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Natural and human impacts on invertebrate communities in Brazilian caves.

Species richness, abundance, distribution and similarity between cave invertebrate communities were compared among seven caves located in the Peruaçu River valley, north of Minas Gerais State, Brazil. Such comparisons aimed to determinate the degree of biological complexity in the sampled caves, calculated by the "Index of Biological Complexity in Caves", presented in this manuscript. The presence of potential or real impacts on the cave fauna was also investigated. A total of 1,468 individuals belonging to 57 families of: Acarina, Pseudoscorpionida, Araneida, Opilionida, Amblypygi, Isopoda, Geophilomorpha, Scutigeromorpha, Spirostreptida, Coleoptera, Collembola, Diptera, Dictyoptera, Ephemeroptera, Ensifera, Heteroptera, Hymenoptera, Lepidoptera, Plecoptera, Psocoptera, and Trichoptera was collected. Caves with higher resource availability (as those hidrologicaly actives) had a higher biological complexity than those with less resource. There are two types of impacts that occur in the area: the natural (geological) and the anthropic, as intense "stepping" and visitation or use of cave entrances as cattle shelters. There are caves with different preservation degrees in the area, with invertebrate communities in varied complexity states. The communities of these caves undoubtedly deserve care, since the area is extremely important in the Brazilian biospeleological context.

Animals↗

Relationship between metabolism and bioaccumulation of benzo[a]pyrene in benthic invertebrates.

The potential influence of polycyclic aromatic hydrocarbon (PAH) metabolism on bioaccumulation is well accepted, but rarely has been examined in many species of benthic invertebrates that commonly are found in contaminated sediments, or used in bioaccumulation or toxicity tests. In this study, the relative ability of 11 species of near-shore benthic invertebrates to metabolize and bioaccumulate a model PAH, benzo[a]pyrene (B[a]P), was evaluated. Species examined included six polychaetes (Clymenella torquata, Nereis virens, Nereis succinea, Nephtys incisa, Spio setosa and Cirriformia grandis), three bivalves, (Macoma balthica, Mya arenaria, and Mulinia lateralis), and two amphipods (Ampelisca abdita and Leptocheirus plumulosus). After 7 d of exposure to sediments spiked with radiolabeled B[a]P, metabolites comprised between 6.1% (Clymenella torquata) to 85.7% (Nereis succinea) of total accumulated B[a]P, with individual species from the same phylogenetic groups showing large differences in their ability to metabolize this PAH. Bioaccumulation factors (B[a]P in tissue/B[a]P in sediment) were inversely related to the species' ability to metabolize PAH, highlighting the importance of considering metabolism when interpreting bioaccumulation across several species. These data argue strongly against the continued use of the large polychaete Nereis virens, one of the species showing the greatest ability to metabolize B[a]P, for bioaccumulation testing when PAHs are being considered. Other commonly used test species had relatively low levels of metabolism (Ampelisca abdita, Leptocheirus plumulosus, and Macoma balthica), supporting their use in evaluation of potential PAH impact on the environment.

Amphipoda↗

Responses of periphyton and invertebrates to a tetradecyl-pentadecyl sulfate mixture in stream mesocosms.

Alkyl sulfates (AS) are anionic surfactants widely used in household and personal cleansing applications. Aquatic toxicity of AS under laboratory conditions indicated effects at relatively low concentrations (50-230 microg/L) for some sensitive species. A comprehensive stream mesocosm study of an AS mixture composed of tetra- (C14) and pentadecyl (C15) chain lengths was conducted to better understand effects on microbial and macroinvertebrate populations and communities. A 56-d exposure of AS was performed at concentrations ranging from 57 to 419 microg/L (analytically confirmed exposures) and was accompanied by detailed investigations of periphyton community function (autotrophy, heterotrophy, and metabolism of test chemical), periphyton structure (algal population and community dynamics based on taxonomic identity), and invertebrate structure (benthic abundance, drift, and insect emergence patterns based on taxonomic identity). A no-observed-effect-concentration (NOEC) of 222 microg/L was concluded for several individual algal and invertebrate species based on univariate statistical analyses. An apparent energetic subsidy from C14-15AS at the highest concentrations of 222 to 419 microg/L was observed and tied to changes in microbial community processing of AS when added at these high concentrations. A multivariate analysis based on principal response curves (PRC) indicated that communities in streams exposed to 222 to 419 microg/L were significantly different from the controls leading to an overall (multivariate and univariate) conclusion that 106 microg/L was the ecosystem NOEC. Exposure to AS in the environment has been demonstrated to be in the range of 5 to 21 microg/L in 100% wastewater treatment plant effluent. Potential environmental effects are at least 5 to 20 times above worst-case environmental exposures; therefore, C14-15AS does not pose a risk to the aquatic environment due to normal use patterns.

Animals↗

Modeling zinc toxicity for terrestrial invertebrates.

Acute and chronic ecotoxicity tests with zinc were performed with the earthworm Eisenia fetida, the potworm Enchytraeus albidus, and the springtail Folsomia candida. To assess the influence of the soil type on zinc toxicity for these soil invertebrates, these tests were carried out in a standard artificial soil, a sandy and a loamy field soil. Based on the results of this experimental work and data taken from literature, models were developed relating the ecotoxicity of zinc to the most important parameters controlling bioavailability: pH and cation exchange capacity. Models were developed for E. fetida and F. candida using the regression technique partial least squares projection to latent structures (PLS). Acute as well as chronic toxicity data of both organisms could be normalized on the basis of the pH and the cation exchange capacity of the test soils. For other terrestrial invertebrates, not enough data were available to develop reliable models.

Animals↗

A comparison of the effect of new and established insecticides on nontarget invertebrates of winter wheat fields.

Six insecticides, a carbamate (pirimicarb), four pyrethroid/mixtures (tao-fluvalinate, zeta-cypermethrin, deltamethrin, and pirimcarb+deltamethrin), and an organophosphate (dimethoate), registered for use within cereals, were field-tested for toxicity to beneficial nontarget invertebrate groups important as food for farmland birds. Compared to the control and the other compounds, pirimicarb was the least toxic for most invertebrate groups examined. The pyrethroids and pyrethroid mixture significantly reduced numbers of many of the nontarget groups compared to the control and pirimicarb. Tao-fluvalinate was the least toxic pyrethroid tested when compared to the other compounds. Toxicity was further tested on larvae of graminivorous sawflies, an important chick- food group, using field-sprayed plants in laboratory and semifield trials. None of the pyrethroid and carbamate compounds and mixtures was as toxic as the organophosphate, but all were more toxic than the control. The semifield experiments provided a useful way of determining potential insecticide impact on this nontarget group where field populations are often too low for field trials to have statistically conclusive results.

Agriculture↗

Behavior and reproduction of invertebrate animals during and after a long-term microgravity: space experiments using an Autonomous Biological System (ABS).

Aquatic invertebrate animals such as Amphipods, Gastropods (pond snails), Ostracods and Daphnia (water flea) were placed in water-filled cylindrical vessels together with water plant (hornwort). The vessels were sealed completely and illuminated with a fluorescent lamp to activate the photosynthesis of the plant for providing oxygen within the vessels. Such ecosystem vessels, specially termed as Autonomous Biological System or ABS units, were exposed to microgravity conditions, and the behavior of the animals and their reproduction capacity were studied. Three space experiments were carried out. The first experiment used a Space shuttle only and it was a 10-day flight. The other two space experiments were carried out in the Space station Mir (Shuttle/Mir mission), and the flight units had been kept in microgravity for 4 months. Daphnia produced their offspring during a 10-day Shuttle flight. In the first Mir experiment, no Daphnia were detected when recovered to the ground. However, they were alive in the second Mir experiment. Daphnia were the most fragile species among the invertebrate animals employed in the present experiments. All the animals, i.e., Amphipods, pond snails, Ostracods and Daphnia had survived for 4 months in space, i.e., they had produced their offspring or repeated their life-cycles under microgravity. For the two Mir experiments, in both the flight and ground control ecosystem units, an inverse relationship was noted between the number of Amphipods and pond snails in each unit. Amphipods at 10 hours after the recovery to the ground frequently exhibited a movement of dropping straight-downward to the bottom of the units. Several Amphipods had their legs bent abnormally, which probably resulted from some physiological alterations during their embryonic development under microgravity. From the analysis of the video tape recorded in space, for Ostracods and Daphnia, a half of their population were looping under microgravity. Such looping animals could be observed still at the end of the 4 month stay in space. No looping behavior was noted for Amphipods and pond snails.

Animals↗

Fertilization success in marine invertebrates: the influence of gamete age.

Gamete age has been postulated to be unimportant to the fertilization ecology of marine invertebrates. However, recent research suggests that, for some species at least, it may have a direct impact upon fertilization success. We present comparative data on the influence of gamete age on fertilization and development success in several marine invertebrates: the polychaetes Arenicola marina and Nereis virens and the asteroid echinoderm Asterias rubens. Oocytes are much longer lived in the polychaetes than in the echinoderm, with A. marina oocytes still capable of fertilizing and developing normally 96 h post-spawning. Developmental abnormalities and failure to reach blastula tend to occur well before the fertilizable life of the oocytes has expired. Sperm are similarly longer lived in the polychaetes; however, fertilizing capacity is markedly reduced following incubation in conspecific egg-conditioned seawater. These results are discussed in terms of the fertilization strategies of the three species. We further suggest that, for A. marina at least, longer-lived sperm and eggs are central to the fertilization strategy of this species.

Animals↗

Molecular and immunological characterization of arginine kinase from the Indianmeal moth, Plodia interpunctella, a novel cross-reactive invertebrate pan-allergen.

IgE recognition of indoor allergens represents a major cause of allergic asthma in atopic individuals. We found that 52 of 102 patients suffering from allergic symptoms indoors contained IgE Abs against allergens from the Indianmeal moth (Plodia interpunctella), a ubiquitous food pest. Using serum IgE from a moth-sensitized patient we screened an expression cDNA library constructed from P. interpunctella larvae. cDNAs coding for arginine kinase (EC 2.7.3.3), a 40-kDa enzyme commonly occurring in invertebrates that is involved in the storage of such high-energy phosphate bonds as phosphoarginine, were isolated. Recombinant moth arginine kinase, designated Plo i 1, was expressed in Escherichia coli as a histidine-tagged protein with enzymatic activity, and purified to homogeneity by nickel chelate affinity chromatography. Purified recombinant arginine kinase induced specific basophil histamine release and immediate as well as late-phase skin reactions. It reacted with serum IgE from 13 of the 52 (25%) moth-allergic patients and inhibited the binding of allergic patients' IgE to an immunologically related 40-kDa allergen present in house dust mite, cockroach, king prawn, lobster, and mussel. Our results indicate that arginine kinases represent a new class of cross-reactive invertebrate pan-allergens. Recombinant arginine kinase may be used to identify a group of polysensitized indoor allergic patients and for immunotherapy of these individuals.

Adolescent↗

The flea, Megabothris abantis: an invertebrate host of Hepatozoon sp. and a likely definitive host in hepatozoon infections of the montane vole, Microtus montanus.

In searching for an invertebrate host for Hepatozoon sp. infecting the montane vole (Microtus montanus), we collected fleas, ticks, and mites from live-trapped voles and searched squash preparations for Hepatozoon oocysts. From 1989 through 1996, we identified six species of fleas in Grand Teton National Park: Megabothris abantis, Megabothris asio megacolpus, Aetheca wagneri, Peromyscopsylla selenis, Peromyscopsylla. hesperomys, and Hystrichopsylla dippiei dippiei. We found Hepatozoon oocysts only in M. abantis; we found no oocysts in mites or ticks. We conclude that M. abantis is an invertebrate host of Hepatozoon sp. and is likely to be the definitive host for theHepatozoon spp. of M. montanus.

Animals↗

Amphetamine elicited potential changes in vertebrate and invertebrate central neurons.

The effects of amphetamine on potential changes in both vertebrate and invertebrate central neurons and factors affecting the potential changes were tested. The animals studied included mice, newborn rat and African snail. Seizure was elicited after lethal doses of d-amphetamine (75 mg/kg, i.p.) administration in mice. Repetitive firing of the action potentials were elicited after d-amphetamine (1-30 microM) administration in thin thalamic brain slices of newborn rat. Bursting firing of action potentials in the giant African central RP4 neuron were also elicited after d-amphetamine or l-amphetamine (0.27 mM) administration. The amphetamine elicited bursting firing of action potentials was not blocked even after high concentrations of d-tubocurarine, atropine, haloperidol, hexamethonium administration. Therefore, the amphetamine elicited potential changes may not be directly related to the activation of the receptors of the neuron. The bursting firing of action potentials elicited by amphetamine occurred 20-30 min after amphetamine administration extracellularly, even after high concentrations of d-amphetamine administration (0.27, 1 mM). However, the bursting firing of potentials occurred immediately if amphetamine was administrated intracellularly at lower concentration. Extracellular application of ruthenium red, the calcium antagonist, abolished the amphetamine elicited bursting firing of action potentials. If intracellular injection of EGTA, a calcium ion chelator, or injection with high concentrations of magnesium, the bursting firing of potentials were immediately abolished. These results suggested that the active site of amphetamine may be inside of the neuron and the calcium ion in the neuron played an important role on the bursting of potentials. In two-electrode voltage clamped RP4 neuron, amphetamine, at 0.27 mM, decreased the total inward and steady outward currents of the RP4 neuron. d-Amphetamine also decreased the calcium, Ia and the steady-state outward currents of the RP4 neuron. Besides, amphetamine elicited a negative slope resistance (NSR) if membrane potential was in the range of -50 to -10 mV. The NSR was decreased in cobalt substituted calcium free and sodium free solution. The effects of secondary messengers on the amphetamine elicited potential changes were tested. The bursting firing of action potentials elicited by amphetamine in central snail neurons decreased following extracellular application of H8 (N-(2-methyl-amino) ethyl-3-isoquinoline sulphonamide dihydrochloride), a specific protein kinase A inhibitor and anisomycin, a protein synthesis inhibitor. However, the bursting firing of action potentials were not affected after extracellular application of H7 (1,(5-isoquinolinesulphonyl)-2-methylpiperasine dihydrochloride), a specific protein kinase C (PKC) inhibitor, or intracellular application of GDPbetaS, a G protein inhibitor. The oscillation of membrane potential of the bursting activity was blocked after intracellular injection of 3'-deoxyadenosine, an adenylyl-cyclase inhibitor. These results suggested that the bursting firing of action potentials elicited by d-amphetamine in snail neuron may be associated with the cyclic AMP second messenger system; on the other hand, it may not be associated with the G protein and protein kinase C activity. It is concluded that amphetamine elicited potential changes in both vertebrate and invertebrate central neurons. The changes are closely related to the ionic currents and second messengers of the neurons.

Action Potentials↗

Cadmium accumulation by invertebrates living at the sediment-water interface.

Benthic animals can take up trace metals both from the sediment compartment in which they burrow and from the water column compartment above their burrows (we define both compartments as containing water and particles). If criteria for the protection of benthic animals are based on metal concentrations in one of these two compartments, then it should first be demonstrated that the majority of the metal taken up by these animals comes from the given compartment. To determine whether benthic animals take up the majority of their cadmium (Cd) from the sediment compartment, we created a Cd gradient in lake sediment and compared Cd accumulation by the invertebrates colonizing these sediments with Cd concentrations in the sediment compartment. On the basis of this relationship and using a bioaccumulation model, we estimate that indigenous benthic invertebrates take up the majority of their Cd from the water column compartment. The results of our experiment are similar to those from a previous study conducted on a different benthic community in a larger lake. Taxa common to both lakes obtained similar proportions of their Cd from the water column compartment, suggesting that Cd accumulation by the same species will be constant across lakes of differing size and chemistry. Our results strengthen the argument that the protection of benthic communities from metal pollution should consider metal in both the water column and sediment compartments. In this regard, the AVS model, which considers only sedimentary metals, was more effective in predicting Cd concentrations in pore waters than those in most animal taxa. We suggest that measurements of vertical chemical heterogeneity in sediments and of animal behavior would aid in predicting the bioaccumulation and effects of sedimentary pollutants.

Animals↗

Selective electrical silencing of mammalian neurons in vitro by the use of invertebrate ligand-gated chloride channels.

Selectively reducing the excitability of specific neurons will (1) allow for the creation of animal models of human neurological disorders and (2) provide insight into the global function of specific sets of neurons. We focus on a combined genetic and pharmacological approach to silence neurons electrically. We express invertebrate ivermectin (IVM)-sensitive chloride channels (Caenorhabditis elegans GluCl alpha and beta) with a Sindbis virus and then activate these channels with IVM to produce inhibition via a Cl- conductance. We constructed a three-cistron Sindbis virus that expresses the alpha and beta subunits of a glutamate-gated chloride channel (GluCl) along with the green fluorescent protein (EGFP) marker. Expression of the C. elegans channel does not affect the normal spike activity or GABA/glutamate postsynaptic currents of cultured embryonic day 18 hippocampal neurons. At concentrations as low as 5 nm, IVM activates a Cl- current large enough to silence infected neurons effectively. This conductance reverses in 8 hr. These low concentrations of IVM do not potentiate GABA responses. Comparable results are observed with plasmid transfection of yellow fluorescent protein-tagged (EYFP) GluCl alpha and cyan fluorescent protein-tagged (ECFP) GluCl beta. The present study provides an in vitro model mimicking conditions that can be obtained in transgenic mice and in viral-mediated gene therapy. These experiments demonstrate the feasibility of using invertebrate ligand-activated Cl- channels as an approach to modulate excitability.

Action Potentials↗

Beauvericin cytotoxicity to the invertebrate cell line SF-9.

The cyclic hexadepsipeptide beauvericin, initially known as a secondary metabolite produced by the entomopathogenic fungus Beauveria bassiana and toxic to Artemia salina larvae, has been more recently recognized as an important mycotoxin synthesized by a number of Fusarium strains, which parasite maize, wheat and rice. Therefore, this mycotoxin may enter the food chain, causing yet unknown effects to the health of both domestic animals and humans. The cytotoxic effects of beauvericin on mammalian cells have been studied. We investigated the cytotoxicity of this compound in an in vitro invertebrate model, viz. the insect cell line SF-9 (immortalized pupal ovarian cells of the lepidopter Spodoptera frugiperda). Cultures of SF-9 cells in the stationary phase were exposed to beauvericin at concentrations ranging from 100 nM to 300 microM, for different periods of time (from 30' to 120 h). The effects on cell viability were assessed by the trypan blue exclusion method. After 4 h of incubation no significant decrease in cell viability was recorded in SF-9 cell cultures exposed to low concentrations of beauvericin, i.e. 100 nM and 300 nM. However, a slight decrease in viability (3.9%) was seen already in cells exposed to the mycotoxin at the 1 microM concentration. This effect became gradually more evident at higher concentrations (approximately equal to 28% at 30 microM, approximately equal to 50% at 100 microM, approximately equal to 68% at 300 microM). An even more pronounced reduction in cell viability was observed after a 24 h exposure. Under these conditions, 1 microM beauvericin caused an approx. 10% decrease in the number of viable cells, which became more significant at higher concentrations approximately equal to 23% at 3 microM, approximately equal to 47% at 10 microM, approximately equal to 65% at 30 microM, approximately equal to 90% at 100 microM, approximately equal to 99% at 300 microM). Therefore, the 50% cytotoxic concentrations (CC50) at 4 h and 24 h could be estimated as 85 microM and 10 microM, respectively. In time-course experiments, no effect of beauvericin (30 microM) on cell viability could be seen after exposure for periods of time as long as 30', 1 h and 2 h, respectively. In contrast, when SF-9 cells were exposed to the mycotoxin for longer periods of time, from 8 h to 120 h, we recorded a strong cytotoxic effect already in the low micromolar concentration range. Thus, the CC50 after both 72 h and 120 h exposure times was assessed as 2.5 microM. Higher concentrations caused a virtually 100% cell death. The data collected suggest that beauvericin exerts a substantial dose- and time-dependent cytotoxic effect on invertebrate cells, comparable to the effects described in mammalian cells.

Animals↗

[An experimental study and a mathematical model of interactions in mixed culture of invertebrates and algae in the "producer-consumer" aquatic biotic cycle].

An experimental investigation was carried out, and a mathematical model of interaction between invertebrates (infusoria Paramecium caudatum and rotifera Brachionus plicatilis) and algae (Chlorella vulgaris and Scenedesmus quadricauda) in the "producer-consumer" aquatic biotic cycle with spatially divided links was constructed. The model describes the dynamics of a mixed culture of infusoria and rotifera in the "consumer" link, when they consume a mixed culture of algae coming from the "producer" link. A negative influence of products of algae Scenedesmus metabolism upon the reproduction of infusoria P. caudatum was revealed. Taking this into account, a qualitative coincidence of the results of mathematical modeling with experimental data was obtained. It was shown that the co-existence of mixed algae culture in the "producer" link with invertebrates in the "consumer" link in the "producer-consumer" aquatic biotic cycle is impossible because of the displacement of infusoria P. caudatum by rotifera Brachionus plicatilis.

Animals↗

Subcellular calcium localization and AT0-dependent Ca2+-uptake by smooth endoplasmic reticulum in an invertebrate photoreceptor cell. An ultrastrucutral, cytochemical and X-ray microanalytical study.

In Hirudo medicinalis an extensive and highly elaborate three dimensional network of smooth endoplasmic reticulum cisternae is found in very close structural relationship to the receptive (microvillar) membrane, as reported for many other invertebrates. A variant of the potassium pyroantimonate technique showed that these submicrovillar endoplasmic reticulum cisternae (SMC) and mitochondria are major intracellular calcium stores. Furthermore, using saponine-skinned photoreceptors for an in situ accumulation experiment, calcium oxalate precipitates in SMC demonstrate that this organelle is able to accumulate Ca2+ from a concentration of 2 x 10(-5) M, when ATP, Mg2+, and oxalate ions are present in the accumulation medium. This result provides direct evidence for the hypothesis that SMC may play a particularly important role in the regulation of intracellular ionized calcium in invertebrate photoreceptor cells. Morphological evidence supports this view.

Animals↗

Natural killer cells in a lower invertebrate, Nereis diversicolor.

Globular, non-adherent coelomocytes, called "G3 granulocytes", of the polychaetous annelid Nereis diversicolor display spontaneous cytotoxicity. These cells were found capable of killing invertebrate as well as vertebrate target cells by a contact-dependent cytolytic process. Cytotoxic activity of G3 granulocytes against foreign cells develops in three steps. At first, the cells become motile and form lamellipodia. In a second step, short, pointed pseudopodia arise from the edge of the lamellipodia and are making contact with the stimulating foreign object. In a third step, the G3 granulocytes release dense granules by exocytosis onto the foreign substrate or cell which finally will undergo lysis. Within few minutes after activation, the G3 granulocyte will alter its polarity, realigning both Golgi apparatus and centrosome towards the target cell. A pore-forming protein may be involved in the cytotoxic activity of the G3 granulocytes. These cells were observed to burst after contact with and release of granules onto an abiotic solid substrate, indicating that under certain circumstances the G3 granulocytes may be sensitive to their own cytotoxic activity. These data support the postulate of Franceschi et al. (Eur. J. Immunol. 21, 489-493 (1991) that a primitive natural killer cell-like activity had been developed early in phylogenesis. A simple method for preparing invertebrate coelomocytes for scanning electron microscopy is described.

Animals↗

Invertebrate predation selects for the loss of a morphological antipredator trait.

Antagonistic selection by different predators has been suggested to underlie variation in morphological antipredator traits among and within species. Direct empirical proof is equivocal, however, given the potential interrelationships of morphological and behavioral traits. Here, we tested whether spines in larvae of the dragonfly Leucorrhinia caudalis, which are selected for by fish predators, are selected against by invertebrate aeshnid predators. Using a manipulative approach by cutting spines instead of making comparisons among species or inducing spines, we were able to decouple the presence of spines from other potentially covarying morphological antipredator traits. Results showed survival selection for the loss of spines imposed by invertebrate predation. Moreover, spined and nonspined larval L. caudalis did not differ in the key antipredator behaviors, activity level, and escape burst swimming speed. The observed higher mortality of spined larvae can therefore be directly linked to selection by aeshnid predation against spines.

Animals↗

[Substitution of the dental root by aquatic invertebrate skeletons in animals and man].

The biocompatibility of mammal bone with aragonite and calcite skeletons of aquatic invertebrates (Corals, Molluscs) led us, after animal experimentation, to implant in humans artificial dental roots derived from such invertebrates. These roots, incorporated but not resorbed, serve as supports for a prosthetic crown; they are equipped to isolate the root from the buccal cavity and to ensure shock absorbtion during mastication. The greater ease of implanting artificial teeth and their excellent acceptance will in many cases modify the strategy of current dental treatment.

Adult↗