DNA-based prenatal diagnosis of the infantile form of neuronal ceroid lipofuscinosis (INCL,CLN1)

Eleven fetuses at risk for the infantile form of neuronal ceroid lipofuscinosis (INCL, CLN1) were studied using DNA markers and the results were compared with the results of electron microscopy (EM) of chorionic villus specimens from pregnancies in the first or early second trimester of pregnancy. In four cases, the prenatal diagnosis was made independently with both methods, and in seven cases, the EM diagnosis was confirmed postnatally or from autopsy material using RFLP analysis. The two methods gave concordant results in all cases. The DNA analysis based on RFLP haplotypes also for the first time facilitates reliable carrier diagnostics. RFLP analysis based on polymorphic markers closely linked to the INCL locus is now available for prenatal diagnosis of this fatal brain disease, whose biochemical background is totally unknown and for which no treatment is available.     

The costs of secondary sexual characters in the lekking great snipe (Gallinago media)

Summary Recent studies of the lekking great snipe (Gallinago media) have shown that male mating success increases with display rate and the amount of white on the tail. An obvious question then arises: what is setting the limits to male display rate and to the evolution of even whiter tails? In this study the costs of these two traits are considered. In the case of the white on the tail, almost no predation was observed during the display period. However, predation on the winter grounds and/or during migration cannot be ruled out. Males differed in the amount of white on the tail between years. It is suggested that increasing the amount of white is costly, though the reason for this is still unknown. In the case of display rate, it is shown that predation cannot be a considerable cost, again because of the near absence of predation on the leks. Instead, it is shown that male displays are energetically costly. The daily energy expenditure of displaying males was estimated to be on average 4 times the basal metabolic rate, which is close to the supposed maximum sustainable work load for birds. Furthermore, males lost on average 1.8 g per h (1.7% of the body mass) which is about 7.2 g (6.8%) during a night of display activity. Males do gain some weight during the day when no displays are performed but loose on average 9 g during an 11-day period in the early part of the breeding season. These results indicate that high energetic demands set the limits to male display performance and that males do not have to trade off the benefit of attracting females by their displays against a cost of attracting predators. Offprint requests to: J. Höglund     

Antigenic characterization of hemocyte subpopulations in the mussel Mytilus edulis by means of monoclonal antibodies

In order to characterize in more detail hemocytes from Mytilus edulis and establish links between histological and cytological featues, monoclonal antibodies (MABs) were prepared using hybridoma technology. Five MABs were identified and their reactivity patterns were analyzed with both immunoperoxidase and ultrastructural immunogold labelling. Four MAB classes were identified by their immunostaining patterns. Optical and ultrastructural examination revealed that Class I MAB reacted specifically with basophilic granulocytes (granulocytes containing small granules). Class II and III MABs differed in the intensity of their indirect immunofluorescence and degree of immunoperoxidase labelling, but all recognized both basophilic and eosinophilic granulocytes. Epitopes recognized by Class IV MABs were not always present in basophilic granulocytes, whereas they were always expressed in the eosinophilic granulocytes (granulocytes containing large granules) and could constitute differentiation antigens which progressively form during the maturation process. Molecular weights of the proteins recognized by these MABs were determined by Western blotting. It is concluded that immunostaining can identify at least three hemocyte types in M. edulis.     

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 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     

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.