Influence of pesticides and abiotic conditions on biochemical biomarkers in Aegla aff. longirostri (crustacea, anomura): Implications for conservation.

Freshwater ecosystems are constantly threatened by the advance of agricultural activities. Abiotic variables (such as temperature, ammonia, and nitrite) and contaminants (e.g. pesticides) can potentially interact, increasing metabolism and the absorption of toxic substances, which can alter the ability of organisms to establish adequate stress responses. This study aimed to verify which pesticides were most frequently found and in the greatest quantities in low-order streams, and whether the combination of these pesticides with the abiotic variables altered the biological metabolism of aeglids. These freshwater crustaceans are important shredders that inhabit low-order streams and are sensitive to disturbances and/or abrupt environmental variations. The animals were exposed in situ in four streams (reference site and sites 1, 2, and 3). The reference site is a preserved stream with no apparent anthropogenic interference where aeglids still occur, while the other sites no longer exhibit populations of these animals and are influenced by agricultural activities. The exposure was performed bimonthly from November 2017 to September 2018 and lasted 96 h. Measured abiotic data and water samples were collected through all days of exposure. The analyzed biochemical parameters were acetylcholinesterase activity in muscle; and glutathione S-transferase, lipid peroxidation, protein carbonylation, non-protein thiols, antioxidant capacity against peroxides, and reactive oxygen species (ROS) in muscle, gills, and hepatopancreas. We found 24 active principles of pesticides, the most frequently being clomazone, atrazine, and propoxur. Bentazone was present at the highest amounts. The parameters evaluated in this study, including biochemical biomarkers and abiotic factors measured from the water, provided a separation of the months as a function of environmental conditions. There was a difference in activity and biomarker levels throughout the year within the same site and in some months between sites. The greater concentration or variety of pesticides associated with extreme abiotic (very high temperatures) data generated increased oxidative stress, with high levels of protein damage and considerable lipid damage in all tissues, as well as elevation in ROS, even with high levels of antioxidant capacity and non-protein thiols. With these data, we intend to warn about the risks of exposure to these environmental conditions by trying to contribute to the preservation of limnic fauna, especially aeglid crabs, because most species are under some degree of threat.

Effect of Cd and Pb Pollutions on Physiological Growth: Wavelet Neural Network (WNN) as a New Approach on Age Determination of Coenobita scaevola.

Environmental pollution of aquatic ecosystems leads to an interference in several fundamental biochemical and physiological functions. In this study the interference of Cd and Pb pollutions on the physiological growth and subsequently on the age determination was investigated. The hermit crab, Coenobita scaevola (C.s) was selected as a bioaccumulator in this study. The direct and indirect age determination methods were carried out using annual band counts and carapace length, respectively. The results showed that, there was very low correlation (R2 < 0.5) between direct and indirect age determination. Wavelet Neural Network (WNN) was applied to take into account the environmental effects such as the accumulation of Cd and Pb elements in the C.s' tissues. It was observed that WNN successfully enhanced the growth rate model and estimated the C.s' age (R2 > 0.95). In addition, it was concluded that the environmental pollution had interaction with the growth physiology such as weight and length.

Low salinity-induced alterations in epithelial ultrastructure, Na+/K+-ATPase immunolocalization and enzyme kinetic characteristics in the gills of the thinstripe hermit crab, Clibanarius vittatus (Anomura, Diogenidae).

Fresh caught Clibanarius vittatus [SW, 31‰ salinity (S)] were acclimated to a dilute medium (15‰ S) for 10 days, employing silver staining to locate gill ion transporting tissue, immunofluorescence to localize the Na+/K+-ATPase α-subunit in the lamellae, and electron microscopy to portray ultrastructural changes in the gill epithelia. Na+/K+-ATPase activity was characterized kinetically in a gill microsomal fraction, including synergistic stimulation by NH4+ plus K+. Silver staining revealed that all 26 phyllobranchiate arthro- and pleurobranchiae participate in ion transport. Na+/K+-ATPase α-subunit staining was weak in SW crabs and distributed exclusively and irregularly within the intralamellar septal cells, particularly at the septal-pillar cell body junctions, and septal cell cytoplasm facing the hemolymph space. In 15‰ S crabs, α-subunit localization was intense, occupying the entire thickened septum. Pillar cells and flanges did not stain. Mitochondria and membrane foldings increased in the pillar cell flanges and intralamellar septal cells, greatly amplifying surface area. Only a single ATP binding site (VM  =  130.8 ± 10.5 nmol min-1 mg protein-1; K0.5  =  55.3 ± 1.7 µmol l-1) obeying Michaelis-Menten kinetics was disclosed. Na+/K+-ATPase activity was modulated by Mg2+, Na+, and NH4+, exhibiting site-site interactions; K+ modulation showed Michaelis-Menten kinetics. K+ plus NH4+ synergistically stimulated activity ≈ 1.7-fold. Ouabain inhibited total ATPase activity by ≈ 70% (KI  =  220-300 µmol l-1), revealing phosphohydrolytic activities other than the Na+/K+-ATPase. Despite ample phylogenetic separation, the phyllobranchiate lamellae of the Anomura and Caridea share many ultrastructural features, that is, an intralamellar septum and opposed abutting pillar cells, similar Na+/K+-ATPase distribution, and comparable kinetic characteristics. These findings suggest either convergent evolution at the structural and biochemical levels, or preservation of traits present in a remote common ancestor.

Effects of distilled water rinsing on stable isotope ratios of acid-treated marine invertebrate (Paguridae) samples.

RATIONALE: Stable isotope ratios are widely used to infer trophic relationships, although a growing number of studies show that sample pre-treatments (such as acidification to remove carbonates) can cause changes in isotope ratios. Samples are often rinsed in distilled water after acidification, and we examine the effects of this step in particular on the isotope ratios of marine invertebrate samples. METHODS: Samples of whole hermit crabs (Paguridae) were subjected to one of three treatments: acidification using dilute hydrochloric acid without subsequent distilled water rinsing; acidification with rinsing; and rinsing with no acidification. Continuous-flow isotope ratio mass spectrometry was used to compare the mean carbon and nitrogen stable isotope ratios of treated and untreated material. RESULTS: Acidification (both with and without subsequent distilled water rinsing) resulted in reductions in mean δ(13)C values (1.939 and 3.146‰, respectively), while rinsing without prior acidification led to a smaller (but still significant) increase. Nitrogen isotope ratios were not affected by acidification, but subsequent rinsing with distilled water caused a decrease of approximately 1‰. CONCLUSIONS: Acidification of samples is clearly necessary in the presence of carbonates to obtain useful carbon isotope ratio data. However, post-acidification rinsing can result in further (potentially undesirable) changes to both carbon and nitrogen isotope ratios. Ideally, rinsing should be avoided, but the impacts are small enough to be of little concern in many studies. Rinsing (or not) should be considered carefully on the basis of the aims of a study.

Two brominated cyclic dipeptides released by the coldwater marine sponge Geodia barretti act in synergy as chemical defense.

The current work shows that two structurally similar cyclodipeptides, barettin (1) and 8,9-dihydrobarettin (2), produced by the coldwater marine sponge Geodia barretti Bowerbank act in synergy to deter larvae of surface settlers and may also be involved in defense against grazers. Previously, 1 and 2 were demonstrated to bind specifically to serotonergic 5-HT receptors. It may be suggested that chemical defense in G. barretti involves a synergistic action where one of the molecular targets is a 5-HT receptor. A mixture of 1 and 2 lowered the EC(50) of larval settlement as compared to the calculated theoretical additive effect of the two compounds. Moreover, an in situ sampling at 120 m depth using a remotely operated vehicle revealed that the sponge releases these two compounds to the ambient water. Thus, it is suggested that the synergistic action of 1 and 2 may benefit the sponge by reducing the expenditure of continuous production and release of its chemical defense substances. Furthermore, a synergistic action between structurally closely related compounds produced by the same bioenzymatic machinery ought to be the most energy effective for the organism and, thus, is more common than synergy between structurally indistinct compounds.

Effects of herbicides and freshwater discharge on water chemistry, toxicity and benthos in a Uruguayan sandy beach.

Environmental water chemistry analysis and microcosm toxicity bioassays (MTB) were performed to assess lethality of herbicides on the mole crab Emerita brasiliensis in a sandy beach affected by a freshwater discharge (Andreoni canal) from rice crops. A 5-yr macrocosm field sampling (MFS) was conducted to evaluate freshwater effects on population abundance. Propanil was only detected at the inner portion of the Andreoni canal (IAC), whereas quinclorac and clomazone were found at the IAC and at the canal mouth (CM). A major propanil metabolite was detected at the CM. Herbicides were undetectable at 13km from the CM. MTB showed an increased susceptibility to propanil with decreasing crab sizes. The MFS showed a drastic decrease in abundance towards the freshwater discharge, concurrently with decreasing salinities. The triad approach that included water analyses, toxicological experiments and long-term field sampling allowed rejecting relationships between herbicide exposure and mole crab lethal effects.

Induction of ovarian growth in Aegla platensis (Crustacea, Aeglidae) by means of neuroregulators incorporated to food

The freshwater crab Aegla platensis was used as a model to induce ovarian growth by adding different neuroregulators to a pellet food formulation. Added compounds were the dopaminergic inhibitor spiperone or the enkephalinergic inhibitor naloxone, both of them at a dose of 10-8 mol/animal. Animals were fed on the enriched pellets twice a week. After 7 wk, the gonadosomatic index (GI) was calculated as (gonad fresh weight / body fresh weight) x 100. GI significantly increased only for those females fed on spiperone pellets, compared to a control group receiving pellets with no compound added. During the assayed period, spiperone would be reverting the arrest exerted by dopamine on the neuroendocrine stimulation of ovarian growth. On the other hand, for both spiperone and naloxone a higher GI was correlated to a higher lipid content of both gonads and/or hepatopancreas, suggesting an increased energetic demand in accordance with an active investment in reproduction. Rev. Biol. Trop. 56 (3): 1201-1207. Epub 2008 September 30.

Se utilizó al anomuro de agua dulce Aegla platensis como modelo para inducir el crecimiento ovárico mediante el agregado de diferentes neuroreguladores a una formulación de alimento pelleteado. Los compuestos agregados fueron el inhibidor dopaminergico spiperona ó el inhibidor encefalinérgico naloxone, ambos a una dosis de 10-8 moles/animal. Los animales fueron alimentados dos veces a la semana con pellets enriquecidos con alguno de los neuroreguladores. Luego de 7 semanas, se calculó el índice gonadomático (IG) como (peso gonadal fresco/ peso corporal fresco) x 100. El IG mostró un incremento significativamente sólo en aquellas hembras alimentadas con pellets enriquecidos con spiperona, en comparación con un grupo control que recibió pellets sin agregado alguno. Durante el período ensayado, la spiperona estaría revirtiendo el arresto ejercido por la dopamina sobre la estimulación neuroendocrina del crecimiento ovárico. Por otro lado, para ambos grupos experimentales (spiperona y naloxone), un mayor valor de IG estuvo correlacionado a un mayor incremento del contenido de lípidos tanto en gonadas como en hepatopáncreas, sugiriendo una demanda energética incrementada en relación con una activa inversión en reproducción.

Induction of ovarian growth in Aegla platensis (Crustacea, Aeglidae) by means of neuroregulators incorporated to food.

The freshwater crab Aegla platensis was used as a model to induce ovarian growth by adding different neuroregulators to a pellet food formulation. Added compounds were the dopaminergic inhibitor spiperone or the enkephalinergic inhibitor naloxone, both of them at a dose of 10(-8) mol/animal. Animals were fed on the enriched pellets twice a week. After 7 wk, the gonadosomatic index (GI) was calculated as (gonad fresh weight/body fresh weight) x 100. GI significantly increased only for those females fed on spiperone pellets, compared to a control group receiving pellets with no compound added. During the assayed period, spiperone would be reverting the arrest exerted by dopamine on the neuroendocrine stimulation of ovarian growth. On the other hand, for both spiperone and naloxone a higher GI was correlated to a higher lipid content of both gonads and/or hepatopancreas, suggesting an increased energetic demand in accordance with an active investment in reproduction.

Toxicological response and sensitivity of estuarine macro-invertebrates exposed to industrial effluents.

The evaluation of toxicological response and sensitivity of some tropical estuarine benthic macro-intertebrates, Clibanarius africanus (Aurivillus) and Tympanotonus fuscatus var radula L. under acute exposure to effluents from brewery, paint and textile industrial establishments in Lagos state, Nigeria was carried out in static laboratory bioassays. On the basis of computed toxicity factors (96h LC50 ratios), the brewery effluent was found to be 6.90 times more toxic than the other test media against the exposed animal. The toxicity sensitivity ranking order for the test animals revealed that C. africanus was more susceptible to the toxicants than T. fuscatus. The relevance of the results in toxicity evaluation and setting realistic environmental standards for aquatic ecosystems in Nigeria is discussed.

Differential tolerance of body fluid dilution in two species of tropical hermit crabs: not due to osmotic/ionic regulation.

The tropical intertidal hermit crabs Clibanarius taeniatus and Clibanarius virescens were examined for differences in survival and physiological responses in low salinity. We found that C. taeniatus survived better in dilute seawater than C. virescens and that these species did not differ in their abilities to regulate haemolymph osmolarity, ionic concentration of the haemolymph or body fluid volume. We also found no difference in oxygen consumption between the species when acutely exposed to a range of temperature and salinity combinations. It is concluded that the greater survival in dilute seawater by C. taeniatus compared to C. virescens is due to a greater tolerance of dilution of body fluids by C. taeniatus. Differences in tolerance to dilute seawater may influence the habitat preferences of these species within the same geographical area.