BACTERIAL RELATIONSHIPS IN STORMWATERS1

ABSTRACT: Data were developed within a three-year period for indicator bacteria and three species of bacterial pathogens following rural storm event hydrographs. The first flush concept was confirmed in all hydrographs. Bacterial density peaking occurred at or before the hydrograph peaks. FC and FS values were higher in more developed areas than the primary rural test site and their numerical ratios followed similar trends. Chlorine demand of storm waters varied between 8 and 16 mg/l and, the ozone requirement was greater than 32 mg/l in the same waters. Aftergrowth of total coliform bacteria occurred following chlorine and ozone doses of 16 mg/l and 32 mg/l respectively. Fecal coliform, fecal streptococci, Salmonella sp., and Pseudomonas sp. all were reduced to near detectable limits by the disinfectants up to 8 days. Staphylococcus sp. demonstrated a propensity to restablish their populations. Multiple regression analysis of the bacterial groups and species in storm waters suggested the fecal streptococci to have been the most useful group in evaluating bacterial storm water quality, with staphylocci have been closely related insofar as their statistical significance was concerned.     

An ecosystem perspective on potential impacts of drilling fluid discharges on seagrasses

Potential effects of oil drilling fluid discharges uponThalassia seagrass ecosystems were examined using seagrass core microcosms. Observed experimental effects, summarized in this article, included changes in both autotrophic (Thalassia and epiphyte) and heterotrophic (dominant benthic macroinvertebrates) species, and the processes of primary productivity and decomposition. The physical disturbance related to greater turbidity and sedimentation caused some effects, while others seemed a direct response to the toxic constituents of drilling fluids. Using these experimental results and the case ofThalassia and drilling fluids as a case study, we explore general methodological and philosophical issues for ecotoxicology and, furthermore, focus upon the challenge of providing a scientific basis for judging acceptability of environmental changes likely to ensue from human activities.     

Chromosomal genes conferring tolerance to heavy metal (Ag) toxicity

Bacterial strains were isolated from sediment samples from the Thames River. Successive transfer growth of the various strains on nutrient agar containing increasing concentrations of AgNO₃ revealed that three of the bacterial isolates were found to be capable of tolerating high concentrations of AgNO₃ ranging from 20 to 80 mM on a solid medium and up to 10 mM AgNO₃ in liquid medium. Molecular characterization and identification based on 16S rDNA gene sequencing of three strains of bacteria that are tolerant to silver nitrate showed that the major tolerant strains include the superbug, Shewanella oneidensis, Pseudomonas sp. and Bacillus sp. Protein extraction and two-dimensional (2D) sodium dodecyl sulfate SDS-polyacrylamide gel electrophoresis (PAGE) of the protein extracts in bacteria exposed to very high concentrations of AgNO₃ revealed a general reduction in the number of expressed proteins, although two protein spots were conspicuously over expressed in the exposed bacteria compared to control. The N-terminal amino acid sequence analysis of the protein spots identified the major up-regulated proteins as the outer membrane protein To1C (45.2 kDa) and the structural protein of the flagellar filament, flagellin (28.34 kDa), encoded for by the to1C and fliC genes, respectively. The roles of these genes in a number of multi-drug resistant pathogen and potentials for biotechnological applications in toxic metal control for treatment of contaminated ecosystems and biomining were discussed.     

钻井液化学剂可生物降解性评定方法

介绍研究钻井液化学剂生物降解性的必要性及开发可生物降解钻井液的进展。阐述了常用的评定有机物生物降解性的试验方法。推荐采用BOD_5／COD_Cr比值来评定我国石油行业钻井液化学剂及油田化学剂的可生物降解性，规范石油行业生物降解研究工作，保证评定结果的可靠性和可比性。     

钻井液化学剂可生物降解性研究

实验测定了16种钻井液化学剂的BOD_5和COD_cr，并计算其BOD_5／COD_cr比值。提出了以BOD_5／COD_cr的比值划分钻井液化学剂可生物降解性的分级标准，并对16种钻井液化学剂的可生物降解性进行分级，为其应用提供依据。     

废弃钻井液微生物降解菌室内筛选研究

针对川渝地区废弃钻井液成分复杂、污染危害大的特点,提出利用微生物法处理废弃钻井液的方法。通过菌株分离、菌株筛选、菌株对废弃钻井液的降解利用实验筛选出8株降解菌株,进行了废弃钻井液的降解利用情况研究,表明筛选出的降解菌株均能够以废弃钻井液为唯一碳源,具备了快速分解废弃钻井液的能力。     

油田化学剂生物毒性的测定及其分级标准

通过试验建立了测定油田化学剂生物毒性的发光细菌法。用发光细菌法测定了几十种钻井/完井液添加剂和钻井／完井液体系的生物毒性，并在此基础上提出了油田化学剂生物毒性的分级标准。     

Re-establishment of the proper injectivity of the CO2-injection well Ktzi 201 in Ketzin, Germany

The onshore CO₂-storage site Ketzin consists of one CO₂-injection well and two observation wells. Hydraulic tests revealed permeabilities between 50 and 100 mD for the sandstone rock units. The designated injection well Ktzi 201 showed similar production permeability. After installation of the CO₂-injection string, an injection test with water yielded a significantly lower injectivity of 0.002 m³/d kPa, while the observation wells showed an injection permeability in the same range as the productivity. Several possible reasons for the severe decline in injectivity are discussed. Acidification of the reservoir interval, injection at high wellhead pressure, controlled mini-fractures and back-production of the well are discussed to remove the plugging material to re-establish the required injectivity of the well. It has been decided to perform a nitrogen lift and analyse the back-produced fluids. Initially during the lift, the back-produced fluids were dark-black. Chemical and XRD analyses proved that the black solids consisted mainly of iron sulphide. Sulphate-reducing bacteria (SRB) were detected in fluid samples with up to 10⁶ cells/ml by fluorescent in situ hybridisation (FISH) indicating that the formation of iron sulphide was caused by bacterial activity. Organic compounds within the drilling mud and other technical fluids were likely left during the well completion process, thus providing the energy source for strong proliferation of bacteria. During the lift, the fraction of SRB in the whole bacterial community decreased from approximately 32% in downhole samples to less than 5%. The lift of Ktzi 201 succeeded in the full restoration of the well productivity and injectivity. Additionally, the likely energy source of the SRB was largely removed by the lifting, thus ensuring the long-term preservation of the injectivity.     

环境可接受的甲酸盐钻井完井液性能探讨

随着环境保护的要求日益严格,一种新型的钻井完井液 ── 环境可接受的甲酸盐钻井完井液,正在国外现场试验和推广应用,目前已引起国内有关研究人员和技术人员的高度重视。分析了甲酸盐钻井完井液的工艺性能,并着重对甲酸盐钻井完井液在环境保护方面的优越性能进行了探讨。     

The Potential Use of Chicken-Drop Micro-Organisms for Oil Spill Remediation

An examination of chicken-drop micro-organisms for oil spill remediation is presented in this work. The chicken droppings contained aerobic heterotrophs (1.2×10⁸ CFU g^–1), total fungi (3.4×10⁴ CFU g^–1) and crude oil (transniger pipeline crude, TNP) degrading bacteria (1.5×10⁶ CFU g^–1). The crude oil degraders were identified as species of Micrococcus, Bacillus, Pseudomonas, Enterobacter, Proteus, Klebsiella, Aspergillus, Rhizopus, and Penicillium. Pseudomonas aeruginosa CDB-06 and species of Bacillus CDB-08 and Penicillium CDF-10 degraded the crude oil at exceedingly high rates. Pseuedomonas aeruginosa CDB-06 degraded 65.5 percent of the crude oil after 16 days, while Bacillus sp. CDB-08, and Penicillium sp. CDF-10 degraded 65.3 percent, and 53.3 percent, respectively of the crude oil over the same period. The chicken droppings also had a pH 7.3, 18.5 percent moisture content, 2.3 percent total nitrogen, and 0.5 percent available phosphorus. Addition of oil polluted soil (10 percent (v/w) pollution level) with chicken droppings enhanced degradation of the crude oil in the soil. 68.2 percent of the crude oil was degraded in the soil amended with chicken droppings, whereas only 50.7 percent of the crude oil was degraded in the unamended soil after 16 days. The amendment raised the acidic reaction (pH 5.7) of the oil-polluted soil to alkaline (pH 7.2) within 16 days. Chicken droppings could, therefore, be used in an integrated oil pollution abatement program.     

Allelopathic Potential of Switchgrass (Panicum virgatum L.) on Perennial Ryegrass (Lolium perenne L.) and Alfalfa (Medicago sativa L.)

This study investigated allelopathy and its chemical basis in nine switchgrass (Panicum virgatum L.) accessions. Perennial ryegrass (Lolium perenne L.) and alfalfa (Medicago sativa L.) were used as test species. Undiluted aqueous extracts (5 g plant tissue in 50 ml water) from the shoots and roots of most of the switchgrass accessions inhibited the germination and growth of the test species. However, the allelopathic effect of switchgrass declined when extracts were diluted 5- or 50-fold. Seedling growth was more sensitive than seed germination as an indicator of allelopathic effect. Allelopathic effect was related to switchgrass ecotype but not related to ploidy level. Upland accessions displayed stronger allelopathic potential than lowland accessions. The aqueous extract from one switchgrass accession was separated into phenols, organic acids, neutral chemicals, and alkaloids, and then these fractions were bioassayed to test for allelopathic potential. Alkaloids had the strongest allelopathic effect among the four chemical fractions. In summary, the results indicated that switchgrass has allelopathic potential; however, there is not enough evidence to conclude that allelopathic advantage is the main factor that has contributed to the successful establishment of switchgrass on China’s Loess Plateau.     

ZONES OF NITRIFICATION1

ABSTRACT: This paper advances the hypothesis that nitrification exists in shallow streams as a result of surface activity and in estuaries due to growth in the water phase. Between these zones no significant levels of nitrification occur. Field measurements of ammonia, nitrite and nitrate nitrogen, enumerations of Nitrosomonas and Nitrobacter, and respirometry on two small streams and two moderately large rivers are presented. The appropriate mathematical models to describe nitrification must be based on the mechanisms involved - bacterial growth kinetics for estuaries and zero kinetics for the surface activity in shallow streams.     

Factors Influencing Prymnesium parvum Population Dynamics During Bloom Initiation: Results from In-lake Mesocosm Experiments1

Roelke, Daniel L., Leslie Schwierzke, Bryan W. Brooks, James P. Grover, Reagan M. Errera, Theodore W. Valenti, Jr., and James L. Pinckney, 2010. Factors Influencing Prymnesium parvum Population Dynamics During Bloom Initiation: Results from In-Lake Mesocosm Experiments. Journal of the American Water Resources Association (JAWRA) 46(1):76-91. DOI: 10.1111/j.1752-1688.2009.00392.x Abstract: The alga Prymnesium parvum forms large fish-killing blooms in many Texas lakes. In some of these lakes, however, P. parvum occurs but does not develop blooms. In this study, we investigated factors that may influence bloom initiation by conducting a series of in-lake experiments involving mixing of waters from Lake Whitney, which has a history of P. parvum blooms, with waters from Lake Waco where no blooms have occurred. In all experiments, the addition of Lake Waco waters resulted in a poorer performance of P. parvum. Various experimental treatments and field data show that differences in grazing, pathogens, nutrients, and salts between the two lakes were not likely factors that contributed to this observation. Industrial and agricultural contaminants, allelochemicals and algicidal chemicals were not measured as a part of this research. However, anthropogenic contaminants other than nutrients were not observed at levels exceeding water quality standards in Lake Waco in recent years. On the other hand, nuisance cyanobacteria are common in Lake Waco, where Microcystis sp. and Anabaena sp. were abundant during the initiation of our experiments, both taxa are known to produce chemicals with allelopathic properties. In addition, the emergent field of algal-heterotrophic bacteria interactions suggests that chemicals produced by heterotrophic bacteria should not be overlooked. Further research focusing on the chemical interactions between cyanobacteria and P. parvum, as well as the potential role of algicidal bacteria, in the initiation of P. parvum blooms is necessary, as it may be important to the management of these blooms.     

EFFECTS OF SUSPENDED SOLIDS ON THE ACUTE TOXICITY OF ZINC TO DAPHNIA MAGNA AND PIMEPHALES PROMELAS1

ABSTRACT: Current procedures for setting site-specific water quality criteria consider abiotic and biotic factors. Suspended solids were shown to be important in reducing zinc toxicity to water column organisms. At zinc concentrations of ～ 1 mg/L in solutions with < 100 mg/L of all suspended solids tested, zinc toxicity to D. magna was reduced. Sorption of zinc to suspended solids and/or changes in water chemistry due to the addition of suspended solids appear to have been the factors causing reductions in zinc toxicity to D. magna. Only suspended solids levels of 483–734 mg/L of a type that increased total alkalinity, total hardness, and total dissolved carbon clearly reduced the toxicity of ～ 20 mg/L zinc to P. promelas. The toxic form of zinc to these organisms appears to reside in the aqueous phase. Characteristics of suspended solids did not influence the partition coefficient of zinc in sorption experiments of 96 h. The slopes of dose-response curves proved to be useful for assessing the potential of an organism to respond to changes in aqueous phase zinc concentrations, and may be a useful biological parameter when considering site-specific water quality criteria for chemicals.     

江汉油田废弃水基钻井液对环境的影响研究

通过对废弃水基钻井液基本组分及生物毒性效应的分析,详细评价了废弃水基钻井液对浮游生物生长、卤虫孵化、土壤植物及糠虾的生物毒性影响。结果表明：江汉油田废弃钻井液对无节生物幼虫的影响不严重,不会影响卤虫的孵化率,但会影响其孵化速度;由于废钻井液中的含盐量高,ｐＨ值较高,因此对土壤及植物的生长有一定影响;其ＬＣ５０均大于３０００ｍｇ／Ｌ,基本上符合美国联邦环保局（ＥＰＡ）颁布的陆相水域排放钻井液和钻屑的条例。     

Bioremediation of cutting fluids contaminated soil by Pleurotus tuber-regium Singer

This study investigated the effect of inoculation of Pleurotus tuber-regium, a Nigerian white rot fungus, period of incubation, different levels of contamination on cutting fluids degradation in contaminated soil over 30, 60, and 90 days. Control for different levels of cutting fluids was also used to compare rates of bioremediation of the contaminant in the soil. At the end of each incubation period, the mycelia-ramified substrate was separated from the soil layer and dried. The soil samples were analyzed for physico-chemical parameters; total petroleum hydrocarbon, lignin content by determining the acid detergent fraction (ADF), heavy metals content of the soil using flame atomic absorption spectrophotometer, and changes in the polyphenol oxidase and peroxidase activities were also determined after 1, 2, and 3 months. P. tuber-regium improved the nutrient status of the soil and increased enzyme activity was recorded. A reduction in the pH and heavy metal contents of the soil at all levels of cutting fluids concentrations was detected. The lignin in the rice straw decreased from 34.50% in the control to 8.06% at 30% cutting fluids concentration after 3 months of incubation. The highest TPH loss of 30.84% was recorded at 20% cutting fluids contamination after 3 months compared to 13.75% at the onset of the experiment. The improvement of the nutrient contents of the soil, bioaccumulation of heavy metals, degradation of TPH, lignin, and increased activity of polyphenol oxidase and peroxidase was due to biodegradation of the cutting fluids.     

RESPONSES OF HETEROTHROPHIC BACTERIAL POPULATIONS TO pH CHANGES IN COAL ASH EFFLUENT1

ABSTRACT: Total culturable heterotrophic bacteria in a coal ash basin and drainage system were monitored over a period of two years. In the first year heavy (bottom) ash was sluiced to the basin resulting in a pH of 6.5. During the second year fly ash was precipitated and added to the sluice lowering the basin pH to 4.6. Sulfate concentrations during 1975 ranged from 16–73 ppm (mean 33) and in 1976 from 44–88 ppm (mean 72). Mean annual basin temperatures were 28.8 and 26.0 C, respectively. Approximately 1500 m in the receiving swamp below the basin, mean pH and temperature were 6.8 and 22.2 C for the first year, and 5.4 and 22.1 C for the second. Total culturable bacteria and diversity (colony types) were reduced at all sampling stations by 44 and 30 percent, respectively, whereas the percentage of the population comprised of chromagenic bacteria increased by 51 percent at the lower pH; Data indicated the pH had a greater effect than did water temperature when temperature was within the range of 15–25 C. The predominant genera within the system in the first year were Bacillus, Sarcina, Achromobacter, Flavobacterium, and Pseudomonas. In the second year, at the lower pH, predominant genera were Pseudomonas, Flavobacterium, Chromobacterium, Bacillus, and Brevibacterium.     

Assessing drilling mud and technical fluid contamination in rock core and brine samples intended for microbiological monitoring at the CO2 storage site in Ketzin using fluorescent dye tracers

The CO₂SINK project in Ketzin represents a field laboratory for the storage of CO₂ in a 650-m deep saline aquifer. The project is accompanied by a microbiological monitoring programme to characterise the composition and activity of the autochthonous microbial community in rock and brine samples and their changes in response to CO₂ storage. A prerequisite of these studies is the acquisition of samples free of contamination from microorganisms and organic and inorganic components. Drilling mud and technical fluids are the main sources of contamination. This study describes the application of the fluorescent dye tracers fluorescein and rhodamine B as contamination controls for rock core and brine samples. Fluorescein was added to drilling mud that was used during the coring phase of the Ketzin wells Ktzi 200, 201 and 202. In addition, total organic carbon (TOC) concentrations, reflecting the carboxymethyl cellulose (CMC) component of the drilling mud, were determined to verify the tracer results. The fluorescence and TOC analyses revealed that drilling mud filtrate penetrated the outer 20 mm of mildly permeable sandstone cores. Rhodamine B was added to brines that were used to displace the drilling mud and to flush the wells after completion. The tracer monitoring during the discharge of drilling mud and displacement brines from the wells during hydraulic tests and nitrogen lifts enabled the quantification of reservoir fluid quality. After the production of 140–190 m³ (16–21 borehole volumes) of fluid, the drilling mud concentration was reduced to about 0.05%. The use of fluorescein emerged as a field-capable, sensitive and reliable method during the sampling of rock core and formation brine samples.     

Effect of Cattle Grazing and Seedling Size on the Establishment ofAraucaria cunninghamiiin a Silvo-pastoral System in Northeast Australia

A study was undertaken to assess the effect of tree age at which grazing commenced and seedling size on the establishment ofAraucaria cunninghamii(hoop pine) trees in a silvo-pastoral system on the Atherton Tablelands of northeast Australia. Grazing with dairy cattle commenced at three ages after tree planting: (1) 2 weeks; (2) 6 months and (3) 18 months. Seedlings of two size classes, small (15 cm tall and 18 months old) and large (45 cm tall and 27 months old), were used. Trees were measured for survival and height, and assessed for physical damage and development of multi-stems at ages 6, 12 and 24 months. Results showed that tree survival and height did not differ significantly with age at which grazing commenced and that grazing did not cause unacceptable mortality, damage or reduction in height growth ofA. cunninghamii. The lack of cattle impact was mainly becauseA. cunninghamiihas a special morphological feature, prickly needles, which discourages cattle from physical contact. Although tree height ofA. cunninghamiivaried significantly between the two seedling sizes, the difference became smaller as the trees grew. It is suggested that for the establishment ofA. cunninghamiiin silvo-pastoral systems, grazing can be allowed almost immediately after tree planting, providing an appropriate stocking rate is used. There are no apparent advantages in using larger, compared with smaller, seedlings.     

Soil properties affecting wheat yields following drilling-fluid application

Oil and gas drilling operations use drilling fluids (mud) to lubricate the drill bit and stem, transport formation cuttings to the surface, and seal off porous geologic formations. Following completion of the well, waste drilling fluid is often applied to cropland. We studied potential changes in soil compaction as indicated by cone penetration resistance, pH, electrical conductivity (EC(e)), sodium adsorption ratio (SAR), extractable soil and total straw and grain trace metal and nutrient concentrations, and winter wheat (Triticum aestivum L. 'TAM 107') grain yield following water-based, bentonitic drilling-fluid application (0-94 Mg ha(-1)) to field test plots. Three methods of application (normal, splash-plate, and spreader-bar) were used to study compaction effects. We measured increasing SAR, EC(e), and pH with drilling-fluid rates, but not to levels detrimental to crop production. Field measurements revealed significantly higher compaction within areas affected by truck travel, but also not enough to affect crop yield. In three of four site years, neither drilling-fluid rate nor application method affected grain yield. Extractions representing plant availability and plant analyses results indicated that drilling fluid did not significantly increase most trace elements or nutrient concentrations. These results support land application of water-based bentonitic drilling fluids as an acceptable practice on well-drained soils using controlled rates.