Recent developments in and intercomparisons of acute and chronic bioassays and bioindicators

The ultimate goal of toxicity testing is to monitor or predict the effects of single compounds, elements or mixtures on the long-term health of individual organisms, populations, communities and ecosystems. Unfortunately, one does not always have all of the information required to determine the long-term or chronic effects of toxicants on the survival, growth or reproduction of aquatic organisms. For this reason, the chronic effects of toxicants are often inferred or estimated from observations made during short-term or or acute field or laboratory studies, which may be conducted at greater concentrations of toxicant. The observations made in the short-term studies are then related to the chronic effects by some statistical relationship. There are basically two approaches: 1) The long-term effects on a parameter, such as survival (lethality.) are predicted from observations on the same parameter, during short-term exposures; 2) Alternatively, the response of one parameter to long-term exposures of a toxicant can be predicted from the short-term responses of a different parameter. In this report we present several different examples of both types of methods for estimating chronic responses from information on more short-term responses and discuss the rationale, advantages and disadvantages of each. We also report on two biochemical indicators; energetic substrates and RNA/DNA ratio. These indicators both act as sensitive, integrative measures of sublethal effects of contaminants during both acute and chronic exposures.     

In vitro mitogenesis of peripheral blood lymphocytes from rainbow trout (Salmo gairdneri)

1. In vitro mitogenesis of rainbow trout peripheral blood lymphocytes (RBT PBL) was investigated to assess the applicability of this procedure in assessment of fish health. The assay variables of media, mitogen type and concentration, serum supplementation, lymphocyte isolation procedure, and duration of incubation were assessed. 2. Concanavalin A (Con A) stimulated greater proliferation of RBT PBL than did lipopolysaccharide (LPS), phytohemagglutinin (PHA), or pokeweed mitogen (PWM). 3. RBT PBL, cultured with 10 micrograms Con A/ml and incubated for four or five days, exhibited greater proliferation than with other treatment combinations. 4. The degree of Con A-induced PBL proliferation varied significantly (P less than 0.05) among fish. The mean was positively correlated with the relative standard deviation and thus exhibited significant heteroscedasticity. 5. Human serum, as an alternative to FBS supplementation of the culture medium, did not enhance RBT PBL proliferation or reduce variation in mean proliferation. 6. Power analysis with variance estimates from this study reveal that sample size requirements of further studies under the given conditions could severely limit the applicability of this procedure for RBT health assessment. Further work in this area should center around standardization of culture conditions pertaining to the source of protein supplementation.     

A strategy for finding regions of similarity in complete genome sequences

MOTIVATION: Complete genomic sequences will become available in the future. New methods to deal with very large sequences (sizes beyond 100 kb) efficiently are required. One of the main aims of such work is to increase our understanding of genome organization and evolution. This requires studies of the locations of regions of similarity. RESULTS: We present here a new tool, ASSIRC ('Accelerated Search for SImilarity Regions in Chromosomes'), for finding regions of similarity in genomic sequences. The method involves three steps: (i) identification of short exact chains of fixed size, called 'seeds', common to both sequences, using hashing functions; (ii) extension of these seeds into putative regions of similarity by a 'random walk' procedure; (iii) final selection of regions of similarity by assessing alignments of the putative sequences. We used simulations to estimate the proportion of regions of similarity not detected for particular region sizes, base identity proportions and seed sizes. This approach can be tailored to the user's specifications. We looked for regions of similarity between two yeast chromosomes (V and IX). The efficiency of the approach was compared to those of conventional programs BLAST and FASTA, by assessing CPU time required and the regions of similarity found for the same data set. AVAILABILITY: Source programs are freely available at the following address: ftp://ftp.biologie.ens. fr/pub/molbio/assirc.tar.gz CONTACT: vincens@biologie.ens.fr, hazout@urbb.jussieu.fr      

Plasma free amino acid profiles and nutrition proteins in chronic renal failure; effect of dialysis treatment

Summary A well preserved nutritional status is beneficial in chronically uremic patients for slowing the pace of deterioration of renal function, and delaying the need for dialysis therapy. The purpose of this study was to assess the nutritional profile of 10 patients in a steady state of advanced CRF, and of 15 patients with terminal renal failure immediately prior to their first hemodialysis session (J0), and 7, 14, 45, 60, days post start of dialysis. Patients were 18 to 65 years old with total plasma proteins 60g/1. Plasma concentrations of amino acids, nutrition proteins, apolipoproteins A1, and B were evaluated. Non inflammatory reaction was evaluated by determination of alpha-1-acid glycoprotein, and C reactive protein. The data (mean ± 1 SD) were compared with mean values of 15 healthy individuals.     

Effects of Naturally Occurring Aquatic Organic Fractions on Am Uptake by Scenedesmus obliquus (Chlorophyceae) and Aeromonas hydrophila (Pseudomonadaceae)

Naturally occurring organics were extracted from water collected from Skinface Pond near Aiken, S.C. Organics were separated into four nominal diameter size fractions (I, >0.0183; II, 0.0183 to 0.0032; III, 0.0032 to 0.0009; IV, <0.0009 mum) by membrane ultrafiltration and introduced into Scenedesmus obliquus and Aeromonas hydrophila cultures to determine their effects on Am availability for uptake. Effects on Am uptake were determined in actively growing S. obliquus cultures after 96 h of growth and in dense cultures of nongrowing cells after 4 h. Uptake by A. hydrophila was determined after 4 and 24 h in actively growing cultures. All organic fractions stimulated S. obliquus growth, with the most pronounced effects due to larger organic fractions, whereas no apparent growth stimulation of A. hydrophila was observed for any organic fraction. For both long-term and short-term studies, cellular Am concentration (picocuries/cell) increased with increasing Am concentration for S. obliquus and A. hydrophila. Fraction IV increased Am uptake by both S. obliquus and A. hydrophila during 4-h incubations. During 96-h incubations fraction I was flocculated and cosedimented, with S. obliquus and A. hydrophila cells causing an apparent increase in Am uptake. Fractions II and III reduced apparent Am uptake by S. obliquus as a result of biological dilution caused by increased algal growth due to the organics. Fraction IV caused a reduction in Am uptake by S. obliquus not attributable to biological dilution. Organics increased Am uptake by A. hydrophila during 4- and 24-h incubations. A. hydrophila also caused flocculation of fraction I during 96-h incubations.