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A review of lysosomal membrane stability measured by neutral red retention: is it a workable earthworm biomarker?

The ability of biomarkers to integrate effects of chemicals on biota has lead to increased calls for their application in assessing the status of polluted ecosystems. In tandem there has been an increase in our knowledge of the ecophysiological responses of keystone species to pollutants, which has allowed the development of a number of promising methods. In contrast to the number of biomarker development studies, the number of biomarker validation studies has remained limited. This paper redresses this imbalance by drawing together data from studies that have used the earthworm lysosomal membrane stability response (measured using the neutral red retention assay). This review first gives a short history of the biomarker's development. Second, it sets published applications of the technique against established criteria for a "good" biomarker (i.e., dose-response relationship, sensitivity, ecological relevance, confounding factors, chemical specificity, species differences, time-response relationship, methodological concerns, and overall public/regulator confidence, and acceptance). Discussion of the biomarker's suitability to each criterion is followed by an overall evaluation of its workability for routine soil quality assessment and caveats for its use.

Animals↗

The effect of environmental pH on sodium balance in the red-spotted newt, Notophthalmus viridescens.

Sodium influx and efflux of the red-spotted newt, Notophthalmus viridescens, at two pHs were investigated in efts and adults in water and on land. The sodium efflux rate constant was not affected by pH. Efflux was generally greater under aquatic than terrestrial conditions, and for efts than for adults. In an aquatic setting, both adults and efts were able to maintain sodium balance at pH 5, but exhibited net sodium loss at pH 3. Sodium influx was greater in efts at pH 5 than in any other group. Adults in all treatments lost mass during the experiment, while efts in all treatments were able to maintain body mass. In their normal terrestrial environment, red efts probably avoid substantial sodium fluxes by maintaining a hydrophobic skin, which decreases contact with soil solutions.

Animals↗

Biodegradation of azo dyes in a sequential anaerobic-aerobic system.

A sequential anaerobic aerobic treatment process based on mixed culture of bacteria isolated from textile dye effluent-contaminated soil was used to degrade sulfonated azo dyes Orange G (OG), Amido black 10B (AB), Direct red 4BS (DR) and Congo red (CR). Under anaerobic conditions in a fixed-bed column using glucose as co-substrate, the azo dyes were reduced and amines were released by the bacterial biomass. The amines were completely mineralized in a subsequent aerobic treatment using the same isolates. The maximum degradation rate observed in the treatment system for OG was 60.9 mg/l per day (16.99 mg/g glucose utilized), for AB 571.3 mg/l per day (14.46 mg/g glucose utilized), for DR 112.5 mg/l per day (32.02 mg/g glucose utilized) and for CR 134.9 mg/l per day (38.9 mg/g glucose utilized).

Aerobiosis↗

Environmental distal renal tubular acidosis in Thailand: an enigma.

Distal renal tubular acidosis is a common health problem in northeastern Thailand, with the population background of the low potassium intake, low urine citrate, and decreased red blood cell Na-K adenosine triphosphatase (ATPase) activity and the environment of the high soil vanadium. The disease is usually seen in the people with low socioeconomic status in summer. The patients have decreased gastric acidity and low urine potassium. There are varying degrees of renal function from normal to impairment. Gastric hypoacidity is an important clue. Defects in H-K ATPase and anion exchange (AE2) mechanism are considered. The urine vanadium is higher in the patients than that of normal rural northeastern villagers. Inhibition of H-K ATPase by vanadium seems possible and requires more supporting evidence. AE1 gene mutation is noted in few patients. The cause of dRTA is not apparent. The AE2 gene and H-K ATPase gene remain to be studied. Both environmental and genetic factors could contribute to the pathogenesis of the disease.

Acidosis, Renal Tubular↗

Beta-thujaplicin: new quantitative CZE method and adsorption to goethite.

Beta-thujaplicin (beta-TH) is a toxic tropolone derivative present in the heartwood of western red cedar (Thuja plicata) and is used as a preservative and antimicrobial additive in a number of commercial goods. beta-TH released from western red cedar timber used outdoor and from other products containing beta-TH may transfer to soil and leach to groundwater and surface waters. The objective of this study was to quantify the adsorption of beta-TH to goethite as a typical model for geosorbents. Adsorption was studied using pH-adjusted goethite suspensions with solid:solution ratios of 1:500, 0.01 M NaNO(3) electrolyte, and 20 degrees C. beta-TH was determined using a new capillary zone electrophoresis (CZE) method providing a detection limit of 0.21 microM. Near-sorption equilibrium was attained within 48 h. beta-TH showed maximum adsorption at low pH (3.8) and a 70% drop in adsorption from pH 6.2 to 8.8. The Langmuir type adsorption isotherm at pH 5.5 approached a maximum adsorption of 220 micromol/g (= 6.2 micromol/m(2)), which is more than twice the amount of phosphate adsorbed under similar conditions. The affinity of beta-TH for goethite is low as compared with organic ligands such as citrate, oxalate, and 2,4-dihydroxybenzoate. The adsorption data and FTIR analyses indicate that beta-TH is most likely adsorbed as monodentate mononuclear surface complexes at the surface of goethite. Hydrophobic adsorption is thought to contribute to the adsorption, in particular at low pH. The strong adsorption of beta-TH to goethite suggests low mobility in most soil environments, the risk of contamination increasing in soils with high pH (calcareous material), low contents of iron and aluminum oxides, phyllosilicates, and organic matter.

Adsorption↗

Influence of ageing on zinc bioavailability in soils.

Currently, soil quality criteria or soil risk assessments of metals are based on laboratory toxicity tests which are carried out in soils freshly spiked with metal salts. With these data, species sensitivity distributions are fitted, from which hazardous concentrations and predicted no effect concentrations are derived. However, due to long-term processes, called ageing, soil metal availability decreases with time. Here we show that pH is the most important parameter determining the effect of ageing on zinc partitioning in soils, with the effect of ageing becoming more important with increasing pH. Furthermore, zinc bioavailability, expressed as the internal zinc concentrations in red clover (Trifolium pratense) is closely related to pore water zinc concentration. In addition, there is a clear dose-response relationship between the survival of the earthworm Eisenia fetida and the calcium chloride-extracted zinc fraction. These results indicate that zinc partitioning can be used to predict zinc bioavailability to terrestrial organisms. However, the use of spiked soils in toxicity assays can result in an over-estimation of the effects of zinc, especially at a high pH.

Animals↗

Soil acidity and manganese in declining and nondeclining sugar maple stands in Pennsylvania.

For decades, the hardwood forests of northern Pennsylvania have been subjected to chronic atmospheric loading of acidifying agents. On marginal, high-elevation, unglaciated sites, sugar maples (Acer saccharum Marsh.) have experienced severe decline symptoms and mortality. Accelerated soil acidification, base cation leaching, and increased availability of toxic metals have been suggested as predisposing factors contributing to this decline. Manganese, an essential micronutrient, is also a potentially phytotoxic metal that may be a factor associated with poor sugar maple health on soils vulnerable to acidification from anthropogenic sources. We measured Mn levels in four compartments of the soil-tree system (soil, foliage, xylem wood, and sap) on three sugar maple stands in northern Pennsylvania. Two stands were classified as declining and one was in good health. Negative correlations were found between soil pH and Mn levels in the soil, foliage, sap, and xylem wood. Levels of Mn in these pools were consistently higher on declining sites, which correspondingly exhibited lower levels of Ca and Mg. Species differences between red maple (Acer rubrum L.) and sugar maple at the two declining sites suggested different tolerances to excessive Mn. Molar ratios of Mg/Mn and Ca/Mn were different among sites and showed potential as indicators of soil acidification. Significant correlations among soil, sap, foliage, and xylem wood Mn were also noted. These results show clear Mn differences among sites and, when viewed with recent Mn toxicity experiments and other observational studies, suggest that excessive Mn may play a role in the observed decline and mortality of sugar maple.

Acer↗

Coccidioidomycosis in Northern California. An outbreak among archeology students near Red Bluff.

An outbreak of coccidioidomycosis occurred among 39 archeology students in the summer of 1972. The students excavated Indian ruins near Red Bluff in Tehama County, California, 20 miles north of the previously recognized northernmost limit of endemicity. At least 17 persons contracted an illness clinically compatible with a diagnosis of coccidioidomycosis. Coccidioidomycosis was documented by skin test conversion as well as by specific serologic reactions. Coccidioides immitis was also isolated from two soil samples taken at the excavation site. In light of its ecological requirements, it is doubtful that C. immitis will be recovered much farther north than Red Bluff. The occupational hazard of coccidioidomycosis to archeologists and others employed in known endemic areas remains a substantial threat to health.

Adolescent↗

Characterization of the coliform and enteric bacilli in the environment of calves with colibacillosis.

In the first part of the present study the coliform and enteric bacilli in the environment of calves with colibacillosis were examined. The occurrence, number, and pathogenic properties of Escherichia coli in barnyard soils were obtained from six cattle ranches. The O and K serogroups of E. coli isolates obtained from the feces of calves with colibacillosis born at these cattle ranches were determined, and their serotypes were compared with the E. coli O and K serotypes found in soils. The results showed a reservoir of potentially pathogenic E. coli in barnyard soils contaminated with bovine feces. For the second part of this study, 6 healthy calves and 51 calves with colibacillosis were studied. The numbers of total aerobic heterotrophic bacteria, total streptococci, fecal streptococci, total coliforms, and fecal coliforms in the feces of calves were determined. In addition, coliform and enteric bacilli from the feces of both healthy and diseased calves were identified, and their indole, methyl red, Voges-Proskauer, citrate (IMViC) types were described. In parallel, the IMViC types of coliform and enteric bacilli isolated from barnyard soils previously contaminated with bovine feces were compared with those isolated from uncontaminated soils. All fecal specimens were also examined for the presence of rotavirus. No significant effect on the numbers of the bacterial types was found. The results suggest that the predominant IMViC types found in the feces of calves with colibacillosis originate from the soil. From this study it is apparent that the occurrence, number, and survival of E. coli in barnyard soils is related to ranch husbandry and sanitary practices.

Animals↗

[Diversity of Sinorhizobium fredii strains].

Rhizobial strains were isolated from soils growing with soybean cultivar Heilong 33 and Willimas. Fifty Sinorhizobium fredii strains were chosen, and their biological characteristics including growth velocity, acid-alkali endurance, resistance of intrinsic antibiotics, utilization of carbon and nitrogen sources, absorption of congo red, ability of melanin production, and plasmid profiles were comparatively researched. The dendrogram was described using the method of cluster analysis, and the biodiversity of Sinorhizobium fredii from different soils was proved.

Carbon↗

Reflected Far-Red Light Effects on Chlorophyll and Light-Harvesting Chlorophyll Protein (LHC-II) Contents under Field Conditions.

The influence of various colors of soil cover (mulch) on the farred/red (FR/R) ratio in upwardly reflected light and on concentrations of chlorophyll (Chl) and light-harvesting Chl protein (LHC-II) were measured under field conditions. The FR/R ratios above green surfaces were higher than over white surfaces. Even though plants (Gossyplum hirsutum L. cv PD-1) were grown in full sunlight, those that received higher FR/R ratios in upwardly reflected light were taller and had thinner leaves with higher concentrations of Chl and LHC-II. A controlled environment experiment showed FR/R control of Chl and LHC-II concentrations. The results illustrate the importance of spectral distribution of reflected light on plant growth and a potential means of altering the chemistry of leaf crops under field conditions.

Journal Article↗

A new antitumor antibiotic, chromoxymycin. I. Taxonomic studies on the producing strain: a new subspecies of the genus Streptomyces.

A new subspecies of the genus Streptomyces, for which the name S. libani subsp. rubropurpureus subsp. nov. is proposed, is described. Soil isolate, strain No. 6362, produces a new antitumor substance chromoxymycin. The strain has a gray or red spore mass color, spiral spore chains with smooth spores, is nonchromogenic, and utilizes most carbon sources. Strain No. 6362 is most closely related to S. libani, although there are differences in colorization of reverse colony and spore chain morphology between each other. Those differences are not enough to establish a new species. It is suitable to consider that strain No. 6362 is identified as a new subspecies in the species of S. libani.

Antibiotics, Antineoplastic↗

Using flow cytometry to determine the viability of Cryptosporidium parvum oocysts extracted from spiked environmental samples in chambers.

Cryptosporidium parvum oocyst viability was determined by a dye permeability assay using a flow cytometric method. Oocysts were inoculated into small chambers with soil and biosolids. Oocysts extracted from soil and biosolids were then stained with propidium iodide (PI) and labeled with a fluorescein isothiocyanate (FITC)-conjugated monoclonal antibody. The oocyst population in each sample was determined using forward and side scatter plots, then further analyzed with fluorescence. A red and green fluorescence detector using gates established single populations of unstained, PI-stained, or FITC-labeled oocysts. No statistical difference was observed between viability of oocysts extracted from soil and biosolids as determined by either flow cytometry or microscopy. The location of excysted oocysts was changed in forward and side scatter plots. Results indicated that, although oocysts are not identified if they excyst, the flow cytometric method could be used to determine oocyst viability from spiked environmental samples.

Animals↗

Microbial counts and characteristics of Streptoverticillium strains isolated from soils in the Jordan valley.

A microbial survey of total bacterial count, agrobacteria, streptomycetes, and fungi was carried out in six locations in the Jordan Valley. The highest microbial counts for the four populations were in spring, the lowest mostly in autumn. No significant differences at p < 0.05 were observed either in the counts between the six locations, or in the seasonal counts of agrobacteria and streptomycetes between the six locations as well as within each location. The total bacterial counts (at p < 0.01) and fungi (at p < 0.05) differed significantly in the six locations. Microbial counts and the studied environmental conditions showed no significant regression. Of 552 streptomycetes strains isolated from soils collected from these locations, 58 belonged to the genus Streptoverticillium. They were distributed in five colour series grey (41%), red (31%), green (17%), blue (7%) and white (3%). Forty-eight strains had biverticillium spore bearing hyphae while only 10 had monoverticillium. While 28 strains produced distinctive reverse side pigment, only 8 and 6 strains produced soluble pigment and melanin pigment, respectively. Most strains utilized arabinose followed by fructose, mannitol, rhamnose, xylose, sucrose, inositol, and raffinose. Sixty-nine per cent of the strains exhibited activity in vitro test against Pseudomonas aeruginosa, and 45, 41, 28, and 7% against Staphylococcus aureus, Bacillus cereus, Agrobacterium tumefaciens and Escherichia coli, respectively.

Soil Microbiology↗

Morphology, chemistry and distribution of neoformed spherulites in agricultural land affected by metallurgical point-source pollution.

Metal distribution patterns in superficial soil horizons of agricultural land affected by metallurgical point-source pollution were studied using optical and electron microscopy, synchrotron radiation and spectroscopy analyses. The site is located in northern France, at the center of a former entry lane to a bunker of World War II, temporarily paved with coarse industrial waste fragments and removed at the end of the war. Thin sections made from undisturbed soil samples from A and B horizons were studied. Optical microscopy revealed the occurrence of yellow micrometer-sized (Ap horizon) and red decamicrometer-sized spherulites (AB, B(1)g horizons) as well as distinct distribution patterns. The chemical composition of the spherulites was dominated by Fe, Mn, Zn, Pb, Ca, and P. Comparison of calculated Zn stocks, both in the groundmass and in spherulites, showed a quasi-exclusive Zn accumulation in these neoformed features. Their formation was related to several factors: (i) liberation of metal elements due to weathering of waste products, (ii) Ca and P supply from fertilizing practices, (iii) co-precipitation of metal elements and Ca and P in a porous soil environment, after slow exudation of a supersaturated soil solution in more confined mineral media.

Agriculture↗

[Iodine levels in food and soil in different regions in Cameroon].

Iodine deficiency diseases (IDD) are a public health problem in many developing countries, such as Cameroon. They are often the consequence of insufficient iodine intake and/or the effect of thiocyanate in foods such as cassava; this chemical inhibits iodine fixation by the thyroid gland. One method of combatting these diseases is to counsel and encourage the consumptions of iodine-rich foods. We accordingly undertook an evaluation of the iodine content of staple foods, the soil, and the salt in some zones of western Cameroon and in the capital. Food (63), salt (04) and soil (05) samples were collected in four villages of western Cameroon, and food (20), water (18) and salt samples (14) were collected in Yaoundé (central province). Iodine content in foods and soils was assessed by the catalytic destructive testing of thiocyanate by nitrate in the presence of iodine. Iodine content in water was determined colorimetrically after mineralization and in salt by volumetric methods. Results show that iodine levels in salt depend on its origin, but was much lower than the 50-100 ppm recommended by the WHO. Soil and water levels depended on the stream and the origin. Several of the more than 20 foods sampled from Yaoundé had iodine concentrations higher than 10 microg/100 g: plantain (22.5 microg/100 g), wheat flour (21 microg/100 g), corn flour (17.75 microg/100 g), groundnuts (12.5 microg/100 g), sweet potatoes (12.25 microg/100 g), zoom (11.34 microg/100 g) and rice (10.45 microg /100 g). In western villages, we noted that iodine levels in food crops depended on the soil levels. Nevertheless, it was high in cocoyam (79.5 microg/100 g), yarns (68 microg/100 g), potatoes (62 microg/100 g), plantains (58.15 microg/100 g), and red beans (53.93 microg/100 g). In general, these crops contain more iodine than the food in Yaoundé; these results can be used to combat IDD, especially among young children, teenagers and pregnant women.

Cameroon↗

[Thallium in wine--trace element vinogram of thallium].

The ultra trace element Thallium is regularly found in very low quantities in wine, in red wines with 0.2 micrograms T1/1 about twice as much as in white wines. Certain hints the origin and behaviour of such low T1-contents can be derived from additional investigations (soil, location, fertilizer, influence of wine making). A correlation between T1 and K is known, but above that it can be shown, that similarly to Rb certain parts of the grape (grape, flesh, peel) are richer in T1 and that thereby the higher content of T1 in red wines can be explained. Slightly higher T1-levels are found in wines which grow next to cement plants or on abandoned mines. The Anodic Stripping Voltammetry in EDTA-containing base solution has proved effective for the determination of very low levels in the investigated samples. Wine and similar samples are decomposed with H2SO4-H2O2. After extraction as T1(III) bromide Thallium is measured by anodic-stripping-voltammetry. The preceding separation of T1 from soil samples or the like is achieved by evaporation analysis.

Electrochemistry↗

Effects of aqueous extracts from leaves and leaf litter on the abundance and diversity of soil gymnamoebae in laboratory microcosm cultures.

The distribution and abundance of microbiota in soil and litter may be significantly affected by the quality and quantity of localized patches of leaf organic matter. This study examined the relative effects of aqueous extracts of shed autumn leaves from American beech (Fagus grandifolia), sugar maple (Acer saccharum), red oak (Quercus rubra), and white oak (Quercus alba) on the density and diversity of gymnamoebae in laboratory cultures. Overall, the beech leaf extract produced the most growth of gymnamoebae followed by white oak with leaf extracts from maple and red oak producing least growth. Cultures using natural leaf litter from beneath beech trees had higher densities and diversity of gymnamoebae than leaf-litter cultures from a maple-oak stand. Soil microcosms confirmed that beech leaf extracts produced a higher density of gymnamoeba growth when added to soil cultures compared with maple and oak leaf extracts. Protein content, CHN (carbon and nitrogen content), and pH of the leaf extracts were assayed, but these alone were not sufficiently different to account for the effects. A dilution experiment indicated that some other concentration-dependent factor in the extract may produce the effects.

Amoebida↗