Sol-gel immobilisation of methyl parathion degrading bacteria isolated from agricultural areas of Pakistan

The following areas are discussed in this paper: immobilisation of bacterial consortium in sol-gel; methyl parathion degradation and bioremediation applications; evaluation of indigenous bacterial isolates of contaminated soils. Bacterial strains were isolated from agricultural areas of Pakistan which were contaminated with methyl parathion. A bacterial consortium of seven (out of 64) Enterobacteriaceae isolates including Citrobacter, Enterobacter and Proteus vulgaris capable of degrading methyl parathion (enzyme activity ranging 410–675 mU mL^?1 for individual isolates and 982 mU/mL for consortium) was selected and subsequently immobilised in tetraethyl orthosilicate (TEOS) and sodium-silicate-based sol-gel matrices. Cell viability of suspended and immobilised bacterial consortium was monitored using a minimal salt medium supplemented with methyl parathion. The results indicate that sol-gel immobilisation can be helpful to increase the shelf life of methyl parathion degrading bacterial strains along with preservation of biological activity for bioremediation applications in field.     

Role of NADH-dependent chromium reductases,exopolysaccharides and antioxidants by Paenibacillus thiaminolyticus PS 5 against damage induced by reactive oxygen species

ABSTRACT

Cr (VI) being used in various activities of industries and its improper treatment lead to contamination of environment. Among different methods, biological is the most efficient method to control pollution from soils affected with metals. Present study was designed to assess the role of Paenibacillus thiaminolyticus PS5 for adsorption, Cr (VI) reduction and mechanism of reduction. Sorption of chromium (VI) by strain PS5 was obtained by batch equilibrium method. Cr (VI) reduction in both free and immobilised cells were evaluated by 1,5-Diphenyl Carbazide method. The formation of biofilm by Paenibacillus thiaminolyticus PS5 was observed for colour change as well as quantified spectrophotometrically. Analysis kits were used to measure the amount of eDNA, superoxide and malondialdehyde (MDA). Metal resistant strain PS5 was characterised as P. thiaminolyticus using 16S rRNA gene sequence. Maximum biosorption of Cr (VI) by strain PS5 was found at pH 6–8 and 100–400 µgCr/mL within 24 hours of incubation. Complete reduction of Cr (VI) by strain PS5 was achieved at pH 6–8 and100–300 µg/mL Cr (VI). Paenibacillus thiaminolyticus PS5 immmobilisation on sodium alginate and polyvinyl alcohol facilitated complete reduction of Cr (VI) within 18 hours due to the formation of more biofilm under metal stress conditions. Strain PS5 reduced almost all Cr (VI) into Cr (III) in supernatant, most of which was immobilised by cell debris. Experiments confirmed the reduction of Cr (VI) by cytosolic cell-free extracts which is a mechanism of detoxification. The release of exopolysaccharides and antioxidants by strain PS5 played a protective role against cell damage by Cr (VI) as Cr (VI) could not release the significant amount of eDNA, superoxide and MDA. The present study proved strain PS5 to be a super bioinoculant which has great capacity for adsorption, biotransformation and can activate cytosolic reductases, exopolysaccharides and antioxidants against oxidative damage.     

Selenium content of rice,mixed plant foods and fish from Bangladesh

Static and dynamic adsorption studies of Co (II) ions have been undertaken at fixed pH and ionic strength taking binary biopolymeric beads of cross-linked calcium alginate and carboxy methylcellulose as biosorbents. The adsorption data were applied to Langmuir and Freundlich isotherm equations and various static parameters were calculated. The dynamic nature of adsorption process was quantified in terms of several kinetic constants such as rate constants for adsorption (K) and Lagergreen rate constant (K _ad). The influence of various experimental parameters such as solid-to-liquid ratio, pH, and temperature, the presence of salts and chemical composition of biopolymeric beads were investigated on the adsorption of cobalt ions.     

Development of chitosan-alginate based biosorbent for the removal of p-chlorophenol from aqueous medium

Removal of p-chlorophenol (pCP) from synthetic aqueous solutions was studied through adsorption on a biosorbent developed from chitosan (CS) and sodium alginate (SA), the natural cationic and anionic polysaccharides, respectively. Chitosan-coated sodium alginate beads were prepared and treated with calcium chloride solution in order to improve the stability as well as hydrophobic character. The resulting beads (CS/CA) were characterized using FTIR spectra, scanning electron microscopy (SEM), and BET surface analysis. The efficiency of this biosorbent in removing pCP from aqueous medium was studied under batch equilibrium and dynamic column flow experimental conditions. The binding capacity of the biosorbent was studied as a function of initial pH, contact time, initial concentration of adsorbate and amount of biomass. The data were fitted to pseudo-first-order, pseudo-second-order, and Weber–Morris models and found that the adsorption process followed pseudo-first-order kinetics. Further, the equilibrium data were fitted to Freundlich, Langmiur, and Dubinin–Radushkevich (D–R) adsorption isotherms and the isotherm constants were evaluated for adsorption of pCP. The maximum monolayer adsorption capacity of CS/CA beads was found to be 127 mg g^?1. Column flow results were used to generate breakthrough curves. The experimental results suggested that the chitosan–calcium alginate blended biosorbent was effective for the removal of pCP from aqueous medium.     

固定化微生物对养殖水体中NH_4~ -N和NO_2~--N的转化作用

采用聚乙烯醇 (PVA) (ρ=gL-1)的方法对沼泽红假单胞菌、诺卡氏菌和假丝酵母 3种菌株进行固定化 ,所得的凝胶颗粒机械强度好 ,经久耐用 .运用这 3种菌株的固定化细胞对养殖水体中NH 4 N和NO-2 N进行转化 ,其最适作用温度范围为 2 5～ 30℃ ,最适pH范围分别为 :6 .5～ 8.5 ,7.0～ 7.5及 5 .5～ 7.0 ,且对水质初始氮浓度有很强的适应性 .3菌株经固定化后 ,其对养殖水体中NH 4 N和NO-2 N的转化效率明显优于其游离细胞 .若将 3菌株按 2 :1:2组合成复合菌株并固定化 ,其对养殖水体中的NH 4 N和NO-2 N转化效果将更佳 .图 3表 3参 9     

Effect of total cadmium and organic complexing on the uptake of cadmium by the brine shrimp,Artemia franciscana

The effect of total cadmium and organic complexing on the rate of cadmium uptake by the brine shrimp, Artemia franciscana has been studied in chemically defined saltwater solutions. The uptake of cadmium from solution by the laboratory-reared brine shrimp displays saturable uptake kinetics. Uptake of cadmium is linear in time up to a total cadmium concentration of 200 moll^-1 and saturates above 800 moll^-1. Complexation of cadmium with organic ligands decreases the uptake of the metal by the brine shrimp. This is in agreement with the view that the availability of cadmium to aquatic organisms is related to the activity of the free cadmium ion in the solution. There is no evidence that the direct uptake of cadmium complexes is important in determining uptake of cadmium. Cadmium uptake is not, however, a mere function of the free cadmium ion activity in the solution, i.e., cadmium uptake rates may differ by an order of magnitude for the same free cadmium ion activity depending on the complexation conditions. In addition to controlling the free cadmium ion activity, the role of organic ligands in metal ion buffering and metal ion masking appear important factors in determining the availability of the metal to the organism.     

固定化菌藻系统及对污水中氮磷营养盐的净化效果

利用海藻酸钙分别包埋固定小球藻、活性污泥及其两者混合物.研究固定态小球藻与悬浮态小球藻对污水中氮磷营养盐的处理效果,实验结果表明固定态藻对氮磷的去除效果明显优于悬浮态藻,其原因主要由于海藻酸钙凝胶对氮磷的吸附,系统pH值提高引起氨的气提及磷酸盐的沉淀作用.研究固定化藻、同定化活性污泥及共固定化菌藻分别对污水中氮磷营养盐的处理效果,在同等条件下,固定化菌藻对氮磷的去除效果优于固定化活性污泥和固定化藻类.这项研究显示菌藻共生在污水处理中具有较大的潜力.     

Removal of cadmium(II), lead(II), and chromium(VI) ions from aqueous solution using clay

Removal of cadmium(II), lead(II), and chromium(VI) from aqueous solution using clay, a naturally occurring low-cost adsorbent, under various conditions, such as contact time, initial concentration, temperature, and pH has been investigated. The sorption of these metals follows both Langmuir and Freundlich adsorption isotherms. The magnitude of Langmuir and Freundlich constants at 30°C for cadmium, lead, and chromium indicate good adsorption capacity. The kinetic rate constants (K _ad) indicate that the adsorption follows first order. The thermodynamic parameters: free energy change (ΔG ^o), enthalpy change (ΔH ^o), and entropy change (ΔS ^o) show that adsorption is an endothermic process and that adsorption is favored at high temperature. The results reveal that clay is a good adsorbent for the removal of these metals from wastewater.     

Effects of dissolved organic matter on the growth of Nodularia spumigena (Cyanophyceae) cultivated under N or P deficiency

We investigated the effects of addition of dissolved organic matter (DOM) on the growth and chemical composition of Nodularia spumigena cells under conditions of P or N deficiency. Semi-continuous monocultures of N. spumigena Mertens were established. The cells were exposed to the following treatments: nutrient-sufficient (+NP), nitrogen-deficient without DOM (−N) and with DOM (−NDOM), and phosphorus-deficient without DOM (−P) and with DOM (−PDOM). The −NDOM treatment had the highest N. spumigena cell density (6.1 × 10⁷ cells l⁻¹) during the experiment, whereas the P-deficient treatments had the lowest cell numbers (0.53 to 0.66 × 10⁷ cells l⁻¹). Both −P and −PDOM treatments yielded P-deficient N. spumigena cells. Dissolved organic nitrogen may provide N and increase N. spumigena yield under conditions of N deficiency. On the other hand, N. spumigena cells did not use DOM as a P source. Received: 19 July 1999 / Accepted: 20 October 1999     

Combined effects of copper and cadmium on Chlorella pyrenoidosa H.Chick: subcellular accumulation,distribution, and growth inhibition

Disposal of waste into aquatic ecosystems may cause microalgae to be exposed to various metals, e.g. copper and cadmium. The effects caused by combinations of metals may be more serious. Evaluations of subcellular fate, bioaccumulation, and biological effects of metals on aquatic organisms are generally derived from experiments with individual metals. The present study aims to evaluate the effects of exposure of Chlorella pyrenoidosa to copper and cadmium in combination on subcellular accumulation, distribution, and growth. The algae were exposed for 72 h to copper at concentrations of 13 – 25 µmol L^?1, cadmium at about 6 µmol L^?1, and combinations thereof. The levels of copper and cadmium in subcellular organelles, heat-denaturated protein, metal-rich granules, and heat-stable protein were determined by atomic absorption spectrometry. Exposure of C. pyrenoidosa to copper and cadmium in combination inhibited growth more strongly than copper and cadmium individually. Highest accumulation was observed in metal-rich granules and heat-stable proteins. Administration of both metals in combination affected their subcellular distribution: copper was mainly distributed into the metal-rich granules (70%–80%) and heat-stable proteins (9%–24%), cadmium in the metal-rich granules (88%–98%).     

Discriminate feeding of the calanoid copepod Acartia clausi in mixtures of phytoplankton and inert particles

Adult female Acartia clausi were allowed to feed in the laboratory on each of three algae, the small green alga Dunaliella tertiolecta (5.6 m ESD), the diatoms Thalassiosira decipiens (13.4 m ESD) and T. nordenskioldii (17.7 m ESD), singly or in mixtures with polystyrene beads (15.7 m ESD). The ingestion rate on beads was much lower than that on cells, even when the supply of beads was one order of magnitude higher than that of cells. Beads offered singly were not ingested. In the experiments with T. nordenskioldii, the more cells adult females ingested, the higher the ingestion rate on beads. In addition, a linear regression equation describing the relationship between bead concentration and ratio of ingested beads to ingested cells was highly significant. These results suggest that beads are ingested by chance in feeding bouts on T. nordenskioldii. The bead interference on the ingestion rate on cells was observed in the experiments with D. tertiolecta and T. decipiens, but not with T. nordenskioldii. In a mixture of beads and T. decipiens, juvenile A. clausi discriminated beads from cells more successfully than adults. The presence of beads did not affect the ingestion rate of juveniles on cells. Starved adult females tended to ingest more beads than well fed individuals. The possible effects of body size and hunger on the discriminate feeding of copepods are discussed.     

The effects of exposure to zinc and cadmium separately and jointly on the free amino acid pool of Gammarus pulex (L.)

Effects of the heavy metals zinc and cadmium on the free amino acid (FAA) pool of Gammarus pulex were studied at different metal concentrations and combinations as well as different exposure times. The dominant effect of these two metals was the reduction of most free amino acids and the whole FAA pool, except in the 10‐day low zinc and cadmium concentration exposures which resulted in a rise of free amino acid pool.

Among the free amino acids, the most sensitive to zinc exposure, were alanine, glutamic acid, arginine, and taurine; valine, leucine, asparagine, and isoleucine were among the most sensitive to cadmium. No predictable changes of individual free amino acids were shown in the mixed metals exposures. Elevation of taurine concentration was constant in seven of the eight treatments, it is suggested that this elevation may be related to the hepatopancreatic damage observed and induced synthesis of metallothioneins.     

Synergistic effects of cadmium and salinity combined with constant and cycling temperatures on the larval development of two estuarine crab species

The developmental stages from megalopa to third crab of the blue crab Callinectes sapidus Rathbun were tested in 12 combinations of cadmium (0, 50, and 150 ppb) and salinity (10, 20, 30, and 40) at 25°C. A reduction in survival and a significant delay in development from megalopa to third crab occurred within each salinity regime in 50 ppb compared with the control. Comparison of the delay in development within each salinity regime revealed that the sublethal effect of cadmium was most pronounced in the salinities normally preferred by C. sapidus. A similar comparison within each cadmium concentration, however, showed that the developmental time from megalopa to third crab was approximately the same irrespective of salinity. The developmental stages from hatch to first crab of the mud-crab Rhithropanopeus harrisii (Gould) were examined in 63 combinations of cadmium (0, 50, and 150 ppb), salinity (10, 20, and 30), constant temperature (20°, 25°, 30°, and 35°C) and cycling temperature (20° to 25°C, 25° to 30°C, and 30° to 35°C). The results indicated that cycling temperatures may have a stimulating effect on survival of the larvae compared to constant temperatures, both in the presence and in the absence of cadmium. Effects of cadmium and salinity and their interaction on the survival of the larvae from zoeae to megalopa were documented at most of the temperatures by analyses of variance. The zoeal larvae were more susceptible to cadmium than the megalopa. Effects of different combinations of cadmium and salinity on the duration of larval development were assessed by a t-test.     

Release of Cd immobilized by soil constituents and its bioavailability

Release of Cd immobilized by soil constituents (clay, humic acids, bacterial cells) in vitro and under soil conditions and its bioavailability to lettuce were studied. The most resistant to extraction with 0.02 M EDTA in vitro as well as under soil conditions was cadmium immobilized by humic acids during decomposition of contaminated plant residues but Cd‐dead cells when 0.1 M NaNO₃ was the extractant.

Cadmium addition (3 mg kg^‐1) to soil and its form were without effect on plant growth and amount of metal accumulated in roots. Tops of plants grown in soil supplemented with Cd‐resting cells contained significantly less metal then other ones.     

Factors affecting enumeration and isolation of actinomycetes from Chesapeake Bay and Southeastern Atlantic Ocean sediments

Studies of the seasonal distribution of heterotrophic bacteria in Chesapeake Bay (USA) led us to the observation that fungizone was selective for actinomycetes without appreciably altering the total viable aerobic, heterotrophic bacterial counts. A study of the actinomycete populations in the Bay was undertaken, with several media compared for ability to enrich for actinomycetes: basal, casein, Czapek, starch-casein, glycerol-glycine, chitin and oil agar No. 2. Between 40 and 100% of the colonies appearing on chitin and oil No. 2 media were found to be actinomycetes, compared with <2% for the other media employed. It was also found that if sediment samples are frozen, a practice followed when laboratory facilities are not available in the field, a decrease in the total counts and in the actinomycete colony counts occurred. However, on a proportional basis, a slight increase in the percent actinomycetes was noted. A minimum of 21 days incubation time is recommended for recovery of actinomycetes. Also, the absence of NaCl from the actinomycete isolation media and incubation temperatures ≥15°C should give good recoveries of actinomycetes. A seasonal distribution of actinomycetes was noted for Chesapeake Bay, with maximal populations of actinomycetes observed in March and November. Actinomycetes were isolated in the ocean from depths ranging from 7 to 7,790 m. Greater numbers of actinomycetes were isolated using an incubation temperature of 25°C for the estuarine stations and 15°C for the deep-ocean stations. Greater numbers of actinomycetes were associated with polluted environments close to shore.     

稻米来源镉对生长期猪的生长特性及不同脏器组织镉蓄积的影响

环境中的镉污染对动植物及人类都带来了极大的危害。目前,多数镉毒理研究关注其无机状态,而来源于稻米的有机态镉的毒害作用报道较少。为评估稻米来源镉对生长期猪的毒性作用,选用初始体重为30 kg左右的健康生长猪28头,随机分为体重无明显差异的2组,即对照组与稻米来源镉组(试验组),研究稻米来源镉对生长期猪(生长及育肥全阶段)的生长特性及不同脏器组织镉蓄积的影响。结果表明,在生长及育肥阶段,试验组与对照组相比:1)生长特性无显著差异(P0.05);2)育肥期组织游离氨基酸含量,除肌肉中的赖氨酸显著降低外(P0.05),血浆、肝脏、肾脏及肌肉中其他游离氨基酸水平均无显著差异(P0.05);3)心脏、肝脏、脾脏、肾脏、肺脏、胃、小肠、大肠、骨骼中镉蓄积量显著增高(P0.05),而肌肉、血液中无显著差异(P0.05),脂肪、皮中未检出镉。日粮使用大比例镉超标稻米后镉在各脏器和组织中的蓄积规律为肾脏肝脏小肠脾肺大肠胃血液骨头心腿肌背肌脂肪、皮。由此可见,镉超标稻米对生长期猪的生长特性未产生显著的影响,稻米来源镉主要蓄积于肝脏和肾脏,而主流食用组织肌肉和血液中镉的蓄积量较少。     

Food-particle size and selection by bivalve larvae in a temperate embayment

Epifluorescence microscopy was used to analyze the stomach contents of bivalve larvae collected in the Baie des Chaleurs (western Gulf of St. Lawrence, Canada) in order to document food-particle sizes, compare feeding among taxa, and compare the diet with the in situ phytoplankton community. Stomach contents were mainly composed of small autotrophic flagellates (<5 μm) and cyanobacteria (<2 μm), reflecting the microbial food web which characterizes these waters. More than half (55%) of all veligers examined contained algal cells of 5 to 15 μm, whereas only 3% had cells of 15 to 25 μm. Differences in the size ranges of ingested algal cells among similar-sized larvae of different species suggests that veligers actively selected food particles. Among the smallest veligers (185 to 260 μm), scallops (Placopecten magellicanus) and mussels (Mytilus edulis) ingested more <5 μm and 5 to 15 μm algae than clams (Mya arenaria). Among larger veligers (261 to 405 μm), clams contained significantly more <5 μm cells than mussels, whereas mussels contained significantly more 5 to 15 μm algae than clams. Algal cells of 15 to 25 μm were preferentially ingested by mussel veligers. Feeding also differed between different-sized veligers within taxa, i.e. the smallest clam veligers ingested fewer of 5 to 15 μm algae than the larger size classes. Mussel veligers ingested significantly more 15 to 25 μm and fewer <5 μm cells as their size increased. The dominance of ultraplankton in the nearshore waters of Baie des Chaleurs and in the stomach contents suggests that veliger larvae may be an important export path for carbon produced by small phytoplankton. Received: 17 July 1996 / Accepted: 20 September 1996     

Enzymes from marine phycophages that degrade cell walls of seaweeds

Agarase, cellulase and alginate lyase activities from crude extracts of Aplysia dactylomela Rang, Haliotis coccinea canariensis Nordsieck, Littorina striata King et Broderip and Diadema antillarum Phillipi were measured in vitro to compare digestive efficiencies against several components of complex seaweed cell walls. Commercial abalone acetone powder (AAP, an extract from Haliotis sp.; Sigma, Ref. A-7514) and purified (Sigma, Ref. A-6306) and non-purified agarases from Pseudomonas atlantica were used with the same objective. Optimum conditions for agarase and cellulase activities were 40°C and pH 6.0. For alginate lyase, optimum temperature and pH were species-dependent. Highest reducing sugar release was shown by crude extracts from A. dactylomela. These crude extracts displayed high agarase activity compared with bacterial agarases at 40°C, and were significantly higher at 25°C. AAP and crude extracts from L. striata and D. antillarum exhibited high specific activities on all the substrates. Cold extract from Gracilaria spp. was the best substrate with which to measure agarase activity.     

Water-soluble extracts of the diatom Thalassiosira rotula induce aberrations in embryonic tubulin organisation of the sea urchin Paracentrotus lividus

Eggs and embryos of the sea urchin Paracentrotus lividus were used as a model to study the effect at the cellular level of potential anti-mitotic compounds extracted from the diatom Thalassiosira rotula. Eggs and embryos incubated in a water-soluble diatom extract, corresponding to 5 × 10⁶ and 10⁷ cells ml⁻¹, were totally blocked (i.e. cell division was blocked) at the one-cell stage. At lower concentrations (2.5 and 1.25 × 10⁶ cells ml⁻¹), the first mitotic division was inhibited in 32 ± 26% and 25 ± 3.5% of the zygotes, respectively, demonstrating the dose-dependent effect of diatom extracts on sea urchin development. Immunofluorescence dyes, specific for DNA and α-tubulin subunits, were used to stain nuclei and microtubules in sea urchin embryos during various phases of development. Images with the confocal laser scanning microscope showed that tubulin was not organised in filaments at the sperm aster and cortex levels, and that the pronuclei were not fused in embryos incubated soon after fertilisation with water-soluble diatom extracts corresponding to 10⁷ cells ml⁻¹. At lower diatom-extract concentrations (4 × 10⁶ cells ml ⁻¹), fusion of the pronuclei occurred but the mitotic spindle was not formed. Microtubules were clearly de-polymerised and the chromatin appeared globular and compacted at the centre of the cell. A similar structure was observed for sea urchin embryos incubated with 0.1 mM colchicine, a potent anti-mitotic compound. When sea urchin embryos were incubated in water-soluble diatom extracts at different times prior to the first mitotic division, microtubules appeared de-polymerised at each step, from pronuclear fusion to telophase, and cell division was blocked. At the histological level, embryos incubated with 4 × 10⁶ cells ml⁻¹ diatom extract showed nuclear fragmentation without cytokinesis. The possible use of sea urchin embryos as a bioassay to test for other unknown compounds with cytotoxic activity in phytoplankton species is discussed. Received: 7 May 1998 / Accepted: 9 December 1998     

Heavy metals uptake from aqueous solutions using marine algae (Colpomenia sinuosa): kinetics and isotherms

The adsorption of copper, zinc, cobalt, lead and cadmium ions onto Colpomenia sinuosa was studied as a function of contact time, initial metal ion concentration and initial pH. In addition, desorption studies were performed. Characterisation of this adsorbent was also confirmed by Fourier transform infrared (FTIR) spectroscopy and scanning electron microscopy (SEM) analysis. Batch adsorption experimental data were analysed using Langmuir, Freundlich and Dubinin–Raduschkevich (D–R) adsorption isotherms. The results indicated that the biosorption equilibrium was well described by both the Freudlich and D–R isotherms. Moreover, sorption kinetics was performed and it was observed that equilibrium was reached in<60 min, which could be described by the pseudo-second-order kinetic model for all heavy metals. The sorption of heavy metals onto the biomass was largely dependent on the initial solution pH. The elution efficiency for heavy metal ions desorption from C. sinuosa was determined for 0.1 M HCl, 1.0 M HCl and 1.0 M HNO₃. Desorption efficiency and also adsorption capacity were highest for Pb(II). The results indicate that C. sinuosa has great potential for the removal of heavy metals in an ecofriendly process.