Liver glutathione content and glutathione‐dependent enzymes of two species of freshwater fish as bioindicators of chemical pollution

Abstract

Glutathione content and glutahione‐dependent enzymes were measured in the liver of two fish species, gudgeon (Gobio gobio) and roach (Rutilus arcasii), from the river Bernesga (Spain) caught downstream and upstream of the waste site of several chemical industries. Animals from contaminated sites display a reduced glutathione concentration and a tendency to the decrease of glutathione S‐transferase activity. Glutathione peroxidase activity was significantly elevated only in the liver of Gobio gobio and glutathione reductase activity in that of Rutilus arcasii. Our data indicate that the glutathione system constitutes a sensitive biochemical indicator of chemical pollution. Relative changes of glutathione and glutathione‐dependent enzymes in both fish species suggest a different susceptibility to toxins.     

Metabolic fate of the 14C-labeled herbicide clodinafop-propargyl in soil

The metabolic fate of ¹⁴C-phenyl-labeled herbicide clodinafop-propargyl (CfP) was studied for 28 days in lab assays using a soil from Germany (A_p horizon, silt loam, and cambisol). Mineralization amounted to 12.40% of applied ¹⁴C after 28 days showing a distinct lag phase until day 7 of incubation. Portions of radioactivity extractable by means of 0.01 M CaCl₂ solution (bioavailable fraction) decreased rapidly and were 4.41% after 28 days. Even immediately after application, only 57.31% were extracted with the aqueous solvent. Subsequent extraction using accelerated solvent extraction (ASE; acetonitrile/water 4:1, v/v) released 39.91% of applied ¹⁴C with day 0 and 26.16% with day 28 of incubation from the samples. Non-extractable portions of radioactivity thus, increased with time amounting to 11.99% (day 0) and 65.00% (day 28). A remarkable increase was observed between 14 and 28 days correlating with the distinct increase of mineralization. No correlation was found throughout incubation with general microbial activity as determined by DMSO reduction. Analysis of the CaCl₂ and ASE extracts by radio-TLC, radio-HPLC and GC/MS revealed that CfP was rapidly cleaved to free acid clodinafop (Cf), which was further (bio-) transformed; DT₅₀ values (based on radio-TLC detection of the parent compound) were far below 1 day (CfP) and about 7 days (Cf). TLC analysis pointed to 2-(4-hydroxyphenoxy)-propionic acid as further metabolite. Due to fractionation of non-extractable residues, most of the ¹⁴C was associated with fulvic and humic acids, portions in humin fractions and non-humics were moderate and low, respectively. Using a special strategy, which included pre-incubation of the soil with CfP and then mineralization of ¹⁴C-CfP as criterion, a microorganism was isolated from the soil examined. The microorganism grew using CfP as sole carbon source with concomitant evolution of ¹⁴CO₂. The bacterium was characterized by growth on commonly used carbon sources and by 16S rDNA sequence analysis. The sequence exhibited high similarity with that of Rhodococcus wratislaviensis (99.56%; DSM 44107, NCIMB 13082).     

连续施用发酵猪粪对土壤中四环素抗性基因数量的影响

为了研究连续施用发酵猪粪之后,土壤中抗性基因的数量以及发酵猪粪的施用量对土壤抗性基因的影响,采用实时荧光定量PCR方法,对稻麦轮作模式下连续6年施用4.5t/hm2和9.0t/hm2发酵猪粪的土壤进行了四环素抗性基因(TRGs)的检测和定量分析.在施用发酵猪粪的土壤中能够检测到9种TRGs.施用发酵猪粪显著增加了土壤中tet G、tet L、tet B(P)、tet O、tet W的绝对数量,其中tet B(P)、tet W、tet O的绝对数量受到了施肥用量的影响;而tet Z、tet C和tet S的绝对数量则没有受到施肥的影响.上述8种TRGs在同一施肥处理的0~5cm、5~10cm以及10~20cm土壤中的相对丰度分布均无显著差异.发酵猪粪的施用也显著增加了tet G、tet L、tet B(P)和tet O的相对丰度,但是仅有tet O的相对丰度受到了施肥用量的显著影响.本研究表明,在稻麦轮作模式下,农田土壤中的TRGs数量会受到发酵猪粪中残留TRGs的影响,连续施用该粪肥会显著增加土壤中tet G、tet L、tet B(P)和tet O的绝对数量和相对丰度.因此,需要优化畜禽粪堆肥发酵工艺,以减少抗性基因在粪肥中的残留.     

微生物聚磷及其酶学调控

本文论述了微生物细胞内磷酸盐转运机制和与聚磷相关的酶—PPK、PPX及两者对微生物聚磷行为的调控,并对今后研究重点做出展望.论文认为对于微生物聚磷的研究应从纯种微生物转移到强化生物除磷系统的混合微生物菌群,运用分子生物学和生物信息学手段分析聚磷优势菌种的PPK和PPX酶的结构特征及其在活性污泥中的变化规律,进一步深入认识生物除磷调控机理,开辟寻求提高系统除磷效率的新途径.     

Bacterial degradation of monocrotophos and phyto- and cyto-toxicological evaluation of metabolites

The bacterium Serratia marsescens strain JAS16 was isolated from agricultural soil which had prior exposure to monocrotophos for three years. The strain JAS16 tolerated up to 1200 mg L^–1 monocrotophos and degraded the insecticide (1000 mg L^–1) at a degradation rate constant of 136 d^?1 (DT₅₀ = 3.7 d). In soil, the degradation rate constant was 105 d^?1 (DT₅₀ = 4.8 d). A schematic pathway is being proposed from the degraded products derived from gas chromatography--mass spectrometry (GC-MS). The phytotoxicity of degradation products to Vigna radiata, Vigna unguiculata, and Macrotyloma uniflorum and the genotoxicity to Allium cepa roots were found to be low. A cost-effective powder-based formulation was achieved with the isolate. The isolate remained viable during the storage and also multiplied with a higher colony forming units (CFU) load g^–1 for over a period of seven weeks of storage.     

Interspecific and intraspecific identification of Dendrobium based on the psbAtrnH intergenic region sequences and the 5S rRNA gene spacer sequences北大核心CSCD

This research aimed to investigate the interspecific and intraspecific identification of Dendrobium by using the multi-locus method so as to provide a molecular basis for Dendrobium identification through the combination of chloroplast psbA-trnH intergenic region sequences and ribosome 5S rRNA gene spacer sequences. PCR direct sequencing was applied to detect the chloroplast psbA-trnH intergenic region sequences as well as the ribosome 5S rRNA gene spacer sequences of 12 Dendrobium species, while the psbA-trnH intergenic region sequences of Dendrobium denneanum dq-2 variety and dq- 5line were cloned and sequenced for single nucleotide polymorphism (SNP) analyzing. The sequences were analyzed by the software Sequencher4.14, Bioedit7.0, MEGA5.2 and Dansp5.0; the interspecific and intraspecific Kimara-2-Parameter(K2P) distances were also calculated. The phylogenetic tree (using Neighbor joining method) was constructed with Bulbophyllum odoratissimum and Bletilla striata as outgroup. The results showed an average length of chloroplast psbA-trnH gene sequences in Dendrobium as 742.3 bp, with 72 variable sites, including 33 information sites; the average length of the ribosome 5S rRNA gene spacer sequences in Dendrobium was 336.4 bp, with 213 variable sites including 139 information sites. Using psbAtrnH intergenic region sequences in combination with ribosome 5S rRNA gene spacer sequences can not only identify D. denneanum, D. hancockil, D. thysiflorum, D. devonianum, D. moniliforme, D. chrysotoxum, D. officinale, D. heterocarpum and D. nobile, but also differentiate D. officinale from different geographical populations, and distinguish the dq-2 variety and dq 5line with SNP in the multi locus of D. denneanum.     

基于esp基因和JCV标记的MST方法在五大流域典型水源地中的应用

非点源污染目前已经成为影响水体环境的重要污染。微生物源示踪技术(microbial source tracking,MST)是解决非点源污染的一项新技术,它可以确定污染的宿主来源。肠球菌esp基因和多瘤病毒JCV可以作为检测水体中人源粪便污染的分子标记,其灵敏度和特异性都很高。为分析五大流域水源地是否受到人源粪便的污染,对辽河、海河、淮河、长江和黄河五大流域典型水源地水样进行采集和检测,选取人源粪便特异病原微生物肠球菌的esp基因和多瘤病毒JCV建立了相应的MST分子检测方法。结果表明,五大流域的典型水源地采样点均有可能受到了人源粪便的污染,可为当地相关部门提供技术支持和数据参考。     

湖南石门雄黄矿区矿样中的砷氧化细菌的分离及鉴定

绿色的生物学方法处理含砷废水,对于缓解工业发展所带来砷污染问题,提高居民健康生活质量具有重要意义。以湖南石门雄黄矿区的尾矿池积水（SY）、地下400 m处的矿坑水（NY）和尾矿池底泥表层（XY）3个样本为研究对象,利用细菌富集和多次传代的方法,从这3个环境样本中获得可氧化三价砷的3个菌群。在砷质量浓度为l~5 g·L-1的培养基中分别检测上述3个菌群的耐受能力,发现在砷质量浓度高达5 g·L-1的条件下,上述菌群均能在2~3 d内达到109 cells·mL-1。利用上述菌群处理l g·L-1亚砷酸钠溶液,SY、XY和NY菌群可分别在25、20和35 h内将三价砷完全氧化,并且使溶液中的总砷质量浓度降低66.7%。进一步对3个菌群进行多次平板分离,利用五价砷和硝酸银的颜色反应,获得11株可氧化三价砷的目的菌株。对这些菌株16S rDNA进行NCBI的BLASTN序列比对发现,菌株SMY24、SMY33、SMY22、SMY32、SMY21和SMY31均属于假单胞菌属（Pseudomonas）,菌株SMY104属于不动杆菌属（Acinetobacter）,菌株SMY17、SMY25、SMY9和SMY1在所有的已知序列中,最大的相似率只有91%,最小的相似率为76%,且都是未曾报道纯培养的细菌。对这些菌株三价砷氧化能力进行测定发现,其中菌株SMY104和SMY21在20 h内能将三价砷几乎完全氧化。通过对这些菌株三价砷氧化酶基因进行初步检测,发现了多个菌株具有三价砷氧化酶基因（AoxB）。     

秸秆添加对厨余垃圾堆肥时H_2S和NH_3排放的影响

厨余垃圾堆肥过程中NH3和H2S的排放不但降低了堆肥的养分含量,而且会引发严重的恶臭。以厨余垃圾为研究对象,以玉米秸秆为膨松剂,设置5%、10%、15%、20%(质量分数)4个添加比例(湿基)的堆肥处理,研究秸秆添加量对厨余垃圾堆肥过程中H2S和NH3排放的影响。结果表明:从温度来看,仅T4处理未达到无害化和腐熟的要求。氧气不足是造成H2S排放的主要原因,4个堆肥处理的H2S主要集中在前2周排放,随着秸秆添加量的增加,H2S的排放量逐渐降低。与T1处理相比,T2、T3、T4的H2S累积排放量分别降低了35.5%、44.7%、64.2%。各处理NH3的排放趋势与H2S类似,高温期为NH3释放的关键时期,NH3累积释放量占总释放量的62.2%~72.2%,与T1处理相比,T2、T3和T4的NH3累积排放量分别降低了36.9%、45.2%、76.3%。由此可见,添加适量的玉米秸秆不但能促进厨余垃圾堆肥的进行,明显降低堆肥过程中H2S和NH3的排放,而且可以实现玉米秸秆的资源化利用。