP in the active site, but are non-reactive and thus competitive inhibitors of the sulfurylase reaction. Chlorate is bound in a crevice between the active site and the intermediate domain III of the complex structure. It forms hydrogen bonds to residues of both domains and stabilizes a "closed" conformation, inhibiting the release of the reaction products APS and PPi. These new observations are evidence for the crucial role of the displacement mechanism for the catalysis by ATPS.

Adenosine Diphosphate↗