Saline-saturated DMSO-EDTA as a storage medium for microbial DNA analysis from coral mucus swab samples

The mucus surface layer of corals plays a number of integral roles in their overall health and fitness. This mucopolysaccharide coating serves as vehicle to capture food, a protective barrier against physical invasions and trauma, and serves as a medium to host a community of microorganisms distinct from the surrounding seawater. In healthy corals the associated microbial communities are known to provide antibiotics that contribute to the coral’s innate immunity and function metabolic activities such as biogeochemical cycling. Culture-dependent (Ducklow and Mitchell, 1979; Ritchie, 2006) and culture-independent methods (Rohwer, et al., 2001; Rohwer et al., 2002; Sekar et al., 2006; Hansson et al., 2009; Kellogg et al., 2009) have shown that coral mucus-associated microbial communities can change with changes in the environment and health condition of the coral. These changes may suggest that changes in the microbial associates not only reflect health status but also may assist corals in acclimating to changing environmental conditions. With the increasing availability of molecular biology tools, culture-independent methods are being used more frequently for evaluating the health of the animal host. Although culture-independent methods are able to provide more in-depth insights into the constituents of the coral surface mucus layer’s microbial community, their reliability and reproducibility rely on the initial sample collection maintaining sample integrity. In general, a sample of mucus is collected from a coral colony, either by sterile syringe or swab method (Woodley, et al., 2008), and immediately placed in a cryovial. In the case of a syringe sample, the mucus is decanted into the cryovial and the sealed tube is immediately flash-frozen in a liquid nitrogen vapor shipper (a.k.a., dry shipper). Swabs with mucus are placed in a cryovial, and the end of the swab is broken off before sealing and placing the vial in the dry shipper. The samples are then sent to a laboratory for analysis. After the initial collection and preservation of the sample, the duration of the sample voyage to a recipient laboratory is often another critical part of the sampling process, as unanticipated delays may exceed the length of time a dry shipper can remain cold, or mishandling of the shipper can cause it to exhaust prematurely. In remote areas, service by international shipping companies may be non-existent, which requires the use of an alternative preservation medium. Other methods for preserving environmental samples for microbial DNA analysis include drying on various matrices (DNA cards, swabs), or placing samples in liquid preservatives (e.g., chloroform/phenol/isoamyl alcohol, TRIzol reagent, ethanol). These methodologies eliminate the need for cold storage, however, they add expense and permitting requirements for hazardous liquid components, and the retrieval of intact microbial DNA often can be inconsistent (Dawson, et al., 1998; Rissanen et al., 2010). A method to preserve coral mucus samples without cold storage or use of hazardous solvents, while maintaining microbial DNA integrity, would be an invaluable tool for coral biologists, especially those in remote areas. Saline-saturated dimethylsulfoxide-ethylenediaminetetraacetic acid (20% DMSO-0.25M EDTA, pH 8.0), or SSDE, is a solution that has been reported to be a means of storing tissue of marine invertebrates at ambient temperatures without significant loss of nucleic acid integrity (Dawson et al., 1998, Concepcion et al., 2007). While this methodology would be a facile and inexpensive way to transport coral tissue samples, it is unclear whether the coral microbiota DNA would be adversely affected by this storage medium either by degradation of the DNA, or a bias in the DNA recovered during the extraction process created by variations in extraction efficiencies among the various community members. Tests to determine the efficacy of SSDE as an ambient temperature storage medium for coral mucus samples are presented here.

Comparison of stabilised/solidified mixes of model drill cuttings based on the North Sea and Red Sea Areas

Following the global stringent legislations regulating the wastes generated from the drilling process of oil exploration and production activities, the management of hazardous drill cuttings has become one of the pressing needs confronting the petroleum industry. Most of the prevalent treatment techniques adopted by oil companies are extremely expensive and/or the treated product has to be landfilled without any potential end-use; thereby rendering these solutions unsustainable. The technique of stabilisation/solidification is being investigated in this research to treat drill cuttings prior to landfilling or for potential re-use in construction products. Two case studies were explored namely North Sea and Red Sea. Given the known difficulties with stabilising/solidifying oils and chlorides, this research made use of model drill cutting mixes based on typical drill cutting from the two case studies, which contained 4.2% and 10.95% average concentrations of hydrocarbons; and 2.03% and 2.13% of chlorides, by weight respectively. A number of different binders, including a range of conventional viz. Portland cement (PC) as well as less-conventional viz. zeolite, or waste binders viz. cement kiln dust (CKD), fly ash and compost were tested to assess their ability to treat the North Sea and Red Sea model drill cuttings. The dry binder content by weight was 10%, 20% and 30%. In addition, raw drill cuttings from one of the North Sea offshore rigs were stabilised/solidified using 30% PC. The characteristics of the final stabilised/solidified product were finally compared to those of thermally treated cuttings. The effectiveness of the treatment using the different binder systems was compared in the light of the aforementioned two contaminants only. A set of physical tests (unconfined compressive strength (UCS)), chemical tests (NRA leachability) and micro-structural examinations (using scanning electron microscopy (SEM), and X-ray diffraction (XRD)) were used to evaluate the relative performance of the different binder mixes in treating the drill cuttings. The results showed that the observed UCS covered a wide range of values indicating various feasible end-use scenarios for the treated cuttings within the construction industry. The teachability results showed the reduction of the model drill cuttings to a stable non-reactive hazardous waste, compliant with the UK acceptance criteria for non-hazardous landfills: (a) by most of the 30% and 20% binders for chloride concentrations, and (b) by the 20% and 30% of compost-PC and CKD-PC binders for the Red Sea cuttings. The 20% and 30% compost-PC and CKD-PC binders successfully reduced the leached oil concentration of the North Sea cuttings to inert levels. Copyright 2007, Society of Petroleum Engineers.

Marketing of value-added products from silver carp and involvement of rural people in the production and marketing chain

Cost-profit analysis and market testing of some value-added products from silver carp such as fish mince block, fish sausage, fish ball, fish stick and fish burger were analyzed during April 2001 to March 2002. The study also explored the possibility to involve rural low-income people in the production and marketing of such products. The production of silver carp was higher in greater Jessore and Mymensingh districts but the price remained low during the peak-harvesting season in October to November. The price varied with size of the fish, season, market characteristics and effective demand of the buyers. Price of about 500 g size fish was found to be Tk. 20-25/kg in the rural markets. The average size of fish in the rural markets was 3S0-550 g while that in the urban markets it was 700-1,200 g. The cost of production of the value added products and profit margin were assessed on the basis of market price of the raw material as well as that of the finished products, transportation, storage and marketing costs. The profit margins of 34%, 39%, 81% and 31% of their sales price were obtained for fish sausage, fish ball, fish stick and fish burger, respectively. Actual production cost could be minimized if the fish is purchased directly from the farmers. Consumer's acceptance and marketability tests showed that both rural and urban people preferred fish ball than fish sausage. However, response towards the taste, flavor and color of fish ball and fish sausage was found to vary with occupations and age of the consumers. A correlation was observed between age group and acceptance of new products. Fish ball, fish stick and fish burger were found to be the most preferable items to the farmers because of easy formulation process with common utensils. Good marketing linkage and requirement of capital had been identified as the prerequisites for operating small-scale business on value-added fish products.

The long road to freedom: the end of apartheid has signalled a new beginning for women in South Africa’s fisheries but real equality is still a far-off dream

Women in fishing communities are increasingly moving from traditional, community based occupations to seeking employment in the labour market. While this is an opportunity for women, their employment is also largely in the male dominated fishing industry, where job segregation into ‘less skilled and low paid’ jobs for women define employment opportunities. However, engagement as members in local non-government networks help women to challenge these stereotypes. In South Africa, for instance, the recent legislation promoting opportunity for women in male dominated sectors of employment is an opportunity for women to earn wages equal to those of men.

Part II: Binders & technologies - Research

The first report of report series I, II and III entitled 'basic principles' presented details of the binders and technologies available and used in the stabilisation/ solidification (S/S) treatment of hazardous waste and contaminated land. This second report entitled 'research' presents an overview of the main research work, both experimental and numerical, carried out in the UK concentrating on the last decade or so but also highlighting earlier significant research work. The research work is reported under the headings of the individual binders and for each binder the work is presented in chronological order. In this work, most of the S/S materials are prepared by manual/mechanical mixing. The latter part of this report presents research work on S/S materials prepared using soil mixing with mixing augers. © 2005 Taylor & Francis Group.

A condensation particle counter insensitive to volatile particles

A High Temperature Condensation Particle Counter (HT-CPC) is described that operates at an elevated temperature of up to ca. 300. °C such that volatile particles from typical combustion sources are not counted. The HT-CPC is functionally identical to a conventional CPC, the main challenge being to find suitable non-hazardous working fluids, with good stability, and an appropriate vapour pressure. Some key design features are described, and results of modelling which predict the HT-CPC counting efficiency. Experimental results are presented for several candidate fluids when the HT-CPC was challenged with ambient, NaCl and diesel soot particles, and the results show good agreement with modelled predictions, and confirm that counting of particles of diameters down to at least 10. nm was achievable. Possible applications are presented, including measurement of particles from a diesel car engine and comparison with a near PMP system. © 2014 Elsevier Ltd.

Degradation of 17 alpha-ethinylestradiol in aqueous solution by ozonation

This study investigates the ozonation of 17 alpha-ethinylestradiol (EE2) in aqueous solution. The affecting factors on the degradation of EE2 were studied and described in details, such as initial EE2 concentration, initial pH value and ozone concentration. In addition, some parameters such as pH. electrical conductivity, mineralization efficiency and degradation products were monitored during the process. The mineralization efficiency of EE2 could reach 53.9%. During the ozonation process the rapid decrease of pH and the sharp increase of electrical conductivity indicated the fort-nation of acidic by-products, small fragments and ions which were confirmed by high performance liquid chromatography (HPLC) and gas chromatography-mass spectrometry (GUMS) analysis. Results showed that there were intermediate products of smaller molecule with higher polarity produced during the course of EE2 degradation. Then a possible reaction pathway for EE2 degradation involving all intermediates detected is proposed. During the ozonation process EE2 was first oxidized into hydroxyl-semiquinone isomers which were subsequently degraded into low molecular weight compounds such as oxalic acid, malonate, glutarate, and so on. Furthermore. these organic acids are easily oxidized by ozone into carbon dioxide (CO2). This work shows that ozonation process is promising for the removal of EE2. The results can provide some useful information for the potential treatment of EE2 by ozonation in aqueous solution. (c) 2005 Elsevier B.V. All rights reserved.

Growth and antioxidant system of the cyanobacterium Synechococcus elongatus in response to microcystin-RR

Microcystins, one type of the cyanobacterial toxins, show a broad range of hazardous effects on other organisms. Most of the researches on the toxic effects of microcystins have involved in animals and higher plants. Little work, however, has been done on evaluating the mechanisms of microcystin toxicity on algae. In this study, the toxicological effects of microcystin-RR (MC-RR) on the cyanobacterium Synechococcus elongatus were investigated. For this purpose, six physio-biochemical parameters (cell optical density, reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH), glutathione peroxidase (GSH-Px) and glutathione S-transferase (GST)) were tested in algal cells when exposed to 100 mug(-1) microcystin-RR. The results showed that the growth of Synechococcus elongatus ( expressed as optical density) was significantly inhibited compared with the control. At the same time, the treated algae exhibited a pronounced increase in production of ROS and MDA after 6 days exposure to microcystin-RR. Signi. cant changes in GSH levels and GSH-Px, GSH activities were also detected in algal cells, with higher values being observed in the toxin treated algae after 6 days exposure. GST activities in the treated algae exhibited a decline after exposure and rapid augmentation on day 3, thereafter, they kept at a high level when compared to the control group. GSH contents and GSH-Px activities were also significantly raised in the toxin-treated algae cells from day 3, but they showed a sharp decrease on day 4, which was the onward of cell proliferation. These results suggested that oxidative stress manifested by elevated ROS levels and MDA contents might be responsible for the toxicity of microcystin to Synechococcus elongatus and the algal cells could improve their antioxidant ability through the enhancement of enzymatic and non-enzymatic preventive substances.

Effects of nitrogen forms on the production of cyanobacterial toxin microcystin-LR by an isolated Microcystis aeruginosa

A cyanobacterial strain, which produced high content of microcystin-LR (MC-LR) but no rnicrocystin-RR (MC-RR), was isolated from the hypertrophic Dianchi Lake in China and identified as Microcystis aeruginosa DC-1. Effects of nitrogen containing chemicals and trace elements on the growth and the production of MC-LR by this strain were Studied. In the presence of bicine, compared with urea and ammonium, nitrate greatly promoted the growth and the production of MC-LR. However, leucine and arginine, which were the constitutional components in the molecular structure of MC-LR or RR, inhibited the production of MC-LR. Iron and silicon up to 10mg/L had little effects on the growth of M. aeruginosa DC-1, but the production of MC-LR was apparently enhanced. Under all conditions studied here, only MC-LR but no RR was detected within the cells of M. aeruginosa DC-1. Thus, chemical forms of nitrogen, rather than the usually concerned the total nitrogen, Lind trace elements played important roles in the production of MC toxins during cyanobacterial blooms.

Variation of microcystins in a lake for water supply

A site investigation was conducted to correlate the relationships between microcystins (MC) concentration and algal growth in Dianchi Lake in China. Laboratory experiments were undertaken to test the effects of sediment adsorption, photoirradiation and biodegradation on microcystins removal. Bioaccumulation of microcystins was also determined using silver carp fish. It was observed that MC concentrations varied in accordance with algae growth in Dianchi Lake. The results obtained in the laboratory demonstrated that the removal of MC with fresh sediments was less than 10%, photoirradiation removed more than 75% MC within two hours, and the biodegradation needed much longer time to produce substantial degradation of MC. The results suggest that bioaccumulation of microcystins in fish was not significant in Dianchi Lake.

Photoluminescence studies on pseudomorphic delta-doped AlGaAs/InGaAs/GaAs quantum wells

Photoluminescence (PL) measurements were performed on several series of single-side Si-doped pseudomorphic high electron mobility transistors (p-HEMTs) quantum well (QW) samples, with different spacer layer widths, well widths and Si delta -doped concentrations , under different temperatures and excitation power densities. The dynamic competitive luminescence mechanism between the radiations of e2-hh1 and e1-hh1 was discussed in detail. The confining potential, subband energies, corresponding envelope functions, subband occupations and transferring efficiency etc., were calculated by self-consistent finite differential method at different temperatures in comparison with the present experiment results. The relative variation of the integrated luminescence intensity of the two transitions (e1-hh1 and e2-hh1) was found to be dependent on the temperature and the structure's properties, e. g. spacer layer width, dopant concentration and well width.

Species transformation of trace elements and their distribution prediction in dyestuff residue incineration

The release of heavy metals from the combustion of hazardous wastes is an environmental issue of increasing concern. The species transformation characteristics of toxic heavy metals and their distribution are considered to be a complex problem of mechanism. The behavior of hazardous dyestuff residue is investigated in a tubular furnace under the general condition of hazardous waste pyrolysis and gasfication. Data interpretation has been aided by parallel theoretical study based on a thermodynamic equilibrium model based on the principle of Gibbs free energy minimization. The results show that Ni, Zn, Mn, and Cr are more enriched in dyestuff residue incineration than other heavy metals (Hg, As, and Se) subjected to volatilization. The thermodynamic model calculation is used for explaining the experiment data at 800 degrees C and analyzing species transformation of heavy metals. These results of species transformation are used to predict the distribution and emission characteristics of trace elements. Although most trace element predictions are validated by the measurements, cautions are in order due to the complexity of incineration systems.