Primary amino acid sequences of decapod (Na+, K+)-ATPase provide evolutionary insights into osmoregulatory mechanisms.

Decapod Crustacea exhibit a marine origin, but many taxa have occupied environments ranging from brackish to fresh water and terrestrial habitats, overcoming their inherent osmotic challenges. Osmotic and ionic regulation is achieved by the gill epithelia, driven by two active ATP-hydrolyzing ion transporters, the basal (Na+, K+)-ATPase and the apical V(H+)-ATPase. The kinetic characteristic of gill (Na+, K+)-ATPase and the mRNA expression of its α subunit have been widely studied in various decapod species under different salinity challenges. However, the evolution of the primary structure has not been explored, especially considering the functional modifications associated with decapod phylogeny. Here, we proposed a model for the topology of the decapod α subunit, identifying the sites and motifs involved in its function and regulation, as well as the patterns of its evolution assuming a decapod phylogeny. We also examined both the amino acid substitutions and their functional implications within the context of biochemical and physiological adaptation. The α-subunit of decapod crustaceans shows greater conservation (∼94% identity) compared to the ß-subunit (∼40%). While the binding sites for ATP and modulators are conserved in the decapod enzyme, the residues involved in the α-ß interaction are only partially conserved. In the phylogenetic context of the complete sequence of (Na+, K+)-ATPase α-subunit, most substitutions appear to be characteristic of the entire group, with specific changes for different subgroups, especially among brachyuran crabs. Interestingly, there was no consistent separation of α-subunit partial sequences related to habitat, suggesting that the convergent evolution for freshwater or terrestrial modes of life is not correlated with similar changes in the enzyme's primary amino acid sequence.

Assessment of the physiological performance of the invasive oriental shrimp Palaemon macrodactylus from an atypical marine population.

Since 2000, a well-established population of the invasive oriental shrimp Palaemon macrodactylus has been present in fully marine conditions in the southwestern Atlantic Ocean (~38° S). To assess the physiological performance of this atypical population restricted to fully marine conditions, we conducted a laboratory experiment in which individuals were transferred from 35 ‰S (local seawater) to 2 ‰S; 5 ‰S; 10 ‰S; 20 ‰S; 50 ‰S and 60‰ for short (6 h), medium (48 h), and long (>504 h) acclimation periods. We measured the time course response of relevant parameters in the shrimp's hemolymph; activity of Na+, K+-ATPase (NKA), and V-H+-ATPase (VHA); and muscle water content. Shrimp showed great osmoregulatory plasticity, being able to survive for long periods between 5 ‰S and 50 ‰S, whereas no individual survived after transfer to either 2 ‰S or 60 ‰S. Shrimp hyper-regulated hemolymph osmolality at 5 ‰S and 10 ‰S, hypo-regulated at 35 ‰S and 50 ‰S, and isosmoticity was close to 20 ‰S. Compared to 35 ‰S, prolonged acclimation to 5 ‰S caused a decrease in hemolymph osmolality (~34%) along with sodium and chloride concentrations (~24%); the NKA and VHA activities decreased by ~52% and ~88%, respectively, while muscle water content was tightly regulated. Our results showed that the atypical population of P. macrodactylus studied here lives in a chronic hypo-osmo-ion regulatory state and suggest that fully marine conditions contribute to its poor performance at the lower limit of salinity tolerance (<5 ‰S).

Molecular Footprints on Osmoregulation-Related Genes Associated with Freshwater Colonization by Cetaceans and Sirenians.

The genetic basis underlying adaptive physiological mechanisms has been extensively explored in mammals after colonizing the seas. However, independent lineages of aquatic mammals exhibit complex patterns of secondary colonization in freshwater environments. This change in habitat represents new osmotic challenges, and additional changes in key systems, such as the osmoregulatory system, are expected. Here, we studied the selective regime on coding and regulatory regions of 20 genes related to the osmoregulation system in strict aquatic mammals from independent evolutionary lineages, cetaceans, and sirenians, with representatives in marine and freshwater aquatic environments. We identified positive selection signals in genes encoding the protein vasopressin (AVP) in mammalian lineages with secondary colonization in the fluvial environment and in aquaporins for lineages inhabiting the marine and fluvial environments. A greater number of sites with positive selection signals were found for the dolphin species compared to the Amazonian manatee. Only the AQP5 and AVP genes showed selection signals in more than one independent lineage of these mammals. Furthermore, the vasopressin gene tree indicates greater similarity in river dolphin sequences despite the independence of their lineages based on the species tree. Patterns of distribution and enrichment of Transcription Factors in the promoter regions of target genes were analyzed and appear to be phylogenetically conserved among sister species. We found accelerated evolution signs in genes ACE, AQP1, AQP5, AQP7, AVP, NPP4, and NPR1 for the fluvial mammals. Together, these results allow a greater understanding of the molecular bases of the evolution of genes responsible for osmotic control in aquatic mammals.

Osmoregulatory responses in the neotropical fish species Astyanax lacustris, exposed to single and combined microplastics, polycyclic aromatic hydrocarbons, and their mixture.

Microplastic (MP) pollution poses a significant environmental threat. These MPs can adsorb toxic compounds such as polycyclic aromatic hydrocarbons (PAH), which are highly lipophilic and carcinogenic. To assess the potential effects of virgin MP, PAH, and MP+PAH in association with osmoregulation and energetic substrate, we conducted experiments with the tetra cardinal Astyanax lacustris. The environmentally relevant concentration of MP (10 mg L-1) and 20 % of the LC50-96 h of crude oil for A. lacustris (2.28 µg L-1) were used during the 96-h exposure. Fish were exposed to virgin MP, PAH, MPC (MP loaded with PAH), PAH+MP (PAH and MP in association), and the control without (CT) and with handling (CH). After 96 h, blood was collected for osmoregulatory parameters (plasma osmolality; Na+, K+, Cl-, Mg2+; glycose and lactate); gills for osmoregulatory enzyme activities (Na+, K+ ATPase, H+ ATPase, and carbonic anhydrase); and white muscle samples were used to determine glycogen as an energetic substrate. The low molecular weight PAH was not detected in PAH-loaded MP (MPC) and PAH in combination with MP (PAH+MP). The PAH concentration of the MPC and PAH+MP was similar and low compared to other works. Virgin MP, PAH, MPC, and PAH+MP were able to cause muscle glycogen depletion. The activity of v-type H+ ATPase and plasma Na+ concentrations were lower in PAH with MP (MPC). However, the hydromineral balance (K+, Mg2+, Cl-, and osmolality) was not affected by any treatment. In this sense, we can conclude that the MPC caused osmoregulatory disturbances not seen in the MP associated with PAH (MP+PAH). However, this seems unrelated to the PAH leaking from the MPC or the PAH absorption to the virgin MP once the PAH concentrations from the MPC and PAH+MP were similar.

Acute metabolic responses of two marine brachyuran crabs to dilute seawater: The aerobic cost of hyper regulation.

Hepatus pudibundus ("flecked box crab") is a stenohaline osmoconfomer, and restricted to marine habitats. Callinectes danae ("swimming crab Dana") lives in coastal/estuarine waters and is a weak hyper regulator. There is no consensus on which strategy is more expensive metabolically face salinity challenges: conformation with higher dependence on cell volume regulation, or hyper regulation, alleviating the need for intense cell volume regulation. Crabs were probed for their acute response to dilute seawater through exposures to salinities 35‰, 30‰, 25‰, and 20‰ for 2, 4, and 6 h. Hemolymph osmolality, lactate, and ions (chloride, sodium, magnesium, potassium) were assayed, as well as muscle water content. Water dissolved oxygen, ammonia, and pH levels were also measured. H. pudibundus conformed for osmolality and displayed increase in muscle hydration along the decrease in salinity down to 25‰, while C. danae efficiently maintained hemolymph osmo ionic stability, consumed more oxygen, acidified more the water, and released more ammonia. In 25‰, both species spent energy: H. pudibundus putatively controlling cell volume, and C. danae regulating hemolymph concentrations. In 20‰, H. pudibundus closed itself, avoiding the contact of the interface epithelia with the external environment and producing much lactate, whereas C. danae spent more energy (aerobic) in extracellular osmo ionic stability. Under these conditions, anisosmotic extracellular regulation (together with additional cell volume regulation) is more oxygen consuming than osmoconformation with a putatively more intense challenge to cell volume. The exposure to hyposalinity limits the occupation of estuarine environments by H. pudibundus in short and middle term.

Can hyper/hypo-osmoregulating fiddler crabs from the Atlantic coast of South America mobilize intracellular free amino acids as osmotic effectors during salinity challenge?

Weakly osmoregulating crustaceans use intracellular free amino acids (FAA) to attenuate cell volume changes consequent to alterations in hemolymph osmolality. Whether semiterrestrial, strong hyper/hypo-osmoregulators exhibit this ability is unknown. We investigate FAA mobilization in muscle tissue of 10 fiddler crabs from the genera Minuca, Leptuca, and Uca distributed along the Atlantic coast of South America. Crabs were subjected to severe hypo- or hyper-osmotic challenge at their lower or upper critical salinity limits for 5 days; reference crabs were held in isosmotic media. Hemolymph osmolality was measured, chela muscle FAA were identified and quantified, and percent contribution to intracellular osmolality (%FAA) was calculated. At isosmoticity, total FAA were nominally twofold higher in Minuca species (≈116 mmol/kg wet mass) compared to Uca (≈60 mmol/kg wet mass). Glycine, alanine, arginine, and taurine constituted >80% of the total FAA pool. On hyperosmotic challenge, hemolymph osmolalities ranged from 843 to 1282 mOsm/kg H2 O. FAA increased, although %FAA remained unaltered. Hypo-osmoregulating crabs thus can mobilize FAA, likely owing to a lesser ability to secrete salt near their upper critical limits. On hypo-osmotic challenge, osmolalities were regulated more tightly, between 475 and 736 mOsm/kg H2 O. Total FAA and %FAA showed little change, probably due to the crabs' strong hyper-osmotic extracellular regulatory ability, FAA consequently playing a diminished role in isosmotic intracellular regulation (IIR). Total FAA responses to hyper/hypo-osmotic challenge are thus asymmetrical. The lack of phylogenetic signal in FAA mobilization suggests that closely related fiddler crabs do not share similar strategies of IIR.

Physiology and production of sugar-apple under water stress and application of proline / Fisiologia e produção de pinheira sob estresse hídrico e aplicação de prolina

The objective of this study was to evaluate the physiology and production of sugar-apple as a function of irrigation intervals and foliar application of proline under the conditions of Paraíba's semi-arid region. A randomized block design was laid out in a 4 × 2 factorial scheme, with treatments resulting from the combination of four irrigation intervals (1, 4, 8 and 12 days) and two concentrations of proline (0 and 10 mM), with four replicates, and the plot consisted of four usable plants. Increase in irrigation intervals reduced the gas exchange of sugar-apple plants at 298 days after transplanting. Exogenous application of proline at concentration of 10 mM increased contents of chlorophyll a, chlorophyll b, total chlorophyll and carotenoids and fruit fresh mass in plants grown under 12-day irrigation intervals.

Objetivou-se com este trabalho avaliar a fisiologia e a produção da pinheira em função dos turnos de rega e aplicação foliar de prolina em condições do semiárido Paraibano. Foi utilizado o delineamento de blocos casualizados em esquema fatorial 4 × 2, cujos tratamentos resultam da combinação de quatro turno de rega (1, 4, 8 e 12 dias) e duas concentrações de prolina (0 e 10 mM), com quatro repetições, cuja a parcela foi constituída de quatro plantas úteis. O incremento nos turnos de rega reduziu as trocas gasosas das plantas de pinheira, aos 298 dias após o transplantio. A aplicação de prolina na concentração de 10 mM aumentou o extravasamento de eletrólitos no limbo foliar, a condutância estomática e diminuiu a concentração interna de CO2 das plantas de pinheira. A aplicação exógena de prolina na concentração de 10 mM aumentou os teores de clorofila a, b, total e carotenoides e a massa fresca de frutos nas plantas cultivadas sob turno de rega de 12 dias.

Arbuscular Mycorrhizal Fungi Promote Physiological and Biochemical Advantages in Handroanthus serratifolius Seedlings Submitted to Different Water Deficits.

Climate change causes increasingly longer periods of drought, often causing the death of plants, especially when they are in the early stages of development. Studying the benefits provided by arbuscular mycorrhizal (AM) fungi to plants in different water regimes is an efficient and sustainable strategy to face climate change. Thus, this study investigated the influence of AM fungi on Handroanthus serratifolius seedlings under different water regimes, based on biochemical, and nutritional growth parameters. The experiment was carried out in H. serratifolius seedlings cultivated with mycorrhizas (+AMF) and without mycorrhizas (-AMF) in three water regimes; a severe water deficit (SD), a moderate water deficit (MD), and a well-watered (WW) condition. AM fungi provided greater osmoregulation under water deficit conditions through the accumulation of soluble sugars, total free amino acids, and proline, as well as by reducing sugar. The increase in the absorption of phosphorus and nitrate was observed only in the presence of fungi in the well-watered regimen. A higher percentage of colonization was found in plants submitted to the well-watered regimen. Ultimately, AM fungi promoted biochemical, nutritional, and growth benefits for H. serratifolius seedlings under the water deficit and well-hydrated conditions, proving that AMF can be used to increase the tolerance of H. serratifolius plants, and help them to survive climate change.

Ontogenetical development of branchial chambers of Litopenaeus vannamei (Boone, 1931) and their involvement in osmoregulation: ionocytes and Na+/K+-ATPase.

Branchial chambers constitute the main osmoregulatory site in almost all decapod crustaceans. However, few studies have been devoted to elucidate the cellular function of specific cells in every osmoregulatory structure of the branchial chambers. In decapod crustaceans, it is well-known that the osmoregulatory function is localized in specific structures that progressively specialize from early developmental stages while specific molecular mechanisms occur. In this study, we found that although the structures developed progressively during the larval and postlarval stages, before reaching juvenile or adult morphology, the osmoregulatory capabilities of Litopenaeus vannamei were gradually established only during the development of branchiostegites and epipodites, but not gills. The cellular structures of the branchial chambers observed during the larval phase do not present the typical ultrastructure of ionocytes, neither Na+/K+-ATPase expression, likely indicating that pleura, branchiostegites, or bud gills do not participate in osmoregulation. During early postlarval stages, the lack of Na+/K+-ATPase immunoreactivity of the ionocytes from the branchiostegites and epipodites suggests that they are immature ionocytes (ionocytes type I). It could be inferred from IIF and TEM results that epipodites and branchiostegites are involved in iono-osmoregulation from PL15, while gills and pleura do not participate in this function.

Modulation of miR-429 during osmotic stress in the silverside Odontesthes humensis.

Silverside fish inhabit marine coastal waters, coastal lagoons, and estuarine regions in southern South America. Although silversides are not fully adapted to freshwater, they can tolerate a wide range of salinity variations. MicroRNAs (miRNAs) are a class of ∼22 nucleotide noncoding RNAs, which are crucial regulators of gene expression at post-transcriptional level. Current data indicate that miRNAs biogenesis is altered by situations of environmental stress, thereby altering the expression of target mRNAs. Foremost, the silversides were acutely exposed to 30 g.L-1 of salt to reveal in which tissue miR-429 could be differentially expressed. Thus, fish were acclimated to freshwater (0 g.L-1) and to brackish water (10 g.L-1), and then exposed to opposite salinity treatment. Here, we reveal that miR-429, a gill-enriched miRNA, emerges as a prime osmoregulator in silversides. Taken together, our findings suggest that miR-429 is an endogenous regulator of osmotic stress, which may be developed as a biomarker to assist silverside aquaculture.

Osmoregulation and the Hypothalamic Supraoptic Nucleus: From Genes to Functions.

Due to the relatively high permeability to water of the plasma membrane, water tends to equilibrate its chemical potential gradient between the intra and extracellular compartments. Because of this, changes in osmolality of the extracellular fluid are accompanied by changes in the cell volume. Therefore, osmoregulatory mechanisms have evolved to keep the tonicity of the extracellular compartment within strict limits. This review focuses on the following aspects of osmoregulation: 1) the general problems in adjusting the "milieu interieur" to challenges imposed by water imbalance, with emphasis on conceptual aspects of osmosis and cell volume regulation; 2) osmosensation and the hypothalamic supraoptic nucleus (SON), starting with analysis of the electrophysiological responses of the magnocellular neurosecretory cells (MNCs) involved in the osmoreception phenomenon; 3) transcriptomic plasticity of SON during sustained hyperosmolality, to pinpoint the genes coding membrane channels and transporters already shown to participate in the osmosensation and new candidates that may have their role further investigated in this process, with emphasis on those expressed in the MNCs, discussing the relationships of hydration state, gene expression, and MNCs electrical activity; and 4) somatodendritic release of neuropeptides in relation to osmoregulation. Finally, we expect that by stressing the relationship between gene expression and the electrical activity of MNCs, studies about the newly discovered plastic-regulated genes that code channels and transporters in the SON may emerge.

Decapod egg membranes: powerful barriers or regulatory structures?

Osmoregulatory abilities and mechanisms of adults and larvae of decapod crustaceans have been extensively investigated. However, how embryos carried by their mothers can deal with changing or extreme salinities is less understood. The egg membranes are believed to isolate embryos from a challenging environment, although osmoregulatory ability has been demonstrated in early developing embryos (naupliar stage) of two crabs. To establish whether embryos are isolated by their membranes and/or are able to osmoregulate, we measured the survival and volume change over 48 h of oocytes and embryos in different stages of three carideans (Betaeus lilianae, Palaemon macrodactylus and P. argentinus) and the brachyuran Neohelice granulata, subjected to different salinities. In addition, we recorded osmolality changes in homogenates of the same stages in P. argentinus and N. granulata after 2 h of exposure and mapped the presence of putative sites of ions exchange in the membrane of all species. High mortality, when it occurred, was associated with low salinity and mortality variation with the stage of development depended on the species. All species precipitated silver salts in or under the egg envelope, with a different pattern between carideans and the brachyuran. Changes in osmolality and egg volume after hypo- or hyper-osmotic salinity challenges indicate that eggs are not fully isolated by their membranes, and that some osmoregulatory mechanisms are in play to maintain developmental homeostasis. We suggest that egg membranes can participate in osmoregulation by selectively transporting ions to an intramembrane space, with differences between carideans and brachyurans.

Metabolic adjustment and regulation of gene expression are essential for increased resistance to severe water deficit and resilience post-stress in soybean.

Background: Soybean is the main oilseed crop grown in the world; however, drought stress affects its growth and physiology, reducing its yield. The objective of this study was to characterize the physiological, metabolic, and genetic aspects that determine differential resistance to water deficit in soybean genotypes. Methods: Three soybean genotypes were used in this study, two lineages (L11644 and L13241), and one cultivar (EMBRAPA 48-C48). Plants were grown in pots containing 8 kg of a mixture of soil and sand (2:1) in a greenhouse under sunlight. Soil moisture in the pots was maintained at field capacity until the plants reached the stage of development V4 (third fully expanded leaf). At this time, plants were subjected to three water treatments: Well-Watered (WW) (plants kept under daily irrigation); Water Deficit (WD) (withholding irrigation until plants reached the leaf water potential at predawn of -1.5 ± 0.2 MPa); Rewatered (RW) (plants rehydrated for three days after reached the water deficit). The WW and WD water treatments were evaluated on the eighth day for genotypes L11644 and C48, and on the tenth day for L13241, after interruption of irrigation. For the three genotypes, the treatment RW was evaluated after three days of resumption of irrigation. Physiological, metabolic and gene expression analyses were performed. Results: Water deficit inhibited growth and gas exchange in all genotypes. The accumulation of osmolytes and the concentrations of chlorophylls and abscisic acid (ABA) were higher in L13241 under stress. The metabolic adjustment of lineages in response to WD occurred in order to accumulate amino acids, carbohydrates, and polyamines in leaves. The expression of genes involved in drought resistance responses was more strongly induced in L13241. In general, rehydration provided recovery of plants to similar conditions of control treatment. Although the C48 and L11644 genotypes have shown some tolerance and resilience responses to severe water deficit, greater efficiency was observed in the L13241 genotype through adjustments in morphological, physiological, genetic and metabolic characteristics that are combined in the same plant. This study contributes to the advancement in the knowledge about the resistance to drought in cultivated plants and provides bases for the genetic improvement of the soybean culture.

Effects of salinity and plant-based diet or animal-plant combination diet on the performance and metabolic status of juvenile Nile tilapia / Efeitos da salinidade e de dietas contendo ingredientes de origem vegetal ou de origem animal e vegetal no desempenho e na condição metabólica de juvenis de tilápia do Nilo

The purpose of this study was to evaluate the effects of salinity and plant-based diet or animal-plant combination diet on the performance and metabolic status of juvenile Nile tilapia (Oreochromis niloticus). The experimental design was completely randomized in a 4 × 2 factorial scheme with four replicates. The treatments were established by the combination of salinities of 0, 10, 20, and 30 g L-1 with an animal-plant combination diet (AP) or plant-based diet (P). The replicates were 60 L tanks with 12 fish per tank. Diets were provided for 32 days, and the fish were fed three times a day (8, 12, and 17 h) until apparent satiety. Daily feed intake (DFI) was measured, body weight (BW) was recorded at the beginning and end of the trial, and total length (TL) and standard length (SL) were measured at the end of the trial. Average daily gain (ADG), specific growth rate (SGR), feed conversion ratio (FCR), and survival rate were calculated. After the biometric measurements were made at the end of the trial, blood samples were collected to determine the plasma concentrations of total protein (TP), glucose, cholesterol, and triglycerides (TG). The fish were euthanized, and the hepatopancreas was collected and weighed; thereafter, the hepatosomatic index (HSI) was calculated. An interaction was detected between salinity and diet type for final BW, ADG, TL, and SL. These traits were not influenced by salinity when it was associated with the AP diet, but reduced linearly with salinity in the P diet. DFI and survival rate were independently affected by salinity: DFI reduced linearly with salinity levels and survival rate was higher at a salinity of 10 g L-1. HSI increased linearly with salinity levels and was lower in the P diet than in the AP diet. Salinity had a quadratic effect on plasma TP, and the maximum value for this metabolite (2.96 g dL-1) is attained at a salinity of 10.26 g L-1. There was an independent effect of diet on the plasma concentrations of cholesterol and TG, which were lower in the P diet than in the AP diet. The salinity of 10 g L-1 associated with diet composed of animal and plant ingredients led to a better performance, higher survival rate, and less stressful environmental conditions for juvenile Nile tilapia.(AU)

O objetivo deste estudo foi avaliar os efeitos da salinidade e de dietas compostas por ingredientes de origem vegetal, ou por ingredientes de origem animal e vegetal no desempenho e na condição metabólica de juvenis de tilápia do Nilo (Oreochromis niloticus). O delineamento foi inteiramente casualizado em esquema fatorial 4 × 2 com quatro repetições. Os tratamentos foram estabelecidos pela combinação das salinidades 0, 10, 20 e 30 g L-1 com dietas contendo apenas ingredientes de origem vegetal (V) ou ingredientes de origem animal e vegetal (AV). As repetições foram caixas de 60 L contendo 12 peixes por caixa. As dietas foram fornecidas durante 32 dias e os peixes foram alimentados três vezes ao dia (8, 12 e 17 h) até saciedade aparente. O consumo de ração foi mensurado diariamente (CRD), o peso corporal (PC) foi registrado no início e no final do experimento, e o comprimento total (CT) e comprimento padrão (CP) foram mensurados no final do experimento. O ganho médio diário (GMD), a taxa de crescimento específico (TCE), a conversão alimentar (CA) e a taxa de sobrevivência também foram calculadas. Após a obtenção das medidas biométricas ao final do experimento, amostras de sangue foram coletadas para determinar as concentrações plasmáticas de proteínas totais (PT), glicose, colesterol e triglicerídeos (TG). Posteriormente, os peixes foram eutanasiados e o hepatopâncreas foi coletado, pesado e o índice hepatossomático (IHS) foi calculado. Houve interação entre salinidade e tipo de dieta para PC final, GMD, CT e CP. Essas variáveis não foram influenciadas pela salinidade quando associada com a dieta AV, mas reduziram linearmente com a salinidade na dieta V. O CRD e a taxa de sobrevivência foram influenciados exclusivamente pela salinidade, onde o CRD reduziu linearmente com a salinidade, e a taxa de sobrevivência foi maior na salinidade de 10 g L-1. O IHS aumentou linearmente com a salinidade, e foi menor da dieta V comparado com a dieta AV. Houve efeito quadrático da salinidade na concentração plasmática de PT, em que o máximo valor para este metabólito (2.96 g dL-1) pode ser alcançado com a salinidade de 10.26 g L-1. As concentrações plasmáticas de colesterol e TG foram exclusivamente influenciadas pelo tipo de dieta, apresentando menores valores na dieta V comparados com a dieta AV. A salinidade de 10 g L-1 associada com dieta composta por ingredientes de origem animal e vegetal proporcionou melhor desempenho, maior taxa de sobrevivência e condições ambientais menos estressantes aos juvenis de tilápia do Nilo.(AU)

Respiratory profile and gill histopathology of Carassius auratus exposed to different salinity concentrations / Perfil respiratório e histopatologia das brânquias de Carassius auratus expostas a diferentes concentrações de salinidades

The aim of this study was to evaluate the chronic salinity tolerance of Carassius auratus and the effects on blood parameters, gill morphology, and survival. In the first test, nine different concentrations (0.0, 0.5, 1.0, 2.5, 5.0, 10, 15, 20, and 25 g L-¹) of NaCl were used with nine repetitions for 96 h. The survival of fish subjected to 15 g L-¹ NaCl was 4 h, and 5 min at a concentration of 25 g L-¹. The mortality of fish with 15g L-¹ NaCl was 100%. Morphological analyses of the gills showed hyperplasia of the coated cells in the interlamellar space and hypersecretion of mucus in fish exposed to 10 g L-¹ of NaCl. At concentrations of 20 and 25 g L-¹, necrosis of the support collagen caused the cells to detach from the lamellar structure mucosa. In the chronic test, two concentrations were used, with four replications containing nine fish in each aquarium for a period of 21 days. Blood samples and gills from the fish were collected, and it was observed that the fish showed a decrease in the concentration of bicarbonate (NaHCO3) in the blood, indicating hypernatremia. Acute exposure of C. auratus to sodium chloride (NaCl) should be at a maximum of 10 g L-¹ of NaCl, after which level there would be a loss in animal performance and/or mortality. Chronic exposure to 5 g L-¹ of NaCl promotes acidemia, ionic imbalance, and pathological changes in the gills; therefore, it is not recommended.

O objetivo deste estudo foi avaliar a tolerância crônica de Carassius auratus à salinidade e os efeitos sobre os parâmetros sanguíneos, morfologia branquial e sobrevivência. No primeiro teste, foram utilizadas nove concentrações de NaCl (0.0, 0.5, 1.0, 2.5, 5.0, 10, 15, 20, e 25 g L-¹) e nove repetições por 96 h. A sobrevivência dos peixes submetidos a até 15 g L-¹ de NaCl foi de 04h00, sendo 00h05 na concentração de 25 g L-¹ de NaCl. A mortalidade dos peixes com 15 g foi de 100%. As análises morfológicas das brânquias mostraram hiperplasia das células revestidas no espaço interlamelar e hipersecreção de muco em peixes expostos a uma concentração de 10 g L-¹ de NaCl. Nas concentrações de 20 e 25 g L-¹, observou-se que a colágeno de suporte perdeu a estrutura das células da mucosa, alterando as lamelas secundárias. Em um segundo experimento, um delineamento inteiramente casualizado foi utilizado com dois tratamentos (0 e 5g L-¹ de NaCl) e quatro repetições com nove peixes por 30 L em 21 dias. Amostras de sangue e brânquias dos peixes foram coletadas e observou-se que os peixes apresentaram diminuição nas concentrações de bicarbonato (NaHCO3) no sangue, indicando hipernatremia. Conclui-se que a exposição aguda de C. auratus ao cloreto de sódio (NaCl) deve ser de no máximo 10 g L-¹, levando à perda de desempenho e/ou mortalidade dos animais. A exposição crônica a 5 g L-¹ de NaCl promove acidemia, desequilíbrio iônico e alterações patológicas nas brânquias, por isso não é recomendado.

Respiratory profile and gill histopathology of Carassius auratus exposed to different salinity concentrations / Perfil respiratório e histopatologia das brânquias de Carassius auratus expostas a diferentes concentrações de salinidades

The aim of this study was to evaluate the chronic salinity tolerance of Carassius auratus and the effects on blood parameters, gill morphology, and survival. In the first test, nine different concentrations (0.0, 0.5, 1.0, 2.5, 5.0, 10, 15, 20, and 25 g L-¹) of NaCl were used with nine repetitions for 96 h. The survival of fish subjected to 15 g L-¹ NaCl was 4 h, and 5 min at a concentration of 25 g L-¹. The mortality of fish with 15g L-¹ NaCl was 100%. Morphological analyses of the gills showed hyperplasia of the coated cells in the interlamellar space and hypersecretion of mucus in fish exposed to 10 g L-¹ of NaCl. At concentrations of 20 and 25 g L-¹, necrosis of the support collagen caused the cells to detach from the lamellar structure mucosa. In the chronic test, two concentrations were used, with four replications containing nine fish in each aquarium for a period of 21 days. Blood samples and gills from the fish were collected, and it was observed that the fish showed a decrease in the concentration of bicarbonate (NaHCO3) in the blood, indicating hypernatremia. Acute exposure of C. auratus to sodium chloride (NaCl) should be at a maximum of 10 g L-¹ of NaCl, after which level there would be a loss in animal performance and/or mortality. Chronic exposure to 5 g L-¹ of NaCl promotes acidemia, ionic imbalance, and pathological changes in the gills; therefore, it is not recommended.(AU)

O objetivo deste estudo foi avaliar a tolerância crônica de Carassius auratus à salinidade e os efeitos sobre os parâmetros sanguíneos, morfologia branquial e sobrevivência. No primeiro teste, foram utilizadas nove concentrações de NaCl (0.0, 0.5, 1.0, 2.5, 5.0, 10, 15, 20, e 25 g L-¹) e nove repetições por 96 h. A sobrevivência dos peixes submetidos a até 15 g L-¹ de NaCl foi de 04h00, sendo 00h05 na concentração de 25 g L-¹ de NaCl. A mortalidade dos peixes com 15 g foi de 100%. As análises morfológicas das brânquias mostraram hiperplasia das células revestidas no espaço interlamelar e hipersecreção de muco em peixes expostos a uma concentração de 10 g L-¹ de NaCl. Nas concentrações de 20 e 25 g L-¹, observou-se que a colágeno de suporte perdeu a estrutura das células da mucosa, alterando as lamelas secundárias. Em um segundo experimento, um delineamento inteiramente casualizado foi utilizado com dois tratamentos (0 e 5g L-¹ de NaCl) e quatro repetições com nove peixes por 30 L em 21 dias. Amostras de sangue e brânquias dos peixes foram coletadas e observou-se que os peixes apresentaram diminuição nas concentrações de bicarbonato (NaHCO3) no sangue, indicando hipernatremia. Conclui-se que a exposição aguda de C. auratus ao cloreto de sódio (NaCl) deve ser de no máximo 10 g L-¹, levando à perda de desempenho e/ou mortalidade dos animais. A exposição crônica a 5 g L-¹ de NaCl promove acidemia, desequilíbrio iônico e alterações patológicas nas brânquias, por isso não é recomendado.(AU)

Physicochemical properties of the dissolved organic carbon can lead to different physiological responses of zebrafish (Danio rerio) under neutral and acidic conditions.

Previous studies have suggested that the capacity of natural dissolved organic carbon (DOC) molecules to interact with biological membranes is associated with their aromaticity (SAC340 ); origin (allochthonous versus autochthonous, FI); molecular weight (Abs254/365 ); and relative fluorescence of DOC moieties (PARAFAC analysis). These interactions may be especially important when fish are challenged by acidic waters, which are known to inhibit the active uptake of Na+ and Cl- , while stimulating diffusive ion losses in freshwater fishes. Therefore, zebrafish were acclimated (7 days, pH 7.0) to five natural DOC sources (10 mg C/L), two from the Amazon Basin and three from Canada, together with a "no-added DOC" control. After the acclimation, fish were challenged by exposure to acidic water (pH 4.0) for 3 h. Osmoregulatory parameters were measured at pH 7.0 and 4.0. Acclimation to the five DOC sources did not disturb Na+ , Cl- and ammonia net fluxes, but resulted in differential elevations in Na+ , K+ ATPase and v-type H+ ATPase activities in fish at pH 7.0. However, after transfer to pH.4.0, the control fish exhibited rapid increases in both enzymes. In contrast the DOC- acclimated animals exhibited unchanged (Na+ , K+ ATPase) or differentially increased (v-type H+ ATPase) activities. Na+ , Cl- and ammonia net fluxes remained unchanged in the control fish, but were differentially elevated in most of the DOC treatments at pH 4.0, relative to the same DOC treatments at pH 7.0. Correlations between the osmoregulatory data the DOCs properties highlight that the DOC properties drive different effects on gill physiology.

Short communication: Effects of acute copper exposure on ionic regulation of the freshwater crab Aegla castro.

Aeglids are unique freshwater decapods whose habitats are being impacted by metallic compounds, such as copper (Cu). Thus, we investigated the effects of acute Cu exposure on ionic regulation of Aegla castro. For this, male specimens in intermolt were collected from a reference stream and acclimated for 5 days in laboratory. After which, crabs were exposed to 11 µg L-1 Cu (Cu11) or only to water (CTR) for 24 h. Hemolymph samples were withdrawn for the determination of Na+, K+, Ca2+, and Mg2+ concentrations and the posterior gills removed for the analysis of Na+/K+-ATPase, Ca2+-ATPase, H+-ATPase, and carbonic anhydrase (CA) activities. Increased Ca2+ and Mg2+ hemolymph concentrations were observed in animals from Cu11, when compared with CTR group. In addition, decreased activity of CA was observed in animals exposed to Cu. In the current study, alterations in Ca2+ and Mg2+concentrations probably indicate that animals activated exoskeleton reabsorption mechanisms, characteristic of the premolt. Therefore, increased Ca2+ and Mg2+ concentrations in hemolymph may indicate that a biochemical signal associated with the molting cycle was triggered by Cu exposure. Despite the known harmful effects of Cu on osmoregulatory enzymes, here we observed decreased activity only in CA. However, decreased activity of CA could trigger both acid-base imbalance and ionic disruption, since CA provides H+ and HCO3- for intracellular pH maintenance, and underpins Na+ and Cl- for ionic regulation. Therefore, understanding how aeglids respond to metal contamination in laboratory conditions is crucial to assess their potential as an alternative biological model for aquatic ecotoxicology.

Effects of copper on an omnivorous (Astyanax altiparanae) and a carnivorous fish (Hoplias malabaricus): A comparative approach.

Copper is an essential metal for life. However, in excess, it can lead to osmoregulatory disorders and oxidative stress in fish and these effects appear to be species specific. In order to evaluate the effects of copper and to compare the sensitivity of two Neotropical fishes that co-occur in nature as prey (Astyaynax altiparanae) and predator (Hoplias malabaricus), the fish were exposed to three concentrations of Cu (5 µg L-1, 10 µg L-1, and 20 µg L-1) for 96 h. At the end of the experimental period, copper concentration in tissues, osmoregulatory parameters, oxidative stress biomarkers, plasma glucose, muscle glycogen and acetylcholinesterase activity were evaluated. Fish mortality (25%) was only observed for A. altiparanae exposed to Cu 20 µg L-1. The results revealed species-specific ionic disturbances. Despite hypocalcemia, H. malabaricus showed an increase in the main gill ATPases, which probably guaranteed the maintenance of plasma Na+.  In A. altiparanae, there was no change in ATPase activity in the gills and hyponatremia was observed at all copper concentrations, as well as a decrease in plasma Cl- in the Cu 20 µg L-1 group. The strategy adopted by H. malabaricus seems to have contributed to the absence of copper accumulation in the tissues, in addition to possibly being related to the absence of oxidative stress in this species. On the other hand, there was an increase in the concentration of copper in the gills, liver, and gastrointestinal tract of A. altiparanae, as well as oxidative stress evidenced by increased lipoperoxidation in the liver and damage to erythrocytes DNA. This work reinforces the idea that copper effects are species specific and that a given concentration may not be safe for different species which can coexist in the same environment.

Comparison of lipids and fatty acids among tissues of two semiterrestrial crabs reveals ecophysiological adaptations in changing coastal environments.

Decapods have successfully colonized changing coastal habitats throughout the world by adapting their behavior, physiology, and biochemistry. Biochemical reserves, such as lipids and fatty acids (FAs), play fundamental roles in this adaptation process. These energy reserves are key for the development of decapods and their composition mainly depends on the type and quality of food available in their habitats. This study evaluated the lipid content and FA composition of three tissues (hepatopancreas, gills, and muscle) in two widely distributed, semi-terrestrial coastal crab species in Chile, Cyclograpsus cinereus from the upper intertidal and Hemigrapsus crenulatus from estuaries. This evaluation aimed to assess the physiological role of the bioenergetic reserves of these crabs, which tolerate fluctuating environmental conditions. Our results showed that both species had a higher lipid content in the hepatopancreas and a lower lipid content in its gills and muscle. All three of the evaluated tissues in C. cinereus showed high contents of saturated fatty acids (SFAs), and its hepatopancreas displayed the highest contents of monounsaturated (MUFAs) and polyunsaturated fatty acids (PUFAs). In turn, H. crenulatus had the highest contents of MUFAs and PUFAs in its gills and muscle tissues, including an important amount of eicosapentaenoic acid (EPA). The FA content of C. cinereus may indicate an adaptive physiological response aimed at maintaining its cellular fluid balance during periods of desiccation in the upper intertidal zone. In contrast, the FAs found in H. crenulatus may be linked to the high activity of the sodium­potassium pump in its gills, in order to maintain osmoregulation in estuaries.