Markers of genetic susceptibility in human environmental hygiene and toxicology : the role of selected CYP, NAT and GST genes

Inherited genetic traits co-determine the susceptibility of an individual to a toxic chemical. Special emphasis has been put on individual responses to environmental and industrial carcinogens, but other chronic diseases are of increasing interest. Polymorphisms of relevant xenobiotic metabolising enzymes may be used as toxicological susceptibility markers. A growing number of genes encoding enzymes involved in biotransformation of toxicants and in cellular defence against toxicant-induced damage to the cells has been identified and cloned, leading to increased knowledge of allelic variants of genes and genetic defects that may result in a differential susceptibility toward environmental toxicants. "Low penetrating" polymorphisms in metabolism genes tend to be much more common in the population than allelic variants of "high penetrating" cancer genes, and are therefore of considerable importance from a public health point of view. Positive associations between cancer and CYP1A1 alleles, in particular the *2C I462V allele, were found for tissues following the aerodigestive tract. Again, in most cases, the effect of the variant CYP1A1 allele becomes apparent or clearer in connection with the GSTM1 null allele. The CYP1B1 codon 432 polymorphism (CYP1B1*3) has been identified as a susceptibility factor in smoking-related head-and-neck squameous cell cancer. The impact of this polymorphic variant of CYP1B1 on cancer risk was also reflected by an association with the frequency of somatic mutations of the p53 gene. Combined genotype analysis of CYP1B1 and the glutathione transferases GSTM1 or GSTT1 has also pointed to interactive effects. Of particular interest for the industrial and environmental field is the isozyme CYP2E1. Several genotypes of this isozyme have been characterised which seem to be associated with different levels of expression of enzyme activity. The acetylator status for NAT2 can be determined by genotyping or by phenotyping. In the pathogenesis of human bladder cancer due to occupational exposure to "classical" aromatic amines (benzidine, 4-aminodiphenyl, 1-naphthylamine) acetylation by NAT2 is regarded as a detoxication step. Interestingly, the underlying European findings of a higher susceptibility of slow acetylators towards aromatic amines are in contrast to findings in Chinese workers occupationally exposed to aromatic amines which points to different mechanisms of susceptibility between European and Chinese populations. Regarding human bladder cancer, the hypothesis has been put forward that genetic polymorphism of GSTM1 might be linked with the occurrence of this tumour type. This supports the hypothesis that exposure to PAH might causally be involved in urothelial cancers. The human polymorphic GST catalysing conjugation of halomethanes, dihalomethanes, ethylene oxide and a number of other industrial compounds could be characterised as a class theta enzyme (GSTT1) by means of molecular biology. "Conjugator" and "non-conjugator" phenotypes are coincident with the presence and absence of the GSTT1 gene. There are wide variations in the frequencies of GSTT1 deletion (GSTT1 *0/0) among different ethnicities. Human phenotyping is facilitated by the GST activity towards methyl bromide or ethylene oxide in erythrocytes which is representative of the metabolic GSTT1 competence of the entire organism. Inter-individual variations in xenobiotic metabolism capacities may be due to polymorphisms of the genes coding for the enzymes themselves or of the genes coding for the receptors or transcription factors which regulate the expression of the enzymes. Also, polymorphisms in several regions of genes may cause altered ligand affinity, transactivation activity or expression levels of the receptor subsequently influencing the expression of the downstream target genes. Studies of individual susceptibility to toxicants and gene-environment interaction are now emerging as an important component of molecular epidemiology.

Tertiary Treatment of Ayr Municipal Wastewater using Bioremediation: A Pilot-Scale Study. A Report Prepared for the Burdekin Shire Council and the Burdekin Rangeland Reef Initiative

This joint DPI/Burdekin Shire Council project assessed the efficacy of a pilot-scale biological remediation system to recover Nitrogen (N) and Phosphorous (P) nutrients from secondary treated municipal wastewater at the Ayr Sewage Treatment Plant. Additionally, this study considered potential commercial uses for by-products from the treatment system. Knowledge gained from this study can provide directions for implementing a larger-scale final effluent treatment protocol on site at the Ayr plant. Trials were conducted over 10 months and assessed nutrient removal from duckweed-based treatments and an algae/fish treatment – both as sequential and as stand-alone treatment systems. A 42.3% reduction in Total N was found through the sequential treatment system (duckweed followed by algae/fish treatment) after 6.6 days Effluent Retention Time (E.R.T.). However, duckweed treatment was responsible for the majority of this nutrient recovery (7.8 times more effective than algae/fish treatment). Likewise, Total P reduction (15.75% reduction after 6.6 days E.R.T.) was twice as great in the duckweed treatment. A phytoplankton bloom, which developed in the algae/fish tanks, reduced nutrient recovery in this treatment. A second trial tested whether the addition of fish enhanced duckweed treatment by evaluating systems with and without fish. After four weeks operation, low DO under the duckweed blanket caused fish mortalities. Decomposition of these fish led to an additional organic load and this was reflected in a breakdown of nitrogen species that showed an increase in organic nitrogen. However, the Dissolved Inorganic Nitrogen (DIN: ammonia, nitrite and nitrate) removal was similar between treatments with and without fish (57% and 59% DIN removal from incoming, respectively). Overall, three effluent residence times were evaluated using duckweed-based treatments; i.e. 3.5 days, 5.5 days and 10.4 days. Total N removal was 37.5%, 55.7% and 70.3%, respectively. The 10.4-day E.R.T. trial, however, was evaluated by sequential nutrient removal through the duckweed-minus-fish treatment followed by the duckweed-plus-fish treatment. Therefore, the 70.3% Total N removal was lower than could have been achieved at this retention time due to the abovementioned fish mortalities. Phosphorous removal from duckweed treatments was greatest after 10.4-days E.R.T. (13.6%). Plant uptake was considered the most important mechanism for this P removal since there was no clay substrate in the plastic tanks that could have contributed to P absorption as part of the natural phosphorous cycle. Duckweed inhibited phytoplankton production (therefore reducing T.S.S) and maintained pH close to neutral. DO beneath the duckweed blanket fell to below 1ppm; however, this did not limit plant production. If fish are to be used as part of the duckweed treatment, air-uplifts can be installed that maintain DO levels without disturbing surface waters. Duckweed grown in the treatments doubled its biomass on average every 5.7 days. On a per-surface area basis, 1.23kg/m2 was harvested weekly. Moisture content of duckweed was 92%, equating to a total dry weight harvest of 0.098kg/m2/week. Nutrient analysis of dried duckweed gave an N content of 6.67% and a P content of 1.27%. According to semi-quantitative analyses, harvested duckweed contained no residual elements from the effluent stream that were greater than ANZECC toxicant guidelines proposed for aquaculture. In addition, jade perch, a local aquaculture species, actively consumed and gained weight on harvested duckweed, suggesting potential for large-scale fish production using by-products from the effluent treatment process. This suggests that a duckweed-based system may be one viable option for tertiary treatment of Ayr municipal wastewater. The tertiary detention lagoon proposed by the Burdekin Shire Council, consisting of six bays approximately 290 x 35 metres (x 1.5 metres deep), would be suitable for duckweed culture with minor modification to facilitate the efficient distribution of duckweed plants across the entire available growing surface (such as floating containment grids). The effluent residence time resulting from this proposed configuration (~30 days) should be adequate to recover most effluent nutrients (certainly N) based on the current trial. Duckweed harvest techniques on this scale, however, need to be further investigated. Based on duckweed production in the current trial (1.23kg/m2/week), a weekly harvest of approximately 75 000kg (wet weight) could be expected from the proposed lagoon configuration under full duckweed production. A benefit of the proposed multi-bay lagoon is that full lagoon production of duckweed may not be needed to restore effluent to a desirable standard under the present nutrient load, and duckweed treatment may be restricted to certain bays. Restored effluent could be released without risk of contaminating the receiving waterway with duckweed by evacuating water through an internal standpipe located mid-way in the water column.

Microencapsulated Termiticides

Testing of the efficacy of a long-acting termite bait toxicant system.

Tracking and identifying wastewater toxicants and assessing their biodegradation products

Screening of wastewater effluents from municipal and industrial wastewater treatment plants with biotests showed that the treated wastewater effluents possess only minor acute toxic properties towards whole organisms (e.g. bacteria, algae, daphnia), if any. In vitro tests (sub-mitochondrial membranes and fish hepatocytes) were generally more susceptible to the effluents. Most of the effluents indicated the presence of hormonally active compounds, as the production of vitellogenin, an egg yolk precursor protein, was induced in fish hepatocytes exposed to wastewater. In addition, indications of slight genotoxic potential was found in one effluent concentrate with a recombinant bacteria test. Reverse electron transport (RET) of mitochondrial membranes was used as a model test to conduct effluent assessment followed by toxicant characterisations and identifications. Using a modified U.S. EPA Toxicity Identification Evaluation Phase I scheme and additional case-specific methods, the main compound in a pulp and paper mill effluent causing RET inhibition was characterised to be an organic, relatively hydrophilic high molecular weight (HMW) compound. The toxicant could be verified as HMW lignin by structural analyses using nuclear magnetic resonance. In the confirmation step commercial and in-house extracted lignin products were used. The possible toxicity related structures were characterised by statistical analysis of the chemical breakdown structures of laboratory-scale pulping and bleaching effluents and the toxicities of these effluents. Finally, the biological degradation of the identified toxicant and other wastewater constituents was evaluated using bioassays in combination with chemical analyses. Biological methods have not been used routinely in establishing effluent discharge limits in Finland. However, the biological effects observed in this study could not have been predicted using only routine physical and chemical effluent monitoring parameters. Therefore chemical parameters cannot be considered to be sufficient in controlling effluent discharges especially in case of unknown, possibly bioaccumulative, compounds that may be present in small concentrations and may cause chronic effects.

Hanging drop: An in vitro air toxic exposure model using human lung cells in 2D and 3D structures

Using benzene as a candidate air toxicant and A549 cells as an in vitro cell model, we have developed and validated a hanging drop (HD) air exposure system that mimics an air liquid interface exposure to the lung for periods of 1 h to over 20 days. Dose response curves were highly reproducible for 2D cultures but more variable for 3D cultures. By comparing the HD exposure method with other classically used air exposure systems, we found that the HD exposure method is more sensitive, more reliable and cheaper to run than medium diffusion methods and the CULTEX (R) system. The concentration causing 50% of reduction of cell viability (EC50) for benzene, toluene, p-xylene, m-xylene and o-xylene to A549 cells for 1 h exposure in the HD system were similar to previous in vitro static air exposure. Not only cell viability could be assessed but also sub lethal biological endpoints such as DNA damage and interleukin expressions. An advantage of the HD exposure system is that bioavailability and cell concentrations can be derived from published physicochemical properties using a four compartment mass balance model. The modelled cellular effect concentrations EC50(cell) for 1 h exposure were very similar for benzene, toluene and three xylenes and ranged from 5 to 15 mmol/kg(dry weight) which corresponds to the intracellular concentration of narcotic chemicals in many aquatic species, confirming the high sensitivity of this exposure method. (C) 2013 Elsevier B.V. All rights reserved.

Destruction of Chemical Agents in a Plasma Reactor with Working Gas of Hydrogen

Survey of sediment quality in Sabine Lake, Texas and vicinity

The toxicity of sediments in Sabine Lake, Texas, and adjoining Intracoastal Waterway canals was determined as part of bioeffects assessment studies managed by NOAA’s National Status and Trends Program. The objectives of the survey were to determine: (1) the incidence and degree of toxicity of sediments throughout the study area; (2) the spatial patterns (or gradients) in chemical contamination and toxicity, if any, throughout the study area; (3) the spatial extent of chemical contamination and toxicity; and (4) the statistical relationships between measures of toxicity and concentrations of chemicals in the sediments. Surficial sediment samples were collected during August, 1995 from 66 randomly-chosen locations. Laboratory toxicity tests were performed as indicators of potential ecotoxicological effects in sediments. A battery of tests was performed to generate information from different phases (components) of the sediments. Tests were selected to represent a range in toxicological endpoints from acute to chronic sublethal responses. Toxicological tests were conducted to measure: reduced survival of adult amphipods exposed to solid-phase sediments; impaired fertilization success and abnormal morphological development in gametes and embryos, respectively, of sea urchins exposed to pore waters; reduced metabolic activity of a marine bioluminescent bacteria exposed to organic solvent extracts; and induction of a cytochrome P-450 reporter gene system in exposures to solvent extracts of the sediments. Chemical analyses were performed on portions of each sample to quantify the concentrations of trace metals, polynuclear aromatic hydrocarbons, and chlorinated organic compounds. Correlation analyses were conducted to determine the relationships between measures of toxicity and concentrations of potentially toxic substances in the samples. Based upon the compilation of results from chemical analyses and toxicity tests, the quality of sediments in Sabine Lake and vicinity did not appear to be severely degraded. Chemical concentrations rarely exceeded effects-based numerical guidelines, suggesting that toxicant-induced effects would not be expected in most areas. None of the samples was highly toxic in acute amphipod survival tests and a minority (23%) of samples were highly toxic in sublethal urchin fertilization tests. Although toxic responses occurred frequently (94% of samples) in urchin embryo development tests performed with 100% pore waters, toxicity diminished markedly in tests done with diluted pore waters. Microbial bioluminescent activity was not reduced to a great degree (no EC50 <0.06 mg/ml) and cytochrome P-450 activity was not highly induced (6 samples exceeded 37.1 ug/g benzo[a]pyrene equivalents) in tests done with organic solvent extracts. Urchin embryological development was highly correlated with concentrations of ammonia and many trace metals. Cytochrome P450 induction was highly correlated with concentrations of a number of classes of organic compounds (including the polynuclear aromatic hydrocarbons and chlorinated compounds). (PDF contains 51 pages)

Regional Fisheries Advisory Committee Meeting 30th January, 1984

This is the report from the Regional Fisheries Advisory Committee meeting, which was held on the 30th January, 1984. The report contains information on the use of fyke nets and an application to use Rotenone fish Toxicant. Also included is information on Dunsop Trout Farm and the establishment of migratory fish rearing facilities, the decline of salmon and sea trout catches in South Cumbria, and the decline of salmon stocks in the River Eden. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

Acute effect of sumithion on certain blood parameters and mortality rate of fish, Heteropneustes fossilis (Bloch)

The acute toxic effect of the toxicant sumithion (50% E.C.) on mortality rate (after 24, 48, 72, and 96 h), total RBC count and haemoglobin content (after 48 and 72 h) on Heteropneustes fossilis was investigated at four concentrations (9.7, 10.7 and 11.1 ppm). The sumithion treated fishes showed lower RBC and Hb levels than the untreated ones. A gradual decrease in the total RBC counts and Hb contents was recorded with increasing concentration of toxicant after 72 h but the blood showed fluctuating values after 48 h of treatment.

Effect of organophosphate, diazinon on some immunophysiological, haematological and histological variable of Rutilus frisii kutum (Kamensky,1901)

The acute toxicity and effects of diazinon on some haematological parameters of kutum (Rutilus frisii kutum, Kamensky, 1901) weighing 613.33 g±157.06 g were studied under static water quality conditions at 15°C ± 2ºC in winter and spring 2009. The effective physical and chemical parameters of water were pH= 7-8.2, dh= 300mg/L (caco3), DO= 7 ppm and T= 15°C±2ºC. The first test was primarily to determine the effects of acute toxicity (LC5096 h) of the agricultural toxicant diazinon (emulsion 60%) on kutum male brood stocks. For this purpose, 4 treatments were used to test toxicity; each treatment was repeated in 3 tanks with 9 fish per treatment and with 180 litres water capacity. After obtaining the final results, the information was analysed statistically with Probit version 1.5 (USEPA, 1985), and we determined the LC10, LC50 and LC90 values at 24 hours, 48 hours, 72 hours and 96 hours; the maximum allowable concentration value (LC5096 h divided by 10) (TRC, 1984); and the degree of toxicity. The second stage of testing consists of four treatments: LC0= 0 as experimental treatment, treatment A with a concentration of LC1= 0.107 mg/L, treatment B with concentration of LC5= 0.157 mg/L, treatment C with concentration of MAC value= 0.04 mg/L. Male brood stocks of kutum were treated with these concentrations for 45 days. Experiments were carried out under static conditions based on the standard TRC, 1984 method over 45 days. Our results show that long-term exposure to diazinon causes a decrease in the erythrocyte count (RBC), haemoglobin (Hb), haematocrit (PCV), mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), leucocyte count (WBC), lymphocyte, testosterone, iron (Fe), sodium (Na), lactate dehydrogenase (LDH), and cholinesterase (CHeS). In addition, diazinon also causes an increase in prolymphocyte, aspartate aminotransferase (AST), cholesterol, alkaline phosphatase (ALP) and adrenaline (P<0.05). There are no significant effects on monocyte, eosinophil, magnesium (Mg), chloride (Cl), glucose (BS), urea (BUN), uric acid (U.A), triglyceride (TG), calcium (Ca), albumin (Alb), total protein (TP), cortisol, noradrenaline and high density lipoprotein (HDL) levels in kutum male brood stocks (P>0.05). Pathology results showed toxin diazinon no effect on average weight and fish body length, the average weight of heart, brain, spleen, liver, kidney and liver index but caueses decrease of gonad weigth and gonad index and also, cause complications of tissue necrosis, vascular congestion, inflammation in the liver, a sharp reduction in the number of glomeruli, necrosis, vascular congestion and haemorage in the kidney, capsule thickening and fibrosis, atrophy, vascular congestion, macrophages release increased, increasing sediment Hemosiderine and thickening of artery walls in the spleen, atrophy, fibrosis and necrosis in testis , vascular congestion, increased distance between the myocardium and fibrous string in heart and neuronal loss, vascular congestion and edema in the brain of kutum male brood stocks.

Involvement of Fas/FasL system in apoptotic signaling in testicular germ cells of male Wistar rats injected i.v. with microcystins

Previous studies have shown that gonads were the second target organ of microcystins (MCs), and that MCs exposure exerted obvious toxic effects on male reproductive system of mammals. However, relevant molecular evidences are still lacking. Fas-signaling pathway plays a key role in toxicant-induced germ cell apoptosis. This study was to evaluate the responses of Fas/FasL system related genes and proteins in testes of rats injected intravenously with MCs. Enhanced apoptosis of germ cells in the testes of MCs-treated rats was detected by the terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick end labeling (TUNEL) associated with up-regulation of the Fas/FasL system. Both Fas and FasL protein expression were induced evidently from I h post-injection, and this high expression level maintained throughout the experiment. In addition, the activation of caspase-8 and caspase-3 protein was also observed, which were indicators of apoptosis. These results suggested the likely involvement of Fas/FasL system in the MCs-induced germ cell apoptosis. It is also suggested that MCs can cause damage to Sertoli cells directly. (C) 2009 Elsevier Ltd. All rights reserved.

Toxicity of cypermethrin to Daphnia magna HB

The acute toxic effect of the pesticide cypermethrin to Daphnia magna HB was examined. D. magna HB was exposed to cypermethrin at concentrations of 0, 1, 3, 5, 7, and 9 mg/L for 24 In. Data showed that the 24 h-LC50 of cypermthrin on D. magna HB was 4.81 mg/L. In contrast, the 24 h-LC50 of K2Cr2O7 (the national standard toxicant) to Daphnia magna was 0.38 mg/L in the current study. Results indicated that the Daphnia magna was very sensitive to pesticides. In addition, the effects of the culture condition(such as hardness, temperature and DO etc.) on Daphnia magna HB was also studied.

A phototaxis inhibition assay using barnacle larvae

The effects of sublethal concentrations of phenol and cadmium on the phototactic responses of the stage II nauplii of the barnacle Balanus amphitrite were investigated. Increased toxicant concentrations caused a reduction in phototactic responses. Balanus amphitrite nauplii exposed to nominal phenol concentrations of 100 ppm and higher for 1-12 h failed to exhibit phototactic responses, while longer exposure times of 24 and 48 h reduced the lowest observable effect concentration (LOECs) to 80 and 60 ppm, respectively. For cadmium, the LOECs, based on nominal concentrations, for B. amphitrite following 1, 6, 12, 24, and 48 h exposures were 20, 4.5, 4.0, 1, and 0.75 ppm, respectively. The LOECs can be significantly reduced by increasing the duration of exposure to the toxicants. A good relationship exists between the phototactic response and toxicant concentration as well as exposure time. Results of this study indicate that the toxicant-induced reduction in phototactic responses of barnacle larvae can be used in a sensitive, rapid screening test for ecotoxicological assessments. (C) 1997 by John Wiley & Sons, Inc.

GENOTOXIC EFFECTS OF LINEAR ALKYL BENZENE SULFONATE, SODIUM PENTACHLOROPHENATE AND DICHROMATE ON TETRAHYMENA-PYRIFORMIS

DNA in macro- and micronuclei of Tetrahymena pyriformis treated with linear alkyl benzene sulfonate (LAS) and sodium pentachlorophenate (PCP-Na) were determined by microspectrophotometry. The effects on rate of formation of macronuclear DNA extrusion bodies were also studied. We found DNA content of micronuclei in 0.14 ppm LAS and 0.9 ppb PCP-Na was lower than in that of the control, and LAS was able to increase the formation rate of macronuclear DNA extrusion bodies (the formation rate was 54% in 11.3 ppm LAS and 25.6% in 16.7 ppm dichromate). We concluded that 0.14 ppm LAS (below the maximum acceptable toxicant concentration) was genotoxic, whereas 0.014 ppm LAS was not. Dichromate 0.05 ppm and 0.9 ppb PCP-Na, equal to and below the maximum acceptable toxicant concentration, respectively, were potentially genetoxic.

功能菌剂的混合发酵研究

随着化工行业的发展，大量有毒有害难降解有机物随工业废水的排放进入环境，这些物质能够在环境中长期存在、积累和扩散，通过食物链对动植物的生存及人类的健康造成不良影响。本文以苯酚、对氯硝基苯、氯苯和十六烷为模拟污染物，以前期研制的功能菌剂为对象，经过紫外线线诱变筛选出优于出发菌株的功能菌，对诱变后功能菌的理化性能进行了研究，对菌种进行了鉴定，在此基础上，就其相互之间的微生态关系进行研究，为混合发酵提供理论基础，并就其最佳发酵条件及发酵参数进行了研究，最后对发酵产品的性能进行了检测。目前，国内外有关功能菌剂的研究还存在多方面的不足，主要包括：①由于多菌种混合发酵过程较为复杂，各菌之间存在复杂的相互作用，影响因素较多，关于菌种之间的相互关系研究得很少，环境功能菌剂的发酵方法大多采用单独发酵后混合的方式。单独发酵对原材料、设备和能源的利用率较低，对于多菌种制剂发酵，在设备、能源和原材料的方面造成的浪费更大，将会大幅增加菌剂的生产成本，影响多菌种功能菌剂的发展；②功能菌剂生产过程的质量控制方面研究得较少；③功能菌剂产品的稳定性、抗冲击性能研究得较少，对环境微生物制剂的研究主要集中在菌种选育和培养条件优化方面。 通过本论文研究，得到以下主要结论。 （1）在紫外线诱变处理中，用紫外线对发生一定程度退化的出发菌株进行诱变处理后，六株具有高效降解性能的菌株被筛选出来，诱变筛选出的菌株形态和ERIC-PCR指纹图谱与出发菌株相比发生了明显改变；而且诱变后的菌株对目标难降解底物的降解能力均得到改善，其中，FPN、FCB、F14、FEm对目标底物的降解率提高了20%以上；诱变后菌株经过7次连续传代接种后，对目标难降解底物的降解率无显著变化，具有一定的遗传稳定性。并对诱变后的功能菌进行了初步的鉴定，这6株菌都分别是芽孢杆菌。 （2）对诱变后的功能菌相互之间的微生态关系进行了研究，通过抑菌实验、生长量以及基质消耗量的比较，确定它们之间的生长关系是无害共栖关系，可以进行混合发酵。 （3）对该功能菌剂进行发酵培养条件研究，结果表明发酵培养基的最佳成分（g/L）：葡萄糖 31.0g/L、玉米粉10.0g/L、磷酸氢二钾1.0g/L、硫酸铵1.1g/L、硫酸镁0.55g/L。通过研究不同的培养条件对菌体生长和降解性能的影响，确定了最佳培养条件：培养基初始pH7.5；最适温度32℃；培养基装液量125mL（250 mL三角瓶），以及培养时间对降解性能的影响，培养20 h的产物对降解最为有利。通过研究添加不同目标污染物对菌体生长和降解性能的影响，确定了添加目标污染物的最佳量以及最佳时间：苯酚投加量：1.125 g/L，对氯硝基苯投加量：0.1 g/L；最佳投加时间为发酵培养开始后4 h。 （4）以摇瓶分批发酵最优条件为基础，对FPN、F10、FCB、FNa、F14 和 FEm进行了摇瓶分批发酵试验。以摇瓶分批发酵试验数据为依据，对功能菌剂分批发酵动力学进行了研究，建立了菌体生长和基质消耗的动力学模型，拟合模型能较好的反映功能菌剂分批发酵过程。 （5）功能菌剂和活性污泥协同作用，可以提高系统的生物降解能力，功能菌剂投加量为2%，新鲜活性污泥3500 mg/L，降解24 h条件下，功能菌剂和活性污泥的协同作用对COD的去除率和对照组相比，最多的提高了36.8%。功能菌剂和活性污泥协同作用以及活性污泥的单独作用，其生物降解过程均符合一级反应动力学过程，功能菌剂和活性污泥协同作用的生物降解动力学方程为：，相关系数97%。采用SBR运行方式，引入功能菌剂的SBR系统明显能够改善和提高生物降解的效率。与仅有活性污泥的系统相比，系统对COD的平均去除率可以提高27.1%，同时，系统的耐负荷冲击以及耐毒害冲击的性能比仅有活性污泥的SBR系统强，特别是负荷冲击对引入功能菌剂的SBR系统影响很小。仅有活性污泥的SBR系统经过负荷冲击和毒害冲击之后，不能恢复到冲击之前的水平，而且系统有效作用时间的周期比引入功能菌剂的SBR系统相比大大缩短，而引入功能菌剂的SBR系统处理效果较为稳定，恢复能力很强。 Along with the development of industries, many recalcitrant organic chemicals have been discharged into natural environments together with wastewaters and can exist in waters, soil and sediments for a long time without degradation. These haz-ardous substances, their byporducts and metabolizabilities can be highly toxic, mu-tagenic and carcinogenic, thereby threatening animals, plants and human health through food chain. Consequently the removal of these compounds is of significant interest in the area of wastewater treatment. In this dissertation, the phenol, hydro-quinone, chlorobenzene and hexadecane treated as the model pollutants, the func-tional microorganism agent was used as the starting strains, they treated with ultra-violet light, and then the mutant strains with high degradation ability were screened out and identified primarily, the relationship between these stains were studied, the medium composition and fermentation conditions were optimized, the degradation ability of the fermented production was tested. The literature survey indicates that the study of the microorganism agent is far from complete and more information is re-quired on following problems. 1, Because of the complexity of relationship in mixed fermentation and the complicated factors, the study is hardly to process.2, There is a lack of information on the quality control of the producing process .3, And there is a lack of information on the stability about the microorganism agent. In this dissertation, the main results of the present study could be summarized as follows: (1)The degenerate starting strains were treated with the ultraviolet light, and six mutant strains with high biodegradation ability were screened out by using the me-dium with selective pressure of model pollutants. The mutant strains had great changes in colonialmorphology and ERIC-PCR fingerprinting. And the mutant strains got obvious advantages over the starting strains in degradation ability and over 20% improvement of removal rates was achieved for FPN、FCB、F14 and FEm. The de-gradation ability of the mutant strains was stable after seven generations. After that, the mutant strains were primarily identified as bacillus respectively. (2) The relationship between these mutant strains was studied. By the compari-son of antibiosis effect, biomass and consumption of substrate, the relationships were neutralism and they could be mixed fermented. (3) The optimized cultivation conditions were as follows: glucose 31.0 g/L, corn power 10 g/L, K2HPO4 1.0 g/L, (NH4)2SO4 1.1 g/L, MgSO4 0.55 g/L, initial pH7.5, temperature 32℃, working volume 125 mL/250 mL, and cultivation time 20h (con-sidering the time effect on degradation ability), adding pollutants phenol (1.125 g/L) and hydroquinone (0.1 g/L) into the broth at 4 h after cultivation. (4) Based on the above optimum condition, the batch fermentation was per-formed with strains FPN, F10, FCB, FNa, F14 and FEm in shake flask. The batch fermentation kinetics was studied based on the experimental data. Two kinetic models were constructed which could reflect the regularity of growth and substrate consump-tion in the process of batch fermentation. (5) The co-operation of functional microorganism agent and activated sludge could raise biodegradation of system by adding some microorganism agent and 3500 mg/L fresh activated sludge. Bioaugumentation by the addition of high effective deg-radation culture enhanced the treatment effect of SBR system and the COD removal rate was increased by 20%-36.8%. Its biodegradation matched first-order dynamical reaction equation, and the reaction equation was ln0.2327.391ct=−+. The micro-organism agent had the effect of optimization to activated sludge micro-ecosystem. The SBR system adding 2% microorganism agent, the average COD removal rate of that was increased by 27.1% and stronger anti-shock ability to load and toxicant were achieved (compared with SBR system just adding activated sludge). Especially the load-shock has barely effect to the SBR system adding microorganism agent. After the load and toxicant shock, the SBR system just adding activated sludge couldn’t come back to original level and the activated sludge micro-ecosystem was frustrated. The applying of microorganism agent increased biological activity and system’s re-sistance ability to load shock and toxicant shock.