Isolation and characterisation of a carbofuran degrading Burkholderia sp. PCL3 from carbofuran-phytoremediated rhizosphere soil

A bacterial strain capable of degrading carbofuran as the sole carbon source was isolated from carbofuran-phytoremediated rhizosphere soil of rice. A 16S rRNA study identified the strain as Burkholderia sp. (isolate PCL3). Free cells of isolate PCL3 possessed inhibitory-type degradation kinetics with a q _max of 0.087 day^?1 and S _m of 248.76 mg·L^?1. Immobilised PCL3 on corncob and sugarcane bagasse possessed Monod-type degradation kinetics with a q _max of 0.124 and 0.098 day^?1, respectively. The optimal pH and temperature with the highest degradation rate coefficient of carbofuran were pH 7.5 and 35 °C, respectively.     

Biodegradation of Hexachlorocyclohexane by two species of Bacillus isolated from contaminated soil

Biodegradation of α, β, γ and δ hexachlorocyclohexane (HCH) isomers was studied in broth medium and soil microcosm by Bacillus circulans and Bacillus brevis isolated from contaminated soil. Degradation of α and γ isomers by both the bacterial isolates was higher than thermodynamically stable β and δ isomers. However, B. circulans was found more effective than B. brevis for β and δ isomers. Maximum rate of degradation was recorded at 150 mg/L followed by 100 and 50 mg/L. Soil microcosm study revealed maximum degradation of HCH isomers in the treatment containing natural soil, pesticide and bacterial inocula than the treatment having sterilised soil, pesticide and bacterial isolates. Chloride release was positively co-related with HCH degradation in broth medium as well as in soil microcosm, suggesting that B. circulans and B. brevis hold promising potential by having efficient enzyme(s) required for dechlorination of HCH from contaminated sites.     

Biodegradation of carbofuran in sequencing batch reactor augmented with immobilised Burkholderia cepacia PCL3 on corncob

The performance of sequencing batch reactors (SBRs) augmented with immobilised Burkholderia cepacia PCL3 on corncob for biodegradation of carbofuran in basal salt medium (BSM) was studied. A 2.0-L SBR with a working volume of 1.5 L was operated for a total cycle of 48 h, consisting of 1.0 h fill phase, 46 h react phase and 1.0 h decant phase. The initial pH of the feed medium was 7.0. Air was fed into the reactor at a controlled flow rate of 600 mL·min ^?1. The effect of hydraulic retention time (HRT) (14 to 6 days) on carbofuran-degradation efficiency was investigated at a carbofuran concentration in the feed medium of 20 mg·L ^?1. The shortest HRT resulting in complete degradation of carbofuran was 8 days. At 75% of the optimum HRT (6 days), the effects of biostimulation using organic amendments, i.e. molasses, cassava pulp, rice bran and spent yeast, and the effect of carbofuran concentration in the feed medium (20–80 mg·L ^?1) were investigated. The optimum conditions for SBRs were an initial carbofuran concentration of 40 mg·L ^?1 and 0.1 g·L ^?1 of rice bran as a biostimulated amendment. Complete degradation of carbofuran with a first-order kinetic constant (k ₁) of 0.044 h^?1 was achieved under these optimum conditions.     

Changes in desert steppe soil culturable bacteria from northwestern China and correlation with physicochemical parameters

Seasonal variations effects on desert steppe soil culturable bacteria in northwestern China and physicochemical parameters were investigated. Soil temperature ranged from 5.37 to 23.73 °C. Moisture content varied from 3.22% to 7.62%. The pH was between 7.98 and 8.72. Soil nutrients were altered in different seasons. Total potassium (K) remained the same, but available K levels ranged from 101.29 to 227.55 mg/kg. The culturable bacterial counts in January, April, July and October were 0.13 × 10⁷, 4.09 × 10⁷, 5.33 × 10⁷ and 1.8 × 10⁷ colony forming units (cfu)/g, respectively. Bacterial populations were most abundant in surface layers and declined with increase of soil depth. Seventy-two bacterial strains were isolated and classified according to 27 genera of 5 phyla groups: Firmicutes, Actinobacteria, Alphaproteobacteria, Gammaproteobacteria and Bacteroidates. The bacterial diversities varied according to seasons. In spring, only Firmicutes and Actinobacteria were isolated. In summer, Firmicutes, Actinobacteria and Bacteroidetes were detected with Firmicutes the dominant species. In autumn and winter, Firmicutes, Actinobacteria, α-Proteobacteria and γ-Proteobacteria predominated. Our results indicated that climatic variations, vegetation coverage and soil physicochemical parameters are critical factors for maintaining culturable bacterial populations and diversity in the desert steppe. Soil moisture content and temperature exerted marked influence on bacterial quantities and diversities.     

Mercuric chloride effects on adult rat oval cells-induced apoptosis

In the present investigation, the toxicity of mercuric chloride (HgCl₂) was evaluated in adult oval cells isolated from rat utilizing the 2-acetylaminofluorene/partial hepatectomy technique. Isolated oval cells were incubated with 5 μM of HgCl₂ for 8 hr to elucidate in vitro cytotoxic responses. Recently, autophagic cell death was found in rat hepatocytes in vitro within 30 min of incubation with 5 μM of mercury (Hg) which triggered apoptosis and necroptosis in a time-dependent manner. Nuclear degradation occurred within 30 min of incubation and progressed with time until 8 hr. Apoptosis evidenced by activation of caspase-dependent pathway between 30 min to 8 hr of incubation was mediated via interchange of death domain signaling pathways. Receptor-interacting protein played a positive role to modulate the death domain receptors in the scenario of apoptotic death of oval cells until 6 hr. Autophagic marker proteins ATG12 and LC3B exerted a significant role in triggering apoptosis in 5 μM Hg-treated oval cells. No apparent expression of apoptosis-inducing-factor (AIF) and HMGB1 indicated absence of caspase-independent apoptosis and necrosis in the rat oval cells between 30 min and 8 hr. Thus a low concentration of Hg modulates programmed cell death in adult rat oval cells by altering expression of proteins involved in the molecular mechanisms of cellular functions.     

Influence of 1,2,4,5-tetrazine derivatives on growth of bacterial consortium isolated from soil

This article presents the results of laboratory studies of the influence of tetrazine derivatives on the growth kinetic parameters of soil bacteria. 3,6-Dihydrazinotetrazine (DHTz), 3,6- bis(3,5-dimethylpyrazol-1-yl)-dihydro-1,2,4,5-tetrazine (DMPDHT) and N,N′-bis(1,2,4,5-tetrazine-6-(3,5-dimethylpirazylo))hydrazine (BDMPT) were applied. 3,6-Dihydrazinetetrazine had the largest influence on the growth of bacteria, reflected in a significant lengthening of the lag-phase and a decrease in the specific growth rate. Dehydrogenase activity was also determined in bacterial cultures exposed to tetrazine derivatives. Dehydrogenases remained active even at DHTz concentrations of 80 mg · L^?1, which completely inhibited bacterial growth. The compounds studied variously influence the kinetics of growth in the bacterial consortium; at the same time, they undergo biodegradation in soil by autochthonous microflora.     

Acute toxicity I: effect of profenofos and triazophos (organophosphates) and carbofuran and carbaryl (carbamates) to Labeo rohita

Profenofos and trizaophos (organophosphates) carbofuran and carbaryl (carbamates) are among the major toxicants polluting freshwater bodies, as well as exerting a significant effect on the health of fish. In this study, the 96 h LC₅₀ of the most commonly used organophosphates and carbamates was determined in one of the indigenous fish species, Labeo rohita fingerlings. Pesticides were applied to fingerlings that had been grown under optimized standard conditions under a maintained static bioassay system. Probit analysis was used for the determination of LC₅₀ values, which were ascertained as 6.64, 0.32, 1.4, and 8.24 mg/L for triazophos, profenofos, carbofuran, and carbaryl, respectively. Acute toxic stress was noted with fish exhibiting behavioral toxicity including suffocation, lying on the bottom, erratic swimming, lethargy and downward movements, and gulping prior to mortality.     

Effect of poultry,cattle, sheep manures and sewage sludge on N mineralisation

The effects of various organic wastes on nitrogen mineralisation in soil were investigated. For this purpose, poultry (pl), cattle (ct), sheep (sh) manures and sewage sludge were used as organic wastes. This study was conducted as a laboratory incubation experiment with a calcareous soil. Organic wastes (3%) were added to pots of soil and incubated at 28°C for 16 weeks under non-leaching conditions. The cumulative mineralised N was then fitted to a single exponential model. Maximum nitrogen mineralisation was determined in the second week of incubation in soil treated with poultry manure. Overall, the results showed that the amount of mineral nitrogen in soil treated with different organic wastes was controlled by the type of manure. The results indicated that among the organic wastes, sewage sludge induced the highest quantities of net N mineralisation. Generally, organic wastes increased the amount of N mineralisation in the studied soil and the values of N₀ and k in treated soil varied depending on the type of organic waste. The highest N₀ and k values were found in poultry-treated soil.     

In vitro genotoxicity of extracellular extract from Nostoc piscinale

The extracellular extract obtained from 3 weeks incubation of the soil isolate cyanobacterium strain Nostoc piscinale GT-319 in BG-11 broth medium showed cytotoxic activity against Chinese Hamster Ovary (CHO) cells. Based on comet assay, a concentration of 333 µg mL^?1 (IC₅₀) produced DNA breakages in CHO cells. The concentration of 481 mg kg^?1 (LD₅₀) produced acute toxicity in mice at 48 h.     

Single-species bacteria in sediments induce larval settlement of the infaunal polychaetes Polydora cornuta and Streblospio benedicti

Larval settlement of the infaunal spionid polychaetes Polydora cornuta and Streblospio benedicti is mediated by sediment-associated microorganisms. To investigate if larval preference for certain sediment is guided by individual sediment-associated bacteria, 13 bacterial isolates (5 phyla) obtained from the natural habitat of adult polychaetes (Wadden Sea, Germany) in 2008 were screened in still-water, no-choice settlement assays. Two isolates (α-Proteobacterium-Strain DF11 and Flavobacterium-Strain 54) significantly triggered larval settlement in comparison with sterile sediment. In still-water, multiple-choice settlement assays comprising natural and sterile sediment and sediment re-inoculated with isolates DF11 and 54, significant preferences for natural sediment and sediment containing bacterial isolates at 10⁸ cells g^?1 were observed. Larval settlement was influenced by bacterial abundance in sediment but the correlation was not strictly positive; thus, maximum larval settlement in response to single bacterial species may occur at certain optimum densities. Non-viable or suspended bacteria and water-soluble bacterial products did not induce larval settlement, suggesting that sediment-associated bacterial settlement cues for P. cornuta and S. benedicti were either produced in situ and/or consisted of heat-labile bacterial products.     

Organochlorine pesticides (HCHs and DDTs) in soils along the north coastal areas of the Bohai Sea,China

A systematic survey of organochlorine pesticides (OCPs) including hexachlorocyclohexane isomers (α-HCH, β-HCH, γ-HCH, δ-HCH and ΣHCH) and dichlorodiphenyltrichloroethane metabolites (p,p′-DDT, p,p′-DDE, o,p′-DDT, p,p′-DDD and ∑DDT) in soils along the north coastal areas of the Bohai Sea, China, has been lacking. In this study, 31 representative surface soil samples were collected along the north coastal and riverine areas of the Bohai Sea to characterise the potential for adverse effects of ∑HCH, ∑DDT and their individual isomers and transformation products. Concentrations of ΣHCH and ΣDDT in soils ranged from less than the limit of detection (1 ng · g^?1 dw (mean: 3.5 ng · g^?1 dw) and2 ng · g^?1 dw (mean: 1.7 × 10¹ ng · g^?1 dw), respectively. Compared with studies of OCPs in soils from other locations, concentrations of HCHs and DDTs observed in this study were moderate. Concentrations of OCPs observed in soils were generally less than proposed reference values. HCH residues were a mixture of historical technical HCH and current lindane sources. The pattern of DDTs was consistent with historical releases of technical DDTs. Selected soil physicochemical properties did not explain the sorption and/or partitioning of HCHs or DDTs.     

Application of eggshell waste for the immobilization of cadmium and lead in a contaminated soil

Liming materials have been used to immobilize heavy metals in contaminated soils. However, no studies have evaluated the use of eggshell waste as a source of calcium carbonate (CaCO₃) to immobilize both cadmium (Cd) and lead (Pb) in soils. This study was conducted to evaluate the effectiveness of eggshell waste on the immobilization of Cd and Pb and to determine the metal availability following various single extraction techniques. Incubation experiments were conducted by mixing 0–5% powdered eggshell waste and curing the soil (1,246 mg Pb kg^?1 soil and 17 mg Cd kg^?1 soil) for 30 days. Five extractants, 0.01 M calcium chloride (CaCl₂), 1 M CaCl₂, 0.1 M hydrochloric acid (HCl), 0.43 M acetic acid (CH₃COOH), and 0.05 M ethylendiaminetetraacetic acid (EDTA), were used to determine the extractability of Cd and Pb following treatments with CaCO₃ and eggshell waste. Generally, the extractability of Cd and Pb in the soils decreased in response to treatments with CaCO₃ and eggshell waste, regardless of extractant. Using CaCl₂ extraction, the lowest Cd concentration was achieved upon both CaCO₃ and eggshell waste treatments, while the lowest Pb concentration was observed using HCl extraction. The highest amount of immobilized Cd and Pb was extracted by CH₃COOH or EDTA in soils treated with CaCO₃ and eggshell waste, indicating that remobilization of Cd and Pb may occur under acidic conditions. Based on the findings obtained, eggshell waste can be used as an alternative to CaCO₃ for the immobilization of heavy metals in soils.     

Arbuscular mycorrhiza confers lead tolerance and uptake in Miscanthus sacchariflorus

The effects of an arbuscular mycorrhizal fungi (AMF) association on the growth, survival capabilities, nutrients and lead (Pb) uptake of Miscanthus sacchariflorus under different Pb concentrations were studied in the form of pot cultures. The treatments comprised inoculation or non-inoculation of the AMF, Gigaspora margarita, and the addition of three Pb concentrations to the soil (0, 100 and 1000?mg?kg^?1). The addition of Pb significantly decreased mycorrhizal colonisation. The inoculation of AMF with Pb increased chlorophyll content, Fv/Fm, total dry mass, indole-3-acetic acid (IAA), total nitrogen, and total phosphorus, whereas H₂O₂ level, indole-3-acetic acid oxidase (IAAO) activity, and peroxidase (POD) activity were low compared to those in the non-inoculated treatments. Moreover, the application of AMF together with Pb doses induces concentrations of Pb in the plant, where the higher dose of Pb (1000?mg?kg^?1) induces a lower content of Pb in the aerial part of the plant but a higher content in the root. G. margarita enhanced the tolerance of M. sacchariflorus against Pb toxicity, and facilitated the accumulation of Pb in the plant roots, whereas translocation to the shoots was inhibited at the highest dose Pb (1000?mg?kg^?1). However, in contaminated soil, the Pb removal capability of M. sacchariflorus with AMF was remarkable.     

Phytoavailability and potential transfer of Pb from a salt-affected soil to Atriplex verucifera,Salicornia europaea and Chenopodium album

The phytoavailability and potential transfer of Pb to Atriplex verucifera, Salicornia europaea and Chenopodium album in two calcareous soils with different salinity/sodicity were compared. The soils were spiked with 0, 250, 500 and 1000 mg Pb kg^?1 soil. Plant shoots were harvested and analysed for total Pb after they had been grown in the contaminated soils. Visual MINTEQ 3.0 was used to calculate the speciation of soluble Pb in the experimental soils. Results showed that although the concentrations of 1 M NH₄NO₃-extractable Pb were relatively similar, speciation of Pb in the soils were not the same. Salicornia europea was found to be the most salinity/sodicity-tolerant plant. When the plants were grown in non-saline soil, the Pb tolerance of the three plants was as follows: A. verucifera > C. album >S. europea, whereas in saline (sodic) soil, Pb tolerance was in the order S. europea > C. album > A. verucifera. Lead phytotoxicity to A. verucifera and C. album was higher in the saline soil, whereas for S. europea, Pb toxicity was higher in the non-saline soil. It could be concluded that the phytoavailability of Pb and its interactions with plants are widely dependent on soil salinity level and type of plant.     

Impact of natural and calcined starfish (<Emphasis Type="Italic">Asterina pectinifera</Emphasis>) on the stabilization of Pb,Zn and As in contaminated agricultural soil

Metal stabilization using soil amendments is an extensively applied, economically viable and environmentally friendly remediation technique. The stabilization of Pb, Zn and As in contaminated soils was evaluated using natural starfish (NSF) and calcined starfish (CSF) wastes at different application rates (0, 2.5, 5.0 and 10.0 wt%). An incubation study was conducted over 14 months, and the efficiency of stabilization for Pb, Zn and As in soil was evaluated by the toxicity characteristic leaching procedure (TCLP) test. The TCLP-extractable Pb was reduced by 76.3–100 and 91.2–100 % in soil treated with NSF and CSF, respectively. The TCLP-extractable Zn was also reduced by 89.8–100 and 93.2–100 % in soil treated with NSF and CSF, respectively. These reductions could be associated with the increased metal adsorption and the formation of insoluble metal precipitates due to increased soil pH following application of the amendments. However, the TCLP-extractable As was increased in the soil treated with NSF, possibly due to the competitive adsorption of phosphorous. In contrast, the TCLP-extractable As in the 10 % CSF treatment was not detectable because insoluble Ca–As compounds might be formed at high pH values. Thermodynamic modeling by visual MINTEQ predicted the formation of ettringite (Ca₆Al₂(SO₄)₃(OH)₁₂·26H₂O) and portlandite (Ca(OH)₂) in the 10 % CSF-treated soil, while SEM–EDS analysis confirmed the needle-like structure of ettringite in which Pb was incorporated and stabilized in the 10 % CSF treatment.     

Carbofuran-induced effects on acetylcholinesterase (AChE) in erythrocytes and liver of Cyprinus carpio

A study was conducted to determine the effects of long-term exposure to carbofuran on erythrocyte and liver acetylcholinesterase (AChE) activity in Cyprinus carpio, and assess which tissue was more sensitive to pesticide exposure examining this enzyme. Fish were allowed to accilimatize in toxicant-free water for 24 days after 60-day exposure. AChE activity was determined with a spectrophotometer using acetylthiocholine iodide as a substrate in erythrocytes and liver. Enzymic activity of erythrocytes and liver decreased over time. A higher degree of enzyme inhibition was observed in erythrocyte compared to liver. The degree of enzyme inhibition was positively correlated with exposure time. When exposed fish were transferred to clean water, recovery was greater in erythrocytes than liver. Erythrocyte AChE activity recovered after 18 days, while it required 21 days in liver. The findings of this study indicate that erythrocyte AChE is more sensitive to carbofuran than liver. The greater sensitivity of erythrocyte AChE suggests that it may be more useful as a biomarker for monitoring status of pollution in freshwater bodies.     

Performance evaluation of isoproturon-degrading indigenous bacterial isolates in soil microcosm

Isoproturon (IPU)-degrading soil bacteria were isolated from herbicide-applied wheat fields. These isolates were identified using cultural, morphological, biochemical and 16S rRNA sequencing methods. 16S rRNA sequences of both the bacterial isolates were compared with NCBI GenBank data base and identified as Bacillus pumilus and Pseudoxanthomonas sp. A soil microcosm study was carried out for 40 days in six different treatments. Experimental results revealed maximum 95.98% IPU degradation in treatment 6 where bacterial consortia were augmented in natural soil, followed by 91.53% in treatment 5 enriched with organic manure as an additional carbon source. However, only 14.03% IPU was degraded in treatment 1 (control) after 40 days. In treatments (2–4), 75.59%, 70.92% and 77.32% IPU degradation was recorded, respectively. IPU degradation in all the treatments varied significantly over the control. 4-Isopropylaniline was detected as IPU degradation by-product in the medium. The study confirmed that B. pumilus and Pseudoxanthomonas sp. performed effectively in soil microcosms and could be employed profitably for field-scale bioremediation experiments.     

Effects of paclobutrazol (PP333) on lead and zinc accumulations in Pseudostellaria maximowicziana

Paclobutrazol (PP333) can enhance the resistance capabilities of plants to stress conditions. In this study, PP333 were sprayed on the lead (Pb) and zinc (Zn) accumulator plant Pseudostellaria maximowicziana, which was planted in Pb–Zn contaminated soil, and the effects of PP333 on Pb and Zn accumulation levels in P. maximowicziana were studied. Spraying 10?mg/L PP333 increased, while 20, 30 and 50?mg/L PP333 decreased, the biomass of P. maximowicziana compared with the control. The 10?mg/L PP333 had no significant effects on the photosynthetic pigment contents of P. maximowicziana compared with the control, while the other doses increased the contents. The activities of antioxidant enzymes (superoxide dismutase, peroxidase and catalase) and the Pb and Zn concentrations in P. maximowicziana were increased by PP333 compared with the control. These items had the increase trend with the increase of PP333 concentrations. Only 10 and 20?mg/L PP333 increased the amount of Pb extracted by P. maximowicziana shoots, while all of the doses increased the amount of Zn extracted by P. maximowicziana shoots. Thus, low concentration of PP333 could promote the growth and heavy metal extraction ability of P. maximowicziana shoots, with the 10?mg/L being the best.     

Effects of mutual intercropping on Pb and Zn accumulation of accumulator plants Rumex nepalensis,Lolium perenne and Trifolium repens

Intercropping is one of the most potential ways to remediate contaminated soil. In this study, three plants, including Rumex nepalensis which is a lead (Pb) accumulator plant, Lolium perenne which is tolerant to zinc (Zn) and a native plant Trifolium repens, were mutually intercropped in pots to study the effects of intercropping on co-remediation of Pb-Zn-contaminated soil from the Tangjia lead-zinc mine. Compared to the respective monoculture, the dry weights of R. nepalensis and T. repens were decreased under intercropping, while that of L. perenne was increased. The concentrations of total Zn in the three plants were increased by intercropping in which roots were higher than shoots. Besides, intercropping increased the accumulation of Zn in R. nepalensis and L. perenne per plant. The accumulations of heavy metals for R. nepalensis and T. repens in a single pot were enhanced by intercropping. Both of the maximum values of heavy metals’ accumulation were produced by three-species intercropping (7819.31?μg/pot for Pb, 12576.05 μg/pot for Zn) in terms of a single pot. These results indicated that three-species intercropping could promote the phytoremediation efficiency for Tangjia lead-zinc mine, and thus further proved intercropping is an effective method to remediate the Pb-Zn-contaminated soil by R. nepalensis and T. repens.     

Effect of light and temperature on the population dynamics of two toxic bloom forming Cyanobacteria – Microcystis ichthyoblabe and Anabaena aphanizomenoides

The effect of light and temperature on the growth of Microcystis ichthyoblabe and Anabaena aphanizomenoides, isolated from the subtropical Oued Mellah lake, Morocco (33°30′N–07°20′W), were investigated in batch culture. Growth rates at 66 light–temperature combinations were determined and fitted with different mathematical models. The results show that the two Cyanobacteria grow at all light intensities and temperatures, except at 10 °C for A. aphanizomenoides, where the growth was strongly limited. The μ_max of M. ichthyoblabe increased with temperature from 0.56 d^?1 at 10 °C to 1.32 d^?1 at 35 °C. At all tested temperatures, a relative photoinhibition within the studied range of irradiance was observed and the photosensitivity was thermodependent. For Anabaena, the obtained μ_max ranged between 0.07 d^?1 at 10 °C and 1.46 d^?1 at 35 °C, and a weak photoinhibition was observed at 15 °C. The positive correlation between μ_max and I_opt (r²≥0.93) indicates a close interaction between light and temperature on the cyanobacteria growth. The results obtained in this work suggest that the growth of these two species is possible under low light and low temperature.