Insights into the Activation of a Crustacean G Protein-Coupled Receptor: Evaluation of the Red Pigment-Concentrating Hormone Receptor of the Water Flea Daphnia pulex (Dappu-RPCH R).

The validation of a previously developed model of the interaction between the red pigment-concentrating hormone of Daphnia pulex and its cognate receptor (Jackson et al., IJBM 106, 969-978, 2018) was undertaken. Single amino acid replacements, noticeably an Ala scan, of the ligand, Dappu-RPCH, were docked to the receptor, and the binding energies calculated and compared to the one with Dappu-RPCH. As a second step, the same molecules were docked using molecular dynamics (MD) in a 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) membrane. Changes in binding energy were compared to previous results on in vitro receptor activation (Marco et al., Sci. Rep. 7, 6851, 2017). Residue scanning and MD simulations both gave comparable results for binding energy. For most mutants, there was a good inverse correlation between in vitro activity and binding. There were, however, exceptions; for example: [Ala4]Dappu-RPCH bound as tightly as the cognate ligand but had little activity. This seeming discrepancy was explained when the MD data were analyzed in detail, showing that, although [Ala4]Dappu-RPCH had multiple interactions with the receptor accounting for the high binding energy, the interacting residues of the receptor were quite different to those of Dappu-RPCH. The MD calculations show clearly that the strong binding affinity of the ligand to the receptor is not sufficient for activation. Interaction of the binding of the ligand to two residues of the receptor, Ser 155 and Gln 237, is also essential. A comparison of our computational results with the experimental results of Marco et al. and comparison with the extensive data on GnRH supports the validity of our Dappu-RPCH R model.

Hexavalent chromium removal by magnetic particle-loaded micro-sized chitinous egg shells isolated from ephippia of water flea.

Modified chitin and magnetic particles are two important materials widely used in heavy metal removal studies. Loading of magnetic particles into conventional adsorbents has emerged as a recent convenient way to improve the properties of adsorptive materials. Compared to its deacetylated form chitosan, chitin has very limited use in removal of contaminants because of its insolubility in aqueous environments. This study reports an easy way to produce micro-sized chitin and gives details on loading of magnetic particles into micro-sized chitin to enhance the interaction of chitin with heavy metal contaminant; hexavalent chromium Cr(VI). Removal of Cr(VI) ions from the aquatic environments is of high importance considering its detrimental effects on human health. Chitin microcages that had been isolated from the ephippial eggs of Daphnia longispina (crustacean, water flea) were incorporated with magnetic particles. Chitin microcages and magnetic particles-loaded chitin microcages were tested in removal of Cr(VI) under different solution and operational parameters; adsorbent dosage, contact time, Cr(VI) ion concentration, pH and temperature. Magnetic particles-loaded chitin microcages yielded higher Cr(VI) removal performance under all the specified conditions; chitin microcages: 0.77 mmol/ g and particles-loaded chitin microcages: 1.29 mmol/ g.

Acute and chronic toxicity of diuron and carbofuran to the neotropical cladoceran Ceriodaphnia silvestrii.

In order to contribute to the increase of the body of knowledge on the sensitivity of tropical indigenous species to pesticides, acute and chronic toxicity tests were conducted with the neotropical cladoceran Ceriodaphnia silvestrii. Tests were carried out with the active ingredients diuron and carbofuran and one of their commercial formulations, the Diuron Nortox® 500 SC and the Furadan® 350 SC, respectively. For carbofuran, the active ingredient was more toxic than the commercial product, whereas for diuron, the commercial product appeared more toxic. In addition, hormetic effects on fertility were recorded for intermediate diuron concentrations. Acute and chronic toxicity data indicated that C. silvestrii was among the most sensitive invertebrate species for both test compounds. Based on concentrations measured in Brazilian water bodies, these compounds represent ecological risks for causing direct and indirect toxic effects on C. silvestrii and other aquatic organisms. Our results support previous claims on the advantages of using native species to better tune ecological risk assessment of chemicals in tropical ecosystems.

The derivation of effects threshold concentrations of lead for European freshwater ecosystems.

The main objective of the present study was to derive ecologically relevant effect threshold concentrations of (dissolved) Pb for selected European Union (EU) freshwater rivers, using the 2008 EU Voluntary Risk Assessment Report as a starting point and more advanced methodologies than those used in the Voluntary Risk Assessment Report. This included 1) implementing more robust quality criteria for selecting chronic toxicity data; 2) the conversion of total to dissolved Pb concentrations using a combination of an empirical equation relating inorganic Pb solubility and geochemical speciation modeling to account for effects of dissolved organic matter; 3) the use of bioavailability models for chronic toxicity for species belonging to 3 different trophic levels; and 4) the use of robust methods for large data set handling (such as species sensitivity distribution [SSD] analysis). The authors used published bioavailability models for an algal species (Pseudokirchneriella subcapitata) and a daphnid (Ceriodaphnia dubia) and developed a new model for the fathead minnow (Pimephales promelas). The research has shown that these models are also useful for, and reasonably accurate in, predicting chronic toxicity to other species, including a snail, a rotifer, midge larvae, and an aquatic plant (read-across). A comprehensive chronic toxicity data set for Pb was compiled, comprising 159 individual high-quality toxicity data for 25 different species. By applying the total dissolved conversion and the bioavailability models, normalized toxicity values were obtained, which were then entered into a SSD analysis. Based on the parametric best-fitting SSDs, the authors calculated that ecological threshold concentrations of Pb protecting 95% of freshwater species for 7 selected European freshwater scenarios were between 6.3 µg dissolved Pb/L and 31.1 µg dissolved Pb/L.

Rare earth element distributions and fractionation in plankton from the northwestern Mediterranean Sea.

Rare earth element (REE) concentrations were measured for the first time in plankton from the northwestern Mediterranean Sea. The REE concentrations in phytoplankton (60-200 µm) were 5-15 times higher than those in four size fractions of zooplankton: 200-500 µm, 500-1000 µm, 1000-2000 µm and >2000 µm. The concentrations within these zooplankton fractions exhibited the same ranges with some variation attributed to differences in zooplankton taxonomy. The REE concentrations in plankton were poorly related to the reported REE concentrations of seawater, but they correlated well with the calculated REE(3+), concentrations especially with regard to middle REE (MREEs) and heavy REEs (HREEs). Plankton and seawater revealed different PAAS-normalised REE distributions, with the greatest differences observed in the light REEs. Interestingly, a comparison of PAAS-normalized sediment particles from the study of Fowler et al. (1992) showed concentrations of the same order of magnitude and a similar REE distribution without MREE enrichment. Based on this comparison, we propose a conceptual model that emphasizes the importance of biological scavenging of REEs (especially LREEs) in surface waters.

Trophic transfer potential of aluminium oxide nanoparticles using representative primary producer (Chlorella ellipsoides) and a primary consumer (Ceriodaphnia dubia).

The transfer of nanoparticles through the food chain can lead to bioaccumulation and biomagnification resulting in a long term negative impact on the ecosystem functions. The primary objective of this study was evaluation of aluminium oxide nanoparticles transfer from primary producers to primary consumers. A simple set up consisting of a primary producer (Chlorella ellipsoides) and a primary consumer (Ceriodaphnia dubia) was used. Here, C. ellipsoides were exposed to the varying concentrations of the nanoparticles ranging from 20 to 120µg/mL (196 to 1176µM) for 48h and the infested algal cells were used as the feed to C. dubia. The bioaccumulation of the nanoparticles into the daphnids was noted and the biomagnification factors were computed. The exposure was noted to cause subtle alterations in the feeding behaviour of the daphnids. This might have long term consequences in the energy flow through the food chain. The reproductive behaviour of the daphnids remained unaffected upon exposure to nanoparticle infested algal feed. Distinct observations at ultra-structural scale using transmission electron microscopy provided visual evidences for the disrupted feeding behaviour upon exposure to nanoparticle treated algae. Internalization of nanoparticle like inclusion bodies in the intracellular space of algae was also detected. The findings were further substantiated by a detailed analysis of hydrodynamic stability, bioavailability and dissolution of ions from the nanoparticles over the exposure period. Altogether, the study brings out the first of its kind of observation of trophic transfer potential/behaviour of aluminium oxide nanoparticles and its probable impacts on the energy flow in the fresh water aquatic ecosystem.

Bioaccumulation of Fe2O3(magnetic) nanoparticles in Ceriodaphnia dubia.

While nano-Fe(2)O(3)(magnetic) is generally considered non-toxic, it could serve as a carrier of other toxic chemicals such as As(V) and enhance their toxicity. The bioaccumulation of nano-Fe(2)O(3)(m) with different exposure times, NP concentrations, and pH conditions was investigated using Ceriodaphnia dubia (C. dubia) as the model organism. Under natural pH conditions, C. dubia significantly accumulated nano-Fe(2)O(3)(m) in the gut, with the maximum accumulation being achieved after 6 h of exposure. The concentration of nano-Fe(2)O(3) also impacted its accumulation, with the maximum uptake occurring at 20 mg/L or more. In addition, the highest bioaccumulation occurred in a pH range of 7-8 where the highest feeding rate was reported, confirming that the ingestion of NPs is the main route of nano-Fe(2)O(3)(m) bioaccumulation. In a clean environment without NPs, depuration of nano-Fe(2)O(3)(m) occurred, and food addition accelerated the depuration process.

Large scale laboratory cultures of Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera) / Cultivo em large escala de Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera) em laboratório

Large-scale lab culture of Ankistrodesmus gracilis and Diaphanososma birgei were evaluated by studying the biology and biochemical composition of the species and production costs. Ankistrodesmus gracilis presented exponential growth until the 6th day, with approximately 144 x 10(4) cells.mL-1, followed by a sharp decrease to 90 x 10(4) cells.mL-1 (8th day). Algae cells tended to increase again from the 11th day and reached a maximum of 135 x 10(4) cells.mL-1 on the 17th day. D. birgei culture showed exponential growth until the 9th day with 140 x 10² individuals.L-1, and increased again as from the 12th day. Algae A. gracilis and zooplankton D. birgei contain 47 to 70 percent dry weight protein and over 5 percent dry weight carbohydrates. The most expensive items in the context of variable costs were labor and electricity. Data suggested that temperature, nutrients, light availability and culture management were determining factors on productivity. Results indicate that NPK (20-5-20) may be used directly as a good alternative for mass cultivation when low costs are taken into account, promoting adequate growth and nutritional value for cultured A. gracilis and D. birgei.

O objetivo deste trabalho foi avaliar o cultivo em larga escala de Ankistrodesmus gracilis e Diaphanososma birgei em laboratório através do estudo da biologia das espécies, composição bioquímica e custo operacional de produção. A. gracilis apresentou um crescimento exponencial até o sexto dia, ao redor de 144 x 10(4) células mL-1. Logo em seguida, sofreu um brusco decréscimo apresentando 90 x 10(4) células mL-1 (oitavo dia). A partir do décimo primeiro dia, as células algais tenderam a crescer novamente, apresentando um máximo de 135 x 10(4) células mL-1 no 17º dia. No cultivo de D. birgei, foi observado o primeiro pico de crescimento no nono dia com 140 x 10² indivíduos L-1, aumentando novamente a partir do décimo segundo dia. A alga clorofícea A. gracilis e o zooplâncton D. birgei possuem aproximadamente 50 e 70 por cento de proteína (PS), respectivamente, com teor de carboidrato acima de 5 por cento. A eletricidade e mão de obra foram os itens mais dispendiosos e, de acordo com os dados obtidos, a temperatura, nutrientes, disponibilidade de luz e manejo do cultivo, foram fatores determinantes sobre a produtividade. Os resultados indicam que o meio NPK (20-5-20) pode ser utilizado diretamente como uma alternativa de cultivo em larga escala, considerando o baixo custo de produção, promovendo adequado crescimento e valor nutricional para A. gracilis e D. birgei.

Large scale laboratory cultures of Ankistrodesmus gracilis (Reisch) Korsikov (Chlorophyta) and Diaphanosoma biergei Korinek, 1981 (Cladocera).

Large-scale lab culture of Ankistrodesmus gracilis and Diaphanososma birgei were evaluated by studying the biology and biochemical composition of the species and production costs. Ankistrodesmus gracilis presented exponential growth until the 6th day, with approximately 144 x 10(4) cells x mL(-1), followed by a sharp decrease to 90 x 10(4) cells x mL(-1) (8th day). Algae cells tended to increase again from the 11th day and reached a maximum of 135 x 10(4) cells x mL(-1) on the 17th day. D. birgei culture showed exponential growth until the 9th day with 140 x 10(2) individuals x L(-1), and increased again as from the 12th day. Algae A. gracilis and zooplankton D. birgei contain 47 to 70% dry weight protein and over 5% dry weight carbohydrates. The most expensive items in the context of variable costs were labor and electricity. Data suggested that temperature, nutrients, light availability and culture management were determining factors on productivity. Results indicate that NPK (20-5-20) may be used directly as a good alternative for mass cultivation when low costs are taken into account, promoting adequate growth and nutritional value for cultured A. gracilis and D. birgei.

Trophic transfer of paralytic shellfish toxins from the cladoceran (Moina mongolica) to larvae of the fish (Sciaenops ocellatus).

The dynamic transmission and transformation of paralytic shellfish toxins (PSTs) from the toxic dinoflagellate Alexandrium tamarense to the cladoceran Moina mongolica and subsequently to the larvae of the fish Sciaenops ocellatus were investigated under laboratory conditions. The results showed that PSTs could be transferred to S. ocellatus when they preyed on PST-containing M. mongolica. During the experimental period, A. tamarense, M. mongolica and the digestive glands of the fish larvae contained C(1/2) toxins, and the viscera of S. ocellatus contained neoSTX. The proportion of beta toxin (C2) in C(1+2) toxins increased when PSTs were transferred from A. tamarense to M. mongolica, but in the subsequent transfer from M. mongolica to S. ocellatus the proportion of alpha toxin (C1) increased. During depuration, the contents of C1 and C2 toxins in fish larvae decreased with the duration of depuration, but neoSTX remained relatively constant. The present results indicated that, using a cladoceran as the vector, PSTs can be transferred from toxic algae to a high trophic level fish and metabolized in the fish. Future work should address the metabolic characteristics of PSTs in cladocerans and the end result when they are transferred to fishes.

Carotenoprotein complexes in entomostracan crustaceans (Streptocephalus dichotomus and Moina micrura).

The carotenoprotein complexes of a freshwater fairy shrimp (Streptocephalus dichotomus) and a daphnid (Moina micrura) were characterised and compared. Based on thin layer chromatography and mass spectral analysis, a variety of cartenoprotein complexes such as astaxanthin, canthaxanthin, antheraxanthin, lutein, beta-cryptoxanthin and violaxanthin were found. Both crustaceans had astaxanthin and canthaxanthin as predominant prosthetic groups. Amino acid analysis of the complexes further revealed high levels of asparagine, glutamine and glycine in both species. Our study highlights the presence of naturally available carotenoid species in both crustaceans and their possible inter-conversion in anostracans.

Role of food partitioning in structuring the zooplankton community in mountain lakes.

Trophic-niche differentiation is often cited as a main factor in structuring zooplankton assemblages, although field evidence for this is rarely presented. The study was based on a survey of 29 Pyrenean lakes with altitudes ranging between 1,875 and 2,990 m carried out during July and August 2000. Because of the oligotrophic nature of these lakes, we aimed to confirm that food partitioning is a major factor in shaping zooplankton assemblages. We analysed the amino acid composition of six cladocera and seven copepod species. A discriminant analysis showed that each species could be distinguished according to its amino acid composition. A negative relationship between amino acid differentiation and co-occurrence among the cladocera and cyclopoid copepod was observed. In contrast, calanoids did not show any relationship and were characterised by a high amino acid differentiation between species. As the differences in the amino acid composition among zooplankton species indicate distinct food sources, the relationship found indicates that trophic-niche differentiation plays a key role in determining the assemblage of these zooplankton communities. Therefore exploitative competition, either at present or in the past by driving co-evolutionary histories, has been a significant factor in structuring the cladocera and cyclopoid communities in these oligotrophic lakes.

Population growth and development of two species of Cladocera, Moina micrura and Diaphanosoma birgei, in laboratory

The objective of the present work was to investigate the influence of four diets on population growth, development, total length, dry weight, and nutritional value of two zooplanktonic species, Moina micrura and Diaphanosoma birgei. The four dietary treatments were: algae alone (A); algae + vitamins (AV); algae + ration (AR); and algae + ration + vitamins (ARV). Growth rate peak for both species occurred faster with AV treatment. In general, AV treatment for M. micrura showed better results for intrinsic rate, fecundity, and embryonic and post-embryonic development. On the other hand, longevity and total spawning number were better with AR treatment (p < 0.05). Vitamin and ration treatments produced the best results in D. birgei species (p < 0.05). The highest percentage of protein and lipids for both cladocerans was verified for ration treatments. Carbohydrate was higher for the treatment containing algae alone (p < 0.05). Generally, diets containing ration and vitamin showed better results in cladocerans development, with water quality adequate for culture systems. Ration and vitamin diets may also be used in high-density cultures in the laboratory

Population growth and development of two species of Cladocera, Moina micrura and Diaphanosoma birgei, in laboratory.

The objective of the present work was to investigate the influence of four diets on population growth, development, total length, dry weight, and nutritional value of two zooplanktonic species, Moina micrura and Diaphanosoma birgei. The four dietary treatments were: algae alone (A); algae + vitamins (AV); algae + ration (AR); and algae + ration + vitamins (ARV). Growth rate peak for both species occurred faster with AV treatment. In general, AV treatment for M. micrura showed better results for intrinsic rate, fecundity, and embryonic and post-embryonic development. On the other hand, longevity and total spawning number were better with AR treatment (p < 0.05). Vitamin and ration treatments produced the best results in D. birgei species (p < 0.05). The highest percentage of protein and lipids for both cladocerans was verified for ration treatments. Carbohydrate was higher for the treatment containing algae alone (p < 0.05). Generally, diets containing ration and vitamin showed better results in cladocerans development, with water quality adequate for culture systems. Ration and vitamin diets may also be used in high-density cultures in the laboratory.