In vitro establishment of continuous cell cultures and cell lines from ten colonial cnidarians

Continuous cell cultures from the ten taxa of sedentary colonial marine cnidariansStylophora pistillata, Porites lutea, Favia favus (Anthozoa, Hexacorallia, Madeporaria),Parerythropodium fulvum fulvum, Dendronephthya hemprichi, Nephthya sp.Heteroxenia fuscescence (Anthozoa, Octocorallia, Alcyonacea),Clathraria rubrinoides, Plexaura A (Anthozoa, Octocorallia, Gorgonacea) andMillepora dichotoma (Hydrozoa) were established in vitro. Primary cultures of various cell types (5 to 20 m) were obtained from colony fragments and/or planula larvae (collected in 1993 from coral reefs at Eilat, Red Sea and from the San Blas islands, Panama) using three dissociation approaches: a mechanical approach, chemical approach and a novel approach, spontaneous dissociation. Cells were cultured in a modified Leibowitz L15 medium, with 5 to 10% heat-inactivated fetal bovine serum, diluted in seawater. Cell proliferation was observed in primary cultures within 7 to 20 d following dissociation. When developed to secondary cultures, spindle-shaped cells (5 to 10 m) gradually replaced all other cell types seen in primary cultures ofS. pistillata, C. rubrinoides, P.f. fulvum andM. dichotoma. The spindle-shaped cells differentiated to several cell types when the diluted medium was replaced with a concentrated one, or when microbial contamination occurred. Secondary cultures of spindle-shaped cells from three species (S. pistillata, C. rubrinoides andM. dichotoma) were cloned. They gave rise to continuously proliferating cell lines (NIO-SPP-1, NIO-CR-1 and NIO-MD-1, respectively). A clone fromPlexaura planula, originated from one rounded cell, started to differentiate and gave rise to a heterogeneous culture. Samples of the above four cultures were frozen for future work. Rounded cells of various sizes (5 to 30 m) dominated secondary cell cultures ofH. fuscescence, D. hemprichi, Nephthya sp.,P. f. fulvum andF. favus. Several cell cultures fromP. lutea developed flattened cell interconnected by ectoplasmic networks characteristic of the eukaryotic unicellular organisms of the phylum Labyrinthulomycota. We propose that this culture methodology may be used as a ubiquitous protocol for producing tissue cultures from other cnidarians. The established cell lines may be used in a variety of disciplines in cnidarian biology and ecology.     

Effect of pH and temperature on the kinetics of odor oxidation using chlorine dioxide

Increasing public concerns over odors and air regulations in nonattainment zones necessitate the remediation of a wide range of volatile organic compounds (VOCs) generated in the poultry-rendering industry. Currently, wet scrubbers using oxidizing chemicals such as chlorine dioxide (ClO2) are utilized to treat VOCs. However, little information is available on the kinetics of ClO2 reaction with rendering air pollutants, limiting wet scrubber design and optimization. Kinetic analysis indicated that ClO2 does not react with hexanal and 2-methylbutanal regardless of pH and temperature and implied that aldehyde removal occurs primarily via mass transfer. Contrary to the aldehydes, ethanethiol or ethyl mercaptan (a model compound for methanethiol or methyl mercaptan) and dimethyl disulfide (DMDS) rapidly reacted with ClO2. The overall reaction was found to be second and third order for ethanethiol and DMDS, respectively. Moreover, an increase in pH from 3.6 to 5.1 exponentially increased the reaction rate of ethanethiol (e.g., k2 = 25-4200 L/mol/sec from pH 3.6 to 5.1) and significantly increased the reaction rate of DMDS if increased to pH 9 (k3 = 1.4 x 10(6) L2/mol2/sec). Thus, a small increase in pH could significantly improve wet scrubber operations for removal of odor-causing compounds. However, an increase in pH did not improve aldehyde removal. The results explain why aldehyde removal efficiencies are much lower than methanethiol and DMDS in wet scrubbers using ClO2.     

Assessment of the phototoxicity of weathered Alaska North Slope crude oil to juvenile pink salmon

Petroleum products are known to have greater toxicity to the translucent embryos and larvae of aquatic organisms in the presence of ultraviolet radiation (UV) compared to toxicity determined in tests performed under standard laboratory lighting with minimal UV. This study assessed the acute phototoxicity of the water accommodated fractions of weathered Alaska North Slope crude oil (ANS) to juvenile pink salmon, which are a heavily pigmented life stage. Fish in the highest ANS treatments exhibited melanosis, less mobility, reduced startle response, erratic swimming, and loss of equilibrium. Gills from fish exposed to ANS had elevated levels of hydroperoxides in oil-only, UV-only, and oil+UV treatments compared to control fish, which was indicative of increased lipid peroxidation in gill tissue. Under the test conditions of moderate salinity, low UV and high short-term oil exposure there were no indications of photoenhanced toxicity as assessed by elevation of mortality, behavioral impairment, or gill lipid peroxidation in oil+UV treatments. The results of this study suggest that pink salmon may be at less risk from photoenhanced toxicity compared to the translucent early-life stages of several other Alaska species.     

Status and trends of fecal indicator bacteria in two urban watersheds.

This paper examines bacterial levels and their causes in two Houston bayous (Texas). Buffalo and Whiteoak bayous are two of the most contaminated water bodies in Texas for indicator bacteria, based on the frequency and magnitude of contact recreation water quality exceedances. Examination of historical data indicates frequent exceedances, although some improvement has been made since the 1970s. Statistical analyses showed some correlation between in-stream fecal coliform concentrations and rainfall and with land use. Differences in fecal coliform concentrations were found between high- and low-flow conditions in Whiteoak Bayou, while reservoir releases confounded this relationship in Buffalo Bayou. Wastewater treatment plant effluent was found to make up two-thirds to three-fourths of the median flow in both bayous. Effluent sampling was conducted at 72 of the approximately 140 wastewater treatment plants (WWTPs) in the watersheds, providing evidence that WWTP effluent could act to maintain low-flow concentrations of fecal coliform in the bayous.     

Decision support tools for collaborative marine spatial planning: identifying potential sites for tidal energy devices around the Mull of Kintyre,Scotland

The expansion of offshore renewable energy production, such as wind, wave and tidal energy, is likely to lead to conflict between different users of the sea. Two types of spatial decision support tools were developed to support stakeholder workshops. A value mapping tool combines regional attributes with local knowledge. A negotiation support tool uses these value maps to support stakeholders in finding acceptable locations for tidal energy devices. Interactive value mapping proved useful to address deficiencies in data and to create credibility for these maps. The negotiation tool helped stakeholders in balancing objectives of the various stakeholders.     

Co-utilisation of CO2 and steelmaking slags for production of pure CaCO3 – legislative issues

The steel industry is characterised by large amounts of CO₂ emissions, but there is no easy means to reduce these emissions. One interesting option for the reduction of CO₂ emissions could be the utilisation of steelmaking slags for carbon dioxide mineralisation. In this option CO₂ is bound with the calcium of the slag material, producing stable carbonate as an end product. The utilisation of steelmaking slags as the raw material for carbon dioxide mineralisation will change the quality of the slags. If, however, this change degrades the slags it could prevent the use of slags in carbon dioxide mineralisation or make it very expensive.The purpose of the research presented here is to evaluate this issue with the help of a case study where the quality of the residual slag from the recently suggested carbonation method was experimentally investigated. The CO₂ mineralisation method, based on steelmaking slags and ammonium salt solutions, was found to change the quality of the slags: the calcium content was reduced, the CaO and Ca(OH)₂ phases were completely dissolved, and the solubility of the V and Cr increased notably. This residual slag would presumably have to be handled as waste. Currently, the steelmaking slag used in the case study is defined as a by-product, but if it is used for CO₂ mineralisation instead of liming its legal status will be re-evaluated. Subsequently, the CO₂ mineralisation process could possibly be defined as an end-of-waste procedure.