Diseases of the gladiolus: I. Control of hard rot, due to Septoria gladioli Passer., by fungicidal treatment of the corms

Losses from hard rot, measured by an arbitrary disease index, were reduced by treating the dehusked corms before planting with mercuric chloride (with or without the addition of 10% hydrochloric acid), mercurous chloride (calomel), three proprietary mercury compounds (Aretan, Uspulun and Ceresan) and one proprietary non-mercury compound (Folosan). Calomel was the least effective. All the treatments were relatively less effective when corms with definite lesions were treated.
The weight of clean corms produced per old corm planted (weight index) was usually increased by all the fungicides tried, but calomel and Ceresan were less satisfactory than the others.
Mercuric chloride (3 hr. steep in a 0.1% solution) was not rendered more effective by the addition of hydrochloric acid nor by a preliminary dip in methylated spirits to facilitate wetting, while the addition of a proprietary wetting compound (Agral) was definitely harmful to the corms and usually less effective than mercuric chloride alone. Increase in time of steeping or concentration of mercuric chloride was not beneficial and was sometimes harmful. Reduction in time of steeping to 1 hr. gave promising results.
Treatment in November had some advantages over treatment in March.
All the mercury compounds tended to delay flowering, this being most marked in the presence of the wetting compound. Stunted foliage and poor quality flowers resulted from the use of Ceresan.     

Uptake of Hg from203Hg-labeled mercury compounds by wheat and beans grown on an oxisol

Summary Studies were conducted to evaluate the uptake of mercury by wheat (Triticum aestivum L. runar) and beans (Phaseolus vulgaris L. marshal) growth on an oxisol with different levels of 2-methoxyethylmercury chloride (Aretan) and mercuric chloride. Dry matter and grain yields of wheat were little affected by either Aretan or mercuric chloride, although Aretan at 50 mg Hg/kg soil delayed germination by four to five days. Germination of beans grown with both compounds at the 50 mg Hg/kg soil failed completely, even after repeated sowing. Yields were somewhat, though not significantly, decreased by mercury chloride up to 5 mg Hg/kg soil.The concentration of Hg in wheat straw and grain increased significantly with increased levels of Aretan and HgCl₂ application, with more Hg taken up by the plants grown with HgCl₂ than with those grown with Aretan. Translocation of Hg to grain was greater in the plants grown with HgCl₂.The concentration of Hg in bean straw, but not grain, increased significantly with increasing levels of Aretan and HgCl₂ application, and was greater in plants grown with HgCl₂. Translocation to grain was low, with little difference between plants grown with Aretan or HgCl₂.     

THE EFFECT OF CERTAIN SOIL TREATMENTS ON DIDYMELLA STEM ROT OF TOMATOES PART I. GLASSHOUSE EXPERIMENTS

When tomatoes were planted in steamed soil inoculated with spores of Didymella lycopersici Kleb., a large proportion became infected, but very few became infected in unsteamed soil similarly inoculated. In soil treated with formalin and subsequently inoculated the number of diseased plants was about half that in steamed soil. Reinfecting steamed soil with soil micro-organisms before inoculation with D. lycopersici reduced the amount of disease. Covering steamed soil with a layer of maiden loam filtered out a large proportion of D. lycopersici spores applied in water suspension and reduced the amount of disease. Addition of stable manure either before or after steaming did not affect the number of diseased plants. Steamed soil remained susceptible to infection by D. lycopersici for 4 months. D. lycopersici survived in a glasshouse soil over the winter, but the number of diseased plants was small. Cultivation may lessen the amount of disease by burying most of the infected surface layers of the soil. The peak of infection under glass occurred in May. The amount of disease and the yield were correlated, but with severe infection the yield varied with the time of infection and the distribution of diseased plants.     

The efficiency of two organomercury compounds in controllingg seed-borne Septoria nodorutn on winter wheat

The influence of various factors on the performance of organomercury seed treatments in controlling seed-borne Septoria nodorum was studied in laboratory experiments. Control was least effective on the most heavily infected seed samples. On heavily infected seed ethyl mercuric chloride was more effective than phenyl mercuric acetate, although 5 wk storage of the treated seed before sowing increased the effectiveness of the latter compound. Effectiveness was not generally influenced by differences in soil moisture. Seedling vigour was significantly improved by levels of mercury below those applied commercially. The significance of seed treatment in the epidemiology of Septoria disease is discussed.     

Biomonitoring of low levels of mercurial derivatives in water and soil by Allium micronucleus assay

The Allium micronucleus (MNC) assay was developed to monitor low levels of mercury in aquatic and terrestrial environments. Four mercurial derivatives namely mercuric chloride (MC), methyl mercuric chloride (MMC), phenyl mercuric acetate (PMA) and a methoxy ethyl mercuric chloride based fungicide, Emisan-6, were tested to assess the sensitivity and versatility of the Allium MNC assay. Allium bulbs were set directly on water and soil contaminated with known levels of mercurial derivatives (0.0001-10.00 ppm). On the 5th day the endpoints measured were root length, mitoses with spindle abnormality and cells with MNC in root meristems. The effective concentrations of the test chemicals that cause 50% of root length as compared to control (EC50) were determined from dose-response curves so obtained. The lowest effective concentration tested (LECT) and highest ineffective concentration tested (HICT) for each of the mercurial derivatives for the induction of spindle malfunction and MNC were determined. It was found that EC50, LECT and HICT values for mercurial derivatives in soil were higher than those in water. The frequencies of cells with MNC and mitoses with spindle abnormality were highly correlated indicating that MNC is a good parameter of spindle malfunction. The present approach increased the sensitivity of the Allium assay by 10-fold, the detection limit being 0.001-0.1 ppm and 0.1-1.0 ppm in aquatic and terrestrial environments respectively, depending on the species of mercury.     

Comparison of methods to measure acute metal and organometal toxicity to natural aquatic microbial communities.

Microbial communities in water from Baltimore Harbor and from the mainstem of Chesapeake Bay were examined for sensitivity to mercuric chloride, monomethyl mercury, stannic chloride, and tributyltin chloride. Acute toxicity was determined by measuring the effects of [3H]thymidine incorporation, [14C]glutamate incorporation and respiration, and viability as compared with those of controls. Minimum inhibitory concentrations were low for all metals (monomethyl mercury, less than 0.05 microgram liter-1; mercuric chloride, less than 1 microgram liter-1; tributyltin chloride, less than 5 micrograms liter-1) except stannic chloride (5 mg liter-1). In some cases, mercuric chloride and monomethyl mercury were equally toxic at comparable concentrations. The Chesapeake Bay community appeared to be slightly more sensitive to metal stress than the Baltimore Harbor community, but this was not true for all treatments or assays. For culturable bacteria the opposite result was found. Thymidine incorporation and glutamate metabolism were much more sensitive indicators of metal toxicity than was viability. To our knowledge, this is the first use of the thymidine incorporation method for ecotoxicology studies. We found it the easiest and fastest of the three methods; it is at least equal in sensitivity to metabolic measurements, and it likely measures the effects on greater portion of the natural community.     

Nutritional imbalance caused by nitrogen excess is correlated with the occurrence of watermark disease in white willow

Willows in the Flemish region are threatened by the watermark disease, a wood disease caused by the bacterium Brenneria salicis (Hauben et al. Syst Appl Microbiol 21:384–397, 1998). Affected trees are observed to grow at specific sites which seem more sensitive to periodical watermark symptom development than other sites. In this study we aim at revealing abiotic site factors that could play a role in triggering the watermark disease. During three consecutive years we analyzed stands of young isogenic willows (Salix alba L. cv. Lichtenvoorde), which were planted on four different sites in an agricultural environment. These test sites were chosen, based on the occurrence/absence of watermark disease in the old willows that grow on these sites. Two sites had old willow pollards with clear watermark symptoms, and on the two other sites the old willows were unaffected. Differences in abiotic characteristics between the sites were analysed and their effects on the nutrient status in willow were measured in the isogenic willows that were newly planted. Brenneria salicis-specific multiplex PCR revealed the presence of this pathogenic bacterium in both the old and the freshly planted young willows. At each site, the physical and chemical characteristics of soil and groundwater were investigated; the young tree growth was monitored and nutritional status of the willow plantings was assessed by foliar analysis. Physical soil properties including soil type and profile water availability, and chemical soil analysis were unable to explain differences between healthy and affected sites. Diseased sites, however, had significantly higher N concentrations and traces of herbicides in the groundwater. In young trees, foliar levels of N and S were always greater at the affected sites, while P contents were lower. Dry mass based N concentration in the leaves of the newly planted trees increased systematically over the years, but more at affected than at healthy sites. Also, chlorophyll content was significantly greater at the affected sites. Growth rate of the young isogenic trees was twice as fast at the diseased sites. The presented data suggest that application of fertilisers in agriculture causes an imbalance in the nutritional status of Salix alba L. Excess nitrogen makes this species fast-growing, reflected by an average foliar N:P ratio >10, and more susceptible for watermark disease, indicated by an average foliar N:Ca ratio exceeding 1.5.     

Mercury incorporation by mature and immature red blood cells

Intact and ghost erythrocytes and reticulocytes were incubated with 0.1 ppm 203-Hg as either mercuric chloride or methyl mercury chloride. Both mature and immature cells accumulated alkyl mercury more avidly than inorganic mercury. Methyl mercury chloride, but not mercuric chloride, readily penetrated the membrane and became incorporated into the intracellular compartment of intact cells. Although uptake of alkyl mercury was approximately the same for intact erythrocytes and reticulocytes, developing cells accumulated inorganic mercury more avidly than did mature cells. Increased uptake of inorganic mercury represented predominantly an increase in stromal binding, illustrating differences in the surface membrane or reticulocytes and erythrocytes.     

Bioaccessibility of Mercury in Soils

The initial risk assessment for the East Fork Poplar Creek (EFPC) floodplain in Oak Ridge, Tennessee, a superfund site heavily contaminated with mercury, was based on a reference dose for mercuric chloride. Mercuric chloride, however, is a soluble mercury compound not expected to be present in the floodplain, which is frequently saturated with water. Previous investigations had suggested mercury in the EFPC floodplain was less soluble and therefore potentially less bioavailable than mercuric chloride, possibly making the results of the risk assessment unduly conservative. A bioaccessibility study, designed to measure the amount of mercury available for absorption in a child's digestive tract (the most critical risk pathway endpoint), was performed on 20 soils from the EFPC floodplain. The average bioac-cessible mercury for the 20 soils was 5.3%, compared with 100% of the mercuric chloride subjected to the same conditions. The alteration of the procedure to more closely mimic conditions in the digestive tract did not significantly change the results. Therefore, the use of a reference dose for mercuric chloride at EFPC, and potentially at other mercury-contaminated sites, without incorporating a corresponding bioavailability adjustment factor may overestimate the risk posed by the site.     

Uptake of mercury and mercury-amino acid complexes by rat renal cortex slices.

This study was undertaken in order to assess the effects of metabolism and complexations with amino acids on the renal uptake of mercury using rat renal cortex slices as the experimental system. Mercury levels attained in the slices after 60 min of incubation were 50% higher with mercuric cysteine than with mercuric chloride. This enhancement of uptake with mercuric cysteine was reduced in the presence of a tenfold molar excess of histidine or lysine, but not by serine. Excess cysteine markedly increased mercury uptake. Incubation at 25 degrees significantly reduced uptake of mercuric cysteine, but not mercuric chloride. Anaerobic conditions and incubation in the presence of DNP each reduced mercuric cysteine uptake to the control level of mercuric chloride without affecting uptake of mercuric chloride. The differential aspects of metabolism on the uptake of mercuric cysteine and mercuric chloride and the competitive effects obtained with amino acids known to compete with cysteine in renal reabsorption support the hypothesis that a portion of the renal uptake of mercury operates through amino acid transport mechanisms acting on mercury-amino acid complexes.     

Metachromasia of basic dyes induced by mercuric chloride. I

Summary Interactions of the cationic dye methylene blue with mercuric chloride have been studied conductometrically, analytically and spectrophotometrically. Methylene blue produces red colored precipitate with mercuric chloride; in presence of large excess of mercuric chloride a strong metachromasia is induced in the dye. Metachromasia induced by mercuric chloride is more hypsochromic as well as hypochromic than that induced by chromotopes like heparin. The complexes formed between methylene blue and mercuric chloride have variable compositions, the complex responsible for the red metachromatic color of the dye has the composition 2 dye: 1 HgCl₂. A model has been proposed for the metachromatic complex consisting hexa-coordinated mercury, dye is coordinated to the mercury by donating the lone pair electrons of terminal nitrogen. The non-metachromatic dye capri blue also interacts with mercuric chloride but without any change in the visible spectrum. Potassium iodide also gives metachromatic reddish blue colored precipitate with methylene blue.University Research Scholar.     

Relationships between alterations in glutathione metabolism and the disposition of inorganic mercury in rats: effects of biliary ligation and chemically induced modulation of glutathione status

Influences of biliary ligation and systemic depletion of glutathione (GSH) or modulation of GSH status on the disposition of a low, non-nephrotoxic i.v. dose of inorganic mercury were evaluated in rats in the present study. Renal and hepatic disposition, and the urinary and fecal excretion, of inorganic mercury were assessed 24 h after the injection of a 0.5-micromol/kg dose of mercuric chloride in control rats and rats pretreated with acivicin (two 10-mg/kg i.p. doses in 2 ml/kg normal saline, 90 min apart, 60 min before mercuric chloride), buthionine sulfoximine (BSO; 2 mmol/kg i.v. in 4 ml/kg normal saline, 2 h before mercuric chloride) or diethylmaleate (DEM; 3.37 mmol/kg i.p. in 2 ml/kg corn oil, 2 h before mercuric chloride) that either underwent or did not undergo acute biliary ligation prior to the injection of mercury. Among the groups that did not undergo biliary ligation, the pretreatments used to alter GSH status systemically had varying effects on the disposition of inorganic mercury in the kidneys, liver, and blood. Biliary ligation caused the net renal accumulation of mercury to decrease under all pretreatment conditions. By contrast, biliary ligation caused significant increases in the hepatic burden of mercury in all pretreatment groups except in theacivicin-pretreated group. Blood levels of mercury also increased as a result of biliary ligation, regardless of the type of pretreatment used. The present findings indicate that biliary ligation combined with methods used to modulate GSH status systemically have additive effects with respect to causing reductions in the net renal accumulation of mercury. Additionally, the findings indicate that at least some fraction of the renal accumulation of inorganic mercury is linked mechanistically to the hepato-biliary system.     

Experiments on the control of white rot (Sclerotium cepivorum Berk.) in onions

Experiments-made during 1943 and 1944 on the use of mercurous chloride (calomel) for the control of Sclerotium cepivorum in spring-sown onions are described. The application of 4% calomel dust to the seed drill at sowing time gave better results than seed treatment. 1 lb. of dust per 25 yd. of seed drill gave good disease control in bulb onions, and 1 lb./so yd. may suffice for salad onions.
The relation between disease control and the effective mercury content of the treated soil is discussed.     

Plasmid Frequency Fluctuations in Bacterial Populations from Chemically Stressed Soil Communities

The frequency of plasmids in chemically stressed bacterial populations was investigated by individually adding various concentration of kanamycin, ampicillin, and mercuric chloride to soil samples. Viable bacterial populations were enumerated, soil respiration was monitored for up to 6 weeks as an indicator of physiological stress, and bacterial isolates from stressed and control soils were screened for the presence of plasmids. Low levels of the chemical stress factors did not for the most part significantly alter population viability, soil respiration, or plasmid frequency. Exposure to high stress levels of mercury and ampicillin, however, resulted in altered numbers of viable organisms, soil respiration, and plasmid frequency. Plasmid frequency increased in response to ampicillin exposure but was not significantly changed after exposure to kanamycin. In mercuric chloride-stressed soils, there was a decrease in plasmid frequency despite an increase in overall mercury resistance of the isolates, suggesting that mercury resistance in these populations is largely, if not completely, chromosome encoded. Chemical stress did not cause an increase in plasmid-mediated multiple resistance. A genetic response (change in plasmid frequency) was not found unless a physiological (phenotypic) response (change in viable cells and respiratory activity) was also observed. The results indicate that a change in plasmid frequency is dependent on both the amount and type of chemical stress.     

Diseases of the gladiolus; note on the incidence and control of scab disease Bacterium marginatum McCull

Stocks of imported gladioli which originally contained a proportion of scabbed corms showed very little carry-over of the disease during the eight growing seasons 1938-45. Carry-over was greatest on ground where gladioli had been planted for several successive years.
The incidence of the disease was greatest in the hot summer of 1940.
Treatment with mercurial fungicides just before planting time gave good control. Two non-mercurial fungicidal dusts were less effective.     

The Relative Importance of Seed-borne Inoculum to Common Scab Disease of Potato and the Efficacy of Seed Tuber and Soil Treatments for Disease Control

The importance of both seed and soil-borne inoculum in the epidemiology of common scab disease under Australian conditions was clearly demonstrated. In field trials the severity of disease in harvested potatoes was directly related to the severity of disease on the planted seed tubers. Chemical seed dressing treatments were assessed for common scab disease control under field conditions in four trials over 5 years. Where seed treatments were applied to both diseased and visibly clean seed pieces significantly more disease was found in tubers harvested from diseased seed than the corresponding clean seed treatment. In all but one trial, the treatments applied to diseased seed significantly reduced the incidence of common scab. Fluazinam, flusulfamide (at elevated rates), fenpiclonil, pentachloronitrobezene and mancozeb seed treatments were particularly effective as seed dressing treatments. Applications to visibly clean seed failed to significantly diminish disease levels below that found on untreated seed. Preliminary investigations of some chemical soil treatments gave disappointing levels of control.     

Comparison of methods to measure acute metal and organometal toxicity to natural aquatic microbial communities

Microbial communities in water from Baltimore Harbor and from the mainstem of Chesapeake Bay were examined for sensitivity to mercuric chloride, monomethyl mercury, stannic chloride, and tributyltin chloride. Acute toxicity was determined by measuring the effects of [3H]thymidine incorporation, [14C]glutamate incorporation and respiration, and viability as compared with those of controls. Minimum inhibitory concentrations were low for all metals (monomethyl mercury, less than 0.05 microgram liter-1; mercuric chloride, less than 1 microgram liter-1; tributyltin chloride, less than 5 micrograms liter-1) except stannic chloride (5 mg liter-1). In some cases, mercuric chloride and monomethyl mercury were equally toxic at comparable concentrations. The Chesapeake Bay community appeared to be slightly more sensitive to metal stress than the Baltimore Harbor community, but this was not true for all treatments or assays. For culturable bacteria the opposite result was found. Thymidine incorporation and glutamate metabolism were much more sensitive indicators of metal toxicity than was viability. To our knowledge, this is the first use of the thymidine incorporation method for ecotoxicology studies. We found it the easiest and fastest of the three methods; it is at least equal in sensitivity to metabolic measurements, and it likely measures the effects on greater portion of the natural community.     

Influence of selenium on mercuric chloride cellular uptake and toxicity indicating protection: studies on cultured K-562 cells

Selenium and mercuric chloride (MC) interactions regarding cellular uptake and selenium protection on MC toxicity have been studied. Human K-562 cells were pretreated or simultaneously treated with either selenite (5 or 50 microM) or selenomethionine (10 or 50 microM) together with MC (35 or 50 microM). Both treatments with selenite showed an increase of mercury uptake with increased selenium dose. In the pretreated or simultaneously treated selenite and 35 microM MC combinations, no inhibition of growth was seen, whereas all 50-microM MC combinations were toxic to the cells. A selenite-dependent protection was obtained for both exposure protocols when considering the cellular uptake of mercury. The cells died when the accumulation on d 4 reached more than about 0.8 x 10(-15) mol/cell of mercury, whereas they survived up to twofold more mercury uptake when exposed to selenite. Selenomethionine gave, with a few exceptions, similar effects as selenite on MC uptake and toxicity.     

Evaluation of methyl mercury chelating agents using red blood cells and isolated hepatocytes

The relative efficacy of thiol-containing mercurial scavengers was assayed by using cellular suspensions of erythrocytes or isolated hepatocytes. The blood cells incubated in a buffer (pH 7.4) containing 1 mM glucose (10% hematocrit) were exposed to 5 μM methyl mercuric chloride. In the absence of extracellular thiols the red blood cells took up more than 90% of methyl mercury from the surrounding medium during 5–10 min. This uptake was almost completely inhibited by dimercaptosuccinic acid (DMSA) (1 mM) and the same chelant could rapidly remove 80% of the mercury from ‘pre-loaded’ erythrocytes. Hepatocytes prepared according to the method of Seglen [11] in a suspension of 10⁶ cells/ml in a buffer containing 5 mM glucose and 5 mg/ml of bovine serum albumin were also exposed to methyl mercuric chloride (4 μM). Almost 50% of the mercurial was taken up by the cells slowly during the incubation period of 240 min. DMSA (1 mM) almost completely blocked the methyl mercury binding by the hepatocytes. 2-Mercaptopropionylglycin (Thiola) or mercaptosuccinic acid (MSA) was almost as effective mercurial scavengers as DMSA in hepatocytes and in red blood cells. Diethyldithiocarbamate (DDC) and dimercaptopropanol (BAL) were considerably less effective than DMSA to inhibit the mercurial binding to hepatocytes. Experiments in vivo have shown that DMSA is a better mercurial chelator than Thiola or MSA, whereas DDC and BAL may both be considered to be inapplicable in methyl mercury poisonings. Our cellular assay provides preliminary information of the efficiency of chelating thiols and may serve as a useful first approximation when planning further experiments.     

Development of a biological mercury removal-recovery system

A mercury removal-recovery system was developed for collection of elemental mercury volatilized by biological mercuric ion reduction. Using the mercury removal-recovery system, removal of mercuric chloride from mercury-containing buffer without nutrients by resting cells of mercury-resistant bacterium, Pseudomonas putida PpY101/pSR134 was tested. Optimum temperature, pH, thiol compounds and cell concentration on removal of mercuric chloride were determined, and 92 to 98% of 40 mg Hg l^–1 was recovered in 24 h. The efficiency of mercuric chloride removal from river water and seawater was as high as that observed when using a buffered solution.