Genetic mapping of variation in dauer larvae development in growing populations of Caenorhabditis elegans.

In the nematode Caenorhabditis elegans, the appropriate induction of dauer larvae development within growing populations is likely to be a primary determinant of genotypic fitness. The underlying genetic architecture of natural genetic variation in dauer formation has, however, not been thoroughly investigated. Here, we report extensive natural genetic variation in dauer larvae development within growing populations across multiple wild isolates. Moreover, bin mapping of introgression lines (ILs) derived from the genetically divergent isolates N2 and CB4856 reveals 10 quantitative trait loci (QTLs) affecting dauer formation. Comparison of individual ILs to N2 identifies an additional eight QTLs, and sequential IL analysis reveals six more QTLs. Our results also show that a behavioural, laboratory-derived, mutation controlled by the neuropeptide Y receptor homolog npr-1 can affect dauer larvae development in growing populations. These findings illustrate the complex genetic architecture of variation in dauer larvae formation in C. elegans and may help to understand how the control of variation in dauer larvae development has evolved.

Type of disturbance and ecological history determine structural stability.

This study aims to reveal whether complexity, namely, community and trophic structure, of chronically stressed soil systems is at increased risk or remains stable when confronted with a subsequent disturbance. Therefore, we focused on a grassland with a history of four centuries of patchy contamination. Nematodes were used as model organisms because they are an abundant and trophically diverse group and representative of the soil food web and ecosystem complexity. In a field survey, a relationship between contaminants and community structures was established. Following, two groups of soil mesocosms from the field that differed in contamination level were exposed to different disturbance regimes, namely, to the contaminant zinc and a heat shock. The zinc treatment revealed that community structure is stable, irrespective of soil contamination levels. This implies that centuries of exposure to contamination led to adaptation of the soil nematode community irrespective of the patchy distribution of contaminants. In contrast, the heat shock had adverse effects on species richness in the highly contaminated soils only. The total nematode biomass was lower in the highly contaminated field samples; however, the biomass was not affected by zinc and heat treatments of the mesocosms. This means that density compensation occurred rapidly, i.e., tolerant species quickly replaced sensitive species. Our results support the hypothesis that the history of contamination and the type of disturbance determine the response of communities. Despite that ecosystems may be exposed for centuries to contamination and communities show adaptation, biodiversity in highly contaminated sites is at increased risk when exposed to a different disturbance regime. We discuss how the loss of higher trophic levels from the entire system, such as represented by carnivorous nematodes after the heat shock, accompanied by local biodiversity loss at highly contaminated sites, may result in detrimental effects on ecosystem functions.

Molecular cloning and characterization of Cpn60 in the free-living nematode Plectus acuminatus.

Heat shock proteins (Hsps) have provoked interest not only because of their involvement in human diseases but also for their potential as biomarkers of environmental pollution. Whereas the former interest is covered by numerous reports, the latter is an exciting new field of research. We report the isolation of the full-length cpn60 messenger ribonucleic acid (mRNA) and partial genomic deoxyribonucleic acid from the free-living, environmental sentinel nematode Plectus acuminatus, a species used in classical ecotoxicity tests. Although the primary sequence displays high identity scores to other nematodes and human Cpn60 (75% and 70%, respectively), the intron-exon structure differs markedly. Furthermore, although mRNA levels remained constant after exposure to ZnCl2 (0-330 microM) under laboratory conditions, protein levels increased significantly in a dose-dependent manner. In conclusion, this first account of molecular genetic similarities and differences of Cpn60 in a neglected nematode taxon provides a valuable insight into its potential uses in gene-based ecotoxicological risk assessment exercises.

Biomarkers in terrestrial invertebrates for ecotoxicological soil risk assessment.

This review has served to present the most recent information on a selected series of biomarker studies undertaken on soil invertebrates during two extensive European-funded scientific consortia, BIOPRINT and BIOPRINT-II. The goals were to develop and validate methods for the analysis of markers of stress in a range of soil-dwelling organisms. We have discussed the potential and limitations of the following invertebrate biomarkers for soil risk assessment purposes: heat shock proteins, histological and ultrastructural markers, metallothioneins and metal-binding proteins, esterases, lysosomal integrity, and the novel biomarker histidine. The hsp response in soil invertebrates is especially suitable to indicate the effects of exposure to comparatively low concentrations for a range of toxicants and can be regarded as a biomarker of general stress. The application of MTs and other metal-binding proteins as biomarkers for exposure in soil invertebrates has been well described, and new methods are being developed for analyzing MT induction both at the protein and molecular level, and reliable and reproducible methods are now available. (Cd)-MT is well characterized for the springtails and its MT concentration is a useful biomarker for exposure as well as for effect. For snails, (Cd)-MT can accumulate in the midgut gland over extended periods of time and therefore its concentration is a biomarker not only for recent intoxication but also for events of cadmium exposure that snails may have experienced a long time before the measurement took place. Cellular and histological alterations can be regarded as reflecting the "health" state of a cell, which may be a measure for the presence of toxicants. Histopathological work on terrestrial invertebrates, however, is still scarce. Isozymes have been poorly studied in soil invertebrates despite their promising role as potential biomarkers in aquatic organisms. Among the large diversity of isozymes, the most well studied are esterases that are frequently used a biomarkers of exposure to various classes of pesticides. Many other isozymes offer potentials for biomarker research, such as glucosephosphate isomerase and phosphoglucomutase, both enzymes necessary for the glycolytic pathway. The lysosomal system has been identified as a particular target for the toxic effects of xenobiotics, although it has yet a limited application in soil invertebrates. This marker is nonspecific, responding equally sensitively to organic or inorganic contamination; however, if used in combination with an earthworm immnunocompetence assay such as total immunoactivity of the coelomocytes, then it is possible to be more specific as to the likely nature of contamination. Free histidine was positively correlated with increasing copper exposure and total copper burden in earthworms from a semifield study. Histidine may thus act as a biomarker of exposure. The transient responses and confounding factors of biomarkers obscure a proper interpretation of biomarker responses under field conditions. These factors are still very poorly understood and require more study. For risk assessment purposes it is recommended that the aforementioned biomarkers may show promise when included in a suite of biomarkers among different soil invertebrate species. It is recommended that a risk assessment protocol draw upon ranking of biomarker responses on a defined scale. It is also hoped that the problems outlined in this review will aid the direction of future research on soil invertebrate biomarkers.

The Effects of Founding Events and Agricultural Practices on the Genetic Structure of Three Metapopulations of Globodera pallida.

To assess the genetic constitution of the Globodera pallida populations in the Netherlands and the effects of agricultural practices, three geographically separated metapopulations, in total consisting of 226 local populations, were analyzed by two-dimensional gel electrophoresis (2-DGE) of total proteins. This technique allows the accurate assessment of allele frequencies in homogenates of mixtures of individuals. Based on the estimated average heterozygosity, the average proportion of polymorphic loci and the average number of alleles per locus, the genetic diversity among 226 local G. pallida populations was small. The small genetic basis of G. pallida in the Netherlands will facilitate the identification of Solanum genotypes with a broad and durable resistance to G. pallida. Instead of clusters of local populations with unique alleles, a continuous range of allele frequencies was observed. Analysis of the three metapopulations by the Shannon-Weaver index and Nei's G(ST) revealed that the metapopulation from a region with sandy-loam soils was clearly distinguishable from the remaining two; the local populations within this metapopulation were more similar and the genetic diversity within the individual local populations was significantly higher than the local populations from the two remaining regions. These regions are characterized by wider crop rotation schemes and a very limited use of nematicides. The less intensive cultivation of potatoes in these regions with sandy-clay soils resulted in relatively little variation within and more variation between local nematode populations. To our knowledge, the effects of agricultural practices on the genetic constitution of potato cyst nematode populations have not been pinpointed before.

Environmental influence on the genetic correlations between life-history traits in Caenorhabditis elegans.

Empirical evidence is mounting to suggesting that genetic correlations between life-history traits are environment specific. However, detailed knowledge about the loci underlying genetic correlations in different environments is scant. Here, we studied the influence of temperature (12 degrees C and 24 degrees C) on the genetic correlations between egg size, egg number and body mass in the nematode Caenorhabditis elegans. We used a quantitative trait loci (QTL) approach based on a genetic map with evenly spaced single nucleotide polymorphism markers in an N2 x CB4856 recombinant inbred panel. Significant genetic correlations between various traits were found at both temperatures. We detected pleiotropic or closely linked QTL, which supported the negative correlation between egg size and egg number at 12 degrees C, the positive correlation across temperatures for body mass, and the positive correlation between body mass and egg size at 12 degrees C. The results indicate that specific loci control the covariation in these life-history traits and the locus control is prone to environmental conditions.

Mapping phenotypic plasticity and genotype-environment interactions affecting life-history traits in Caenorhabditis elegans.

Phenotypic plasticity and genotype-environment interactions (GEI) play an important role in the evolution of life histories. Knowledge of the molecular genetic basis of plasticity and GEI provides insight into the underlying mechanisms of life-history changes in different environments. We used a genomewide single-nucleotide polymorphism map in a recombinant N2 x CB4856 inbred panel of the nematode Caenorhabditis elegans to study the genetic control of phenotypic plasticity to temperature in four fitness-related traits, that is, age at maturity, fertility, egg size and growth rate. We mapped quantitative trait loci (QTL) for the respective traits at 12 and 24 degrees C, as well as their plasticities. We found genetic variation and GEI for age at maturity, fertility, egg size and growth rate. GEI in fertility and egg size was attributed to changes in rank order of reaction norms. In case of age at maturity and growth rate, GEI was caused mainly by differences in the among-line variance. In total, 11 QTLs were detected, five QTL at 12 degrees C and six QTL at 24 degrees C, which were associated with life-history traits. Five QTL associated with age at maturity, fertility and growth rate showed QTL x environment interaction. These colocalized with plasticity QTL for the respective traits suggesting allelic sensitivity to temperature. Further fine mapping, complementation analyses and gene silencing are planned to identify candidate genes underlying phenotypic plasticity for age at maturity, fertility and growth.

Patterns of sensitivity to cadmium and pentachlorophenol among nematode species from different taxonomic and ecological groups.

The variation of acute toxicity data among nematode species belonging to different taxonomic and ecological groups was investigated. Twelve different nematode species were extracted from the soil and directly exposed to cadmium and pentachlorophenol. LC50-values were estimated after 24, 48, 72, and 96 h of exposure in aqueous solutions. The species exhibited large differences in sensitivity. LC50-values (72 h) for pentachlorophenol ranged from 0.5 to more than 34.5 mumol/L and for cadmium from 29 to more than 800 mumol/L. These toxicity data could be described by a log-logistic distribution function. LC50-values for cadmium were not correlated with those for pentachlorophenol. Species of the subphylum Secernentia were less sensitive to pentachlorophenol than species of the subphylum Penetrantia, while no differences were observed for cadmium. In addition, no relationship was found between toxicity data and life-history strategies. Slow colonizers (K-strategists, sensu lato) were not more sensitive to cadmium and pentachlorophenol than opportunistic species (r-strategists, sensu lato). Nematodes appeared to be as sensitive to pentachlorophenol as other soil invertebrates. Nematodes were generally tolerant to cadmium.

The toxic effect of deltamethrin on linyphiid and erigonid spiders in connection with ambient temperature, humidity, and predation.

The first part of this study concerns the effect of temperature and air humidity on the toxicity of deltamethrin to the erigonid Oedothorax apicatus. The second part concerns the effect of deltamethrin on behavior of linyphiids and erigonids with respect to their ability to escape from predators and to select between unfavorable (dry) and favorable (moist) habitat conditions. The toxic effect of deltamethrin was highest at the combination of high temperature and low air humidity. It was concluded that the spider's sensitivity to drought is increased by this pyrethroid. Affected spiders, however, are less able to select moist habitat conditions than unaffected ones. Walking speed of spiders was decreased by exposure to deltamethrin and their predation by carabid beetles was increased. It was concluded that the effect observed under field conditions is the result of a combination of neurological, physiological, and behavioral disturbance.