An optimized method for automated analysis of algal pigments by HPLC

A recent development in algal pigment analysis by high-performance liquid chromatography (HPLC) is the application of automation. An optimization of a complete sampling and analysis protocol applied specifically in automation has not yet been performed. In this paper we show that automation can only be successful if the various methodological aspects of the sampling and analysis protocol are considered in coherence. We introduce an optimized protocol that involves freeze-drying of the sample, subsequent extraction in 90% acetone and the application of water-packing during analysis. The method was evaluated on both natural plankton populations and a broad spectrum of microalgal cultures: Thalassiosira weisflogii (Bacillariophyceae), Emiliania huxleyi (Prymnesiophyceae), Phaeocystis globosa and Phaeocystis antarctica (Prymnesiophyceae) and Pyramimonas sp. (Prasinophyceae). Whereas pigment extracts were unstable in methanol, with recorded chlorophyll a losses from 10% to 60% per day, pigment degradation rates in acetone were generally less than 1% over 18 h storage in the autosampler (4 °C). In addition, it was found that the extraction efficiency of acetone significantly increased upon freeze-drying prior to extraction. Increases as high as 50–60% were measured in P. antarctica. The application of water-packing of the sample during injection resulted in improved peak shape and peak separation, without diluting the pigment concentrations. Automation is especially beneficial for application in the field, when mixed algal assemblages and low biomass put a high demand on the sensitivity as well as reproducibility of the method.     

Developing a Marine Information System by Integrating Existing Ocean Models Using Object-Oriented Technology

Information systems developed for different applications within the environmental domain have common characteristics, which can potentially be abstracted for sharing and reuse of design and software modules. This article presents an approach to designing for reuse by abstracting commonalities in the design of a Marine Information System (MIS), facilitating data management in a prototype operational monitoring, forecasting, and management system for the North Atlantic and the Nordic Seas. A detailed study of the requirements and data analysis was carried out, and Object-Oriented Technology (OOT) is employed to encapsulate abstractions and to promote reuse of code and design. This article identifies the Object-Oriented Frameworks (OOFW) required to build the MIS. It also provides guidelines to environmental scientists for restructuring legacy software and employing modern programming techniques.     

Inhomogeneous substrate analysis using EM300 backscatter imagery

Backscatter reflectivity from multibeam echo-sounders provides a powerful tool to efficiently characterize seafloor substrates. A comprehensive EM300 bathymetric and backscatter survey has been completed of Cook Strait, in central New Zealand. This paper presents a detailed analysis of the realtime corrections applied to the raw EM300 multibeam data and additional corrections required to compute angular variations of the backscatter strength. The corrections, including the local absorption coefficient, the influence of seafloor topography and sound refraction in the water column, are determined for different Cook Strait seafloor substrates. Modifying MB-System software code, we extracted the backscatter signal parameters in order to quantify the raw backscatter strength and apply additional processing. Profiles of backscatter strength versus incidence angle were computed for a variety of sites characterized by flat seafloor and homogeneous substrates, and for which ground-truth data were available. For each homogeneous site, different but characteristic backscatter profiles are observed that can be interpreted in terms of sediment facies. To analyze heterogeneous substrates, we present a statistical technique, based on a 3-dimensional distribution of (incidence angle, backscatter strength) couples that preserves geological information of the substrate components. This analysis, using backscatter data acquired on a submarine volcano, north of New Zealand, clearly differentiates soft sediments and lava flows within a heterogeneous substrate.     

Partitioning of polycyclic aromatic hydrocarbons between water and particles compared to bioaccumulation in mussels: a harbour case

Water and particles from three sites located in Halifax Harbour, near and further away from combined municipal sewage effluents (CMSE) were analysed for parental and alkylated polycyclic aromatic hydrocarbons (par and alkPAH). Bioavailability of PAH was compared for inter-tidal mussels collected at the same sites in April and November. The PAH fingerprint determined over 9 sampling times covering a period of 19 months differed more between phases (water and particles) and seasons, than between sites. In the spring, more alkPAH associated with diesel and gasoline were detected in the soluble phase, along with more bioaccumulation of alkPAH in inter-tidal mussels. A broader number of parPAH were detected in mussels collected in the fall. The mean sum of dissolved alkPAH concentrations was higher in water at the site closer to raw CMSE than at the other two sites and particles of that site. However, lowest bioconcentration factors (BCF) were determined in mussels of this more contaminated site. Similar biota-particle accumulation factors (BPAF) were determined for parPAH in mussels from the three sites, all lower than the BCF of alkPAH. The study indicates that sewage treatment plants will reduce the amount of parPAH especially larger than fluoranthene and pyrene from being deposited in the harbour; that alk naphthalenes and fluorenes present in water will continue to be discharged; that the disturbance of sediments can make particle-bound PAH available to mussels.     

Estradiol and estriol suppress CYP1A expression in rainbow trout primary hepatocytes

Hepatic levels of the pollutant inducible enzyme, CYP1A, are strongly suppressed in spawning female fish, a phenomenon attributed to high plasma levels of the female sex steroid hormone, estradiol. To evaluate the contribution of estrogen metabolites to estradiol-mediated CYP1A regulation, we treated primary hepatocytes isolated from juvenile rainbow trout (Oncorhynchus mykiss) with vehicle, 17beta-estradiol, or the estrogen metabolite, estriol, alone and in combination with each other and with the potent CYP1A inducer, benzo[a]pyrene (B[a]P). We found dose-dependent suppression of B[a]P-induced CYP1A activity by both steroids relative to controls. At 10(-7) M doses, estradiol and estriol suppressed B[a]P-induced CYP1A activity by 3- and 2-fold, respectively. Although not statistically significant, mean basal CYP1A activity levels were 15- and 13-fold lower in estradiol and estriol treated hepatocytes, respectively, relative to vehicle treated controls. Combining doses of estradiol and estriol failed to produce synergistic suppression of either basal or B[a]P-induced CYP1A activity relative to treatment with either steroid alone. The observed suppression is well below the often strong suppression observed in spawning female fish. We conclude that factors in addition to estradiol and estriol are likely involved in producing sexual dimorphism in CYP1A expression observed in spawning fish.     

Variations in wave direction estimated using first and second order Fourier coefficients

In most design applications such as alignment of the berthing structure and breakwater alignment, it becomes necessary to determine the direction of design wave. There are two different approaches to determine wave direction. One involves the use of first order Fourier coefficients (mean wave direction) while the other uses second order Fourier coefficients (principal wave direction). Both the average wave direction over the entire frequency range (0.03–0.58 Hz) and the direction corresponding to the peak frequency are used in practice. In the present study, comparison is made on wave directions estimated based on first and second order Fourier coefficients using data collected at four locations in the west and east coasts of India. Study shows that at all locations, the mean and principal wave directions for frequencies ranging from 0.07 to 0.25 Hz (±0.5 times peak frequency) co-vary with a correlation coefficient of 0.99 but at lower and higher frequencies, difference between the parameters is large. Average difference between the mean wave direction at peak frequency and the average over the frequency related to spectral energy more than 20% of maximum value is less, around 13°. Study shows that average difference in the sea and swell directions is around 39°.