Toxicity of cadmium to schistosoma mansoni cercariae: effects on vitality and developmental ability in white mice

Time-until-death studies were run on cercariae of Schistosoma mansoni in 8 concentrations of cadmium (from cadmium sulfate) ranging from 100 ppm to 0.0001 ppm. All concentrations used were found to be toxic, and at 10 ppm all cercariae were dead within 4 hours, which coincides with their period of maximum infectability following emergence from the snail host. At 2 ppm, all cercariae died within 8 hours, and at 1 ppm all died within 16 hours. In addition, groups of cercariae were exposed to cadmium concentrations of 10, 1, and 0.1 ppm for periods of 30, 20, and 10 minutes. Thereafter, cercariae from these groups were allowed to penetrate the tails of white mice or were injected subcutaneously into mice. After 8 weeks, these mice were autopsied and the adult worms collected by perfusion. Maturation of cercariae from both methods of invasion was seriously impaired. Statistical analysis using a 3 × 4 × 2 factoral design for analysis of variance showed both time of exposure and concentration of cadmium ion to be significant factors in determining number of worms developing at p = 0.01. A significant interaction between time of exposure and concentration was found to exist. The two methods of infection did not have a significant effect on the number of worms recovered. Therefore, it appears that those toxicant-exposed cercariae capable of maturing do not need assistance in traversing the skin barrier but can penetrate and migrate to reach the mesenteric venules for maturation.     

Effect of 6-mercaptopurine on mineral and metallothionein metabolism in the mouse

The effect of the anticancer drug 6-mercaptopurine (6-MP) on mineral metabolism was investigated in mice. C57Bl/6J female mice were injected intraperitoneally with 6-MP at 100 mg/kg body wt for one, two, four, or five consecutive days. On d 6 of the study, liver, kidney, and intestine were removed, and concentrations of zinc, copper, iron, manganese, magnesium, and calcium were measured. Hepatic concentrations of zinc, copper, iron, and calcium became higher as the number of drug injections increased. To determine if the altered mineral metabolism was a function of a drug-induced, acute-phase response, liver metallothionein and plasma ceruloplasmin were measured. Metallothionein concentrations in the liver became higher with increased number of injections, correlating with the stepwise increase in hepatic zinc. Gel filtration chromatography showed that most of the increase in liver zinc concentration was associated with a protein of mol wt of 6000–8000, the approximate weight of metallothionein. Ceruloplasmin concentrations were not affected by 6-MP injection. These results suggested that 6-MP alters zinc metabolism by sequestering zinc into the liver via induction of metallothionein synthesis and that the drug may induce an acute-phase response with an atypical acute-phase protein profile.     

Studies on the role of iron in zinc toxicity in chicks

The interaction of dietary iron and zinc was studied in chicks. Zinc was found to be more toxic in iron-deficient animals than iron-supplemented animals as measured by hemoglobin concentrations and growth. Analyses of the kidney and liver for iron and zinc were carried out. As the level of iron was increased from 0-1000 ppm supplementation, the concentration of liver zinc increased. The organ levels of iron were decreased as the dietary zinc levels were increased from 0-5000 ppm. Radioisotope studies using⁶⁵Zn revealed that the iron content of the diet did not affect absorption of zinc. Administration of the isotope, either in an intestinal segment or intravenously, resulted in more zinc being taken up by the liver in the iron supplemented animals. This was especially noted when the ratio of the isotope in liver to that in the blood was compared. Gel chromatography of kidney and liver homogenates revealed that iron deficiency resulted in less zinc being eluted in a volume characteristic of metallothionein compared to homogenates of organs from iron supplemented animals. The results indicate that iron-supplemented animals have a greater capacity for sequestering zinc on metallothionein than do iron-deficient animals. Conversely, iron-deficient chicks were more susceptible to the effects of zinc toxicity than are iron-adequate chicks.     

Chromium tolerance,oxidative stress response,morphological characteristics,and FTIR studies of phytopathogenic fungus <Emphasis Type="Italic">Sclerotium rolfsii</Emphasis>

Sclerotium rolfsii is one of the most destructive fungal plant pathogens that can infect over 500 plants and can adapt to diverse environmental conditions. The present research work was carried out to evaluate the impact of both hexa- and trivalent chromium (Cr) on growth, morphology, enzymatic characteristics, and metal accumulation in S. rolfsii under laboratory conditions. Experiments were performed in both malt extract broth and agar growth medium amended with six different concentrations (10, 20, 40, 60, 80, and 100 ppm) of each Cr(III) and Cr(VI) ions inoculated with fungus and incubated for 6–7 days at 25 ± 3 °C. In broth medium, the total protein content was declined and activities of antioxidant enzymes were increased with an increase in metal concentrations. Lower concentrations (10 ppm) of the metal ions stimulated the growth of fungus and higher concentrations (60–100) inhibited it. The Fourier transform infrared spectroscopy (FTIR) assessment showed hydroxyl, carboxyl, and amine groups as major metal binding sites. In agar medium, tolerance index was decreased up to 0.56 at 10–80 ppm of Cr(III) and up to 0.62 at 10–60 ppm of Cr(VI). Considerable modifications were observed in hyphal and sclerotial morphology with an increase in concentration of metal ions. The current study concluded that interference of Cr with growth and physiological process of S. rolfsii could affect its infection level on its host plant. This study provides important information regarding cultivation of susceptible plant varieties in Cr-polluted soil as evidenced by pathogen growth up to 50 ppm of Cr(III) and Cr(VI) ions.     

Ethrel-induced ripening of immature and mature-green tomato fruits

Ethrel accelerated the ripening of both immature and mature-green ‘2029’ tomato fruits. The best concentrations and time of dipping used were 2500 to 10,000 ppm and 5 minutes, respectively. Addition of surfactants and DMSO did not markedly influence the effectivity of Ethrel. Ethrel-treated fruits harvested 21 and 28 days after fruit set ripened normally but 14 day old fruits did not.     

Reduction of aflatoxin B1 levels by sheep saliva

This study determined the decrease of aflatoxin B₁ by sheep saliva at concentrations of 150 and 300 μg aflatoxin B-₁/L saliva. Analyses for aflatoxins B₁, M₁, and aflatoxicol (R₀) were performed after 2, 4, 6, 24, and 48 hours of incubation. Aflatoxin M₁ and R₀ were not detected and only residues of aflatoxin B₁ were found. 4 to 13% of aflatoxin B₁ were decomposed by sheep’s saliva within 2 hrs and 33 to 43% of aflatoxin B₁ after 24 hrs. Decomposition was affected by the aflatoxin concentration. Decrease of aflatoxin B₁ at 2, 4, 6 hrs was nearly three times higher at the low concentration (150 ppb) compared to the high concentration (300 ppb). After 48 hrs incubation more than 80% of the initial aflatoxin B₁ had been decomposed by the saliva.     

Amniotic fluid zinc in risk pregnancies

The zinc concentration of amniotic fluid (AF) of 129 pregnant women was analyzed by the flame atomic absorption spectrometry. This prospective study was performed in order to find out whether the determination of the AF zinc concentration can be used to monitor the growth and development of the fetus. There were two groups of patients: early stage (15th–19th gestation wk) in which the amniocentesis was performed as a prenatal genetic examination, and late stage (26th–40th wk) in which the amniocentesis was performed due to obstetric reasons. The average AF zinc concentrations were 1.2 and 1.0 μmol/L in the early and late gestation group, respectively. The AF zinc concentration did not correlate with the weight, height, or Apgar scores of the newborn nor with the maternal diseases, age, or parity. The AF zinc concentration in the late gestation group was significantly lower if the fetus was male than if it was female. If the AF was greenish the zinc concentration was elevated. One malformation, congenital nephrosis, with an exceptionally high zinc concentration (9.0 μmol/L), was found.     

Field observations on the influence of low water velocities on drifting of Bulinus globosus

The influence of water velocities of less than 30 cm/s on drifting of Bulinus globosus in a natural stream was investigated. It is shown that although a low water velocity does not sweep away adhering snails, it carries away snails releasing their hold and crawling along the water surface in search of new food sources. A low water velocity therefore assists considerably to the spreading of a snail species. Vegetation was found to have a decisive impact on local water velocity conditions by reducing the current and creating protected pockets. Upstream migration was found not to be common among Bulinus globosus. In addition, evidence for snail drifting under natural conditions within well established Schistosoma haematobium transmission sites is presented.     

Callus formation and plant regeneration from protonemal protoplasts of the mossAnoectangium thomsonii Mitt.

The protoplasts of the mossAnoectangium thomsonii, isolated from protonemal cells underwent divisions in 48–72 hours on modified MS medium enriched with growth regulators, 2,4-D and kinetin, 10 % sucrose and coconut water (5 %). Subculture of protoplast derived cells in a medium of relatively low osmotic potential (5 % sucrose) produced dark green calli which could be maintained completely undifferentiated.     

Conditional rifampicin sensitivity of arif mutant ofEscherichia coli: rifampicin induced changes in transcription specificity

Arif mutantof Escherichia coli that exhibits medium and temperature-dependent sensitivity to rifampicin is described. In the absence of rifampicin, this strain grows in minimal and rich media at 30°C and 42°C. In its presence it is viable in rich medium at both temperatures, but in minimal medium only at 30°C. In minimal-rifampicin medium at the higher temperature, RNA synthesis is decreased. The addition of certain divalent salts (MgSO₄, CaCl₂, BaCl₂) in excess, or chelators (EDTA, EGTA, o-phenanthrolein) greatly increase viability in minimal-rifampicin medium at 42°C. Excess MgSO₄ (10 mM) also increases the rate of RNA synthesis in the same medium. A model is proposed wherein therif mutation is suggested to cause a structural change in RNA polymerase that allows the binding of rifampicin and other ligands at 42°C. Rifampicin-binding is suggested to alter the conformation of RNA polymerase, impairing its ability to express genes required for growth in minimal medium. Implicit in this view is the assumption that these genes are structurally different from those expressed in rich medium in respect of certain template features recognized by RNA polymerase.     

Native isolate of <Emphasis Type="Italic">Trichoderma</Emphasis>: a biocontrol agent with unique stress tolerance properties

Species of Trichoderma are widely recognized for their biocontrol abilities, but seldom studied collectively, for their plant growth promotion, abiotic stress tolerance and bioremediation properties. Our study is a concentrated effort to establish the potential of native isolate Trichoderma harzianum KSNM (T103) to tolerate biotic (root pathogens) and abiotic stresses [high salt (100–1000 mM); heavy metal (chromium, nickel and zinc: 1–10 mM); pesticides: malathion (100–600 ppm), carbofuran (100–600 ppb)], along with its ability to support plant growth. In vitro growth promotion assays with T103 treated Vigna radiata, Vigna mungo and Hordeum vulgare confirmed ‘non-species specific’ growth promotion effects of T103. At lower metal concentration, T103 treatment was found to completely negate the impact of metal stress [60 % increase in radicle length (RL) with no significant decrease in %germination (%G)]. Even at 10 mM metal, T103 inoculation gave 80 % increase in %G and >50 % increase in RL. In vitro experiments confirmed high metal reduction capacity (47 %-Cr, 35 %-Ni and 42 %-Zn) of T103 at concentrations as high as 4 mM. At maximum residual concentrations of malathion (440 ppm) and carbofuran (100 ppb) reported in agricultural soils, T103 maintained 80 and 100 % survivability, respectively. T103 treatment has improved %G and RL in all three hosts challenged with pesticide. Isolate T103 was found to effectively suppress growth of three major root pathogens: Macrophomina phaseolina (65.83 %) followed by Sclerotium rolfsii (19.33 %) and Fusarium oxysporum (19.18 %). In the light of these observations, native T. harzianum (T103) seems to be a competent biocontrol agent for tropical agricultural soils contaminated with residual pesticides and heavy metals.     

Buoyant density of EMT6 fibrosarcoma cells

EMT6 fibrosarcoma cells were grown to the exponential phase in tissue culture and incubated at 37°C under hypoxic conditions. Buoyant density was determined as a function of the time in hypoxia. Hypoxia was produced in two ways. The first involved incubation of the cells in sealed aluminum chambers containing 95% N₂, 5% CO₂ gas, and <10 ppm oxygen, resulting in the cells rapidly becoming exposed to the hypoxic environment. After incubation at 37°C, they were centrifuged in linear Ficoll gradients to their isopycnic density. A significant decrease in density was found after 4 h, and prolonged incubation up to 24 h did not result in further change. This density change was reversible on transfer back to aerobic conditions, with the hypoxic cells reverting to their aerobic density after about 10 h reincubation in air. The second method of producing hypoxia involved growing about 8×10⁶ cells in a medium-filled air-tight container. Hypoxia was produced gradually as the oxygen in the medium was consumed by cellular respiration. Similar results were obtained; that is, hypoxic cells became significnatly less dense. However, when the level of hypoxia was varied between 4000 and <10 ppm at 2-h intervals after the cells had depleted all of the original oxygen, no significant difference in density was found between hypoxic and aerobic cells.     

Effect of natural antioxidants on the lipase activity in the course of batch and continuous glycerolysis of babassu oil

The effects of several natural antioxidants (copaiba oil, buriti oil, cocoa butter, tucuman butter, oregano and white thyme) were assessed in the enzymatic synthesis of monoglycerides (MAG) from the glycerolysis of babassu oil. The reactions were catalyzed by Burkholderia cepacia lipase immobilized on SiO₂–PVA and the assays carried out in batch and continuous runs. Results were compared with those attained in the control reactions (without any strategy to avoid oxidation), and the best approach was tested in a continuous packed-bed reactor. The best performance was obtained using N₂ in the reaction medium (60 % of MAG) followed by buriti oil (57.6 % of MAG) and cocoa butter (56.6 % of MAG), preventing the oxidation of babassu oil in batch reaction. However, the incorporation of buriti oil in the medium influenced the MAG profile, leading to the largest formation of monoolein, unlike other runs. Similar results were obtained in continuous reactions, using inert atmosphere and cocoa butter (24–25 % of MAG). Thereby, among the tested antioxidant agents, cocoa butter was the most effective in both systems, because it did not interfere in the MAG profile and also reduced the cost of the process.     

Effect of temperature on embryonic development and reproduction of the freshwater snailLymnaea luteola Troshel (Gastropoda), a vector of schistosomiasis

The effect of temperature on embryonic development and reproduction ofLymnaea luteola was studied. This snail did not develop completely and failed to reproduce at 15 °C and 40 °C. The temperature range of 25 °C–35 °C was observed to be optimum for development and reproduction of this snail. The utility of this study in predicting seasonal fluctuations of snail population in nature is discussed.     

Characterization of a class II 5-enopyruvylshikimate-3-phosphate synthase with high tolerance to glyphosate from Sinorhizobium fredii

5-Enopyruvylshikimate-3-phosphate synthase (EPSP synthase) is an important enzyme in the shikimate pathway mediating the biosynthesis of aromatic compounds in plants and microorganisms. A novel class II EPSP synthase AroA _{S. fredii} from Sinorhizobium fredii NGR234 was overexpressed in Escherichia coli BL21. It was purified to homogeneity and its catalytic properties were studied. The enzyme exhibited optimum catalytic activity at pH 8.0 and 50 °C. It was stable below 40 °C, and over a broad range of pH 5.0–9.0. The EPSP synthase was increasingly activated by 100 mM of the chlorides of NH₄ ⁺, K⁺, Na⁺ and Li⁺. Kinetic analysis of AroA _{S. fredii} suggested that the enzyme exhibited a high glyphosate tolerance and high level of affinity for phosphoenolpyruvate, which indicates the enzyme with a high potential for structural and functional studies and its potential usage for the generation of transgenic crops resistant to the herbicide.     

Characterization of an organic solvent-tolerant lipase from Haloarcula sp. G41 and its application for biodiesel production

A haloarchaeal strain G41 showing lipolytic activity was isolated from the saline soil of Yuncheng Salt Lake, China. Biochemical and physiological characterizations along with 16S rRNA gene sequence analysis placed the isolate in the genus Haloarcula. Lipase production was strongly influenced by the salinity of growth medium with maximum in the presence of 20 % NaCl or 15 % Na₂SO₄. The lipase was purified to homogeneity with a molecular mass of 45 kDa. Substrate specificity test revealed that it preferred long-chain p-nitrophenyl esters. The lipase was highly active and stable over broad ranges of temperature (30–80 °C), pH (6.0–11.0), and NaCl concentration (10–25 %), with an optimum at 70 °C, pH 8.0, and 15 % NaCl, showing thermostable, alkali-stable, and halostable properties. Enzyme inhibition studies indicated that the lipase was a metalloenzyme, with serine and cysteine residues essential for enzyme function. Moreover, it displayed high stability and activation in the presence of hydrophobic organic solvents with log P _ow?≥?2.73. The free and immobilized lipases from strain G41 were applied for biodiesel production, and 80.5 and 89.2 % of yields were achieved, respectively. This study demonstrated the feasibility of using lipases from halophilic archaea for biodiesel production.     

Regeneration of the cell wall of protoplasts of the fission yeastSchizosaccharomyces versatilis in liquid media and their reversion to cells

Regeneration of the cell wall and reversion of protoplasts with a completely regenerated cell wall to cells were studied by light and electron microscopy in protoplasts of the fission yeastsSchizosaccharomyces versatilis. On their surface the protoplasts regenerated a complete new wall even m liquid media The wall regeneration began with the formation of a thin irregular net of flat bundles of long microfibrils and the net was gradually filled with aggregates of short straight microfibrils and small piles of amorphous material. Osmotically resistant organisms with regenerated walls were detected after a 4–6 h cultivation Depending on the nutrient medium used 10–80 % of protoplasts with the regenerated wall were obtained that reverted subsequently to cells. The high percentage of the wall regeneration and reversion to cells was reached by combining cultivation in a poor medium with that in a rich medium Reversion to cells could only occur after the protoplasts had regenerated rigid cell walls These walled protoplasts underwent septation, and, by polar growth, produced cylindrical cells, further dividing by fission.     

Effect of tortilla-preparation process on aflatoxins B1 and B2 in corn

Naturally contaminated corn containing 450 and 54 ppb aflatoxins Bi and B2, respectively was treated with Ca(OH)₂ for making tortillas. The cleaned corn and tortillas were analyzed for aflatoxins B1 and B2 by high performance liquid chromatography (HPLC) and confirmed by thin layer chromatography (TLC). The average concentrations of aflatoxins B1 and B2 in the final products (tortillas) were only 40% and 28% lower than that in starting materials (corn kernels), respectively. Aflatoxins G1 and G2 were not detected in either corn or tortilla samples.