Investigation of acute toxicity of chlorpyrifos-methyl on Nile tilapia (Oreochromis niloticus L.) larvae

Chlorpyrifos-methyl, a wide-spectrum organophosphorus insecticide and potential toxic pollutant contaminating aquatic ecosystems, was investigated for acute toxicity. Larvae of the freshwater fish Nile tilapia (Oreochromis niloticus L.) were selected for the bioassay experiments. The experiments were repeated three times and the 96 h LC50 was determined for the larvae. The static test method for assessing acute toxicity was used. Water temperature was maintained at 25+/-1 degrees C. In addition, behavioral changes at each chlorpyrifos-methyl concentration were observed for the individual fish. Data obtained from the chlorpyrifos-methyl acute toxicity tests were evaluated using Finney's probit analysis statistical method. The 96 h LC50 value for Nile tilapia larvae was calculated to be 1.57 mg/l.     

The evaluation of dioxin and dioxin-like contaminants in selected food samples obtained from the Belgian market: comparison of TEQ measurements obtained through the CALUX bioassay with congener specific chemical analyses

A limited number of different foods were analysed for dioxin-like compounds by the CALUX bioassay which is an in vitro luciferase reporter gene assay measuring chemical activation of the aryl hydrocarbon receptor. Sixty-two milk samples were obtained from a surveillance campaign, 34 meat samples and 34 fishery products were purchased from the Belgian market. Bio-analytical and chemo-analytical dioxin toxicity equivalents (TEQ) values of the same milk samples were compared. Spearman's Rank correlation coefficients of 0.72, 0.67, 0.73 were obtained respectively between CALUX-TEQ and PCDD/F-TEQ, DL-PCB-TEQ and PCDD/F+DL-PCB-TEQ. The bioassay limit of detection was 0.1 pg TEQ from 1 g animal lipid, the limit of quantification was 0.4 pg TEQ. The repeatability of the CALUX bioassay (variability of butter fat samples analysed in the same run) showed a coefficient of variation (CV) of 10%, intra laboratory reproducibility based on independent runs of the same butter fat samples showed more variation (CV of 26% for samples above 2 pg TEQ/g lipid). All milk samples with a chemical TEQ value above the current limit value in Belgium showed an elevated CALUX-TEQ concentration, above 6 pg TEQ/g lipid. No false negative results were obtained. Based on the good correlation between CALUX-TEQ and chemically measured TEQ levels, the CALUX bioassay can be recommended as a screening tool for routine measurement of potentially toxic PHAHs in milk samples. Chemical analyses could then largely be restricted to positive samples, in order to identify the nature and to quantify the concentration of the chemicals that give the positive signal. Meat samples showed lower CALUX-TEQ values per gram lipid compared to fish samples. The fish samples showed a wider range of CALUX-TEQ values than the meat samples.     

The effects of endosulfan on the great ramshorn snail Planorbarius corneus (Gastropoda, Pulmonata): a histopathological study

In this study the great ramshorn snail (Planorbarius corneus), one of the most abundant gastropod of Turkish limnic systems, was investigated to determine the histopathological effects of endosulfan on the digestive gland, foot and mantle under laboratory conditions. Samples were collected from small artificial pools in Karaot at Gelendost-Isparta (southwest of Turkey), where agricultural activities are widespread. The snails were exposed to two sublethal concentrations of endosulfan (0.4 and 0.8 mg/l) for periods of 10, 20 and 30 days. Fifteen snails were kept in 2.5 l glass jars containing dechlorinated tap water and exposed under semi-static test (daily exchange of test water). All the testing was carried out on adult specimens, and snails were maintained on a photothermal period with 16 light hours at 22 +/- 2 degrees C. The histopathological examinations revealed the following changes: amoebocytes infiltration, dilatation in hemolymphatic spaces between the tubules, degeneration of cells, abnormal lumen, necrosis of cells and atrophy in the connective tissue of digestive gland; desquamation of the epithelium cells, changes in the number of mucocytes and protein gland cells, lipid vacuolus and atrophy of the columnar muscle fibers of the foot and mantle tissues. Pycnotic state of cells was also seen in the mantle tissues. Endosulfan caused significant histopathological alterations in the digestive gland, foot and mantle tissues of the snail, irrespective of concentrations of the pesticide and its exposure periods. The results are discussed, particularly in comparison to those of other aquatic organisms.     

Relating cadmium concentrations in three macrophyte-associated freshwater invertebrates to those in macrophytes,water and sediments

We found that Cd concentrations in three species of macrophyte-associated invertebrates (the gastropods Bithynia tentaculata and Physa gyrina, and the amphipod Gammarus fasciatus) collected at twenty sites along the St. Lawrence River were correlated with Cd concentrations in their main food source, i.e. macrophytes and associated periphyton. Cd in these invertebrates was not significantly correlated with Cd concentrations in the sediments (even when corrections for iron oxide or organic carbon content were applied) or to calculated free-cadmium concentrations in the water. Cd levels in juveniles of B. tentaculata were very tightly linked to Cd concentrations in macrophytes, which is consistent with the close relationship that these organisms have developed with macrophytes. Linear models predicting Cd levels in juveniles of B. tentaculata from Cd levels in different macrophyte species were strong (R2 = 0.69-0.90). Analysis of covariance on these models showed no statistical difference of slope or intercept for any of the macrophyte species, except Vallisneria americana. We suggest that the macrophyte-periphyton complex is a key link in the transfer of cadmium to some aquatic invertebrates in the littoral zone of the St. Lawrence River and that macrophytes and their associated epiphytes should also be used as biomonitors.     

Acute toxicity of alpha-cypermethrin to guppy (Poecilia reticulata, Pallas, 1859)

Alpha-cypermethrin is a synthetic pyrethroid insecticide used to control pests in domestic, industrial and agricultural situations. Adult male guppy fish (Poecilia reticulata standard test species) were selected for the bioassay experiments. The experiments were performed in four series and the 96-h LC50 value was determined for guppies. The acute toxicity experiments were carried out by static method and behavioral changes in guppies were determined for each alpha-cypermethrin (98% technical grade) concentration. Data obtained from the alpha-cypermethrin investigation were evaluated by the use of probit analysis statistical method and the 96-h LC50 value for guppy was estimated as 9.43 microg/l.     

Crinoid phylogeny: a preliminary analysis (Echinodermata: Crinoidea)

We describe the first molecular and morphological analysis of extant crinoid high-level inter-relationships. Nuclear and mitochondrial gene sequences and a cladistically coded matrix of 30 morphological characters are presented, and analysed by phylogenetic methods. The molecular data were compiled from concatenated nuclear-encoded 18S rDNA, internal transcribed spacer 1, 5.8S rDNA, and internal transcribed spacer 2, together with part of mitochondrial 16S rDNA, and comprised 3,593 sites, of which 313 were parsimony-informative. The molecular and morphological analyses include data from the bourgueticrinid Bathycrinus; the antedonid comatulids Dorometra and Florometra; the cyrtocrinids Cyathidium, Gymnocrinus, and Holopus; the isocrinids Endoxocrinus, and two species of Metacrinus; as well as from Guillecrinus and Caledonicrinus, whose ordinal relationships are uncertain, together with morphological data from Proisocrinus. Because the molecular data include indel-rich regions, special attention was given to alignment procedure, and it was found that relatively low, gene-specific, gap penalties gave alignments from which congruent phylogenetic information was obtained from both well-aligned, indel-poor and potentially misaligned, indel-rich regions. The different sequence data partitions also gave essentially congruent results. The overall direction of evolution in the gene trees remains uncertain: an asteroid outgroup places the root on the branch adjacent to the slowly evolving isocrinids (consistent with palaeontological order of first appearances), but maximum likelihood analysis with a molecular clock places it elsewhere. Despite lineage-specific rate differences, the clock model was not excluded by a likelihood ratio test. Morphological analyses were unrooted. All analyses identified three clades, two of them generally well-supported. One well-supported clade (BCG) unites Bathycrinus and Guillecrinus with the representative (chimaeric) comatulid in a derived position, suggesting that comatulids originated from a sessile, stalked ancestor. In this connection it is noted that because the comatulid centrodorsal ossicle originates ontogenetically from the column, it is not strictly correct to describe comatulids as unstalked crinoids. A second, uniformly well-supported clade contains members of the Isocrinida, while the third clade contains Gymnocrinus, a well-established member of the Cyrtocrinida, together with the problematic taxon Caledonicrinus, currently classified as a bourgueticrinid. Another cyrtocrinid, Holopus, joins this clade with only weak molecular, but strong morphological support. In one morphological analysis Proisocrinus is weakly attached to the isocrinid clade. Only an unusual, divergent 18S rDNA sequence was obtained from the morphologically strange cyrtocrinid Cyathidium. Although not analysed in detail, features of this sequence suggested that it may be a PCR artefact, so that the apparently basal position of this taxon requires confirmation. If not an artefact, Cyathidium either diverged from the crinoid stem much earlier than has been recognised hitherto (i.e., it may be a Palaeozoic relic), or it has an atypically high rate of molecular evolution.Electronic Supplementary Material Supplementary material is available in the online version of this article at Communicated by J.P. Thorpe, Port Erin     

Variation in adult annual survival probability and remigration intervals of sea turtles

We analyzed a large dataset to quantify adult annual survival probability and remigration intervals for the Tortuguero, Costa Rica green turtle population. Annual survival probability was estimated at 0.85 (95% CI 0.75–0.92) using a recovery model and at 0.85 (95% CI 0.83–0.87) using an open robust design model. The two most common modes of remigration are 2 and 3 years. Annual survival probability is lower and remigration intervals are shorter than for other green turtle populations. Explanations for short remigration intervals include reproductive compensation due to historic population declines, availability of better quality food items, favorable environmental conditions, and short distance to the main foraging grounds. Variation in survival and remigration intervals have profound consequences for management and life history evolution. The short remigration intervals of Tortuguero green turtles partly offset mortality caused by turtle fishing in Nicaragua and mean that low juvenile survival represents a more urgent threat to the population than low adult survival. Low adult survival probability could result in selective pressure for earlier age at maturity.     

Modulation of individual behavior and collective decision-making during aggregation site selection by the ant<Emphasis Type="Italic"> Messor barbarus</Emphasis>

Insect societies are often confronted with choices among several options such as food sources of different richness or potential nest sites with different qualities. The mechanisms by which a colony as a whole evaluates these situations and takes the appropriate decision are of crucial importance for its survival. Here we studied how collective decisions arise from individual behaviors when a group of workers of the ant Messor barbarus is given a choice between two aggregation sites. Two hundred ants were introduced into an arena and given a choice between two tubes connected to the arena. The tubes had different physical properties: dry and transparent (termed as dry), humid and transparent (termed as humid), or dry and dark (termed as dark). After 30 min, most ants were found to be aggregated in a humid tube when paired with a dry tube, or in a dark tube when paired with a humid one. When two humid tubes were in competition, ants aggregate more in one of the sites. The choice of ants was consistent throughout experiments. An analysis of individual behaviors shows that the probability of an ant recruiting and the intensity of its trail-laying behavior strongly depend on the quality of the tubes. Our study suggests that the selection of an aggregation site does not require that individual ants directly compare sites, but rather relies on the synergy between amplification processes involving recruitment by chemical trails, and a modulation of the individual resting time in a site as a function of its population.Communicated by L. Sundström     

Determination of selected neonicotinoid insecticides by liquid chromatography with thermal lens spectrometric detection

We studied the analysis of trace amounts of neonicotinoid insecticides by liquid chromatography coupled with a thermal lens spectrometric detector (TLS). This multi-residue analysis method is based on the reversed phase separation on C₁₈ column, isocratic elution and collinear dual beam TLS detection. The insecticides thiamethoxam, imidacloprid, acetamiprid and thiacloprid were detected with retention times of 4.4, 5.7, 6.5 and 8.5 min and limits of quantifications of 50, 89, 10, and 25 μg/L, respectively. The retention times agreed well with those obtained by the same chromatographic method but using a diode-array detector (DAD). The limits of quantifications for imidacloprid were identical in both techniques. However, the limits of quantifications for thiamethoxam, acetamiprid and thiacloprid were up to 8.5 times lower using the TLS detector compared to the diode-array detector. The applicability of the developed procedure was tested on spiked river water and potato samples.     

Chemical availability of arsenic and antimony in industrial soils

Total concentrations and extractable fractionations of As and Sb were determined in soil samples from former mining sites in Scotland and Italy. Pseudo-total levels of As and Sb in the sample were between 50–17,428 mg/kg and 10–1,187 mg/kg (Scotland), and 16–691 mg/kg and 1.63–11.44 mg/kg (Italy). Between 0.001–0.63% and <0.001−8.82% of the total soil As and Sb, were extractable using, a single extraction bioavailability estimate. Data from an As-specific extraction procedure revealed that up to 60% of As was associated to amorphous Fe-Al oxyhydroxide phase in all soils. A non-specific-sequential extraction test also showed As to be strongly associated with Fe (and Al) oxyhydroxides at both locations. In the case of Sb, in addition to the crystalline Fe-oxide bound Sb the Al-silicate phase also appeared to be significant. At both sites Sb appears to be chemically more accessible than As with consistent availability despite the varied origin and host soil properties.