Biological studies and phenology of the slug Deroceras reticulatum (Müller, 1774) (Pulmonata: Stylommatophora)

Summary

In the southeast of Buenos Aires Province (Argentina), slugs cause reductions in crop yield by killing seeds and seedlings, by destroying stems and growing points, and by reducing leaf area. Deroceras reticulatum (Müller, 1774) was introduced into Argentina a long time ago, but the biology and ecology of this species in this country are still unknown. The aim of this study was to determine how temperature affects growth rate, reproduction and survival of D. reticulatum and also to assess the seasonal phenology of this species. The life cycle of D. reticulatum was investigated at 12°C, 20°C and at a temperature which alternated between 12°C for 16 h and 20°C for 8 h. The light/dark regime during the experiment was LD: 8/16 h. Slug populations were sampled from grassland located at the Experimental Station of INTA Balcarce. Eggs hatched after 16.3 to 39.8 days and fertility varied between 83.9% and 91.1%. The mean time from hatching to oviposition was longer and fecundity lower at 20°C than 12°C and 12/20°C (p <0.05). The net reproductive rate was 1.49 to 70.53 and the mean generation time varied between the 27 and 30 weeks. In natural conditions, eggs of D. reticulatum were found from early winter until the end of the spring, when soil temperature was 8–15°C and humidity was above 15%. These results indicate that D. reticulatum has one generation per year, univoltine phenology, with peak densities of slugs from the beginning of winter to the end of spring, which coincides with the sowing and emergence of winter and summer crops.     

The influence of nutritional status on the feeding behaviour of the field slug, Deroceras reticulatum (Müller)

The field slug, Deroceras reticulatum, a common pest of agricultural and horticultural crops, is a generalist herbivore with distinct preferences for particular food items. However, these preferences are not fixed, but are influenced by the recent dietary history of the slugs. In particular, slugs tend to select novel food items ('neophilia'). We investigated the basis of such influences, using artificial diets in which protein and carbohydrate composition could be controlled and non-nutritional ('secondary') chemicals added as flavours. The slugs showed no general inclination for neophilia based on taste alone. There was a possible learned association between protein content and taste, but this was weakly expressed. However, the slugs selected food items, when given a choice, containing nutrients that were deficient in earlier diets, even though all the food items contained the same familiar secondary taste chemicals. Injections of missing nutrients into the haemocoel of slugs inhibited such changes in food preference, indicating that slugs' feeding preferences are influenced directly by their internal nutritional status. We suggest that neophilia in D. reticulatum is a physiological response to a nutritional imbalance arising from a suboptimal diet. Copyright 2000 The Association for the Study of Animal Behaviour.     

Benthic flattened cells of the phylogenetically related marine dinoflagellates Protoceratium reticulatum and Ceratocorys mariaovidiorum (Gonyaulacales): a new type of cyst?

A planktonic‐benthic relationship has been described for many dinoflagellate species as part of their ecological strategy to overcome highly variable aquatic environments. Here, the phylogenetically and morphologically related marine dinoflagellates Protoceratium reticulatum and Ceratocorys mariaovidiorum were studied in relation to an unknown benthic life form. In vivo and fixed samples from cultures were analyzed in detail by light and scanning electron microscopy. In both species, a cell type with a morphology different from that of vegetative cells was observed in cultures grown until stationary phase. This cell type was always benthic, swimming sporadically only when it was disturbed. Its main feature included a strong dorsoventral compression. These cells originated from vegetative cells whose protoplasm underwent a progressive flattening, resulting in a gradual detachment of the reticulate and thick thecal plates and the formation of very thin non‐reticulated new plates with pores. When returned to fresh full‐strength medium, the cells recovered their spherical vegetative‐like morphology, including new reticulated thick plates and subsequent cell divisions. The kinetics of flattened cell formation showed that in both species, this cell type increased exponentially until the onset of the culture stationary phase and then decreased. The results of this study are discussed in the context of the planktonic–benthic coupling in dinoflagellate life cycles, including those newly appreciated to be well adapted to the benthic environment.     

Phylogeny of the Alismatales (Monocotyledons) and the relationship of Acorus (Acorales?)

A phylogenetic analysis of the early branching lineages of the monocotyledons is performed using data from two plastid genes (rbcL and matK), five mitochondrial genes (atp1, ccmB, cob, mttB and nad5) and morphology. The complete matrix includes 93 terminals representing Acorus, the 14 families currently recognized within Alismatales, and numerous lineages of monocotyledons and other angiosperms. Total evidence analysis results in an almost completely resolved strict consensus tree, but all data partitions, genomic as well as morphological, are incongruent. The effects of RNA editing and potentially processed paralogous sequences are explored and discussed. Despite a decrease in incongruence length differences after exclusion of edited sites, the major data partitions remain significantly incongruent. The 14 families of Alismatales are all found to be monophyletic, but Acorus is found to be included in Alismatales rather than being the sister group to all other monocotyledons. The placement is strongly supported by the mitochondrial data, atp1 in particular, but it cannot be explained as an artifact caused by patterns of editing or by sampling of processed paralogues.     

Does the concentration of DNA (Co) and the time of incubation (t) as parameters of Cot influence the thermal stability of the DNA duplexes?

It has been shown in a previous paper (8) that the prime product of reassociation of related DNA sequences under open experimental conditions are mismatched duplexes which undergo maturation upon further incubation. Due to this feature, the Tm value of the duplexes of a large number of DNAs is strongly dependent on the Cot value. Here we present data showing that the Tm of the duplexes of such type of DNAs depends also on the concentration of DNA in the range of one and the same Cot value. The significance of this finding in studying the taxonomic relationship by DNA-DNA hybridisation is discussed.Abbreviations Co = initial concentration of single-stranded DNA in moles of nucleotides per liter - t = time of incubation in seconds - Cot = the product of Co and t (mol. sec. 1¹) - PB = an equimolar mixture of NaH₂PO₄ and Na₂HPO₄, pH 6.8 - HAP = hydroxyapatite - Ti = incubation temperature - Tm = melting temperature - Te = elution temperature, i.e. the temperature at which one half of the DNA is eluted as single strands by HAP-thermal chromatography     

Molecular Evolution and Nucleotide Sequences of the Maize Plastid Genes for the α Subunit of Cf1 (atpA) and the Proteolipid Subunit of Cf0 (atpH)

The nucleotide sequences of the maize plastid genes for the alpha subunit of CF1 (atpA) and the proteolipid subunit of CF0 (atpH) are presented. The evolution of these genes among higher plants is characterized by a transition mutation bias of about 2:1 and by rates of synonymous and nonsynonymous substitution which are much lower than similar rates for genes from other sources. This is consistent with the notion that the plastid genome is evolving conservatively in primary sequence. Yet, the mode and tempo of sequence evolution of these and other plastid-encoded coupling factor genes are not the same. In particular, higher rates of nonsynonymous substitution in atpE (the gene for the epsilon subunit of CF1) and higher rates of synonymous substitution in atpH in the dicot vs. monocot lineages of higher plants indicate that these sequences are likely subject to different evolutionary constraints in these two lineages. The 5'- and 3'-transcribed flanking regions of atpA and atpH from maize, wheat and tobacco are conserved in size, but contain few putative regulatory elements which are conserved either in their spatial arrangement or sequence complexity. However, these regions likely contain variable numbers of "species-specific" regulatory elements. The present studies thus suggest that the plastid genome is not a passive participant in an evolutionary process governed by a more rapidly changing, readily adaptive, nuclear compartment, but that novel strategies for the coordinate expression of genes in the plastid genome may arise through rapid evolution of the flanking sequences of these genes.     

Characterization of the flanking regions of the S-RNase genes of Japanese pear (Pyrus serotina) and apple (Malus×domestica)

Genomic sequences of the self-incompatibility genes, the S-RNase genes, from two rosaceous species, Japanese pear and apple, were characterized. Genomic Southern blot and sequencing of a 4.5-kb genomic clone showed that the S⁴-RNase gene of Japanese pear is surrounded by repetitive sequences as in the case of the S-RNase genes of solanaceous species. The flanking regions of the S²- and S^f-RNase genes of apple were also cloned and sequenced. The 5′ flanking regions of the three alleles bore no similarity with those of the solanaceous S-RNase genes, although the position and sequence of the putative TATA box were conserved. The putative promoter regions of the Japanese pear S⁴- and apple S^f-RNase genes shared a stretch of about 200 bp with 80% sequence identity. However, this sequence was not present in the S²-RNase gene of apple, and thus it may reflect a close relationship between the S⁴- and S^f-RNase genes rather than a cis-element important in regulating gene expression. Despite the uniform pattern of expression of the rosaceous S-RNase genes, sequence motifs conserved in the 5′ flanking regions of the three alleles were not found, implying that the cis-element controlling pistil specific gene expression also locates at the intragenic region or upstream of the analyzed promoter region.     

pH-dependence of the specific binding of Cu(II) and Zn(II) ions to the amyloid-β peptide

Metal ions like Cu(II) and Zn(II) are accumulated in Alzheimer's disease amyloid plaques. The amyloid-β (Aβ) peptide involved in the disease interacts with these metal ions at neutral pH via ligands provided by the N-terminal histidines and the N-terminus. The present study uses high-resolution NMR spectroscopy to monitor the residue-specific interactions of Cu(II) and Zn(II) with (15)N- and (13)C,(15)N-labeled Aβ(1-40) peptides at varying pH levels. At pH 7.4 both ions bind to the specific ligands, competing with one another. At pH 5.5 Cu(II) retains its specific histidine ligands, while Zn(II) seems to lack residue-specific interactions. The low pH mimics acidosis which is linked to inflammatory processes in vivo. The results suggest that the cell toxic effects of redox active Cu(II) binding to Aβ may be reversed by the protective activity of non-redox active Zn(II) binding to the same major binding site under non-acidic conditions. Under acidic conditions, the protective effect of Zn(II) may be decreased or changed, since Zn(II) is less able to compete with Cu(II) for the specific binding site on the Aβ peptide under these conditions.     

Adriano Buzzati-Traverso and the foundation of the International Laboratory of Genetics and Biophysics in Naples (1962–1969)

Despite a long tradition of research in applied genetics, particularly in agricultural research, in Italy the transition to the new knowledges and techniques of molecular biology was long and difficult. Political and financial constraints made academic institutions very slow to grasp the importance of molecular approaches to biology and medicine. In fact, the main studies concerning problems of molecular biology took place inside non-academic institutions. We reconstruct the complex paths leading to the birth of the International Laboratory of Genetics and Biophysics (LIGB) in Naples, and describe its work and activities in the period in which it was directed by its creator, Adriano Buzzati-Traverso, between 1962 and 1969. The origins of the LIGB are inextricably bound to the growth of biophysical research at the University of Pavia and at the Higher Health Institute in Rome. For a short period, with the aid provided by Italian and European physicists to biological research on the effects of nuclear radiation, LIGB became a focal point in European molecular biology. In 1968 the scientific and educational activities of the LIGB dramatically fell victim to the contradictions of the Italian academic and scientific system, and to the political climate which emerged in Italy at the time.     

Structure and ultrastructure of the silk glands and spinneret of Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae)

This study provides comprehensive documentation of silk production in the pest moth Helicoverpa armigera from gland secretion to extrusion of silk thread. The structure of the silk glands, accessory structures and extrusion apparatus are reported. The general schema of the paired silk glands follows that found for Lepidoptera. Morphology of the duct, silk press, muscle attachments and spigot are presented as a three-dimensional reconstruction and the cuticular crescent-shaped profile of the silk press is demonstrated in both open and closed forms with attendant muscle blocks, allowing advances in our knowledge of how the silk press functions to regulate the extrusion of silk. Growth of the spigot across instars is documented showing a distinctive developmental pattern for this extrusion device. Its shape and structure are related to use and load-bearing activity.     

Effect of bis(acetato)tetrakis(imidazole) copper(II) in delaying the onset and reducing the mortality rate of strychnine- and thiosemicarbazide—Induced convulsions

The anticonvulsant activity of bis(acetato)tetrakis(imidazole) copper(II), Cu(OAc)₂(Im)₄, was studied in normal mice using chemical convulsions induced by strychnine, thiosemicarbazide, picrotoxin, and pentelenetetrazol. Intraperitoneal administration of Cu(OAc) ₂(Im)₄, 50 mg/kg body mass, has delayed the onset of strychnine (3 mg/kg)-induced convulsion by 204% (p≤0.005) and thiosemicarbazide (20 mg/kg)-induced convulsant by 61% (p≤0.005). The changes in the onset of picrotoxin-(6 mg/kg) and pentelenetetrazol (50 mg/kg)-induced convulsions were not significant. The same dosage of the copper compound was effective in delaying the lethal time and reducing the mortality rate of treated animals. The anticonvulsant activity of Cu(OAc) ₂(Im)₄ complex against strychnine was not related to its constituents because the inorganic form of copper such as copper chloride, copper acetate, and the parent imidazole has no anticonvulsant activity. Other copper(II) complexes like copper(II)aspirinate and bis(acetato)bis(2-methyl imidazole) copper(II) were less effective.     

Palatability of weeds from different European origins to the slugs Deroceras reticulatum Müller and Arion lusitanicus Mabille

As part of a study on the significance of seed provenances in schemes to enhance biodiversity in agricultural habitats, juvenile plants of Cichorium intybus, Daucus carota, Leucanthemum vulgare and Silene alba of different European origins were exposed to grazing by two slug species, Deroceras reticulatum and Arion lusitanicus. Living plants were offered in trays, either in a glasshouse (Deroceras) or outdoors (Arion). The amount of herbivory was origin-dependent, with higher losses for all four species from German and Hungarian provenances compared with English and Swiss plants. The main trend was similar for both slug species except in the case of Daucus, and there was a significant ‘origin × plant species’ interaction. We found strong correlations between provenance-specific herbivory and certain climatic characteristics of the corresponding regions, i.e. winter minimum temperatures, and dryness in spring and late summer, which are crucial for the development of slugs. The results can be interpreted in terms of a SW-NE European climatic gradient and may be a consequence of differences in the need for plant defences against herbivory by slugs. Additionally, the data on palatability were compared with susceptibility towards two parasites which occurred in a field experiment, a leaf miner on Leucanthemum vulgare and a rust fungus on Silene alba. While specific leaf mining frequencies on Leucanthemum contrasted with the palatability of the different provenances to slugs, the rust infection on Silene was low on local and German plants, and higher on the more distant provenances from England and Hungary.     

Detection of the genotoxicity of air pollutants in and around the city of São Paulo (Brazil) with the Tradescantia-micronucleus (Trad-MCN) assay

Tradescantia pallida cv. purpurea, a popular garden plant in Brazil, was used for the Tradescantia micronucleus (Trad-MCN) assay. In situ monitoring of the genotoxicity of air pollutants was carried out by sentinel approach, using the plant grown in the field or using the plants in pots which were carried to the monitoring sites. Two highly polluted sites, in S?o Paulo city (Cerqueira Cesar and Congonhas) and two rural sites (the cities of Pirassununga, 200 km and Caucaia do Alto, 50 km from S?o Paulo, respectively) were chosen for this study, in order to determine the gradient difference of the air pollution levels. Sentinel plants in Congonhas site presented the highest frequency of micronuclei (4.4%), in comparison with 2.2 and 2.3% found in plants from Pirassununga and Cerqueira Cesar sites, respectively (Kruskal-Wallis; P<0.020). Significant increases (F test; P<0.0001) in the frequency of micronuclei were observed in plants exposed in the polluted urban sites (Cerqueira Cesar: 5.7%; Congonhas: 7.1% and Caucaia do Alto: 2.3%). The increase in the frequency of micronuclei observed indicates the potential risk of mutagenicity in presence of high concentrations of pollutants.     

Importance of algae in the diet of the oligochaetes Lumbriculus variegatus (MÜller) and Rhyacodrilus sodalis (Eisen)

Summary The importance of algae in the diet of the oligochaetes Lumbriculus variegatus and Rhyacodrilus sodalis was determined from cellections made in a eutrophic bay from April 1977 to April 1978. During the summer, algae represented 70–85% of the gut contents of both species. The most frequently ingested algae were Cymatopleura elliptica, Cymbella spp., Epithemia turgida, Pinnularia spp., and Synedra ulna. Almost all species were consumed in proportion to their abundance in the environment. However, Nostoc pruniforme and Nostoc verrucosum were not eaten due to size selection. The high level of algal consumption exhibited by the oligochaetes was primarily related to the large standing crop of algae on the sediments. During the winter, when algal densities were low in the environment, detritus and associated bacteria were the major food source.     

Checklist of the fly families Chyromyidae and?Heleomyzidae (Diptera) of Finland

A Finnish checklist of the sphaeroceroid fly families Chyromyidae and Heleomyzidae is provided.     

Checklist of the Diptera superfamilies Tephritoidea and?Sciomyzoidea of Finland (Insecta)

A revised checklist of the flies of superfamilies Tephritoidea and Sciomyzoidea of Finland is provided. The following families are covered: Eurygnathomyiidae, Lonchaeidae, Neottiophilidae, Pallopteridae, Piophilidae, Platystomatidae, Tephritidae, Ulidiidae (Tephritoidea); Coelopidae, Dryomyzidae, Heterocheilidae, Phaeomyiidae, Sciomyzidae, Sepsidae (Sciomyzoidea).     

Larval development and morphogenesis of the sea spider Pycnogonum litorale (Ström, 1762) and the tagmosis of the body of Pantopoda

Aspects of pantopod ontogeny have been known for a long time, but specific information is available for only a few species. Our account of the postembryonic development of Pycnogonum litorale is based on laboratory-reared individuals and SEM studies. We documented particularly all early developmental stages, with emphasis on morphogenetic changes of head structures and appendages. In P. litorale the protonymphal limbs, the chelicerae and two more uniramous legs, degenerate already during the larval phase; only the third one, the ovigers, reappears in male juveniles. Other Pantopoda vary in this aspect from retention of all three protonymphal appendages to their complete reduction, as in P. litorale. Accordingly, the two post-cheliceral larval appendages are separate legs in front of the walking legs in the adults, the ‘parapalps’ and the ‘ovigers’, but they do not occur in all pantopods. The scarcity of studies of the ontogeny of Pantopoda prevents us from a more conclusive picture, but our data are promising to state that additional such studies will increase the usability of ontogenetic data for a phylogenetic analysis of Pantopoda, the crown group of the Pycnogonida. We also discuss the phylogenetic implications of our data in the light of new information from Hox genes and developmental-biological data on body segmentation and tagmosis of the Chelicerata. These suggest the homology of chelicerae and antenn(ul)ae of other euarthropods. Accepting this, we conclude that the adult pycnogonid/pantopod head, the cephalosoma, corresponds to the euarthropod head and that the protonymph with three appendage-bearing segments may represent an even shorter, possibly phylogenetically older larval type than the euarthropod ‘head larva’ bearing four pairs of appendages. In further consequence, the fourth walking legs of Pycnogonida/Pantopoda should correspond to the first opisthosomal appendages, the chilaria, of euchelicerates. This implies that within Pycnogonida the post-prosomal region became compacted during evolution to a single leg-bearing segment plus a tubular end piece. Accordingly, neither the anterior nor the posterior functional boundaries of the walking-leg region correspond to the original tagma borders.     

Binding of α-synuclein with Fe(III) and with Fe(II) and biological implications of the resultant complexes

Parkinson’s disease (PD) is hallmarked by the abnormal intracellular inclusions (Lewy bodies or LBs) in dopaminergic cells. Amyloidogenic protein α-synuclein (α-syn) and iron (including both Fe(III) and Fe(II)) are both found to be present in LBs. The interaction between iron and α-syn might have important biological relevance to PD etiology. Previously, a moderate binding affinity between α-syn and Fe(II) (5.8 × 10³ M⁻¹) has been measured, but studies on the binding between α-syn and Fe(III) have not been reported. In this work, electrospray mass spectrometry (ES-MS), cyclic voltammetry (CV), and fluorescence spectroscopy were used to study the binding between α-syn and Fe(II) and the redox property of the resultant α-syn-Fe(II) complex. The complex is of a 1:1 stoichiometry and can be readily oxidized electrochemically and chemically (by O₂) to the putative α-syn-Fe(III) complex, with H₂O₂ as a co-product. The reduction potential was estimated to be 0.025 V vs. Ag/AgCl, which represents a shift by −0.550 V vs. the standard reduction potential of the free Fe(III)/Fe(II) couple. Such a shift allows a binding constant between α-syn and Fe(III), 1.2 × 10¹³ M⁻¹, to be deduced. Despite the relatively high binding affinity, α-syn-Fe(III) generated from the oxidation of α-syn-Fe(II) still dissociates due to the stronger tendency of Fe(III) to hydrolyze to Fe(OH)₃ and/or ferrihydrite gel. The roles of α-syn and its interaction with Fe(III) and/or Fe(II) are discussed in the context of oxidative stress, metal-catalyzed α-syn aggregation, and iron transfer processes.