Various laboratory-scale process configurations were tested for the biological treatment of a combined wastewater stream of several chemical factories. The untreated wastewaters, rich in halogenated organics (1250±389 mg/l DOC), were also highly saline (32±11 g/liter TDS 550°C) and toxic (Microtox™ EC50 = 1.5±2.0%). Biphasic (anaerobic/aerobic) laboratory bench-scale reactor systems yielded reduction of dissolved organic carbon by 70 to 84%, in the absence and presence of powdered activated carbon, respectively. The anaerobic phase proved to be essential in all systems, both for dissolved organic carbon removal and for detoxification. Similar efficiencies were obtained in either activated sludge or aerated lagoon type reactors, but in the latter case, longer hydraulic retention times were required. DOC removal was found to decrease with increased salt concentration; however, a 50% efficiency was achieved even at 90 g/l TDS. Toxicity elimination as judged by the Microtox™ assay was highly variable in the absence of activated carbon but stable and efficient in its presence.
Ever since the introduction of the Salmonella typhimurium mammalian microsome mutagenicity assay (the 'Ames test') over three decades ago, there has been a constant development of additional genotoxicity assays based upon the use of genetically engineered microorganisms. Such assays rely either on reversion principles similar to those of the Ames test, or on promoter-reporter fusions that generate a quantifiable dose-dependent signal in the presence of potential DNA damaging compounds and the induction of repair mechanisms; the latter group is the subject of the present review. Some of these assays were only briefly described in the scientific literature, whereas others have been developed all the way to commercial products. Out of these, only one, the umu-test, has been fully validated and ISO- and OECD standardized. Here we review the main directions undertaken in the construction and testing of bacterial-based genotoxicity bioassays, including the attempts to incorporate at least a partial metabolic activation capacity into the molecular design. We list the genetic modifications introduced into the tester strains, compare the performance of the different assays, and briefly describe the first attempts to incorporate such bacterial reporters into actual genotoxicity testing devices.
This chapter provides basic instructions in the use of genetically engineered Escherichia coli strains that luminesce in response to the presence of oxidants. These strains carry plasmid dertvatives of pUCD615 (1) in which Vibrio fischeri 1uxCDABE is driven by selected promoters of genes responstve to oxidative stress, The construction of such plasmids has been reported before (2–5) and will not be detailed here.
Objectives: The overall objective as defined in the approved proposal was the development of a whole-cell sensor panel for the detection of endocrine disruption activities of agriculturally relevant chemicals. To achieve this goal several specific objectives were outlined: (a) The development of new genetically engineered wholecell sensor strains; (b) the combination of multiple strains into a single sensor panel to effect multiple response modes; (c) development of a computerized algorithm to analyze the panel responses; (d) laboratory testing and calibration; (e) field testing. In the course of the project, mostly due to the change in the US partner, three modifications were introduced to the original objectives: (a) the scope of the project was expanded to include pharmaceuticals (with a focus on antibiotics) in addition to endocrine disrupting chemicals, (b) the computerized algorithm was not fully developed and (c) the field test was not carried out. Background: Chemical agents, such as pesticides applied at inappropriate levels, may compromise water quality or contaminate soils and hence threaten human populations. In recent years, two classes of compounds have been increasingly implicated as emerging risks in agriculturally-related pollution: endocrine disrupting compounds (EDCs) and pharmaceuticals. The latter group may reach the environment by the use of wastewater effluents, whereas many pesticides have been implicated as EDCs. Both groups pose a threat in proportion to their bioavailability, since that which is biounavailable or can be rendered so is a priori not a threat; bioavailability, in turn, is mediated by complex matrices such as soils. Genetically engineered biosensor bacteria hold great promise for sensing bioavailability because the sensor is a live soil- and water-compatible organism with biological response dynamics, and because its response can be genetically “tailored” to report on general toxicity, on bioavailability, and on the presence of specific classes of toxicants. In the present project we have developed a bacterial-based sensor panel incorporating multiple strains of genetically engineered biosensors for the purpose of detecting different types of biological effects. The overall objective as defined in the approved proposal was the development of a whole-cell sensor panel for the detection of endocrine disruption activities of agriculturally relevant chemicals. To achieve this goal several specific objectives were outlined: (a) The development of new genetically engineered wholecell sensor strains; (b) the combination of multiple strains into a single sensor panel to effect multiple response modes; (c) development of a computerized algorithm to analyze the panel responses; (d) laboratory testing and calibration; (e) field testing. In the course of the project, mostly due to the change in the US partner, three modifications were introduced to the original objectives: (a) the scope of the project was expanded to include pharmaceuticals (with a focus on antibiotics) in addition to endocrine disrupting chemicals, (b) the computerized algorithm was not fully developed and (c) the field test was not carried out. Major achievements: (a) construction of innovative bacterial sensor strains for accurate and sensitive detection of agriculturally-relevant pollutants, with a focus on endocrine disrupting compounds (UK and HUJ) and antibiotics (HUJ); (b) optimization of methods for long-term preservation of the reporter bacteria, either by direct deposition on solid surfaces (HUJ) or by the construction of spore-forming Bacillus-based sensors (UK); (c) partial development of a computerized algorithm for the analysis of sensor panel responses. Implications: The sensor panel developed in the course of the project was shown to be applicable for the detection of a broad range of antibiotics and EDCs. Following a suitable development phase, the panel will be ready for testing in an agricultural environment, as an innovative tool for assessing the environmental impacts of EDCs and pharmaceuticals. Furthermore, while the current study relates directly to issues of water quality and soil health, its implications are much broader, with potential uses is risk-based assessment related to the clinical, pharmaceutical, and chemical industries as well as to homeland security.
The benefits of bioluminescence for nonsymbiotic marine bacteria have not been elucidated fully. One of the most commonly cited explanations, proposed more than 30 y ago, is that bioluminescence augments the propagation and dispersal of bacteria by attracting fish to consume the luminous material. This hypothesis, based mostly on the prevalence of luminous bacteria in fish guts, has not been tested experimentally. Here we show that zooplankton that contacts and feeds on the luminescent bacterium Photobacterium leiognathi starts to glow, and demonstrate by video recordings that glowing individuals are highly vulnerable to predation by nocturnal fish. Glowing bacteria thereby are transferred to the nutritious guts of fish and zooplankton, where they survive digestion and gain effective means for growth and dispersal. Using bioluminescence as bait appears to be highly beneficial for marine bacteria, especially in food-deprived environments of the deep sea.
