RESUMO
Ingestion of microplastics can lead to deleterious consequences for organisms, as documented by numerous laboratory studies. The current knowledge is based on a multitude of effect studies, conducted with conventional fossil-based and non-degradable plastics. However, there is a lack of information about the acceptance and the effects of novel bio-based and biodegradable plastics. Biodegradable plastics are considered an alternative to conventional plastics and are showing rapidly growing production rates. Biodegradable plastics can disperse into the environment in the same way as conventional plastics do, becoming available to marine organisms. This study aims to provide new insights into the uptake and effects of biodegradable microplastics on marine invertebrates. Rockpool shrimp, Palaemon elegans, were fed with algal flakes coated with polylactic acid (PLA), polyhydroxybutyrate-co-valerate (PHBV) and conventional low-density polyethylene (LDPE) microparticles. Live observations showed that all of the different types of microplastics were ingested. After dissection of the shrimp, less LDPE particles were found in the stomachs than PLA and PHBV particles. This indicates a longer retention time of biodegradable microplastics compared to conventional microplastics. Presumably, less LDPE particles were ingested or evacuated from the stomach, probably by regurgitation. The ingestion of microparticles of all types of plastics induced enzymatic activity of short-chain carboxylesterases in the midgut glands of the shrimp. However, only PLA induced enzymatic activity of medium-chain carboxylesterases. Palaemon elegans showed no oxidative stress response after ingestion of microparticles, irrespective of polymer type. From our results we conclude that biodegradable plastics might have different effects than conventional plastics. The longer retention times of biodegradable plastics might enhance exposure to leaching additives and other harmful substances. Our study provides new insights into how biodegradable plastics might affect aquatic fauna and indicate that the use of biodegradable plastics needs to be reconsidered to some extent.
Assuntos
Plásticos Biodegradáveis , Palaemonidae , Animais , Microplásticos/toxicidade , Plásticos/toxicidade , Polietileno , Poliésteres/toxicidade , Organismos Aquáticos , Hidrolases de Éster CarboxílicoRESUMO
Lipids play essential roles in cell-structuring, cell-signaling, and as efficient metabolic energy stores. Lipid storage capacities determine life history traits of organisms and, thus, their ecological function. Among storage lipids, triacylglycerols (TAGs) are widespread in marine invertebrates. However, abilities to accumulate TAGs can vary even between closely related species, such as the caridean shrimps Crangon crangon and Pandalus montagui. The first species shows low TAG levels throughout the year in the main storage organ, the midgut gland, while the latter accumulates high TAG-levels, peaking in summer. TAGs synthesis is facilitated by the terminal step of the Kennedy-pathway, where the enzyme diacylglycerol-acyltransferase (DGAT) catalyzes the esterification of diacylglycerols with activated fatty acids. We investigated DGAT activity in the midgut gland using a fluorescent enzyme assay. Sequence information was extracted from whole transcriptome shotgun assembly data, that is publicly available on NCBI, and catalytic properties were deduced from molecular structure analysis. C. crangon showed significantly lower TAG synthesis rates than P. montagui, which explains the native TAG levels. Transcriptome data yielded several isoforms of DGAT enzymes in both species. C. crangon DGAT showed point mutations, which are capable of obstructing the catalytic capacity. The consequences are limited starvation resistance and, thus, presumably restricting C. crangon to a habitat with year-round sufficient food. In contrast, higher TAG synthesis rates presumably enable P. montagui to extend into northern subarctic habitats with limited food availability in winter. Moreover, the limited TAG synthesis and accumulation in the midgut gland may force C. crangon to direct energy into the ovaries, which results in multiple spawnings.
Assuntos
Crangonidae , Características de História de Vida , Pandalidae , Animais , Diacilglicerol O-Aciltransferase/genética , Diacilglicerol O-Aciltransferase/metabolismo , Pandalidae/metabolismo , Crangonidae/metabolismo , Ácidos Graxos/metabolismo , Triglicerídeos/metabolismoRESUMO
Many invertebrate species inhabit coastal areas where loads of plastic debris and microplastics are high. In the current case study, we exemplarily illustrate the principal processes taking place in the Atlantic ditch shrimp, Palaemon varians, upon ingestion of microplastics. In the laboratory, shrimp readily ingested fluorescent polystyrene microbeads of 0.1-9.9 µm, which could be tracked within the widely translucent body. Ingested food items as well as micro-particles cumulate in the stomach where they are macerated and mixed with digestive enzymes. Inside the stomach, ingested particles are segregated by size by a complex fine-meshed filter system. Liquids and some of the smallest particles (0.1 µm) pass the filter and enter the midgut gland where resorption of nutrients as well as synthesis and release of digestive enzymes take place. Large particles and most of the small particles are egested with the feces through the hindgut. Small particles, which enter the midgut gland, may interact with the epithelial cells and induce oxidative stress, as indicated by elevated activities of superoxide dismutase and cellular markers of reactive oxygen species. The shrimp indiscriminately ingest microparticles but possess efficient mechanisms to protect their organs from overloading with microplastics and other indigestible particles. These include an efficient sorting mechanism within the stomach and the protection of the midgut gland by the pyloric filter. Formation of detrimental radical oxygen species is counteracted by the induction of enzymatic antioxidants.
RESUMO
Marine invertebrates inhabiting estuaries and coastal areas are exposed to natural suspended particulate matter (SPM) like clay or diatom shells but also to anthropogenic particles like microplastics. SPM concentrations may reach 1 g per liter and more, comprising hundreds of millions of items in the size range of less than 100 µm. Suspension feeders and deposit feeders involuntarily ingest these particles along with their food. We investigated whether natural and anthropogenic microparticles at concentrations of 20 mg L-1, which correspond to natural environmental SPM concentrations in coastal marine waters, are ingested by the brown shrimp Crangon crangon and whether these particles induce an oxidative stress response in digestive gland tissue. Shrimp were exposed to clay, silica, TiO2, polyvinyl chloride (PVC), or polylactide microplastics (PLA) for 6, 12, 24, and 48 h, respectively. The activities of the anti-oxidative enzymes superoxide dismutase (SOD), glutathione peroxidase (GPx), and glutathione reductase (GR) were measured. All five particle types were ingested by the shrimp along with food. The presence of the particles in the shrimp stomach was verified by scanning electron microscopy. The activities of the anti-oxidative enzymes did not vary between animals exposed to different types of microparticles and control animals that did not receive particles. The temporal activity differed between the three enzymes. The lack of a specific biochemical response may reflect an adaptation of C. crangon to life in an environment where frequent ingestion of non-digestible microparticles is unavoidable and continuous maintenance of inducible biochemical defense would be energetically costly. Habitat characteristics as well as natural feeding habits may be important factors to consider in the interpretation of hazard and species-specific risk assessment.
Assuntos
Crangonidae , Penaeidae , Animais , Microplásticos , PlásticosRESUMO
BACKGROUND: Trophic interactions are key processes, which determine the ecological function and performance of organisms. Many decapod crustaceans feed on plant material as a source for essential nutrients, e.g. polyunsaturated fatty acids. Strictly herbivorous feeding appears only occasionally in marine decapods but is common in land crabs. To verify food preferences and to establish trophic markers, we studied the lipid and fatty acid composition of the midgut glands of two marine crab species (Grapsus albolineatus and Percnon affine), one semi-terrestrial species (Orisarma intermedium, formerly Sesarmops intermedius), and one terrestrial species (Geothelphusa albogilva) from Taiwan. RESULTS: All species showed a wide span of total lipid levels ranging from 4 to 42% of the dry mass (%DM) in the marine P. affine and from 3 to 25%DM in the terrestrial G. albogilva. Triacylglycerols (TAG) were the major storage lipid compound. The fatty acids 16:0, 18:1(n-9), and 20:4(n-6) prevailed in all species. Essential fatty acids such as 20:4(n-6) originated from the diet. Terrestrial species also showed relatively high amounts of 18:2(n-6), which is a trophic marker for vascular plants. The fatty acid compositions of the four species allow to clearly distinguish between marine and terrestrial herbivorous feeding due to significantly different amounts of 16:0, 18:1(n-9), and 18:2(n-6). CONCLUSIONS: Based on the fatty acid composition, marine/terrestrial herbivory indices were defined and compared with regard to their resolution and differentiating capacity. These indices can help to reveal trophic preferences of unexplored species, particularly in habitats of border regions like mangrove intertidal flats and estuaries.
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Bio-based polymers have been suggested as one possible opportunity to counteract the progressive accumulation of microplastics in the environments. The gradual substitution of conventional plastics by bio-based polymers bears a variety of novel materials. The application of bioplastics is determined by their stability and bio-degradability, respectively. With the increasing implementation of bio-based plastics, there is also a demand for rapid and non-elaborate methods to determine their bio-degradability. Here, we propose an improved pH Stat titration assay optimized for bio-based polymers under environmental conditions and controlled temperature. Exemplarily, suspensions of poly(lactic acid) (PLA) and poly(butylene succinate) (PBS) microparticles were incubated with proteolytic and lipolytic enzymes. The rate of hydrolysis, as determined by counter-titration with a diluted base (NaOH), was recorded for two hours. PLA was hydrolyzed by proteolytic enzymes but not by lipase. PBS, in contrast, showed higher hydrolysis rates with lipase than with proteases. The thermal profile of PLA hydrolysis by protease showed an exponential increase from 4 to 30 °C with a temperature quotient Q10 of 5.6. The activation energy was 110 kJ·mol-1. pH-Stat titration proved to be a rapid, sensitive, and reliable procedure supplementing established methods of determining the bio-degradability of polymers under environmental conditions.
RESUMO
Microplastic pollution is an emerging threat to marine biota. Uptake of microplastics can impair nutrition and affect the performance of organisms. However, the vulnerability to microplastics seems to vary between species for yet widely unexplored reasons. We investigated the stomach content of the brown shrimp, Crangon crangon, from the southern North Sea and performed feeding experiments and anatomical studies of the digestive organs to comprehend the distribution of fluorescent microparticles within the shrimp. Shrimp collected in their natural environment contained between 51 and more than 3,000 sand grains and fragments of bivalve shells in their stomachs. Sand grains may have been ingested to exploit the associated biofilm or to support maceration of food. Bivalve shell fragments were particularly abundant in summer when shrimp fed on freshly settled mussels. Shrimps' stomach can be cleaned from ingested particles by regurgitation. In an experimental approach, we administered fluorescent microbeads of 0.1, 2.1, and 9.9 µm diameter. Only the smallest particles (0.1 µm) entered the midgut gland, which is the principal site of nutrient resorption in crustaceans. A fine-meshed chitinous filter system in the stomach of the shrimp prevents the passage of particles larger than about 1 µm. C. crangon appears well adapted to handle natural microscopic particles. This trait might also be advantageous in coping with microplastic pollution.
Assuntos
Crangonidae/efeitos dos fármacos , Crangonidae/fisiologia , Microplásticos , Poluentes Químicos da Água/toxicidade , Animais , Crangonidae/anatomia & histologia , Conteúdo Gastrointestinal/química , Análise Espectral Raman , Poluentes Químicos da Água/químicaRESUMO
The brown shrimp, Crangon crangon, is well adapted to the variable environmental conditions in the southern North Sea. It is very abundant, has high reproduction rates, and holds a key position in coastal ecosystems. This species has very low lipid deposits in the midgut gland, suggesting that the main function of the midgut gland is metabolic turnover rather than energy storage. Based on seasonal gene expression studies and established transcriptome data, we investigated key components of lipid metabolic pathways. Gene expression of triacylglycerol lipase, phospholipase, and fatty acid desaturase were analyzed and compared with that of other digestive enzymes involved in lipid, carbohydrate, and protein catabolism. Our results suggest that gene expression of digestive enzymes involved in lipid metabolism is modulated by the lipid content in the midgut gland and is related to food availability. Brown shrimp seem to be capable of using cellular phospholipids during periods of food paucity but high energetic (lipid) requirements. Two of three isoforms of fatty acid binding proteins (FABPs) from the midgut gland involved in fatty acid transport showed specific mutations of the binding site. We hypothesize that the mutations in FABPs and deficiencies in anabolic pathways limit lipid storage capacities in the midgut gland of C. crangon. In turn, food utilization, including lipid catabolism, has to be efficient to fulfill the energetic requirements of brown shrimp.
Assuntos
Proteínas de Artrópodes/metabolismo , Crangonidae/metabolismo , Animais , Proteínas de Artrópodes/genética , Crangonidae/genética , Proteínas de Ligação a Ácido Graxo/genética , Proteínas de Ligação a Ácido Graxo/metabolismo , Trato Gastrointestinal/metabolismo , Perfilação da Expressão Gênica , Metabolismo dos Lipídeos , Estações do Ano , TranscriptomaRESUMO
Sediment samples were randomly taken in March and August 2017 at the strandlines of nine locations along the coast of Slovenia (Adriatic Sea, Mediterranean). Microparticles were isolated by density separation in saturated aqueous NaCl-solutions and analysed by infrared spectroscopy (ATR-FTIR). 11.3% of these particles were unambiguously confirmed as microplastics. Another 8.2% showed plastic characteristics but failed ATR-FTIR validation. 4.3% were naturally organic. The rest was unidentified material (76.2%). The average microplastic densities were 0.5⯱â¯0.5â¯MPâ¯kg-1 in March and 1.0⯱â¯0.8â¯MPâ¯kg-1 in August. The microplastics comprised fragments, fibres, films, and foams. The characteristics of the microplastics suggest origin from single-used plastic products and from aquaculture. Compared to other studies and sites, the microplastic pollution of the Slovenian coast appeared low. The validity of the results is discussed with respect to microplastic distribution and patchiness, sampling strategies, methodology, and scientific claims.
Assuntos
Monitoramento Ambiental , Microplásticos/análise , Poluentes Químicos da Água/análise , Aquicultura , Monitoramento Ambiental/métodos , Poluição Ambiental/análise , EslovêniaRESUMO
Microplastic fibers represent a significant share of the global marine micrcroplastic pollution, particularly in coastal areas. In controlled laboratory experiments, we offered fluorescent microplastic fibers (40-4400⯵m lengths, median 150⯵m) and spherical microplastic beads (9.9⯵m Ø) together with commercial fish food to the Atlantic ditch shrimp Palaemonetes varians. The shrimps ingested fibers and beads along with the food. Upon ingestion, the beads and the shortest fibers (up to 100⯵m) passed from the stomach into the gut and were egested within the fecal strings. The longer fibers first remained in the stomach but were regurgitated, i.e. extruded through the esophagus, within 12-14â¯h. Regurgitation is an evolutionary adaptation of particular crustacean species and other invertebrates to remove large and indigestible food particles from the stomach. Accordingly, the process of regurgitation attained a new task nowadays, i.e. the elimination of anthropogenic filamentous microplastic debris from the stomach to avoid harm. This behavioral feature may represent a selective advantage in view of the continuously increasing environmental plastic pollution.
Assuntos
Palaemonidae/fisiologia , Plásticos/análise , Poluentes Químicos da Água/análise , Animais , Ingestão de Alimentos , Monitoramento Ambiental , Poluição Ambiental , Peixes , Invertebrados , Alimentos MarinhosRESUMO
Ingestion of microplastics can impair nutrition of marine invertebrates. In a laboratory study, we tested whether microplastics affect ingestion rates and gastrointestinal enzyme activities in the marine isopod Idotea emarginata. Isopods were fed for eight days with one out of four different food formulations: natural food (the brown alga Fucus vesiculosus) or synthetic diet consisting of freeze-dried algal powder embedded in agarose, both, with or without microplastic particles (fluorescent polymethyl methacrylate, 10-100⯵m) at a concentration of 40 items per mg of food. The isopods accepted both types of food but consumed significantly more (average 3.1-fold) of the agar based synthetic food. I. emarginata responded to the reduced content of digestible organic matter in the synthetic food by a compensatory adjustment of the ingestion rates. Addition of microplastics had no effect on ingestion rates in natural food whereas the feeding rates for synthetic food varied in response to microplastics. Similarly, activity patterns of digestive enzymes, particularly those of esterases, changed significantly in the treatment with synthetic food. Isopods fed with synthetic food alone showed elevated esterase activities in the gut while those isopods fed with synthetic food and microplastics showed elevated esterase activities in the midgut gland but not in the gut. Apparently, not the exposure to microplastic alone, but the combined effects of reduced nutrient availability and microplastic ingestion caused considerable biochemical reactions in the digestive organs of the isopods.
Assuntos
Ração Animal , Sistema Digestório/enzimologia , Esterases/metabolismo , Isópodes/fisiologia , Microplásticos , Poluentes Químicos da Água/toxicidade , Animais , Organismos Aquáticos/fisiologia , Ingestão de Alimentos , Monitoramento AmbientalRESUMO
The rapid dissemination of microplastics in many habitats of the oceans has raised concerns about the consequences for marine biota and ecosystems. Many adverse effects of microplastics on marine invertebrates are consequences of ingestion. Accordingly, the identification of mechanisms that facilitate the uptake of microplastics is essential for the evaluation of possible implications for marine organisms and food webs. Gastropods produce mucus for locomotion. Gastropod pedal mucus naturally retains formerly suspended micro-organisms, such as bacteria, microalgae, and seaweed spores. The retained organisms are consumed by gastropods that forage on pedal mucus. Here, we investigated the potential of gastropod pedal mucus to retain suspended microplastic particles and make them available for ingestion by periwinkles that forage on the contaminated mucus. In laboratory experiments, mucus of the periwinkles Littorina littorea and Littorina obtusata efficiently retained microplastics. Retention of microplastics varied between mucus from conspecifics of different size but not between mucus from either species. The density of microplastics in mucus trails increased concomitantly with the experimental particle concentration but was independent of incubation time. Aging of mucus and, particularly, desiccation affected the retention of microplastics. Periwinkles ingested microplastics when foraging on the contaminated mucus. Our results reveal a functional link between biogenic accumulation of microplastics and their trophic transfer by marine benthic herbivores into marine food webs.
Assuntos
Gastrópodes/efeitos dos fármacos , Muco/metabolismo , Plásticos/análise , Alga Marinha/metabolismo , Vinca/metabolismo , Poluentes Químicos da Água/análise , Animais , Sistema Digestório/metabolismo , Cadeia Alimentar , Gastrópodes/metabolismo , Modelos Teóricos , Mar do Norte , Plásticos/metabolismo , Poluentes Químicos da Água/metabolismoRESUMO
Tolerance of organisms towards heterogeneous and variable environments is highly related to physiological flexibility. An effective strategy to enhance physiological flexibility is the expression of polymorphic enzymes. This seems to be the case in the brown shrimp Crangon crangon. It shows high reproduction rates, feeds opportunistically on endo- and epibenthic organisms, and is apparently well adapted to variable environmental conditions. Previous electrophoretic studies revealed a high level of polymorphism and no consistent phenotype of digestive enzymes between individuals. In order to understand the underlying biochemical processes, we carried out a transcriptome-based study of digestive enzymes of C. crangon. Detailed sequence analyses of triacylglycerol lipase, phospholipase A2, alpha amylase, chitinase, trypsin and cathepsin L were performed to identify putative isoforms. The number of isoforms, and thus the degree of polymorphism varied among enzymes: lipases and carbohydrases showed higher numbers of isoforms in enzymes that besides their extracellular function also have diverse intracellular functions. Furthermore, cysteine proteinases showed a lower polymorphism than serine proteinases. We suggest that the expression of enzyme isoforms improves the efficiency of C. crangon in gaining energy from different food sources.
Assuntos
Proteínas de Artrópodes/genética , Crangonidae/genética , Polimorfismo Genético , Animais , Proteínas de Artrópodes/análise , Proteínas de Artrópodes/química , Crangonidae/enzimologia , Glândulas Exócrinas/metabolismo , Trato Gastrointestinal/enzimologia , Perfilação da Expressão Gênica , Análise de Sequência de ProteínaRESUMO
Cathepsin D is an aspartic endopetidase with typical characteristics of lysosomal enzymes. Cathepsin D activity has been reported in the gastric fluid of clawed lobsters where it acts as an extracellular digestive enzyme. Here we investigate whether cathepsin D is unique in clawed lobsters or, instead, common in decapod crustaceans. Eleven species of decapods belonging to six infraorders were tested for cathepsin D activity in the midgut gland, the muscle tissue, the gills, and when technically possible, in the gastric fluid. Cathepsin D activity was present in the midgut gland of all 11 species and in the gastric fluid from the seven species from which samples could be taken. All sampled species showed higher activities in the midgut glands than in non-digestive organs and the activity was highest in the clawed lobster. Cathepsin D mRNA was obtained from tissue samples of midgut gland, muscle, and gills. Analyses of deduced amino acid sequence confirmed molecular features of lysosomal cathepsin D and revealed high similarity between the enzymes from Astacidea and Caridea on one side, and the enzymes from Penaeoidea, Anomura, and Brachyura on the other side. Our results support the presence of cathepsin D activity in the midgut glands and in the gastric fluids of several decapod species suggesting an extracellular function of this lysosomal enzyme. We discuss whether cathepsin D may derive from the lysosomal-like vacuoles of the midgut gland B-cells and is released into the gastric lumen upon secretion by these cells.
Assuntos
Proteínas de Artrópodes , Catepsina D , Decápodes , Regulação Enzimológica da Expressão Gênica/fisiologia , Animais , Proteínas de Artrópodes/biossíntese , Proteínas de Artrópodes/genética , Catepsina D/biossíntese , Catepsina D/genética , Decápodes/enzimologia , Decápodes/genética , Especificidade de Órgãos/fisiologiaRESUMO
Crustaceans are intensively farmed in aquaculture facilities where they are vulnerable to parasites, bacteria, or viruses, often severely compromising the rearing success. The ubiquitin-proteasome system (UPS) is crucial for the maintenance of cellular integrity. Analogous to higher vertebrates, the UPS of crustaceans may also play an important role in stress resistance and pathogen defense. We studied the general properties of the proteasome system in the hemocytes of the whiteleg shrimp, Penaeus vannamei, and the European brown shrimp Crangon crangon. The 20S proteasome was the predominant proteasome population in the hemocytes of both species. The specific activities of the trypsin-like (Try-like), chymotrypsin-like (Chy-like), and caspase-like (Cas-like) enzymes of the shrimp proteasome differed between species. P. vannamei exhibited a higher ratio of Try-like to Chy-like activities and Cas-like to Chy-like activities than C. crangon. Notably, the Chy-like activity of P. vannamei showed substrate or product inhibition at concentrations of more than 25 mmol L-1. The K M values ranged from 0.072 mmol L-1 for the Try-like activity of P. vannamei to 0.309 mmol L-1 for the Cas-like activity of C. crangon. Inhibition of the proteasome of P. vannamei by proteasome inhibitors was stronger than in C. crangon. The pH profiles were similar in both species. The Try-like, Chy-like, and Cas-like sites showed the highest activities between pH 7.5 and 8.5. The proteasomes of both species were sensitive against repeated freezing and thawing losing ~80-90% of activity. This study forms the basis for future investigations on the shrimp response against infectious diseases, and the role of the UPS therein.
Assuntos
Citoplasma/genética , Penaeidae/genética , Complexo de Endopeptidases do Proteassoma/genética , Estresse Fisiológico/genética , Animais , Citoplasma/química , Hemócitos/metabolismo , Ubiquitina/genéticaRESUMO
The ingestion of microplastics has been shown for a great variety of marine organisms. However, benthic marine mesoherbivores such as the common periwinkle Littorina littorea have been largely disregarded in studies about the effects of microplastics on the marine biota, probably because the pathway for microplastics to this functional group of organisms was not obvious. In laboratory experiments we showed that the seaweed Fucus vesiculosus retains suspended microplastics on its surface. The numbers of microplastics that adhered to the algae correlated with the concentrations of suspended particles in the water. In choice feeding assays L. littorea did not distinguish between algae with adherent microplastics and clean algae without microplastics, indicating that the snails do not recognize solid nonfood particles in the submillimeter size range as deleterious. In periwinkles that were feeding on contaminated algae, microplastics were found in the stomach and in the gut. However, no microplastics were found in the midgut gland, which is the principle digestive organ of gastropods. Microplastics in the fecal pellets of the periwinkles indicate that the particles do not accumulate rapidly inside the animals but are mostly released with the feces. Our results provide the first evidence that seaweeds may represent an efficient pathway for microplastics from the water to marine benthic herbivores.
Assuntos
Cadeia Alimentar , Plásticos/química , Plásticos/metabolismo , Alga Marinha/metabolismo , Caramujos/metabolismo , Animais , Biota , Sistema Digestório , Fezes , Herbivoria , Oceanos e Mares , Alga Marinha/químicaRESUMO
To identify the gene responsible for the production of a ß-1,3-glucanase (laminarinase) within crustacea, a glycosyl hydrolase family 16 (GHF16) gene was sequenced from the midgut glands of the gecarcinid land crab, Gecarcoidea natalis and the freshwater crayfish, Cherax destructor. An open reading frame of 1098 bp for G. natalis and 1095 bp for C. destructor was sequenced from cDNA. For G. natalis and C. destructor respectively, this encoded putative proteins of 365 and 364 amino acids with molecular masses of 41.4 and 41.5 kDa. mRNA for an identical GHF16 protein was also expressed in the haemolymph of C. destructor. These putative proteins contained binding and catalytic domains that are characteristic of a ß-1,3-glucanase from glycosyl hydrolase family 16. The amino acid sequences of two short 8-9 amino acid residue peptides from a previously purified ß-1,3-glucanase from G. natalis matched exactly that of the putative protein sequence. This plus the molecular masses of the putative proteins matching that of the purified proteins strongly suggests that the sequences obtained encode for a catalytically active ß-1,3-glucanase. A glycosyl hydrolase family 16 cDNA was also partially sequenced from the midgut glands of other amphibious (Mictyris platycheles and Paragrapsus laevis) and terrestrial decapod species (Coenobita rugosus, Coenobita perlatus, Coenobita brevimanus and Birgus latro) to confirm that the gene is widely expressed within this group. There are three possible hypothesised functions and thus evolutionary routes for the ß-1,3-glucanase: 1) a digestive enzyme which hydrolyses ß-1,3-glucans, 2) an enzyme which cleaves ß-1,3-glycosidic bonds within cell walls to release cell contents or 3) an immune protein which can hydrolyse the cell walls of potentially pathogenic micro-organisms.
Assuntos
Celulases/genética , Decápodes/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Celulases/química , Celulases/metabolismo , Decápodes/classificação , Decápodes/enzimologia , Hemócitos/enzimologia , Hemócitos/metabolismo , Dados de Sequência Molecular , Filogenia , Alinhamento de SequênciaRESUMO
Plastic pollution is an emerging global threat for marine wildlife. Many species of birds, reptiles, and fishes are directly impaired by plastics as they can get entangled in ropes and drown or they can ingest plastic fragments which, in turn, may clog their stomachs and guts. Microplastics of less than 1 mm can be ingested by small invertebrates, but their fate in the digestive organs and their effects on the animals are yet not well understood. We embedded fluorescent microplastics in artificial agarose-based food and offered the food to marine isopods, Idotea emarginata. The isopods did not distinguish between food with and food without microplastics. Upon ingestion, the microplastics were present in the stomach and in the gut but not in the tubules of the midgut gland which is the principal organ of enzyme-secretion and nutrient resorption. The feces contained the same concentration of microplastics as the food which indicates that no accumulation of microplastics happens during the gut passage. Long-term bioassays of 6 weeks showed no distinct effects of continuous microplastic consumption on mortality, growth, and intermolt duration. I. emarginata are able to prevent intrusion of particles even smaller than 1 µm into the midgut gland which is facilitated by the complex structure of the stomach including a fine filter system. It separates the midgut gland tubules from the stomach and allows only the passage of fluids and chyme. Our results indicate that microplastics, as administered in the experiments, do not clog the digestive organs of isopods and do not have adverse effects on their life history parameters.
Assuntos
Sistema Digestório/metabolismo , Isópodes/metabolismo , Plásticos/metabolismo , Poluentes Químicos da Água/metabolismo , Animais , Fezes/química , Conteúdo Gastrointestinal/químicaRESUMO
Marine viruses are ubiquitous, extremely diverse, and outnumber any form of life in the sea. Despite their ecological importance, viruses in marine environments have been largely ignored by the academic community, and only those that have caused substantial economic losses have received more attention. Fortunately, our current understanding on marine viruses has advanced considerably during the last decades. These advances have opened new and exciting research opportunities as several unique structural and genetic characteristics of marine viruses have shown to possess an immense potential for various biotechnological applications. Here, a condensed overview of the possibilities of using the enormous potential offered by marine viruses to develop innovative products in industries as pharmaceuticals, environmental remediation, cosmetics, material sciences, and several others, is presented. The importance of marine viruses to biotechnology should not be underestimated.
Assuntos
Organismos Aquáticos/fisiologia , Biotecnologia/métodos , Biotecnologia/tendências , Fenômenos Fisiológicos Virais , VírusRESUMO
The intracellular ubiquitin-proteasome system is a key regulator of cellular processes involved in the controlled degradation of short-living or malfunctioning proteins. Certain diseases and cellular dysfunctions are known to arise from the disruption of proteasome pathways. Trace metals are recognized stressors of the proteasome system in vertebrates and plants, but their effects on the proteasome of invertebrates are not well understood. Since marine invertebrates, and particularly benthic crustaceans, can be exposed to high metal levels, we studied the effects of in vitro exposure to Hg(2+), Zn(2+), Cu(2+), and Cd(2+) on the activities of the proteasome from the claw muscles of lobsters (Homarus gammarus) and crabs (Cancer pagurus). The chymotrypsin like activity of the proteasome of these two species showed different sensitivity to metals. In lobsters the activity was significantly inhibited by all metals to a similar extent. In crabs the activities were severely suppressed only by Hg(2+) and Cu(2+) while Zn(2+) had only a moderate effect and Cd(2+) caused almost no inhibition of the crab proteasome. This indicates that the proteasomes of both species possess structural characteristics that determine different susceptibility to metals. Consequently, the proteasome-mediated protein degradation in crab C. pagurus may be less affected by metal pollution than that of the lobster H. gammarus.