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Speciation and antibiotic susceptibility patterns of coagulase-negative staphylococci.

During a six month period, 191 isolates of coagulase-negative staphylococci from blood, urine, cerebrospinal fluid and heart valves were identified to species level and tested for antimicrobial susceptibility. Seventy-one percent of isolates were Staphylococcus epidermidis, 8% Staphylococcus warneri, 7% Staphylococcus hominis, 7% Staphylococcus haemolyticus, 4% Staphylococcus capitis, 2% Staphylococcus saprophyticus and 1% Staphylococcus cohnii. Approximately 4% of isolates were felt to be associated with infection. Overall, 18% of isolates were susceptible to penicillin G, 61% oxacillin, 98% cephalothin, 98% cefamandole, 72% cefotaxime, 95% cefsulodin, 76% gentamicin, 64% clindamycin and 98% rifampicin. All isolates were susceptible to vancomycin. Vancomycin, rifampicin, cephalothin and cefamandole showed excellent activity against oxacillin-resistant isolates. With one exception, speciation was not helpful in determining whether or not an isolate was associated with infection.

Anti-Bacterial Agents↗

The speciation of coliform genera from above and below a sewer outfall and their susceptibilities to antimicrobial agents.

The occurrence of coliforms in a small water course was shown to increase by a factor of thirty six below the outfall of a sewage treatment plant. Speciation of the bacteria from above and below the sewer outfall showed that Escherichia coli and Enterobacter species predominated. Drug resistance levels were significant in microorganisms from both sampling sites and the occurrence of a significant number of multiple-resistant microorganisms, particularly E. coli, is reported. Both E. coli and Enterobacter species from below the sewer outfall show a statistically significant increase in resistance to ampicillin and E. coli from below the outfall also shows a statistically significant increase in resistance to sulpha-methoxazole as compared with isolates from above the outfall.

Anti-Bacterial Agents↗

Reduced meiotic fitness in hybrids with heterozygosity for heterochromatin in the speciating Mus terricolor complex.

Mus terricolor I, II and III are the three chromosomal species which differ in stable autosomal short-arm heterochromatin variations established in homozygous condition. Analysis of meiosis in the laboratory-generated F1 male hybrids from crosses (both ways) between M. terricolor I and II and between M. terricolor I and III shows high frequencies of pairing abnormalities at pachytene. The backcross (N3 generation) male hybrids between M. terricolor I and II have meiotic abnormalities as in the F1 male hybrids, though to a lesser extent. They show difference in pairing abnormalities in the different karyotypic forms; the backcross hybrids heterozygous for the heterochromatic short arms have more anomalies compared to the homokaryotypic hybrids. This suggests a negative influence of the heterochromatin heterozygosity in meiotic pairing. The results indicate a role for heterochromatin variations in the development of a reproductive barrier in the speciating M. terricolor complex.

Animals↗

Radiotracer analysis of cadmium speciation in soil solutions using electrophoresis, dialysis, centrifugation, and ultrafiltration.

Speciation of carrier-free 109Cd, added in cationic form to pre-filtered extracts obtained by leaching forest soil samples with distilled water, was analyzed using electrophoresis, dialysis, centrifugation, and ultrafiltration. Rapid establishment of isotopic equilibrium between the added 109Cd and stable cadmium present in the extracts was observed. All the data obtained indicated that 109Cd and also stable cadmium were present in the analyzed extracts in the form of neutral or negatively charged organic complexes or small colloids. The results of electrophoresis enabled the characterization, at least semiquantitative, of the abundance and electrophoretic mobility of the forms present. The incomplete dialysis of 109Cd from the soil extracts through cellophane membrane against water proved the presence of organic associates with molecular weights higher than 10(4). Dialysis against the same, but unlabeled extract was always complete, indicating the reversible (labile) nature of the organic forms of cadmium. Assessment of the stability constants of the organic forms using a simple discrete two-site model suggested that humate and/or fulvate complexes of cadmium were formed.

Cadmium↗

Speciation of inorganic arsenic and selenium in leachates from landfills in relation to water quality assessment.

Speciation of arsenic and selenium was carried out on water samples taken from rivers used as water intake points in the vicinity of landfill areas used for land-based waste disposal system. Leachates from these landfill areas may contaminate the river water through underground seepage or overflowing, especially after a heavy downpour. Preconcentration of the chemical species was done using a mixture of ammonium pyrrolidinethiocarbamate-chloroform (APDTC-CHCl3). Because only the reduced forms of both arsenic and selenium species could be extracted by the preconcentrating mixture, suitable reducing agents such as 25% sodium thiosulfate for As(III) and 6M HCl for Se(IV) were used throughout the studies. Care was taken to exclude the interfering elements such as the alkali and alkali earth metals from the inorganic arsenic and selenium species by introducing 12% EDTA solution as the masking agent. The extracted mixture was irradiated in a thermal neutron flux of 4 x 10(12)/cm/s from a TRIGA Mk.II reactor at the Malaysia Institute of Nuclear Technology Research (MINT). Gamma rays of 559 keV and 297 keV from 76As and 75Se, respectively, were used in the quantitative determination of the inorganic species. Mixed standards of As(III) and Se(IV) used in the percentage efficiency procedure were prepared from salts of Analar grade. The water quality evaluation was viewed from the ratio of the inorganic species present.

Arsenic↗

Chemical speciation of arsenic in urine of patients with blackfoot disease.

Blackfoot disease is a peripheral vascular disease resulting in gangrene of the lower extremities. Although extensive epidemiological study has implicated high arsenic content in artesian well water of the endemic area bears some important connection with the disease, the etiology of the disease is still not clarified. In this study, attention is paid to chemical speciation of arsenic in order to find out whether the concentrations of arsenic species in urine of Blackfoot disease patients are different from those of controls. Experimental results indicate that the total arsenic, inorganic arsenic, monomethylarsonic acid, and other forms of arsenic in the urine of patients are significantly higher than those of the controls. The possible connection of those arsenic species with the etiology of the disease is discussed.

Arsenic↗

Trace element concentration and arsenic speciation in the well water of a Taiwan area with endemic blackfoot disease.

Blackfoot disease is a peripheral vascular disease resulting in gangrene of the lower extremities. Although extensive epidemiological study has implicated high arsenic content in artesian well water in the endemic area, there is more to learn about the etiology of the disease. In this study, effort is paid on multielement determination and arsenic speciation in order to find out whether the trace element concentration pattern in well water in the Blackfoot disease endemic area is different from those of two control areas. Experimental results indicate that the concentrations of Fe, P, Na, and Ba in well water in the Blackfoot disease endemic area are found to be significantly higher than those of the controls, but they are still below the drinking water standard. The total arsenic in well water in the endemic area (671 +/- 149 ppb) is much higher than that of one normal control area of Hsin-Chu (< 0.7 ppb), but is a similar level as that of other control areas of I-Lan (653 +/- 71 ppb) where no Blackfoot disease has ever been found. It was also found that the insoluble arsenic in the endemic area (21.9 ppb) is much higher than that in two control areas (< or = 1.8 ppb), and the concentration ratio between As(III) and As(V) species in the endemic area (2.6) is much lower than that in one of the control areas, where the total arsenic is also high (14.7). The possible connection of Blackfoot disease with trace elements, arsenic species, and possibly other as yet undefined environmental factors in the artesian well water, is discussed.

Arsenic↗

Aluminum speciation and morphological differentiation in murine neuroblastoma cells.

Aluminum is known as a neurotoxic metal ion in experimental animals as well as in humans. The present study was carried out to determine whether and how the physicochemical properties of the metal coordination sphere (metal speciation) can influence the differentiation of murine neuroblastoma cells as has been observed previously in this laboratory (1). Results herein reported indicate that while the aluminum lipophilic species--particularly aluminum acetylacetonate (Alacac3) and aluminum maltolate (Almalt3), both hydrolitically stable and differently lipophilic--are both rather cytotoxic, metal hydrophilic species show different neuritogenic properties indicating the ability of Al(III), when diversely coordinated, to produce different biological effects.

Aluminum↗

Phyto-availability and speciation change of heavy metals in soils amended with lignin as micro-fertilizer.

Lignin is a primary byproduct from the black liquor treatment in paper making industries, its application as micro-fertilizer in agricultural land might provide a promising alternative to sewage discharge. However, application of such a micro-fertilizer might affect the soil properties and result in soil pollution. In this study, the effects of lignin application on phyto-availability and speciation change of heavy metals in soils were investigated. Greenhouse experiments showed that lignin application improved the growth of winter wheat (Triticum aestivum L.) in all three soils investigated. The increase of the biomass for wheat shoot was 59.7%, 39.8% and 12.3% for Beijing soil, Jiangxi soil and Dongbei soil, respectively. In contrast, lignin amendment decreased the concentrations of heavy metals in wheat shoots from 2.2% to 61.0%. Sequential extraction procedure of a three-step BCR was used to investigate the fraction distribution. The extractable fractions were specified as fraction B1: water soluble, exchangeable and carbonate bound or weakly specifically adsorbed; B2: Fe-Mn oxide bound; B3: organic matter and sulfide bound. The results showed that lignin application led to the redistribution of heavy metals in each fraction. Generally, heavy metals decreased in B1 and B2 fractions and increased in B3 fraction. Upon the results short-term application of lignin in agricultural land not only improves the growth of wheat but also reduces the phyto-abailability of heavy metal in wheat.

Biomass↗

Trace cobalt speciation in bacteria and at enzymic active sites using emission Mössbauer spectroscopy.

57Co emission Mössbauer spectroscopy (EMS) allows the chemical state of cobalt, as influenced by its coordination environment, to be monitored in biological samples at its physiological (trace) concentrations. To draw attention to EMS as a valuable tool for speciation of cobalt in biocomplexes, the process of cobalt(II) metabolism in cells of the plant growth-promoting rhizobacterium Azospirillum brasilense Sp245 was investigated using EMS of 57CoII-doped bacterial cells. EMS measurements also showed 57CoII-activated glutamine synthetase (GS, a key enzyme of nitrogen metabolism, isolated from this bacterium) to have two different cobalt(II) forms at its active sites, in agreement with data available on other bacterial GSs. Chemical after-effects following electron capture by the nucleus of the parent 57CoII during the 57Co-->57Fe transition, which contribute to the formation of a stabilised daughter 57FeIII component along with the nucleogenic 57FeII forms, are also briefly considered.

Azospirillum brasilense↗

Speciation of organotin compounds in marine sediments by capillary column gas chromatography-atomic absorption spectrometry coupled with hydride generation.

A method for speciation of organotin compounds in marine sediments by solvent extraction combined with hydride generation gas chromatography-atomic absorption spectrometry has been developed. Sediment samples spiked with tributyltin and triphenyltin chlorides were homogenized in hydrochloric acid. The chlorides were extracted twice into toluene. Recoveries of the organotin compounds from the spiked sediment samples were improved by the addition of 8-quinolinol. Tributyltin and triphenyltin chlorides form ion-associates with 8-quinolinol in aqueous hydrochloric acid. The method was optimized with respect to derivatization reactions and extraction conditions. Interferences from Sn(II/IV) and additional 13 ions were investigated. Recoveries of 84-100% for tributyltin and 86-100% for triphenyltin were achieved using this method. The detection limits obtained for tributyltin and triphenyltin chlorides were 95 and 145 pg, respectively, corresponding to a relative detection limit of 95 and 145 ng kg(-1) in the sediment.

Chromatography, Gas↗

Speciation analysis of trace elements in the brains of individuals with Alzheimer's disease with special emphasis on metallothioneins.

A speciation analysis of protein-bound elements in the cytosol of human brain was achieved by size exclusion chromatographical separation of the biomolecules and on-line detection of the metal profiles in the eluate by hyphenated inductively coupled plasma-mass spectrometry. Post-mortem samples from Alzheimer's disease brains and from brains of a control group were investigated to elucidate changes in the trace element distribution during the pathological process. Special attention was paid to the metallothioneins (MT) - cysteine-rich, metal-binding proteins of low molecular weight, existing in several isoforms. The isoform MT-3 is found especially in the brain and has a growth inhibition function on neurons. The MT peaks were identified in the element profiles. For this purpose, the metal binding capability and the heat stability of MT were taken into consideration. For verification, a comparison with pure MT-3 was carried out and further biochemical and analytical methods were applied to the fractions of the chromatographical run. A comparison between Alzheimer's disease and control brains showed a significant difference concerning the MT-1/-2 and MT-3 metal levels, leading to the assumption that there were oxidative processes having taken place in the Alzheimer's brain samples.

Alzheimer Disease↗

Speciation and subcellular location of se-containing proteins in human liver studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and hydride generation-atomic fluorescence spectrometric detection.

Speciation of Se-containing proteins in the subcellular fractions of human liver was studied by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) followed by hydride generation-atomic fluorescence spectrometric (HG-AFS) detection. It was found that about 24 kinds of Se-containing proteins existed in subcellular fractions of normal human liver. The molecular weights (MW) of the subunits were mostly in the range 20-30 kDa and 50-80 kDa. Major Se-containing protein fractions at 61 kDa and 21 kDa are probably selenoprotein P and glutathione peroxidase, respectively. The 54 kDa protein is probably a thioredoxin reductase, which is presented in nuclei, mitochondria, lysosome, microsome and cytosol. We noticed that the Se-containing protein with the lowest MW of 9.3 kDa only existed in lysosome. Most of the proteins have not been identified and would require further investigation to characterize them. The specific subcellular distributions of different Se-containing proteins suggest that they could play important biological roles in each organelle.

Electrophoresis, Polyacrylamide Gel↗

Speciation of Cr(III) and Cr(VI) in potable waters by using activated neutral alumina as collector and ET-AAS for determination.

The speciation of Cr(III) and Cr(VI) has been performed by using activated neutral alumina as adsorbent. Both species were quantitatively adsorbed on a small column filled with neutral alumina. The adsorbed Cr(III) was eluted with 4 mol L(-1) HNO(3) and Cr(VI) with 1.0 mol L(-1) ammonia solution. Recoveries of Cr(III) and Cr(VI) were 99% and 100%, respectively. Using ET-AAS for Cr determination the limit of detection in the sample was 0.01 microg L(-1). The combined procedure is fast and sensitive. It can be applied for routine analysis of water samples at sub-microg L(-1) levels with a relative standard deviation (RSD) of 2-10% (three determinations).

Aluminum Oxide↗

Modern trends in the speciation of selenium by hyphenated techniques.

The complexity of selenium speciation in the environment and in living organisms results in broad analytical challenges. The importance of the selective determination of the particular species of this element, to understand its metabolism and biological significance in clinical chemistry, biology, toxicology, and nutrition, calls for state-of-the-art analytical techniques. In this paper hyphenated techniques are evaluated with particular emphasis on interfaced separation with element-selective detection and identification of the selenium compounds detected.

Chromatography, Gas↗

Solid-phase reagent for molecular spectroscopic determination of heavy metal speciation in natural water.

A solid-phase reagent based on 1-(4-adamantyl-2-thiasolylazo)-2-naphthol adsorbed onto silica gel was prepared for Co(II) recovery and preconcentration prior to its sorption-spectroscopic detection. The immobilized reagent was applied to the determination of free cobalt ions in natural water. The solid-phase reagent and chemiluminescent method coupled with membrane filtration, gel-permeation and ion-exchange chromatography were applied to the study of the speciation of iron and cobalt in water from the Dnieper reservoirs and lakes of Kyiv City; their predominant forms were complexes of Fe(III) and Co(II) with dissolved organic matter and fulvic acids play a main role in their complexation.

Journal Article↗

Application of capillary electrophoresis for inorganic selenium speciation in the frame of high-level waste management.

Capillary electrophoresis (CE) with direct UV detection is proposed for speciation of inorganic Se in high-level liquid waste. In this aim, the optimal conditions of measurements (pH, electrolyte buffer concentration) and the influence of nitrate excess on the quantitative determination of Se(IV) and Se(VI) were studied. Different electrolyte buffers were considered: carbonate, phosphate and citrate. It was found, that citrate buffer is the most suitable for the application under consideration. Under the chosen optimal conditions (20 mmol L(-1) citrate buffer, pH 2.5), calibration curves for Se(IV) and Se(VI) are linear in the concentration range 10(-4)-10(-3) mol L(-1). The detection limits are 4x10(-6 )for Se(IV) and 2x10(-5) for Se(VI). The accuracy of the procedure was checked by calculating the recovery by spiking simulation solutions. Relative standard deviation (S(r)) is less than 10%.

Buffers↗

Speciation analysis of tellurium by solid-phase extraction in the presence of ammonium pyrrolidine dithiocarbamate and inductively coupled plasma mass spectrometry.

Under acidic conditions tellurium(IV) formed a complex with ammonium pyrrolidine dithiocarbamate (APDC). The tellurium(IV) complex was completely retained on a non-polar Isolute silica-based octadecyl (C(18)) sorbent-containing solid-phase extraction (SPE) cartridge, while the uncomplexed Te(VI) passed through the cartridge and remained as a free species in the solution. Only partial Te(IV) was retained on the SPE cartridge for samples without addition of APDC. On the basis of different retention behaviours of the complexed Te(IV) and uncomplexed Te(VI), a simple and highly sensitive method is proposed for the determination of total tellurium and Te(VI) by SPE separation and inductively coupled plasma mass spectrometry (ICP-MS) detection. The Te(IV) concentration was calculated as the difference between total tellurium and Te(VI) concentrations. The detection limit (3 sigma) is 3 ng L(-1) tellurium. Factors affecting the separation and detection of tellurium species were investigated. Coexisting ions did not show significant interferences with the Te(IV)-APDC complex retention and the subsequent ICP-MS detection of Te. The method has been successfully applied to the tellurium speciation analysis in waters with spiked recoveries for Te(IV) and Te(VI) of 86.0-108% and 87.1-97.4%, respectively.

Mass Spectrometry↗