Fractions composition study of the pyrolysis oil obtained from sewage sludge treatment plant

In this work the parameters of Low Temperature Conversion - LTC were applied in a centrifuged sludge from a sewage treatment plant located in Rio de Janeiro, Brazil. Before the conversion, the sludge was dried and analyzed by TGA to observe its behavior with increasing temperature. The chemical composition of the crude pyrolysis oil was analyzed by FTIR, ¹H NMR and GC-MS. The results showed that the oil is a mixture of hydrocarbons, oxygenated and nitrogenated compounds. Using a catalytic treatment it was possible to fractionate the oil where the predominant constituents were hydrocarbons showing that the cracking was effective. An important result was the difference between the calorific value of dry sludge (10 MJ kg⁻¹), the pyrolysis oil (36 MJ kg⁻¹) and one of the fractions separated by catalytic cracking (40 MJ kg⁻¹) when compared with commercial diesel (45 MJ kg⁻¹).     

Bio-separation, speciation and determination of chromium in water using partially pyrolyzed olive pomace sorbent

Speciation of Cr(III)/Cr(VI) from water using olive pomace (OP) was improved by partial pyrolysis of OP. The sorbents were characterized by physicochemical techniques. Sorption of Cr(III) on raw and partially pyrolyzed OP sorbents followed Freundlich isotherm and second-order rate kinetics. OP pyrolyzed at 150 °C (sorbent OP-150) exhibited maximum sorption capacity, favorability and the lowest sorption energy. Sorption was exothermic and spontaneous for the raw-OP and OP pyrolyzed at 100 or 150 °C; but endothermic and non-spontaneous for OP pyrolyzed at 200, 250, 300 or 400 °C. A speciation method of chromium was proposed, in which Cr(III) was selectively retained at pH 3 on sorbent OP-150; while total Cr was determined after reduction of Cr(VI). The method was selective with a detection limit for Cr(III) of 1.58 μg L⁻¹. The method was applied on natural and industrial waters (recoveries >97.7%, RSD’s <9%) and on tobacco leaves certified reference material (INCT-PVTL-6).     

Effects of Cr(VI) on the performance and kinetics of the activated sludge process

The substrates removal performance, removal kinetics and the electron transport system (ETS) of sludge were investigated by sequencing batch reactors (SBR) and batch assays, respectively. Compared to the control system, significant decreases were observed in substrate removal efficiency with the Cr(VI)-feeding concentration up to 5 mg L⁻¹ in SBR system. And the recovery for NH₄⁺-N removal were more difficult than that of COD after the termination of Cr(VI)-feeding. Significant inhibitory effects of Cr(VI) on the ETS activity and substrate removal kinetics were observed in the batch assays. The inhibitory effects of Cr(VI) would be overestimated on COD removal and underestimated on NH₄⁺-N removal by the short-term batch assay as compared to the long-term operations. Additionally, significant correlations between the ETS activity and the inhibitory rates of Cr(VI) on substrate removal indicated the ETS activity can provide effective predictions on the potential performance of substrate removal in activated sludge.     

Exposure to Cr(VI) induces organ dependent MSI in two loci related with photophosphorylation and with glutamine metabolism

Chromium (Cr), as a mutagenic agent in plants, has received less attention than other metal pollutants. To understand if Cr induces microsatellite instability (MSI), Pisum sativum seedlings were exposed for 28 days to different concentrations of Cr(VI) up to 2000 mg L⁻¹, and the genetic instability of ten microsatellites (SSRs) was analyzed. In plants exposed to Cr(VI) up to 1000 mg  L⁻¹, MSI was never observed. However, roots exposed to 2000 mg L⁻¹ displayed MSI in two of the loci analyzed, corresponding to a mutation rate of 8.3%. SSR2 (inserted in the locus for plastid photosystem I 24 kDa light harvesting protein) and SSR6 (inserted in the locus for P. sativum glutamine synthetase) from Cr(VI)-treated roots presented alleles with, respectively, less 6 bp and more 3 bp than the corresponding controls. This report demonstrates that: (a) SSRs technique is sensitive to detect Cr-induced mutagenicity in plants, being Cr-induced-MSI dose and organ dependent (roots are more sensitive); (b) two Cr-sensitive loci are related with thylakoid photophosphorylation and with glutamine synthetase, respectively; (c) despite MSI is induced by Cr(VI), it only occurs in plants exposed to concentrations higher than 1000 mg L⁻¹ (values rarely found in real scenarios). Considering these data, we also discuss the known functional changes induced by Cr(VI) in photosynthesis and in glutamine synthetase activity.     

Detoxification and accumulation of chromium from tannery effluent and spent chrome effluent by Paecilomyces lilacinus fungi

The tannery industry process involves chromium (Cr) salts as a main constituent of the process. The Cr recovery is a part of the process where other salts are used to achieve separation and recovery for using Cr back in the process. The process steps may contain both forms of Cr [Cr(VI): hexavalent and Cr(III): trivalent]. The recovery of Cr from tannery industry effluent through biological systems is much needed. The diverse physicochemical characteristics of these effluents may limit the growth of microorganisms and hence the limitation towards possible practical application of microorganisms in real industrial effluent conditions. The present study attempted the ability of the Cr-resistant fungus Paecilomyces lilacinus [isolated through an enrichment culture technique at 25 000 mg l⁻¹ of Cr(III)] to grow and remove Cr [Cr(VI) and Cr(III)] from two physicochemically different undiluted tannery industry effluents (tannery effluent and spent chrome effluent) in the presence of cane sugar as a carbon source. Such attempts are made keeping in view the potential integration of biological processes in the overall Cr removal and recovery processes to improve its efficiency and environmental sustainability. The fungus has broad pH tolerance range and can reduce Cr(VI) both in acidic (pH 5.5) and alkaline (pH 8.0) conditions. The fungus showed the ability to remove Cr(VI) (1.24 mg l⁻¹) and total Cr (7.91 mg l⁻¹) from tannery effluent below the detection level within 18 h and 36 h of incubation, respectively, and ability to accumulate 189.13 mg Cr g⁻¹ of dry biomass within 600 h of incubation from spent chrome effluent [containing 3731.4 mg l⁻¹ of initial Cr(III) concentration].At 200 mg l⁻¹ of Cr(VI) in growth media, with 100% detoxification and with only 10.54% of total Cr accumulation in the biomass, P. lilacinus showed Cr(VI) reduction as a major mechanism of Cr(VI) detoxification. The time-course study revealed the log phase of the growth for the maximum specific reduction of Cr(VI) and stationary phase of the growth for its maximum specific accumulation of both the forms of Cr [Cr(III) and Cr(VI)] in its biomass. In growth media at 50 mg l⁻¹ and 200 mg l⁻¹ of Cr(VI), P. lilacinus showed 100% reduction within 36 h and 120 h of incubation, respectively. The high degree of positive correlation and statistically high degree of relationship (r² = 0.941) between the fungal growth and % Cr(VI) reduction by the fungus support the role of metabolically active cellular growth in Cr(VI) reduction by the fungus. Results indicate that expanded solid (sludge) retention times (SRTs) (stationary phase) can be recommended for the removal of Cr(III) through accumulation. In case of Cr(VI), reduction needs a priority; therefore, a non-expanded SRT is recommended for designing a continuous-flow completely stirred bioreactor so that a log phase of cellular growth can be maintained during the reduction process. This study reveals the strong potential of P. lilacinus fungi for the removal of Cr from tannery effluent and spent chrome effluent.     

Accumulation, speciation, and coordination of arsenic in an inbred line and a wild type cultivar of the desert plant species Chilopsis linearis (Desert willow)

This study investigated the absorption of arsenic (As), sulfur (S), and phosphorus (P) in the desert plant Chilopsis linearis (Desert willow). A comparison between an inbred line (red flowered) and wild type (white flowered) plants was performed to look for differential responses to As treatment. One month old seedlings were treated for 7 days with arsenate (As₂O₅, As^V) at 0, 20, and 40 mg As^V L⁻¹. Results from the ICP-OES analysis showed that at 20 mg As^V L⁻¹, red flowered plants had 280 ± 11 and 98 ± 7 mg As kg⁻¹ dry wt in roots and stems, respectively, while white flowered plants had 196 ± 30 and 103 ± 13 mg As kg⁻¹ dry wt for roots and stems. At this treatment level, the concentration of As in leaves was below detection limits for both plants. In red flowered plants treated with 40 mg As^V L⁻¹, As was at 290 ± 77 and 151 ± 60 mg As kg⁻¹ in roots and stems, respectively, and not detected in leaves, whereas white flowered plants had 406 ± 36, 213 ± 12, and 177 ± 40 mg As kg⁻¹ in roots, stems, and leaves. The concentration of S increased in all As treated plants, while the concentration of P decreased in roots and stems of both types of plants and in leaves of red flowered plants. X-ray absorption spectroscopy analyses demonstrated partial reduction of arsenate to arsenite in the form of As-(SX)₃ species in both types of plants.     

Simultaneous Cr(VI) reduction and phenol degradation in pure cultures of Pseudomonas aeruginosa CCTCC AB91095

Simultaneous Cr(VI) reduction and phenol degradation were investigated in a reactor containing Pseudomonas aeruginosa CCTCC AB91095. Phenol was used as carbon source. P.aeruginosa utilized metabolites formed during phenol degradation as energy source for Cr(VI) reduction. Cr(VI) inhibited both Cr(VI) reduction and phenol degradation when Cr(VI) concentration exceeded the optimum value (20 mg/L), whereas phenol enhanced both Cr(VI) reduction and phenol degradation below the optimum initial concentration of 100 mg/L. Cr(III) was the predominant product of Cr(VI) reduction in cultures after incubation for 24 h. Both Cr(VI) reduction and phenol degradation were influenced by the amount of inocula. The concentration of Cr(VI) and phenol declined quickly from 20, 100 to 3.36, 29.51 mg/L in cultures containing of 5% (v/v) inoculum after incubation for 12 h, respectively. The whole study showed that P. aeruginosa is promising for the reduction of toxic Cr(VI) and degradation of organic pollutants simultaneously in the mineral liquid medium.     

Effect of Organic Amendments on the Oxygen Uptake of Pseudomonas putida—PAPs-1 in Chromium-Contaminated Pond Ash

In this study a Cr (VI) resistant bacterium Pseudomonas putida was isolated from pond ash and its oxygen consumption potential at different concentrations of Cr (VI) viz., 0, 100 and 200 mg kg^?1 was studied using Electrolytic Respirometry. Oxygen consumption by the bacterium was noticed up to 200 mg kg^?1 Cr (VI) concentration. To the pond ash (inoculated with and without Pseudomonas) 200 mg kg^?1 Cr (VI) was added and incorporated with different organic amendments such as farmyard manure (FYM), coir pith, paddy straw and press mud and the cumulative oxygen consumption was studied. The cumulative oxygen consumption by the bacterium was higher when the pond ash was incorporated with organic amendments. The highest oxygen consumption of 205 mg l^?1 was observed when press mud was used, which was followed by FYM (198 mg l^{? 1} ). Furthermore, the enrichment with press mud increased the nutrient content of N (57.28 mg kg^?1 ), P (5.5 mg kg^?1 ) and K (42.7 mg kg^?1 ) of the pond ash. The maximum dehydrogenase enzyme activity of 0.63 μ g TPF formed g^?1 sample h^?1 was measured when the pond ash was inoculated with Pseudomonas and enriched with press mud. The results also indicated that maximum reduction of Cr (VI) (42.5%) was observed when Pseudomonas and press mud were used. This study evaluated the possibilities of toxicity reduction and nutrient enrichment of the ash pond using a Cr (VI) resistant bacterium and organic amendments.     

Influence of magnetic field on Cr(VI) adsorption capability of given anaerobic sludge

To provide beneficial guide for the application of the magnetic field in the bio-treatment of the Cr(VI)-contained wastewater, sludge samples from the control bio-system A (absent of magnetic field) and the contrast bio-system B (present of magnetic field) were used to adsorb the synthetic wastewater with 100 mg l⁻¹ Cr(VI). Influences of two adsorption modes, single adsorption and once continuous adsorption, on the Cr(VI) adsorption capacities of both sludge samples were compared. And the influence of regeneration on the Cr(VI) adsorption capacities were also studied. The results of adsorption experiments showed that the Cr(VI) adsorption capacities of the first single adsorption for sludge sample A and B were pretty nearly, which were 9.79 and 9.93 mg, respectively. And after 5 single adsorption periods, the total Cr(VI) adsorption capacity and efficiency of the sample B were 25.88 and 55.66 mg Cr(VI) g⁻¹VSS, while those of the control were 14.95 and 33.98 mg Cr(VI) g⁻¹VSS, respectively. For the sludge sample A and B after a single adsorption, both functions of regeneration were remarkable. But after 13 cycles of the single adsorption-regeneration, the Cr(VI) adsorption capacity and efficiency of the sample B were 110.15 and 189.91 mg Cr(VI) g⁻¹VSS, while those of the control were 70.89 and 140.38 mg Cr(VI) g⁻¹VSS, respectively. Though the Cr(VI) adsorption capacity of a once continuous adsorption period was more than that of a single adsorption period obviously, the Cr(VI) removal rates of the sludge sample A and B in the third period of once continuous adsorption-regeneration were only 8.12 and 33.51%, respectively. It was concluded that the weak magnetic field did improve the Cr(VI) bio-removal efficiency and the sludge stability, the batch treatment was an ideal operation mode for the bio-treatment of the Cr(VI)-contained wastewater, as compared with the continuous operation mode, but regeneration and enough sludge content were two necessary conditions to ensure the efficiency of batch treatment.     

Biodegradation by activated sludge and toxicity of tetracycline into a semi-industrial membrane bioreactor

Much attention has been devoted recently to the fate of pharmaceutically active compounds such as tetracycline antibiotics in soil and water. Tetracycline (TC) biodegradability by activated sludge derived from membrane bioreactor (MBR) treating swine wastewater via CO₂-evolution was evaluated by means of modified Sturm test, which was also used to evaluate its toxicity on carbon degradation. The impact of tetracycline on a semi-industrial MBR process was also examined and confronted to lab-scale experiments. After tetracycline injection in the pilot, no disturbance was detected on the elimination of organic matters and ammonium (nitrification), reaching after injection 88% and 99% respectively; only denitrification was slightly affected. Confirming the ruggedness and the superiority of membrane bioreactors over conventional bioreactors, no toxicity was observed at the considered level of TC in the pilot (20 mg TOC L⁻¹), while at lab-scale sodium benzoate biodegradation was completely inhibited from 10 mg TOC L⁻¹ TC. The origin of the activated sludge showed a significant impact on the performances, since the ultimate biodegradation was in the range −50% to −53% for TC concentrations in the range 10–20 mg TOC L⁻¹ with conventional bioreactor sludge and increased to 18% for 40 mg TOC L⁻¹ of TC with activated sludge derived from the MBR pilot. This confirmed the higher resistance of activated sludge arising from membrane bioreactor.     

Zinc incorporation in the otoliths of juvenile pink snapper (Pagrus auratus Forster): The influence of dietary versus waterborne sources

Separate laboratory experiments were conducted to examine if incorporation of Zn into the otoliths of juvenile pink snapper (Pagrus auratus Forster) was related to levels in the food and/or water. In the first experiment, fish were fed a regular diet (600 mg Zn kg^− 1 dw, control group) or a Zn-enriched diet (6000 mg Zn kg^− 1 dw or 9000 mg Zn kg^− 1 dw) for 35 days. In the second experiment, fish were exposed to waterborne Zn concentrations of < 0.005 µg L^− 1 (control), 50 µg L^− 1, 100 µg L^− 1 and 200 µg L^− 1 for 35 days. The sagittal otoliths were analysed using laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS). Juvenile fish exposed to higher concentrations of waterborne Zn did not display increased Zn levels in their otoliths. However, Zn levels in the otoliths of fish consuming the Zn-enriched diet were significantly higher relative to control fish. This study clearly demonstrated that dietary Zn was the major source of Zn incorporated into the otoliths by this marine fish.     

Effect of freezing and dehydration on ion and cryoprotectant distribution and hemolymph volume in the goldenrod gall fly, Eurosta solidaginis

Extracellular freezing and dehydration concentrate hemolymph solutes, which can lead to cellular injury due to excessive water loss. Freeze tolerant larvae of the goldenrod gall fly, Eurosta solidaginis, may experience extreme cold and desiccation in winter. To determine whether larvae employ protective mechanisms against excessive cellular water loss we examined the effect of extracellular freezing and dehydration on hemolymph volume, and cryoprotectant and ion levels in the hemolymph. Dehydrated larvae or ones that had been frozen at −5 or −20 °C had a significantly smaller proportion of their body water as hemolymph (26.0-27.4%) compared to controls (30.5%). Even with this reduction in water content, hemolymph osmolality was similar or only slightly higher in frozen or dehydrated individuals than controls (908 mOsm kg⁻¹), indicating these stresses led to a reduction in hemolymph solutes. Hemolymph and intracellular content of ions remained largely unchanged between treatment groups; although levels of Mg⁺⁺ in the hemolymph were lower in larvae subjected to freezing (0.21 ± 0.01 μg mg⁻¹ dry mass) compared to controls (0.29 ± 0.01 μg mg⁻¹ dry mass), while intracellular levels of K⁺ were lower in groups exposed to low temperature (8.31 ± 0.21 μg mg⁻¹ dry mass). Whole body glycerol and sorbitol content was similar among all treatment groups, averaging 432 ± 25 mOsm kg⁻¹ and 549 ± 78 mOsm kg⁻¹ respectively. However, larvae subjected to dehydration and freezing at −20 °C had a much lower relative amount of cryoprotectants in their hemolymph (∼35%) compared to controls (54%) suggesting these solutes moved into intracellular compartments during these stresses. The correlation between reduced hemolymph volume (i.e. increased cellular water content) and intracellular movement of cryoprotectants may represent a link between tolerance of dehydration and cold in this species.     

Biogeochemical processes in intensive zero-effluent marine fish culture with recirculating aerobic and anaerobic biofilters

The biogeochemical processes that drive nutrient transformations and recycling in organic marine sediment-water environments were studied for 17 months in a zero-effluent intensive recirculating culture system. The system consisted of a 10 m³ gilthead seabream (Sparus aurata) tank coupled to aerobic and anaerobic water treatment elements. Nutrients and alkalinity were measured in the system to quantify the main biogeochemical processes. Fractions of the carbon fed in feed were found in fish (18.3%) and in sludge (11%); the missing carbon was respired by fish (45%) and by aerobic (8.4%) and anaerobic (7.7%) microorganisms. Fractions of the nitrogen fed in feed were found in fish (15.4%) and in sludge (14.3%); the missing nitrogen was eliminated by nitrification-denitrification. Most of the phosphorus and ash fed in feed and not found in fish accumulated within the sludge in the system. The rates of nitrification, denitrification and sulphate reduction increased with time, reaching 0.3 g N m^− 2 d^− 1, 53 g N m^− 2 d^− 1 and 145 g S m^− 2 d^− 1, respectively. Nitrification developed more rapidly than denitrification, leading at first to nitrate accumulation (to 20 mmol NO₃ l^− 1 by day 200) and a decrease in alkalinity. Once denitrification surpassed nitrification, nitrate concentrations decreased, eventually being reduced to < 0.3 mmol NO₃ l^− 1 by day 510, and alkalinity stabilized. Toxic hydrogen sulphide, generated within the anaerobic sludge, was oxidized by oxygen and nitrate as it diffused through the anaerobic-aerobic sediment-water interface. When nitrate levels in the water above the sludge dropped below 2 mmol l^− 1, sulphide was also oxidized in the fluidized bed reactor. Denitrification reduced nitrate in the water, respired (jointly with sulphate reduction) carbon in the sludge, oxidized the hydrogen sulphide, and contributed to stabilization of alkalinity and accumulation of polyphosphate in bacteria as a major sink of labile P.     

Effect of chitosan on UASB treating POME during a transition from mesophilic to thermophilic conditions

The effects of chitosan addition on treatment of palm oil mill effluent were investigated using two lab-scale upflow anaerobic sludge bed (UASB) reactors: (1) with chitosan addition at the dosage of 2 mg chitosan per g volatile suspended solids on the first day of the operation (R1), (2) without chitosan addition (the control, R2). The reactors were inoculated with mesophilic anaerobic sludge which was acclimatized to a thermophilic condition with a stepwise temperature increase of 5 °C from 37 to 57 °C. The OLR ranged from 2.23 to 9.47 kg COD m⁻³ day⁻¹. The difference in biogas production rate increased from non-significant to 18% different. The effluent volatile suspended solids of R1 was 65 mg l⁻¹ lower than that of R2 on Day 123. 16S rRNA targeted denaturing gradient gel electrophoresis (DGGE) fingerprints of microbial community indicated that some methanogens in the genus Methanosaeta can be detected in R1 but not in R2.     

Soil chromium bioremediation: Synergic activity of actinobacteria and plants

The aim of this work was to evaluate a strategy to reduce the bioavailable chromium fraction in soil, using a Cr(VI) resistant microorganism, Streptomyces sp. MC1, under non sterile conditions, with maize plants as bioindicator and/or bioremediator.Soil samples were contaminated with 100, 200 and 400 mg kg⁻¹ of Cr(VI) or Cr(III). Bioavailable chromium (35%) was only detected in samples with Cr(VI). Soil samples with Cr(VI) 200 mg kg⁻¹ were inoculated with Streptomyces sp. MC1, and bioavailable chromium decreased up to 73%.Zea mays seedlings were planted in soil samples contaminated with chromium. Plantlets accumulated chromium mainly as Cr(III), and biomass decreased up to 88%. Streptomyces sp. MC1 was inoculated in soil samples contaminated with 200 mg kg⁻¹ of Cr(VI) and Z.mays seedlings were planted.Streptomyces sp. MC1 caused Z.mays biomass increase (57%), chromium accumulation and bioavailable chromium decreased up to 46% and 96%, respectively.This work constitutes the first contribution of cooperative action between actinobacteria and Z.mays in the bioremediation of Cr(VI) contaminated soil. The large removal capacity of bioavailable chromium by Streptomyces sp. MC1 and Z.mays infers that they could be successfully applied together in bioremediation of soils contaminated with Cr(VI).     

Reduction of high concentrations of chromate by Leucobacter sp. CRB1 isolated from Changsha, China

In recent years, more and more attentions are put on the remediation of Cr(VI) contamination with chromate resistant bacteria. Leucobacter sp. CRB1 was a novel chromate reducing bacteria isolated from the soil of chromite ore processing residue (COPR) disposal site in Changsha, China. The objectives of this study were to evaluate the Cr(VI) tolerance of Leucobacter sp. CRB1 as well as its tolerant mechanism, and Cr(VI) reduction ability. The results showed that Leucobacter sp. CRB1 was able to tolerate 4,000 mg/l of hexavalent chromium with 34.5% reduction efficiency. At the optimum pH 9.0, the maximum concentration of chromate be reduced completely was 1,818 mg/l in growing cells and 2,100 mg/l in resting cells. Scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDAX) showed that extracellular Cr(VI) reduction of Leucobacter sp. CRB1 contributed to its high tolerance and high reduction ability. With repeating spiking, 2,490 mg/l hexavalent chromium was reduced totally within 17 h. The results suggest Leucobacter sp. CRB1 has potential application for remediation of high concentration of Cr(VI) contamination.     

Radionuclides in plants growing on sludge and water from uranium mine water treatment

Chemical treatment of uranium mine acid drainage generates sludge containing high radioactivity levels with, for example, ²³⁸U and ²²⁶Ra concentrations at about 18 and 9 kBq kg⁻¹ dry weight, respectively. Spontaneous vegetation, such as grass (Polygonum sp.), reeds (Phragmites australis), and bullrush (Typha latifolia) growing in sludge dewatering ponds concentrated uranium and uranium daughter radionuclides. However, bullrush growing in natural wetlands by the stream receiving treated mine water discharges contained even higher radionuclide concentrations, e.g., 21 Bq kg⁻¹ dry weight of ²³⁸U, about four times higher than bullrush growing on sludge, probably because the uranium in water is more bioavailable to Typha than uranium in the chemical sludge. It is suggested that wetlands with plant species could be used as a secondary treatment to further reduce radioactivity in chemically treated uranium mine water and to improve water quality in streams receiving treated water discharges. Furthermore, it is concluded that vegetation both from sludge drying ponds and from streams receiving treated water discharges, due to the high radionuclide concentrations in the vegetation, is not suitable as cattle feeding.     

Synthesis and characterization of a molecularly imprinted polymer and its application as SPE enrichment sorbent for determination of trace methimazole in pig samples using HPLC-UV

A novel molecularly imprinted polymer that could be applied as enrichment sorbent was prepared using methimazole (MMZ) as the template molecule, methacrylic acid as functional monomer, ethylene glycol dimethacrylate as cross-linker. Though evaluated by static, kinetic and competitive adsorption tests, the polymer exhibited high adsorption capacity, fast kinetics and good selective ability. A method for determination of trace MMZ was developed using this polymer as enrichment sorbent coupled with high performance liquid chromatography focusing on complex biological matrices. Under the optimum experimental conditions, the MMZ standard is linear within the concentration range studied, that is, from 0.5 μg L⁻¹ to 150 μg L⁻¹ (r² = 0.9941). Lower limits of detection (LOD, at S/N = 3) and quantification (LOQ, at S/N = 10) in pig samples were 0.63 μg kg⁻¹ and 2.10 μg kg⁻¹ for kidney, 0.51 μg kg⁻¹ and 1.70 μg kg⁻¹ for liver, 0.56 μg kg⁻¹ and 1.86 μg kg⁻¹ for muscle, respectively. Recoveries and relative standard deviation (RSD, n = 9) values for precision in the developed method were from 71.14% to 88.41% and from 2.53% to 6.18%.     

Molecular cloning,characterization and mRNA expression of selenium-dependent glutathione peroxidase from abalone Haliotis discus hannai Ino in response to dietary selenium,zinc and iron

A novel selenium-dependent glutathione peroxidase (Se-GPX) was cloned from abalone Haliotis discus hannai Ino (HdhGPx) by homology cloning with degenerate primers and RACE techniques. The full length of HdhGPx cDNA was 963 bp with a 669 bp open reading frame (ORF) encoding 222 amino acids and a 101 bp eukaryotic selenocysteine insertion sequence (SECIS) in 3′ untranslated region (UTR). It was showed that HdhGPx has a characteristic codon at ²³⁵TGA²³⁷ that corresponds to selenocysteine (SeC) as U₇₂. Sequence characterization revealed that HdhGPx contains a characteristic GPx signature motif 2 (₉₆LGLPCNQF₁₀₃), an active site motif (₁₇₉WNFEKF₁₈₄). In addition, two potential N-glycosylation sites (₁₁₂NGTE₁₁₅ and ₁₃₂NLTQ₁₃₅) were identified in HdhGPx. 3D modeling analysis showed that the overall structure of HdhGPx monomer had more similarity to human GPx3 than human GPx1. Relatively higher-level mRNA expression was detected in hepatopancreas, mantle and gonad by real-time PCR assays. The relative expression levels of HdhGPx mRNA in hepatopancreas and haemocytes were detected by real-time PCR in abalone fed with nine different diets containing graded levels of selenium (0.15, 1.32 and 48.7 mg kg^− 1), zinc (6.69, 33.85 and 710.63 mg kg^− 1) and iron (29.17, 65.7 and 1267.2 mg kg^− 1) for 20 weeks, respectively. The results showed that the expressions of HdhGPx mRNA were statistically higher at adequate dietary selenium (1.32 mg kg^− 1), zinc (33.85 mg kg^− 1) and iron (65.7 mg kg^− 1) than those in low dietary minerals, respectively. But HdhGPx mRNA expression levels were down-regulated by high contents of dietary selenium (48.7 mg kg^− 1), zinc (710.63 mg kg^− 1) and iron (1267.2 mg kg^− 1), respectively. These results indicated that adequate dietary minerals could increase the mRNA expression of HdhGPx, and then to increase the total antioxidant capacities in abalone.