RESUMO
Physiological knowledge gained from questions focused on the challenges faced and strategies recruited by organisms in their habitats assumes fundamental importance about understanding the ability to survive when subjected to unfavorable situations. In the aquatic environment, salinity is particularly recognized as one of the main abiotic factors that affects the physiology of organisms. Although the physiological patterns and challenges imposed by each occupied environment are distinct, they tend to converge to osmotic oscillations. From a comparative perspective, we aimed to characterize the osmoregulatory patterns of the bivalve mollusks Corbicula largillierti (purple Asian cockle), Erodona mactroides (lagoon cockle), and Amarilladesma mactroides (white clam) - inhabitants of different osmotic niches - when submitted to hypo- and/or hyperosmotic salinity variations. We determined the hemolymph osmotic and ionic concentrations, tissue hydration, and the intracellular isosmotic regulation (IIR) from the use of osmolytes (organic and inorganic) after exposure to species-specific salinity intervals. Additionally, we incorporated phylogenetic perspectives to infer and even broaden the understanding about the patterns that comprise the osmoionic physiology of Bivalvia representatives. According to the variables analyzed in the hemolymph, the three species presented a pattern of osmoconformation. Furthermore, both ionic regulation and conformation patterns were observed in freshwater, estuarine, and marine species. The patterns verified experimentally show greater use of inorganic osmolytes compared to the participation of organic molecules, which varied according to the osmotic niche occupied in the IIR for the mantle, adductor muscle, and gills. This finding widens the classic vision about the preferential use of certain osmolytes by animals from distinct niches. Our phylogenetic perspective also indicates that environmental salinity drives physiological trait variations, including hemolymph osmolality and the ion composition of the extracellular fluid (sodium, chloride, magnesium, and calcium). We also highlight the important role played by the shared ancestry, which influences the interspecific variability of the hemolymph K+ in selected representatives of Bivalvia.
Assuntos
Evolução Biológica , Hemolinfa/fisiologia , Homeostase , Moluscos/fisiologia , Osmorregulação , Salinidade , Animais , Água Doce , Moluscos/classificação , Filogenia , Especificidade da EspécieRESUMO
Survival, osmoregulatory pattern, oxygen consumption, energy spent on metabolism, ammonia excretion, type of oxidized energy substrate, and hepatosomatic index were evaluated in decapods (an osmoregulating crab, Callinectes danae, and an osmoconforming seabob shrimp, Xiphopenaeus kroyeri) exposed to carbon dioxide-induced water acidification (pH 7.3, control pH 8.0) and different salinities (20, 25, 30, 35, and 40) for 3 days. Compared to the animals kept at controlled pH, exposure to reduced pH resulted in the loss of osmoregulatory capacity in C. danae at all salinities, except for some hyporegulation at 40, and reduced oxygen consumption and ammonia excretion at 20 and 40. Xiphopenaeus kroyeri remained an osmoconformer in all evaluated conditions, except for some hyporegulation at 40, and when exposed to the reduced pH, it presented changes in oxygen consumption at all salinities and reductions in ammonia excretion at 20 and 35 compared to the control animals. Both species use protein as the main energy substrate and decrease the hepatosomatic index when exposed to reduced pH relative to the control. The observed changes may be associated with changes in the activity of enzymes related to osmoregulation, the use of amino acids as osmotic effectors of cell volume control and recovery, and the Bohr effect, and, because the gills are multifunctional organs related to osmoregulation, the changes may be related to acid-base control, nitrogen excretion, and respiration, with a change in one of these functions bringing about changes in the others.
Assuntos
Braquiúros/fisiologia , Osmorregulação , Penaeidae/fisiologia , Água do Mar/química , Amônia/metabolismo , Animais , Dióxido de Carbono/química , Mudança Climática , Hemolinfa/fisiologia , Concentração de Íons de Hidrogênio , Masculino , Concentração Osmolar , Consumo de OxigênioRESUMO
Insects are organisms extremely well adapted to diverse habitats, primarily due to their innate immune system, which provides them with a range of cellular and humoral responses against microorganisms. Lepidoptera hemolymph proteins involved in humoral responses are well known; however, there is a lack of knowledge about the sugarcane borer Diatraea saccharalis. In this present work, the hemolymph proteins of this pest insect were studied by applying proteomic methodologies. Two-dimensional electrophoresis (2-DE) gels of proteins extracted from naive larvae and larvae challenged with Escherichia coli (ATCC 11224) and Bacillus subtilis (ATCC 6623) showed an average of 300 spots, and 92 of these spots corresponded in all three 2-DE gels. Forty-one spots were excised and digested with trypsin and analyzed using mass spectrometry. After analysis, 10 proteins were identified, including some proteins of the immune system: ß-defensin-like protein, Turandot A-like protein, attacin-like protein, peptidoglycan recognition protein and cyclophilin-like protein. Nine proteins were present in both experimental conditions; however, ß-defensin-like protein was present only in hemolymph challenged by B. subtilis. Notably, attacin-like protein was strongly induced by challenge with E. coli, suggesting an immune response against the infection. However, antimicrobial activity was observed in the test zone of microbial growth inhibition of B. subtilis solely with the hemolymph extract of the larvae challenged with B. subtilis. We made for the first time a proteomic profile of the hemolymph of D. saccharalis in which it was possible to identify the presence of important proteins involved in the immune response.
Assuntos
Peptídeos Catiônicos Antimicrobianos/análise , Hemolinfa/microbiologia , Proteínas de Insetos/análise , Lepidópteros/microbiologia , Animais , Peptídeos Catiônicos Antimicrobianos/metabolismo , Bacillus subtilis/fisiologia , Proteínas de Transporte/análise , Proteínas de Transporte/metabolismo , Escherichia coli/fisiologia , Hemolinfa/fisiologia , Interações Hospedeiro-Patógeno , Proteínas de Insetos/metabolismo , Lepidópteros/fisiologia , ProteômicaRESUMO
Hypoxia is a common and widespread phenomenon in aquatic ecosystems, imposing a significant challenge for the animals that inhabit such waters. In different habitats, however, the characteristics of these hypoxic events may differ, therefore imposing different challenges. We investigated the tolerance of adult ghost shrimp Neotrypaea uncinata (an intertidal mudflat dweller) to different partial pressures of oxygen (pO2), severe hypoxia (2 kPa) and recovery from hypoxia after different exposure times, mimicking the natural tidal cycle (6 h and 12 h). We calculated critical oxygen tension and categorize the adult ghost shrimps as oxyregulators (R value=75.27%). All physiological measurements (metabolic rate, oxyhemocyanin, hemolymph protein and lactate concentrations) were affected by exposure to low partial pressures of oxygen, but most of them recovered (with exception of metabolic rate) control values (21 kPa) after 6h under normoxic conditions. Low metabolic rate, high release of hemolymphatic proteins and anaerobic metabolism are suggested as response mechanisms to overcome hypoxic events during low tide.
Assuntos
Decápodes/fisiologia , Meio Ambiente , Consumo de Oxigênio/fisiologia , Oxigênio/metabolismo , Adaptação Fisiológica/fisiologia , Animais , Metabolismo Basal/fisiologia , Chile , Decápodes/metabolismo , Ecossistema , Geografia , Hemolinfa/metabolismo , Hemolinfa/fisiologia , Ácido Láctico/metabolismo , Masculino , Pressão ParcialRESUMO
Decapod crustaceans have independently invaded freshwater habitats from the sea/estuaries. Tissue hydration mechanisms are necessary for the initial stages of habitat transitions but can be expected to diminish, as the capacity for extracellular homeostasis increases in hololimnetic species. Six decapod species have been compared concerning the maintenance of muscle hydration in vitro: Hepatus pudibundus (marine); Palaemon pandaliformis (estuarine resident), Macrobrachium acanthurus (freshwater diadromous), and the three hololimnetic Macrobrachium potiuna, Dilocarcinus pagei, and Aegla parana. The effects of inhibitors of potassium channels (barium chloride) and NKCC (furosemide) were evaluated under isosmotic, and respectively hypo- (50% below iso) or hyper- (50% above iso) conditions. There was high muscle hydration control in H. pudibundus with a possible role of NKCC in isosmotic conditions. Shrimps consistently showed small deviations in muscle hydration under anisosmotic conditions; P. pandaliformis has shown evidence of the presence of NKCC; M. potiuna was the species less affected by both inhibitors, under iso- or anisosmotic conditions. In the two hololimnetic crab species, both independent long-time inhabitants of freshwater, while the capacity to deal with hyper-osmotic shock is decreased, the capacity to deal with hyposmotic shock is retained, possibly because of hemolymph dilution during molting in fresh water. D. pagei apparently depends on potassium channels for volume recovery after swelling, whereas A. parana shows some dependence on NKCC to minimize volume loss in hyper-osmotic conditions. Although no molecular screening techniques have been tried here, data point to distinct cell/tissue transport mechanisms acting upon hydration/volume challenges in decapods of different habitats and lineages.
Assuntos
Decápodes/fisiologia , Músculos/metabolismo , Pressão Osmótica , Equilíbrio Hidroeletrolítico/fisiologia , Animais , Ecossistema , Água Doce , Hemolinfa/metabolismo , Hemolinfa/fisiologia , Músculos/fisiologiaRESUMO
Several physiological processes can induce daily variations in aerobic metabolism. Lithodes santolla is a decapod crustacean of special concern since it is a sub-Antarctic species of commercial interest. The aim of this work was to study in L. santolla the daily variations in levels of enzymatic and non-enzymatic antioxidants, lipid peroxidation and protein oxidation, and haemolymphatic pH. Males of L. santolla of commercial size were randomly dissected every 4 h during a period of 24 h. Enzymatic activities of superoxide dismutase (SOD), catalase (CAT), glutathione-S-transferase and glutathione peroxidase were determined in samples of gills, muscle, hepatopancreas and haemolymph. Ascorbic acid, total glutathione, lipid peroxidation and protein oxidation were also determined in all tissues. Gills showed the highest enzymatic activity and hepatopancreas the highest concentration of non-enzymatic antioxidants. Maximum antioxidant activity was during the dark phase in gills and during the photophase in the haemolymph. Muscle showed significant daily variations, with peaks during the photophase and scotophase. Overall, an antioxidant protective mechanism is present in all tissues, where SOD and CAT represent the first line of defense. The defense mechanism in L. santolla seems to be more active during the dark phase, with slight differences among the analyzed tissues, indicating a higher metabolic rate.
Assuntos
Antioxidantes/metabolismo , Decápodes/fisiologia , Animais , Ácido Ascórbico/metabolismo , Catalase/metabolismo , Decápodes/enzimologia , Decápodes/metabolismo , Brânquias/enzimologia , Brânquias/metabolismo , Brânquias/patologia , Glutationa/metabolismo , Glutationa Peroxidase/metabolismo , Glutationa Transferase/metabolismo , Hemolinfa/enzimologia , Hemolinfa/metabolismo , Hemolinfa/fisiologia , Hepatopâncreas/enzimologia , Hepatopâncreas/metabolismo , Hepatopâncreas/fisiologia , Concentração de Íons de Hidrogênio , Peroxidação de Lipídeos , Masculino , Músculos/enzimologia , Músculos/metabolismo , Músculos/fisiologia , Oxirredução , Superóxido Dismutase/metabolismoRESUMO
Insect disease vectors show diminished fecundity when infected with Plasmodium. This phenomenon has already been demonstrated in laboratory models such as Aedes aegypti, Anopheles gambiae and Anopheles stephensi. This study demonstrates several changes in physiological processes of A. aegypti occurring upon infection with Plasmodium gallinaceum, such as reduced ecdysteroid levels in hemolymph as well as altered expression patterns for genes involved in vitellogenesis, lipid transport and immune response. Furthermore, we could show that P. gallinaceum infected A. aegypti presented a reduction in reproductive fitness, accompanied by an activated innate immune response and increase in lipophorin expression, with the latter possibly representing a nutritional resource for Plasmodium sporozoites.
Assuntos
Aedes/parasitologia , Plasmodium gallinaceum/crescimento & desenvolvimento , Aedes/genética , Aedes/imunologia , Aedes/fisiologia , Animais , Galinhas/parasitologia , Modelos Animais de Doenças , Ecdisteroides/sangue , Feminino , Fertilidade , Regulação da Expressão Gênica , Aptidão Genética , Hemolinfa/parasitologia , Hemolinfa/fisiologia , Interações Hospedeiro-Parasita , Imunidade Inata , Proteínas de Insetos/genética , Metabolismo dos Lipídeos , Lipoproteínas/genética , Malária Aviária/parasitologia , Plasmodium gallinaceum/fisiologia , Esporozoítos/crescimento & desenvolvimento , Esporozoítos/fisiologia , VitelogêneseRESUMO
Decapod crustaceans occupy various aquatic habitats. In freshwater they are osmoregulators, while marine species are typically osmoconformers. Freshwater crustaceans are derived from marine ancestors. The hypothesis tested here was that osmoregulators, which can rely on salt transport by interface epithelia to prevent extracellular disturbance, would have a lower capacity of tissue water regulation when compared with osmoconformers. Four species of decapod crustaceans (the marine osmoconformer crab Hepatus pudibundus, and three osmoregulators of different habitats) have been exposed in vivo to a salinity challenge, for up to 24 hr. Osmoregulators were: the estuarine shrimp Palaemon pandaliformis, the diadromous freshwater shrimp Macrobrachium acanthurus, and the hololimnetic red crab Dilocarcinus pagei. H. pudibundus displayed hemolymph dilution already after 0.5 hr in 25 per thousand, reaching approximately 30% reduction in osmolality, but its muscle degree of hydration did not increase. To make the different in vivo salinity challenges directly comparable, the ratio between the maximum change in muscle hydration with respect to the control value measured for the species and the maximum change in hemolymph osmolality was calculated (x 1,000): H. pudibundus (25 per thousand, 8.1% kg H(2)O/mOsm x 10(3))>P. pandaliformis (2 per thousand, 9.2)>M. acanthurus (30 per thousand, 12.6)>P. pandaliformis (35 per thousand, 16.7)>D. pagei (7 per thousand, 60.4). Muscle slices submitted in vitro to a 30% osmotic challenge confirmed in vivo results. Thus, the estuarine/freshwater osmoregulators displayed a lower capacity to hold muscle tissue water than the marine osmoconformer, despite undergoing narrower variations in hemolymph osmolality.
Assuntos
Decápodes/fisiologia , Líquido Extracelular/fisiologia , Músculo Esquelético/fisiologia , Equilíbrio Hidroeletrolítico/fisiologia , Animais , Ecossistema , Feminino , Água Doce , Hemolinfa/fisiologia , Masculino , Distribuição Aleatória , Salinidade , Água do MarRESUMO
Ergosan an algal product containing 1% alginic acid, developed for use in aquaculture and reported to have immunomodulatory activity, was administered orally to intermoult adult white shrimp (Litopenaeus vannamei) for 15 days. Examination of haemolymph proteins using SDS-PAGE did not reveal any obvious differences between control and Ergosan treated shrimp. Similarly, total haemocyte counts were found to be roughly equivalent for both the control and experimental samples. However, differential analysis of haemocyte populations revealed marked changes in terms of the relative levels of hyaline, semi-granular, and particularly granular haemocytes between the two groups. Moreover, enhancement of the in vitro antimicrobial activity of haemolymph towards two shrimp pathogenic Vibrio isolates was recorded for shrimp fed with Ergosan. Finally, shrimp fed with Ergosan showed a significant increase in relative growth when compared with control groups.
Assuntos
Alginatos/administração & dosagem , Decápodes/fisiologia , Hemócitos/química , Hemolinfa/química , Imunidade Inata/efeitos dos fármacos , Animais , Antibacterianos , Decápodes/efeitos dos fármacos , Ácido Glucurônico/administração & dosagem , Hemolinfa/fisiologia , Ácidos Hexurônicos/administração & dosagem , Imunização , Vibrio/efeitos dos fármacos , Vibrio/crescimento & desenvolvimento , Vibrio parahaemolyticusRESUMO
Veneza zonata (Hemiptera Coreidae) is an insect which causes losses in several crops, and it is also an important vector of lower trypanosomatids. V. zonata specimens were collected on rural properties in Londrina, state of Paraná, Southern Brazil. Inoculation of Leptomonas 563DT into V. zonata hemocoel caused insect death within approximately 24 h, with large bacterial proliferation into their hemocoels. Some bacteria which were found in the digestive tract of those insects, such as Escherichia coli, Providencia rettgeri, and Kluyveria ascorbata, were also found in their hemolymph, which suggests that trypanosomatid crossing into hemocoel caused mechanical lesions in the digestive tract that allowed intestinal bacteria to infect the hemolymph, thereby leading to lethal septicemia. In this study we analysed proteolytic activities from the 563DT Leptomonas strain, which is pathogenic for V. zonata, aiming at evaluating the potential use of this Leptomonas strain for the biocontrol of the insect. The proteolytic action was evaluated on cells and on culture supernatants of trypanosomatids. We also evaluated the gelatinolytic activities, the action over natural and synthetic substrates for aminopeptidases, and the action of protease inhibitors during all trypanosomatid growth stages. A significant reduction in the number of insect deaths was observed when Leptomonas 563DT were incubated with inhibitors of proteases and phospholipases before being inoculated into the insects, which suggests that those enzymes are involved in the pathogenic mechanism.