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Biomarker response and biomass change of earthworms exposed to chlorpyrifos in microcosms.

Background levels of chlorpyrifos and earthworm abundance were determined in an orchard and adjacent areas on a farm in the Western Cape, South Africa before these areas were again sprayed with this organophosphate. The background concentrations ranged from 0.2 microg/kg dm in the spray drift area adjacent to the orchard to 10.18 microg/kg dm on the slope in the run off area. In the target area the chlorpyrifos concentrations varied from a mean of 15.25 +/- 10.0 microg/kg directly after spraying to a mean of 7.0 +/- 0.9 microg/kg 6 months later and in the nontarget area they varied from a mean of 55.0 +/- 35 microg/kg to 12.0 +/- 5 microg/kg after 6 months. Chlorpyrifos was therefore still present in the field soils, but at lower concentrations, up to 6 months after the last spraying event. Earthworm abundance and population densities were very low. Only Aporrectodea caliginosa was found and the densities were much lower in the orchards (22 per m(2)) than in the nontarget areas (98.3 per m(2)). Microcosm studies were undertaken to relate biomarker responses to chlorpyrifos with biomass changes. Microcosms were filled with soil from the same areas and earthworms of the species A. caliginosa were introduced. The microcosms were treated with a series of concentrations of chlorpyrifos in the laboratory under controlled conditions. These concentrations were chosen to fall within the background ranges found in the soils. The biomass of the worms was determined regularly for a period of 5 weeks and worms in a state of estivation were noted. Earthworms were removed from the microcosms for biomarker tests: for cholinesterase (ChE) inhibition assays every week and for a neutral red retention determination 2 weeks after the exposures started. The most prominent biomass loss was noted in earthworms exposed to the highest pesticide concentration of 8.0 microg/kg. Estivation was higher among earthworms exposed to higher exposure concentrations. Inhibition of ChE increased with higher exposure concentrations and with time but there was no clear dose-related response. A clear dose-related response with exposure concentration was established for the neutral red retention assay. A correlation between ChE inhibition and biomass change existed directly after the second application of chlorpyrifos.

Animals↗

Strategies for nematode transmission: selective migration of Trichostrongylus tenuis infective larvae.

Successful transmission of macroparasites is dependent on exposure of susceptible hosts to free-living infective stages. When these hosts are herbivores that feed mostly on a single food plant then natural selection should favour those infective larvae that selectively ascend this main food plant. Red grouse feed predominantly on heather, Calluna vulgaris, so we predict that the infective larvae (L3) of the caecal nematode Trichostrongylus tenuis selectively locate and ascend heather plants. To determine whether the presence of heather influences the horizontal dispersal of T. tenuis L3 across soil, the movement of L3 across trays of soil with and without heather was investigated in the laboratory. More T. tenuis L3 were recovered from soil when heather was present, implying that larval migration may be influenced by chemical cues produced by heather plants. This was investigated in a second experiment, in which the horizontal dispersal of T. tenuis larvae was examined in the presence of heather and grass vegetation. This trial was repeated with larvae of a second species, Haemonchus contortus, a nematode whose hosts feed on a wide range of grass and shrub species. Significantly more larvae of both nematode species were recovered in the region of the heather than the grass or controls. This implies that T. tenuis and H. contortus L3 exhibit selective migration towards heather, perhaps reflecting a general response to plant cues which may be stronger for heather than for grass.

Animals↗

Major Element, REE, and Other Trace Element Behavior in Amphibolite Weathering under Semiarid Conditions in Southern India.

A body of komatiitic amphibolite, an enclave within the Archean high-grade orthogneisses in southern India, shows mild chemical weathering under semiarid conditions. Along fractures, chemical weathering has advanced (Chemical Index of Alteration &sqbl0;CIA&sqbr0;=53; CIA of fresh rock approximately 26) to the extent that secondary Mg-Fe-Al clay minerals have formed and the rock has turned brownish red, soft, and fine grained. The weathering process has resulted in the mobilization and redistribution of the so-called immobile elements Fe, Al, Ti, and REE effected by the nature of secondary mineral formation (talc vs. aluminous clay minerals) and also possibly by soil microbes. In the initial stages of secondary mineral formation, there is a small loss of Fe, Al, and REE (noticeably Eu). However, in the fracture zone as well as in the incipiently altered zone, there is significant REE enrichment, probably affected by a different precipitation mechanism. Mobilized REE may have come from a minor alteration of clinopyroxene.

Journal Article↗

Development and survival of the free-living stages of Trichostrongylus tenuis, a caecal parasite of red grouse Lagopus lagopus scoticus.

In the laboratory, yields of infective 3rd-stage larvae of Trichostrongylus tenuis were determined by temperature when moisture was adequate. On the moor, eggs in caecal faeces did not develop to infective larvae during the winter. In summer, development did occur and yields of infective larvae were related to temperature: greatest yields were obtained when mean monthly soil surface temperatures ranged between 7.6 and 10.3 degrees C. Rainfall was unimportant, as was desiccation. During summer the mean monthly mortality of infective larvae was 44% but over winter survival was negligible.

Animals↗

Influence of growth regulators and explant type on in vitro shoot propagation and rooting of red sandal wood (Pterocarpus santalinus L.).

In vitro shoot regeneration in Pterocarpus santalinus L. was achieved when detached cotyledons from in vitro germinated seedlings were cultured on MS medium containing NAA (0.1 mg/L), BA (1 mg/L) and kinetin (1 mg/L). The regenerated shoots rooted on 1/4 strength MS medium with IAA (1 mg/L) and the fully developed plantlets were successfully established in the soil.

Fabaceae↗

Role of iron in controlling speciation and mobilization of arsenic in subsurface environment.

Widespread arsenic contamination of groundwater has been reported of late in Bangladesh and West Bengal state of India. On the basis of arsenic geochemistry, three probable mechanisms have been cited for arsenic mobility in aquifers of West Bengal and Bangladesh. First, mobilization of arsenic due to the oxidation of arsenic-bearing pyrite minerals. Second, dissolution of arsenic-contaminated iron oxy-hydroxides (FeOOH) due to onset of reducing conditions in the subsurface. Third, due to the release of arsenic sorbed to aquifer minerals by competitive exchange with phosphate ions, that migrates into aquifers due to application of fertilizer to surface soil. Based on the review of field data from the affected region, it appears that the second mechanism described above is the most probable. Two reduction processes associated with this mechanism were investigated, viz., reduction of iron oxy-hydroxide to iron (II), which results in the mobilization of arsenic, and reduction of arsenic (V) to arsenic (III), which may enhance mobility of arsenic under certain conditions. These reactions, in the opinion of some researchers, are possible in subsurface environments mainly through microbial intervention. However, through the data presented in this paper, it has been demonstrated that above red-ox reactions involving iron and arsenic are also possible through predominantly abiotic pathways. While these results do not necessarily imply that abiotic red-ox processes are dominant in all subsurface environments containing iron and arsenic, it is entirely possible that abiotic interactions as described here may be responsible for a substantial amount of transformations involving iron and arsenic in anoxic subsurface environments.

Adsorption↗

Red list plants: colonization by arbuscular mycorrhizal fungi and dark septate endophytes.

Since information concerning the mycorrhization of endangered plants is of major importance for their potential re-establishment, we determined the mycorrhizal status of Serratula tinctoria (Asteraceae), Betonica officinalis (Lamiaceae), Drosera intermedia (Droseraceae) and Lycopodiella inundata (Lycopodiaceae), occurring at one of two wetland sites (fen meadow and peat bog), which differed in soil pH and available P levels. Root colonization by arbuscular mycorrhizal fungi (AMF) and dark septate endophytes (DSE) was quantified. Colonization by AMF appeared to be more frequent in the fen meadow than in the peat bog, and depended on the host plant. Roots of S. tinctoria and B. officinalis were well colonized by AMF in the fen meadow (35-55% root length) and both arbuscules and vesicles were observed to occur in spring as well as in autumn. In the peat bog, L. inundata showed a low level of root colonization in spring, when vesicles were found frequently but no arbuscules. In roots of D. intermedia from the peat bog, arbuscules and vesicles were observed, but AMF colonization was lower than in L. inundata. In contrast, the amount of AMF spores extracted from soil at the peat bog site was higher than from the fen meadow soil. Spore numbers did not differ between spring and autumn in the fen meadow, but they were higher in spring than in autumn in the peat bog. Acaulospora laevis or A. colossica and Glomus etunicatum were identified amongst the AMF spores extracted from soil at the two sites. S. tinctoria and B. officinalis roots were also regularly colonized by DSE (18-40% root length), while L. inundata was only rarely colonized and D. intermedia did not seem to be colonized by DSE at all.

Asteraceae↗

[Proposed neotype Streptomyces ruber (Krainsky, 1914) Waksman et Henrici, 1948].

Culture 78 was proposed as a neotype of Streptomyces ruber. It was isolated from the soils of the Baikal region and was closest, in its taxonomic properties, to the original description of the species [13] whose representative had been lost. Cultures from different microbial collections designated as S. ruber were shown to be unlike the original description. The neotype had the following taxononic properties: the cell wall of type I; spiral sporophores with extended spirals having 2-3 coils; oval spores with a smooth envelope; greyish pink aerial and dark-red substrate mycelia; a red pigment not passing into the medium; slow gelatin liquefaction and milk peptonization; weak starch hydrolysis; assimilation of glucose, xylose, rammose, fructose, and inositol; weak growth on arabinose, raffinose and mannitol, but not on sucrose; no formation of melanoid pigments; synthesis of riboflavin and prodigiosin pigments; inhibition of Gram-positive bacterial and acid-resistant mycobacterial growth; no inhibition of yeast and fungal growth. The culture was sensitive to streptomycin, neomycin, gentamycin, monomycin, tetracycline,erythromycin, oleandomycin, lincomycin, ristomycin, levomycetin, polymyxin and fusidin, but resistant in penicillin. The population was composed of six variants [3]: main, faded, asporogenic red, asporogenic yellow, asporogenic white and nocardia-like. The latter two were not capable of riboflavin and prodigiosin formation. The asporogenic yellow variant was a monosynthetic organism: it formed riboflavin, but could not synthesize prodigiosin. The neotype of S. ruber 78 is deposited withthe national Collection of Microorganisms (the reference number is VKM A-611).

Anti-Bacterial Agents↗

[White blood stains?].

Following to bullet wounds of the upper maxillary region and of the lungs great quantities of blood were expectorated by the victim soiling the clothes of the perpetrator. (These cloth consisted of a mixture of wool and synthetic fibers producing a rough surface.) The suit was washed out in cold water and dried. The only macroscopic findings were intensively stained white to grey spots missing any brown or red color portion. These were identified to consist of fibrin. Extensive examination proved the existence of blood spots of group A. Experimental stains applied to the same cloth showed identical changes after washing out.

ABO Blood-Group System↗

Characteristics of Streptomyces strains isolated from soils in two landfill areas in north Jordan.

A total of 114 Streptomyces strains were isolated and purified from soils collected from Al Akider (53) and from Al Kafeer (61) landfill areas in north Jordan. The Al Akider strains were classified into 5 colour series gray (27), white (18), yellow (3), violet (3) and green (2); while the Al Kafeer strains were classified into 6 colour series white (26), gray (16), yellow (9), red (2), green (7) and blue (1). Melanin pigment was produced by 21 and 25% of the Al Akider and Al Kafeer strains, respectively, while soluble pigment was produced by 15% of the A; Akider and 21% of the Al Kafeer strains; 17% of Al Akider strains and 23% of Al Kafeer strains showed distinctive reverse side pigment. Forty, 32, 19, and 9% of the Al Akider strains had spiral, rectus, flexous and retinaculum apertum spore-chains, respectively, while the distribution of the Al Kafeer strains in the same spore-chain forms was equally 25% each. Arabinose, xylose, inositol, fructose, rhamnose, sucrose, raffinose, and mannitol were utilized by 91, 53, 89, 60, 30, 25 and 79% of the Al Akider strains and by 84, 59, 62, 64, 70, 75, 41 and 26% of the Al Kafeer strains, respectively, while only 2% of the Al Kafeer strains were found to utilize cellulose. Thirty-six percent of the Al Akider strains showed, in vitro activity against Staphylococcus aureus followed by Aspergillus niger (26%), Bacillus cereus and Saccharomyces cerevisiae (25% each), Fusarium monilliforme (21%), Candida albicans (15%), Escherichia coli (8%) and Pseudomonas aeruginosa (4%), while 51, 49, 46, 34, 25, 15, 7 and 2% of the Al Kafeer strains exhibited activity against A. niger, S. cerevisiae, B. cereus, S. aureus, F. monilliforme, C. albicans, P. aeruginosa and E. coli, respectively.

Antibiosis↗

Retardation of Senescence in Red Clover Leaf Discs by a New Antiozonant, N-[2-(2-Oxo-1-imidazolidinyl)ethyl]-N'-phenylurea.

Dark-induced senescence in leaf discs from O(3)-sensitive red clover trifoliates (Trifolium pratense L. cv. ;Pennscott') was markedly retarded by treatment with N-[2-(2-oxo-1-imidazolidinyl)ethyl-N'-phenylurea (EDU). EDU also protects against acute and chronic foliar O(3) injury when sprayed on intact leaves or supplied to the plants through soil application. Senescence retardation was measured by time-dependent analyses of chlorophyll, protein, and RNA in discs floated on aqueous EDU solutions ranging from 0 to 500 micrograms per milliliter EDU. Chlorophyll degradation, total protein, and nucleic acids were followed over 10-day test periods.EDU at 500 micrograms per milliliter (50 milligrams per pot), a concentration known to provide optimal protection to intact leaves against O(3) injury, was most effective in preventing chlorosis and in maintaining high concentrations of protein and RNA in the discs. In discs treated with 500 micrograms per milliliter EDU 90% of the chlorophyll was retained after 10 days in the dark. In contrast, lower concentrations (0, 125, and 250 micrograms per milliliter) showed the complete loss of chlorophyll or an intermediate retardation. The intermediate concentrations were similarly less effective in maintaining protein and RNA levels in the dark stressed leaf discs. It is suggested that EDU retards senescence and mitigates O(3) injury through the induction of specific free radical scavenging enzymes and in sustaining RNA and protein synthesis.

Journal Article↗

Assessing the Rhizotoxicity of the Aluminate Ion, Al(OH)(4).

Dissolved aluminum (III) in acidic soils or culture media is often rhizotoxic (inhibitory to root elongation). Alkaline solutions of Al are also sometimes rhizotoxic, and for that reason toxicity has been attributed to the aluminate ion, Al(OH)(4) (-). In the present study, seedlings of wheat (Triticum aestivum L. cv Tyler) and red clover (Trifolium pratense L. cv Kenland) were cultured in aerated aluminate solutions at pH 8.0 to 8.9. The bulk phases of these solutions were free of reactive polynuclear hydroxy-Al (including the extremely toxic species AlO(4)Al(12)[OH](24)[H(2)O](7+) (12) [Al(13)]) according to the ferron (8-hydroxy-7-iodo-5-quinolinesulfonic acid) assay. At an aluminate concentration of 25 micromolar (23 micromolar activity) and a pH of 8, root elongation was less than 40% of Al-free controls, but at pH 8.9 elongation was 100% of controls. The hypothesis is offered that aluminate is nontoxic and that the inhibition at lower pH values is attributable to Al(13) postulated to have formed in the acidic free space of the roots where the ratio /{Al(3+)/}//{H(+)/}(3) may rise above 10(10). At this value hydroxy-Al in over-saturated, alkaline solutions begins to undergo rapid conversion to polynuclear species.

Journal Article↗

Metal blood levels and hematological characteristics in wood mice (Apodemus sylvaticus L.) along a metal pollution gradient.

The present study evaluated whether blood could be used as a nondestructive tool for monitoring metal exposure and related hematological effects in wood mice (Apodemus sylvaticus L.) living along a metal pollution gradient. Soil concentrations of arsenic, cadmium, copper, lead, silver, and zinc decreased with distance from the emission source. Blood levels of cadmium and lead differed significantly among sites, whereas those of the other metals did not. Blood levels of cadmium and lead correlated with soil concentrations of cadmium and lead, respectively. No such significant relationships were found for the other measured metals. Hematocrit levels decreased in wood mice from the most polluted site (45.96% +/- 0.53% [mean +/- standard error]) compared to the reference site (48.04% +/- 0.47%). A negative correlation between hematocrit and blood levels of cadmium and lead was found. Erythrocyte count, leukocyte count, hemoglobin concentration, mean corpuscular hemoglobin (average wt of hemoglobin in a red blood cell in pg), and lysozyme activity did not differ among study sites. Mean corpuscular volume (average volume of a red blood cell in microm3) and mean corpuscular hemoglobin concentration (average proportion of hemoglobin in a red blood cell as a %) differed among study sites but showed no relationship with metal exposure. We conclude that whole blood from mice can be used for nondestructive monitoring of the exposure to nonessential metals under natural conditions. The present results indicate that decreased hematocrit levels may be an early warning signal for a negative impact of metal exposure on the oxygen-transport capacity of blood in wood mice in their natural environment.

Animals↗

Australian research on ochratoxigenic fungi and ochratoxin A.

The presence of the mycotoxin, ochratoxin A (OTA), has been reported in Australian grape products. Comprehensive surveys of Australian wines have determined that the frequency and level of OTA contamination are low. Aspergillus carbonarius is the primary OTA-producing species associated with grapes in Australia, and all isolates tested to date produce OTA. Aspergillus niger is isolated more frequently from vineyards, however, few strains produce OTA. A. carbonarius and A. niger exist as saprophytes in the top layer of soil beneath vines, from where they are thought to be blown onto bunches. The level of A. carbonarius in soil may be reduced by temperatures above or below the optimum temperature for survival (25 degrees C), by high soil moisture content, and by modifications to tillage and mulching practices. A. carbonarius is an opportunistic pathogen of damaged berries. In the absence of damage, spores may exist on berry surfaces without causing visible rots. Aspergillus rots are associated with black Aspergillus species, primarily A. niger, A. carbonarius and A. aculeatus. The potential for such rots is increased with berry damage, inoculum coverage and berry maturity. Susceptibility to berry splitting is related, in part, to bunch structure, and may be variety-dependent or influenced by rainfall, irrigation and canopy management. Black Aspergillus spp. are closely associated with berries near the main stem of the bunch. During winemaking, around 80% of the OTA initially present in grapes is removed, primarily with the skins and pulp during pressing. Additional reductions occur with the removal of precipitated grape and yeast solids. Bentonite in white wine and yeast hulls in red wine were the most effective non-carbonaceous fining agents for the removal of OTA.

Aspergillus↗

Isolation of antibiotics turbomycin a and B from a metagenomic library of soil microbial DNA.

To access the genetic and biochemical potential of soil microorganisms by culture-independent methods, a 24,546-member library in Escherichia coli with DNA extracted directly from soil had previously been constructed (M. R. Rondon, P. R. August, A. D. Bettermann, S. F. Brady, T. H. Grossman, M. R. Liles, K. A. Loiacono, B. A. Lynch, I. A. MacNeil, M. S. Osburne, J. Clardy, J. Handelsman, and R. M. Goodman, Appl. Environ. Microbiol. 66:2541-2547, 2000). Three clones, P57G4, P89C8, and P214D2, produced colonies with a dark brown melanin-like color. We fractionated the culture supernatant of P57G4 to identify the pigmented compound or compounds. Methanol extracts of the acid precipitate from the culture supernatant contained a red and an orange pigment. Structural analysis revealed that these were triaryl cations, designated turbomycin A and turbomycin B, respectively; both exhibited broad-spectrum antibiotic activity against gram-negative and gram-positive organisms. Mutagenesis, subcloning, and sequence analysis of the 25-kb insert in P57G4 demonstrated that a single open reading frame was necessary and sufficient to confer production of the brown, orange, and red pigments on E. coli; the predicted product of this sequence shares extensive sequence similarity with members of the 4-hydroxyphenylpyruvate dioxygenase (4HPPD) family of enzymes. Another member of the same family of genes, lly, which is required for production of the hemolytic pigment in Legionella pneumophila, also conferred production of turbomycin A and B on E. coli. We further demonstrated that turbomycin A and turbomycin B are produced from the interaction of indole, normally secreted by E. coli, with homogentisic acid synthesized by the 4HPPD gene products. The results demonstrate successful heterologous expression of DNA extracted directly from soil as a means to access previously uncharacterized small organic compounds, serving as an example of a chimeric pathway for the generation of novel chemical structures.

Anti-Bacterial Agents↗

Hydraulic conductivity and embolism in the mangrove tree Laguncularia racemosa.

We measured xylem pressure potentials, soil osmotic potentials, hydraulic conductivity and percent loss of conductivity (PLC) due to embolism, and made microscopic observations of perfused dye in the white mangrove tree, Laguncularia racemosa (L.) Gaertn. f., (1) to determine its vulnerability to air embolism compared with published results for the highly salt-tolerant red mangrove tree, Rhizophora mangle L., and (2) to identify possible relationships between air embolism, permanent blockage of vessels and stem diameter. Laguncularia racemosa was more vulnerable to embolism than reported for R. mangle, with 50 PLC at -3.4 MPa. Narrow stems (5-mm diameter) had higher PLC than larger stems (8.4- or 14-mm diameter) of the same plants. Basic fuchsin dye indicated that up to 89% of the vessels, especially in the narrow stems, had permanent blockage that could not be reversed by high pressure perfusion. Air embolism could lead to permanent vessel blockage and eventual stem mortality. Such vulnerability to embolism may restrict the growth of L. racemosa and limit its distribution to less salty areas of mangrove communities.

Combretaceae↗

The Almadén district (Spain): anatomy of one of the world's largest Hg-contaminated sites.

We present data from an early reconnaissance survey (stream sediments, soil, and water Hg chemistry; plants and water crustaceans Hg intake) of the Almadén district (central Spain), that was carried out to establish the potential environmental hazards derived from the anomalous mercury concentrations measured in this realm. The Almadén mercury district (approximately 300 km2) can be regarded as the largest geochemical anomaly of mercury on Earth. The district includes a series of mercury mineral deposits, having in common a simple mineralogy (dominant cinnabar: HgS, and minor pyrite: FeS2). The ore deposits have been mined for more than 2000 years, and the main mine of the district (Almadén), has been active from Roman times to present day with almost no interruptions. The mercury distribution in soils of the district reveals the existence of high, and extremely high mercury values (up to 8889 microg g(-1)), whereas concentrations in stream sediments and waters reach exceptional values of up to 16,000 microg g(-1) and 11,200 ng l(-1) respectively. On the other hand, very high concentrations of methylmercury (MeHg) have been detected in calcines (up to 3100 ng g(-1)), sediments (0.32-82 ng g(-1)), and waters (0.040-30 ng l(-1)). Mercury gets incorporated to edible river crustaceans and plants. The red swamp crayfish Procambarus clarkii, has Hg concentrations of up to 9060 ng g(-1) (muscle) and 26,150 ng g(-1) (hepatopancreas). Regarding plants, the local wild asparagus (Asparagus acutifolius) yields values of up to 298 microg g(-1) Hg. Mercury also escapes to the atmosphere, and mineral deposits, together with metallurgical activities, generate strong anomalies of atmospheric Hg. The most important concentrations relate to the emissions from the Almadén metallurgical roaster, in the order of 14,000 ng Hg m(-3). Additionally, large open pit operations also contribute to the district atmospheric pool of mercury, with high concentrations above 1000 ng Hg m(-3). Thus, no system (rocks, soils, sediments, waters, atmosphere, biota) in the Almadén district is free from strong Hg contamination.

Environmental Monitoring↗

Phosphorus Distribution in Red Pine Roots and the Ectomycorrhizal Fungus Hebeloma arenosa.

Red pines (Pinus resinosa Ait.) were grown in a pasteurized sandy loam either unamended with phosphate or fertilized with one of two levels of phosphate (34 or 136 mg/kg) as superphosphate, and with and without addition of Hebeloma arenosa inoculum. Shoot and total dry weights of mycorrhizal seedlings grown in soil unamended with P were greater than those for nonmycorrhizal seedlings grown in the same soil, but less than the dry weights of seedlings grown in soil amended with middle to high levels of P. Mycorrhizal infection was inhibited at the highest level of P amendment. (31)P nuclear magnetic resonance spectra of intact mycorrhizal roots showed the presence of two dominant peaks, orthophosphate (Pi) and polyphosphate (polyP). The polyP peak was absent in spectra of nonmycorrhizal roots. The ratio for areas under the two peaks, Pi/polyP, was 1.8 for mycorrhizal roots grown in both unamended soil and soil that had received middle levels of superphosphate. Apparently, the fungus strongly mediates the supply of phosphate to the tree through the production of polyP, even at growth-limiting levels of soil P, and regulates compartmentalization of P in the mycorrhizal roots.

Journal Article↗