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PubMed2001

Effects and risk assessment of linear alkylbenzene sulfonates in agricultural soil. 3. Sublethal effects on soil invertebrates.

Sewage sludge applied to agricultural soils often contains considerable amounts of linear alkylbenzene sulfonates (LAS). Toxic effects of LAS on soil organisms should, therefore. be evaluated to ensure safe use of sewage sludge as a fertilizer. In this study, dose-response relationships for the toxicity of Na-LAS to six species of soil invertebrates (survival, reproduction, and growth) were established using a sandy, agricultural soil as test substrate. In general, toxic effects on reproduction and growth appeared when the concentration in soil exceeded 40 to 60 mg/kg. Reproduction was approximately fourfold more sensitive in earthworms and enchytracids than in springtails and mites. It is argued that this difference in sensitivity is related to the dependency of soil pore water, which is high in the annelids but comparatively low in the arthropods.

Agriculture↗
PubMed2001

Effect of varying pesticide exposure duration and concentration on the toxicity of carbaryl to two field-collected stream invertebrates, Calineuria californica (Plecoptera: Perlidae) and Cinygma sp (Ephemeroptera: Heptageniidae).

The effect of exposure duration on the toxicity of a forest insecticide (carbaryl) was assessed under environmentally realistic exposure regimes against two stream invertebrates indigenous to the United States Pacific Northwest, Calineuria californica (Plecoptera: Perlidae) and Cinygma sp. (Ephemeroptera: Heptageniidae). Laboratory bioassays were conducted to evaluate the relationship between pulsed exposures of 15, 30, and 60 min and toxicity for a range of chemical concentrations (10.2-1,730 microg/L). For Cinygma sp., the 50% lethal concentration (LC50) values were calculated as 848 microg/L (15 min), 220 microg/L (30 min), and 165 microg/L (60 min). The C. californica consistently had lower mortality at a given concentration compared with Cinygma sp. Fifteen- and 30-min exposures did not elicit 50% mortality with C. californica, and it had a 60-min LC50 of 1,139 microg/L. Time to 50% mortality over 96 h after a 15-, 30-, or 60-min exposure, with the rest of the test period in freshwater (PLT50), was a function of exposure duration and concentration. Analysis of symptomology throughout the test period for C. californica gave evidence of recovery from the knockdown and moribund states, but this was not the case for Cinygma sp. The pulse duration resulting in 50% mortality was calculated as 43 min for Cinygma sp. exposed at 204 microg/L and 16 min at 408 microg/L. A three-dimensional probit plane model [Y = -10.86 + 4.83(ln C) + 3.0(ln T)], where Y is probit mortality, C is concentration in microg/L and T is time in hours, was used to explain the interaction between concentration (microg/L) and duration of exposure (hours) for Cinygma sp.

Animals↗
PubMed1998

Characterization of Aplysia attractin, the first water-borne peptide pheromone in invertebrates.

Although animals in the genus Aplysia are solitary during most of the year, they form breeding aggregations during the reproductive season. The aggregations contain both mating and egg-laying animals and are associated with masses of egg cordons. The egg cordons are a source of pheromones that establish and maintain the aggregation, but none of the pheromonal factors have been chemically characterized. In these studies, specimens of Aplysia were induced to lay eggs, the egg cordons collected and eluted, and the eluates fractionated by C18 reversed-phase HPLC. Four peak fractions were bioassayed in a T-maze. All four increased the number of animals attracted to a nonlaying conspecific and were thus subjected to compositional and microsequence analysis. Each contained the same NH2-terminal peptide sequence. The full-length peptide ("attractin") was isolated from the albumen gland, a large exocrine organ that packages the eggs into a cordon. The complete 58-residue sequence was obtained, and it matched that predicted by an albumen gland cDNA. Mass spectrometry showed that attractin is 21 wt.% carbohydrate as the result of N-linked glycosylation. T-maze bioassays confirmed that the full-length peptide is attractive. Attractin is the first water-borne peptide pheromone characterized in molluscs, and the first in invertebrates.

Amino Acid Sequence↗
PubMed2001

Competition for space among sessile marine invertebrates: changes in HSP70 expression in two Pacific cnidarians.

The role of stress proteins-either constitutive (HSC) or inducible (HSP)-of the HSP70 family in intra- and interspecific competition for space was examined in two sessile Pacific cnidarians. Anthopleura elegantissima, an intertidal anemone, and Corynactis californica, a subtidal corallimorpharian, express HSP70 in the absence of apparent physical stress. HSP70 protein expression is concentrated in the tentacles of A. elegantissima when the animal is exposed to contact with other benthic organisms. Under the same conditions, however, HSP concentrations are similar in the body and tentacles of C. californica. When two different clones of A. elegantissima interact in the field, the outside polyps (warriors) express more HSP70 than the inside ones (2.4 versus 0.6 ng HSP70/microg Protein). When different C. californica clones interact, HSP70 expression in the outside and inside polyps is similar (1.5 versus 1.8 ng HSP70/microg P) and is fairly constant in the corallimorpharian in the different interspecific encounters. HSP70 expression is related to the different kinds of aggression encountered by both cnidarians. HSP70 expression may be involved in the recovery of tissues damaged by the allelochemical, cytotoxical, or corrosive substances produced by different enemies. C. californica clones appear prepared for war, as evidenced by the high constant expression of HSP70 in the polyps. A. elegantissima exhibits differential HSP70 expression depending on the identity of each neighboring intra- or interspecific sessile competitor. We propose that stress proteins can be used to quantify space competition or aggression among sessile marine invertebrates.

Agonistic Behavior↗
PubMed2003

CiC3-1a-mediated chemotaxis in the deuterostome invertebrate Ciona intestinalis (Urochordata).

Deuterostome invertebrates possess complement genes, and in limited instances complement-mediated functions have been reported in these organisms. However, the organization of the complement pathway(s), as well as the functions exerted by the cloned gene products, are largely unknown. To address the issue of the presence of an inflammatory pathway in ascidians, we expressed in Escherichia coli the fragment of Ciona intestinalis C3-1 corresponding to mammalian complement C3a (rCiC3-1a) and assessed its chemotactic activity on C. intestinalis hemocytes. We found that the migration of C. intestinalis hemocytes toward rCiC3-1a was dose dependent, peaking at 500 nM, and was specific for CiC3-1a, being inhibited by an anti-rCiC3-1a-specific Ab. As is true for mammalian C3a, the chemotactic activity of C. intestinalis C3-1a was localized to the C terminus, because a peptide representing the 18 C-terminal amino acids (CiC3-1a(59-76)) also promoted hemocyte chemotaxis. Furthermore, the CiC3-1a terminal Arg was not crucial for chemotactic activity, because the desArg peptide (CiC3-1a(59-75)) retained most of the directional hemocyte migration activity. The CiC3-1a-mediated chemotaxis was inhibited by pretreatment of cells with pertussis toxin, suggesting that the receptor molecule mediating the chemotactic effect is G(i) protein coupled. Immunohistochemical analysis with anti-rCiC3-1a-specific Ab and in situ hybridization experiments with a riboprobe corresponding to the 3'-terminal sequence of CiC3-1, performed on tunic sections of LPS-injected animals, showed that a majority of the infiltrating labeled hemocytes were granular amebocytes and compartment cells. Our findings indicate that CiC3-1a mediates chemotaxis of C. intestinalis hemocytes, thus suggesting an important role for this molecule in inflammatory processes.

Amino Acid Sequence↗
PubMed2006

First identification of a chemotactic receptor in an invertebrate species: structural and functional characterization of Ciona intestinalis C3a receptor.

In mammals, the bioactive fragment C3a, released from C3 during complement activation, is a potent mediator of inflammatory reactions and exerts its functional activity through the specific binding to cell surface G protein-coupled seven-transmembrane receptors. Recently, we demonstrated a Ciona intestinalis C3a (CiC3a)-mediated chemotaxis of hemocytes in the deuterostome invertebrate Ciona intestinalis and suggested an important role for this molecule in inflammatory processes. In the present work, we have cloned and characterized the receptor molecule involved in the CiC3a-mediated chemotaxis and studied its expression profile. The sequence, encoding a 95,394 Da seven-transmembrane domain protein, shows the highest sequence homology with mammalian C3aRs. Northern blot analysis revealed that the CiC3aR is expressed abundantly in the heart and neural complex and to a lesser extent in the ovaries, hemocytes, and larvae. Three polyclonal Abs raised in rabbits against peptides corresponding to CiC3aR regions of the first and second extracellular loop and of the third intracellular loop react specifically in Western blotting with a single band of 98-102 kDa in hemocyte protein extracts. Immunostaining performed on circulating hemocytes with the three specific Abs revealed that CiC3aR is constitutively expressed only in hyaline and granular amoebocytes. In chemotaxis experiments, the Abs against the first and second extracellular loop inhibited directional migration of hemocytes toward the synthetic peptide reproducing the CiC3a C-terminal sequence, thus providing the compelling evidence that C. intestinalis expresses a functional C3aR homologous to the mammalian receptor. These findings further elucidate the evolutionary origin of the vertebrate complement-mediated proinflammatory process.

Amino Acid Sequence↗
PubMed1999

Cellular analysis of appetitive learning in invertebrates.

In recent years significant progress has been made in the analysis of the cellular mechanisms underlying appetitive learning in two invertebrate species, the pond snail Lymnaea stagnalis and the honeybee Apis mellifera. In Lymnaea, both chemical (taste) and tactile appetitive conditioning paradigms were used and cellular traces of behavioural classical conditioning were recorded at several specific sites in the nervous system. These sites included sensory pathways, central pattern generator and modulatory interneurones as well as motoneurones of the feeding network. In the honeybee, a chemical (odour) appetitive conditioning paradigm resulted in cellular changes at different sites in the nervous system. In both the pond snail and the honeybee the activation of identified modulatory interneurones could substitute for the use of the chemical unconditioned stimulus, making these paradigms even more amenable to more detailed cellular and molecular analysis.

Animals↗
PubMed2000

Evidence of opiates and opioid neuropeptides and their immune effects in parasitic invertebrates representing three different phyla: Schistosoma mansoni, Theromyzon tessulatum, Trichinella spiralis.

Studies done in our laboratories have demonstrated that the parasitic trematode, Schistosoma mansoni is capable of producing several proopiomelanocortin (POMC) peptides including beta-endorphin, adrenocorticotropin (ACTH), melanocyte stimulating hormone (alphaMSH) and enkephalin as well as morphine. Some of these opioids have been demonstrated to be immunosuppressive and may play an important part in immune evasion by these parasites. The parasitic nematode Trichinella spiralis also produces immune suppressive substances in vitro as well as causes immune suppression in its encysted stage in vivo. We recently have demonstrated the presence of morphine in both infected mice and in the nematode by HPLC and RIA. In a recent study of the leech Theromyzon tessulatum, we demonstrated the presence of proopiomelanocortin (POMC) and its derived peptides, ACTH and alphaMSH, in the immune tissues. The peptide was cloned and extensively purified by HPGPC and reversed-phase HPLC, and then sequenced. The 25.4 kDa protein was purified by gel permeation chromatography, anti-ACTH-affinity column separation followed by reversed-phase HPLC. Its amino acid determination was performed by Edman degradation, enzymatic treatments and electrospray mass spectrometry. The structure of the leech POMC-like precursor and its derived peptides demonstrates considerable amino acid sequence similarity with mammalian POMC. Taken together, these studies demonstrate that opiates and opioid neuropetides are present in invertebrates and their immunoregulatory actions have been conserved during evolution. The role of opiates and opioid peptides in immune and behavior modification of hosts is also discussed.

Adrenocorticotropic Hormone↗
PubMed2001

[Use of immunomodulators isolated from marine invertebrates for reducing the toxic effects of thermostable toxin and lipopolysaccharides from Yersinia pseudotuberculosis on a macroorganism].

The possibility to use immunomodulators isolated from marine invertebrates for the lowering of the toxic effects caused by Yersinia pseudotuberculosis thermoresistant toxin and lipopolysaccharide was investigated. Effects were evaluated by the animals survival rate in per cent and mice average lifetime after toxin lethal dose injection. It was shown that polypeptide gangleen when compared to timalin as well as glycanes mitilane and strombus had dose-dependent protective effect. These substances increased animals survival rate to 15-17 per cent and prolonged life period for about two times when compared to control group. These results demonstrates the possibility to use investigated immunomodulators is clinical practice at the treatment of the patients with pseudotuberculosis.

Adjuvants, Immunologic↗
PubMed2002

Physiological responses to acute silver exposure in the freshwater crayfish (Cambarus diogenes diogenes)--a model invertebrate?

Adult crayfish (Cambarus diogenes diogenes) exposed to 8.41 +/- 0.17 microg silver/L (19.4% as Ag+) in moderately hard freshwater under flow-through conditions for 96 h exhibited ionoregulatory disturbance, elevated metabolic ammonia (T(amm)) production and substantial silver accumulation in the gills, hemolymph, and hepatopancreas. The ionoregulatory disturbance included both a generally reduced unidirectional Na+ influx and an increased unidirectional Na+ efflux, leading to a substantial net loss of Na+ from the silver-exposed crayfish. The Na+ uptake in silver-exposed crayfish differed overall from controls, while the increased Na+ efflux recovered to control values 48 h into the 96 h of exposure. The general inhibition of Na+ uptake could be explained by a reduced sodium/potassium-adenosine triphosphatase (Na/K-ATPase) activity in terminally obtained gill samples from the silver-exposed crayfish. The silver-induced effect on Na+ uptake and loss translated to reduced hemolymph Na+ concentrations but not significantly reduced hemolymph Cl- concentrations. Hemolymph T(anim) and T(amm) efflux both increased in silver-exposed crayfish, indicating an increased metabolic T(amm) production. The present study demonstrates that the toxic mechanism of waterborne silver exposure in freshwater crayfish resembles that of freshwater teleost fish. The crayfish might therefore be a useful model system for extending current environmental regulatory strategies, currently based on teleost fish, to invertebrates.

Ammonia↗
PubMed2002

Ecological regulation of development: induction of marine invertebrate metamorphosis.

In the marine environment a wide range of invertebrates have a pelagobenthic lifecycle that includes planktonic larval and benthic adult phases. Transition between these morphologically and ecologically distinct phases typically occurs when the developmentally competent larva comes into contact with a species-specific environmental cue. This cue acts as a morphogenetic signal that induces the completion of the postlarval/juvenile/adult developmental program at metamorphosis. The development of competence often occurs hours to days after the larva is morphologically mature. In the non-feeding--lecithotrophic--larvae of the ascidian Herdmania curvata and the gastropod mollusc Haliotis asinina, gene expression patterns in pre-competent and competent stages are markedly different, reflecting the different developmental states of these larval stages. For example, the expression of Hemps, an EGF-like signalling peptide required for the induction of Herdmania metamorphosis, increases in competent larvae. Induction of settlement and metamorphosis results in further changes in developmental gene expression, which apparently is necessary for the complete transformation of the larval body plan into the adult form.

Animals↗
PubMedDate not supplied

Interactions between environmental pollution and cold tolerance of soil invertebrates: a neglected field of research.

The adaptations to climatic stress of invertebrates have been extensively studied and the underlying physiology is relatively well understood. It is therefore possible to predict critical lower temperature and moisture limits at which a species will suffer increased mortality. Environmental pollution can reduce the tolerance to climatic stress considerably but it is not known how significant these synergistic effects are in polluted areas. Results are presented showing examples of synergism between toxic and climatic stress. The physiological mechanisms involved in the synergistic effects are discussed.

Journal Article↗
PubMed1992

First non-vertebrate member of the myc gene family is seasonally expressed in an invertebrate testis.

We have cloned and characterized the first non-vertebrate member of the myc gene family, pAv-myc, from testes of the Northern sea star, Asterias vulgaris (Echinodermata). We have used an oligonucleotide that is complementary to the virtually 100% conserved vertebrate c-myc box A in the second exon and a cDNA library constructed from spermatogenically active testes. Relatives of this echinoderm existed approximately 100 million years prior to the origin of the earliest known organism for which c-myc sequence is currently available (the trout). Nonetheless, our cDNA encodes a protein with approximately 30% amino acid identity and 46% overall conservation to human c-myc. Regions of substantially higher conservation (63-95%) correspond in order to the transcriptional activation (boxes A, B and C), casein kinase II phosphorylation, nuclear-targeting, basic DNA-binding and oligomerization domains in the second and third exons of human c-myc. Sea star c-myc cDNA detects a 2.7-kb transcript on Northern blots of monthly samples of testicular tissue from field-collected individuals (n = 6-8), indicating peak expression during active spermatogenesis. This is also the first example of seasonal variation in expression of c-myc during spermatogenesis in laboratory or natural populations of any animal. It is intriguing to speculate about the potential oncogenic character of c-myc in this invertebrate, in which tumors have not yet been observed, and about the possibility that c-myc is more widely present in eucaryotes than has been anticipated.

Amino Acid Sequence↗
PubMed2003

The impact of wetland vegetation drying time on abundance of mosquitoes and other invertebrates.

Vegetation management for constructed treatment wetlands often involves knocking down emergent vegetation with heavy equipment and inundating the dead vegetation after a period of drying. Such practices create favorable conditions for larval mosquitoes. We studied the relationship between length of the drying period for an emergent macrophyte, Typha sp., and the abundance of aquatic invertebrates in replicated 0.18-m3 wading pools. The mosquito, Culex tarsalis, was significantly more abundant in pools containing vegetation aged for 2 wk before inundation compared to pools containing vegetation aged 5 wk, freshly cut vegetation, or without vegetation. Potential larval mosquito food resources (particles between 2 and 61 microm in equivalent spherical diameter) in the 2-wk aging treatment did not differ significantly from the other treatments during the 5-wk experiment. The abundance of other larval culicids, nonculicine Diptera, and potential mosquito predators (i.e., Dytiscidae and Aeshnidae) did not differ significantly among the vegetation aging treatments.

Animals↗
PubMed1977

Distribution of sulfated mucopolysaccharides in invertebrates.

The sulfated mucopolysaccharide composition of 22 species of invertebrates belonging to the phyla Arthropoda, Mollusca, Annelida, Tunicata, Echinodermata, Coelenterata, and Porifera was analyzed. It is shown that all the species contain variable amounts of one or more types of sulfated mocopolysaccharides, most of which similar to the ones found in vertebrates. It is shown also that each species has a characteristic composition, differing from each other regarding the relative amount and type of chondroitin sulfates A, B, and C, heparitin sulfate, and heparin. The possible biological role of the sulfated mucopolysaccharides in cell recognition or aggregation or both is discussed in view of the present findings.

Animals↗
PubMed2004

Invertebrate disease models in neurotherapeutic discovery.

Models that reproduce many of the cellular and molecular aspects of various human neurodegenerative disorders have been developed in the fruit fly Drosophila melanogaster and the nematode Caenorhabditis elegans. An understanding of the underlying molecular and genetic mechanisms of disease pathogenesis is being gained from studies utilizing the wealth of genetic and molecular tools available for these invertebrate model organisms. This review focuses on recent studies that lay a foundation for utilizing these disease models in drug discovery and for continued genetic dissection of disease mechanisms.

Animals↗
PubMedDate not supplied

[Comparative properties of myogenesis in invertebrates and in lower and higher vertebrates].

Development of the muscle system in invertebrates (on the example of nematode Caenorhabditis elegans and Drosophila) and vertebrates (fish, birds, and mammals) demonstrates a number of common patterns at the level of molecular and genetic control mechanisms but also a number of distinctive features. C. elegans muscle is formed from several cells of different origin at the earliest developmental stages, while inductive interactions play a critical role in Drosophila: the mesoderm, which is a source of muscle formation, receives the inductive signals from the neighboring ectoderm as expression of Dpp and Hh genes. In vertebrates, the induction and commitment of cells to the myogenic lineage are promoted by morphogenetic signals from the neighboring notochord (as expression of Shh gene) and neural tube (as expression of Wnt and Shh genes). The inductive signals entering the mesoderm are related to the subsequent activation of genes encoding protein transcription factors of bHLH gene family and analogs. These include hlh-1 and CeTwi in C. elegans; Twi and nau in Drosophila; and MyoD, Myf5, myogenin, and MRF4 in vertebrates. The diversity of myosin isoforms in different animals is provided by either gene duplication or alternative splicing of particular genes.

Animals↗
PubMed2005

[Human mortality due to invertebrate and vertebrate envenomation in Monagas State, Venezuela].

Human mortality due to envenomation by animals in Monagas state, Venezuela was evaluated. In this state 47 deaths due to envenomations produced by vertebrates and invertebrates (scorpions = 21, snakes = 20, bees = 6) during the period 1980-2000 were registered. The mountainous area, at the north of the state (Turimiquire sub-region) and its piedmonts reported most of the mortality by scorpionism (57.2%) and ophidism (70.0%). The age group of the patients was related with the sort of envenomation causing the death. During the period of dry season the highest percentage of deaths by scorpions and bees occurred; and the deaths produced by snakes were more frequent in the median raining season. The most frequent causes of death (61.9%) by scorpionism were the acute lung edema and congestive heart failure. In 40% of the deaths caused by snakes, alterations of the blood coagulability were observed. Bee envenomation was responsible of 50% of the deaths produced by anaphylactic shock. In none of the deaths, the species responsible of the accident was identified. The annual mean mortality rate/100,000 inhabitants, in the 21 years registered was 0.20 for the scorpion envenomation, 0.19 for ophidism and 0.06 for bee envenomation. The data suggest the importance of the mortality produced by scorpions and snakes, particularly at the north of the state.

Adolescent↗