Hypersalinity tolerance of mummichogs (Fundulus heteroclitus): A branchial transcriptomic analysis.

Along the east coast of North America, mummichogs (Fundulus heteroclitus) are subjected to a broad range of salinities in their nearshore habitats. However, there is a paucity of information regarding the molecular and cellular processes that mummichogs (and other highly osmotolerant fishes) engage to survive environmental salinities greater than seawater (SW). To reveal branchial processes underlying their extraordinarily broad salinity tolerance, we performed an RNA-Seq analysis to identify differentially expressed genes (DEGs) in mummichogs residing in 3, 35, and 105 ppt conditions. We identified a series of DEGs previously associated with both freshwater (FW)- and SW-type ionocytes; however, the heightened expression of anoctamin 1a, a Ca2+-activated Cl- channel, in 35 and 105 ppt indicates that an undescribed Cl--secretion pathway may operate within the SW-type ionocytes of mummichogs. Concerning FW-adaptive branchial processes, we identified claudin 5a as a gene whose product may limit the diffusive loss of ions between cellular tight junctions. Further, in response to hypersaline conditions, we identified DEGs linked with myo-inositol synthesis and kinase signaling. This study provides new molecular targets for future physiological investigations that promise to reveal the mechanistic bases for how mummichogs and other euryhaline species tolerate hypersaline conditions.

Aryl phosphate ester-induced pericardial edema in zebrafish embryos is influenced by the ionic composition of exposure media.

Pericardial edema - fluid accumulation within the pericardium - is a frequently observed malformation in zebrafish embryo-based chemical toxicity screens. We recently discovered that the severity of triphenyl phosphate (TPHP)-induced pericardial edema was dependent on the ionic strength of exposure media. TPHP is an aryl phosphate ester (APE) widely used as a plasticizer and flame retardant. APEs are characterized by having one or more aryl groups bound to a phosphate center, with TPHP containing only unsubstituted aryl groups. Therefore, the objective of this study was to begin investigating whether, similar to TPHP, pericardial edema induced by other structurally related APEs is dependent on the ionic composition of exposure media. We first mined the peer-reviewed literature to identify other APEs that 1) induced pericardial edema in zebrafish embryos within a minimum of three peer-reviewed publications, and 2) demonstrated a statistically significant induction of pericardial edema in at least 70 % of the studies evaluated. Based on this meta-analysis, we identified four other APEs that caused pericardial edema in zebrafish embryos: isopropylated triphenyl phosphate (IPTPP), cresyl diphenyl phosphate (CDP), tricresyl phosphate (TMPP), and 2-ethylhexyl diphenyl phosphate (EDHPHP). Using TPHP as a positive control and pericardial edema as a readout, we developed concentration-response curves for all four APEs based on static exposure from 24 to 72 h post-fertilization (hpf). We then conducted co-exposures with D-Mannitol (an osmotic diuretic) and exposures within reverse osmosis (RO) water determine whether the ionic composition of exposure media mitigated APE-induced pericardial edema at 72 hpf. Using pericardial edema as an endpoint, the approximate EC50s for TPHP (positive control), IPTPP, CDP, TMPP, and EDHPHP were 6.25, 3.125, 3.125, 25, and 100 µM, respectively, based on exposure from 24 to 72 hpf. Interestingly, similar to our findings with TPHP, co-exposure with D-Mannitol and exposure within ion-deficient water significantly mitigated IPTPP- CDP-, TMPP-, and EDHPHP-induced pericardial edema in zebrafish embryos, suggesting that chemically-induced pericardial edema may be 1) dependent on the ionic composition of exposure media and 2) driven by a disruption in osmoregulation across the embryonic epidermis. Therefore, similar to other assay parameters, our findings underscore the need to standardize the osmolarity of exposure media in order to minimize the potential for false positive/negative hits in zebrafish embryo-based chemical toxicity screens conducted around the world.

Physiological responses of euryhaline marine fish to naturally-occurring hypersalinity.

Hypersaline habitats are generally defined as those with salinities in excess of 40 ppt. Well-known hypersaline regions (e.g. salt and soda lakes) have a well-earned reputation for being among the most inhospitable habitats in the world, and fish endemic to these areas have been the subject of much research related to extremophile physiology. Yet, marine coastal hypersalinity is both a common occurrence and a growing consideration in many marine coastal ecosystems, in part owing to human influence (e.g. evaporation, river diversion, desalination effluent). Importantly, any increase in salinity will elevate the osmoregulatory challenges experienced by a fish, which must be overcome by increasing the capacity to imbibe and absorb water and excrete ions. While great attention has been given to dynamic osmoregulatory processes with respect to freshwater to seawater transitions, and to the extreme hypersalinity tolerance that is associated with the adoption of an osmo-conforming strategy, relatively little focus has been placed on the physiological implications of moderate hypersalinity exposures (e.g. ≤ 60 ppt). Importantly, these exposures often represent the threshold of osmoregulatory performance owing to energetic constraints on ion excretion and efficiency limitations on water absorption. This review will explore the current state of knowledge with respect to hypersalinity exposure in euryhaline fishes, while placing a particular focus on the physiological constraints, plasticity and downstream implications of long-term exposure to moderate hypersalinity.

The mayfly Neocloeon triangulifer senses decreasing oxygen availability (PO2) and responds by reducing ion uptake and altering gene expression.

Oxygen availability is central to the energetic budget of aquatic animals and may vary naturally and/or in response to anthropogenic activities. Yet, we know little about how oxygen availability is linked to fundamental processes such as ion transport in aquatic insects. We hypothesized and observed that ion (22Na and 35SO4) uptake would be significantly decreased at O2 partial pressures below the mean Pcrit (5.4 kPa) where metabolic rates (MO2) are compromised, and ATP production is limited. However, we were surprised to observe marked reductions in ion uptake at oxygen partial pressures well above the Pcrit, where MO2 was stable. For example, SO4 uptake decreased by 51% at 11.7kPa, and 82% at the Pcrit (5.4kPa) while Na uptake decreased by 19% at 11.7kPa, and 60% at the Pcrit. Nymphs held for longer time periods at reduced PO2 exhibited stronger reductions in ion uptake rates. Fluids from whole body homogenates exhibited a 29% decrease in osmolality in the most hypoxic condition. The differential expression of atypical guanylyl cyclase (gcy-88e) in response to changing PO2 conditions provides evidence for its potential role as an oxygen sensor. Several ion transport genes (e.g., chloride channel and sodium-potassium ATPase) and hypoxia-associated genes (e.g., ldh and egl-9) were also impacted by decreased oxygen availability. Together, our work suggests that N. triangulifer can sense decreased oxygen availability and perhaps conserves energy accordingly, even when MO2 is not impacted.

Molecular analysis of the reactions in Salicornia europaea to varying NaCl concentrations at various stages of development to better exploit its potential as a new crop plant.

Freshwater scarcity demands exploration of alternative resources like saline water and soils. Understanding the molecular mechanisms behind NaCl regulation in potential crop plants becomes increasingly important for promoting saline agriculture. This study investigated the euhalophyte Salicornia europaea, analyzing its gene expression, yield, and total phenolic compounds under hydroponic cultivation. We employed five salinity levels (0, 7.5, 15, 22.5, and 30 g/L NaCl) across five harvests at 15-day intervals, capturing plant development. Notably, this design deviated from conventional gene expression studies by recording organ-specific responses (shoots and roots) in plants adapted to long-term salinity treatments at various developmental stages. The highest fresh mass of S. europaea was observed four months after germination in 15 g/L NaCl. Identifying a reliable set of reference genes for normalizing gene expression data was crucial due to comparisons across shoots, roots, developmental stages, and salinity levels. A set of housekeeping genes - ubiquitin c (SeUBC), actin (SeActin) and dnaJ-like protein (SeDNAJ) - was identified for this purpose. Interestingly, plants grown without NaCl (0 g/L) displayed upregulation of certain genes associated with a NaCl deficiency related nutritional deprivation. These genes encode a tonoplast Na+/H+-antiporter (SeNHX1), a vacuolar H+-ATPase (SeVHA-A), two H+-PPases (SeVP1, SeVP2), a hkt1-like transporter (SeHKT), a vinorine synthase (SeVinS), a peroxidase (SePerox), and a plasma membrane Na+/H+-antiporter (SeSOS1). Other genes encoding an amino acid permease (SeAAP) and a proline transporter (SeProT) demonstrated marginal or dispersing salinity influence, suggesting their nuanced regulation during plants development. Notably, osmoregulatory genes (SeOsmP, SeProT) were upregulated in mature plants, highlighting their role in salinity adaptation. This study reveals distinct regulatory mechanisms in S. europaea for coping with varying salinity levels. Identifying and understanding physiological reactions and sodium responsive key genes further elucidate the relationship between sodium tolerance and the obligate sodium requirement as a nutrient in euhalophytes.

Unveiling morphophysiological and metabolic adaptive strategies of the CAM epiphytic bromeliad Acanthostachys pitcairnioides to drought.

Ongoing climate changes are expected to intensify drought periods in tropical regions, directly impacting epiphytic bromeliads that depend on intermittent water availability. This study aimed to elucidate if Acanthostachys pitcairnioides, an epiphytic bromeliad of Atlantic Forest, tolerates extended drought periods and the potential strategies involved in its tolerance and recovery capacity. We suppressed irrigation for 42 days, rehydrated plants for four days, and evaluated leaf water status, and photochemical, metabolic, and anatomical changes. During the initial 28 days of drought, translocation of water from hydrenchyma to chlorenchyma, higher chlorophyll content, and accumulation of abscisic and salicylic acid and antioxidants contributed to maintaining the cell turgor and functionality of photosynthetic apparatus. At 42 days, a significant reduction in leaf water content to 45.5% was accompanied by a 2.5-fold increase in non-photochemical quenching and enhanced levels of carotenoids, anthocyanins, osmoregulators (proline, myo-inositol, and trehalose), and phytohormones (abscisic acid and jasmonates). After rewatering, water storage in the hydrenchyma and almost all pigments, hormones, and metabolites were restored to pre-stress conditions. Leaf succulence, carbohydrate and organic acid accumulation, and carbon isotope data (δ13C-14.5‰) provide evidence of induction of CAM metabolism by water limitation in A. pitcairnioides. Our findings indicate the prevalence of water accumulation strategy during the first half of the drought stress. At the end of the drought period, the complete depletion of water from the hydrenchyma favored the osmotic adjustment. Considering this set of tolerance strategies and the rapid recovery after rehydration, A. pitcairnioides can successfully withstand environments with restricted water availability.

The Genome Architecture of the Copepod Eurytemora carolleeae, the Highly Invasive Atlantic Clade of the E. affinis Species Complex.

Copepods are among the most abundant organisms on the planet and play critical functions in aquatic ecosystems. Among copepods, populations of the Eurytemora affinis species complex are numerically dominant in many coastal habitats and are food sources for major fisheries. Intriguingly, certain populations possess the unusual capacity to invade novel salinities on rapid time scales. Despite their ecological importance, high-quality genomic resources have been absent for calanoid copepods, limiting our ability to comprehensively dissect the genome architecture underlying the highly invasive and adaptive capacity of certain populations. Here, we presented the first chromosome-level genome of a calanoid copepod, from the Atlantic clade (Eurytemora carolleeae) of the E. affinis species complex. This genome was assembled using high-coverage long-read and high-throughput chromosome conformation capture sequences of an inbred line, generated through 30 generations of full-sib mating. This genome, consisting of 529.3 megabase (Mb) (contig N50 = 4.2 Mb, scaffold N50 = 140.6 Mb), was anchored onto four chromosomes. Genome annotation predicted 20,262 protein-coding genes, of which ion transporter gene families were substantially expanded based on comparative analyses of 12 additional arthropod genomes. Also, we found genome-wide signatures of historical gene body methylation of the ion transporter genes and the significant clustering of these genes on each chromosome. This genome represents one of the most contiguous copepod genomes to date and among the highest quality marine invertebrate genomes. As such, this genome provides an invaluable resource to help yield fundamental insights into the ability of this copepod to adapt to rapidly changing environments.

Biochemical and molecular responses to long-term salinity challenges in northern quahogs Mercenaria mercenaria.

Salinity is a key environmental factor for aquatic organisms for survival, development, distribution, and physiological performance. Salinity fluctuation occurs often in estuary and coastal zones due to weather, tide, and freshwater inflow and thus heavily affects coastal marine aquaculture. The northern quahog Mercenaria mercenaria is an important aquaculture species along the Atlantic coast in the US, but information on the effects of salinity stress on physiological, immunological, and molecular responses is still scarce. The goal of this study was to investigate cellular and molecular responses through challenges of long-term hypo- and hyper-salinities in northern quahogs. The objectives were to: 1) measure the survival of market-sized quahogs under a three-month salinity challenge at 15 (hyposalinity), 25 (control), and 35 ppt (hypersalinity); 2) determine cellular changes of hemocytes through analysis of immune functions; 3) determine changes of the total free amino acid concentration in gills, and 4) evaluate the molecular responses in gills using RNAseq technology with qPCR verification. After a three-month salinity challenge, no mortality was observed, and increases in body weight were identified with a significantly higher increase in the hypersalinity group. Northern quahogs equilibrated their hemolymph osmolality with the ambient seawater and were verified to be osmoconformers. Significant differences were observed in total hemocyte concentration, lysosomal presence, ROS production, and phagocytic rate, but no differences were found in cell viability. The total free amino acid concentration within gills was positively correlated to water salinity, indicating amino acids were critical organic osmolytes. The transcriptome of gills using RNAseq revealed differential expression genes (DEG) encoding amino acid transporters (SLC6A3, SLC6A6, SLC6A13, SLC25A38), ion channel proteins (T38B1, GluCl, ATP2C1), and water channel protein (AQP8) in hyposalinity or/and hypersalinity groups, indicating these genes play critical roles in intracellular isosmotic regulation. Overall, the findings in this study provided new insights into osmoregulation in northern quahogs.

Regulation of seed soaking with indole-3-butyric acid potassium salt (IBA-K) on rapeseed (Brassica napus L.) seedlings under NaCl stress.

The growth and yield of rapeseed are significantly hampered by salt stress. Indole-3-butyric Acid Potassium Salt (IBA-K) has been found to alleviate the impact of salt stress on plant growth. However, the regulatory effect of IBA-K dipping on salt-stressed rapeseed remains unclear. To explore the implications of IBA-K on the growth and development of rapeseed during the seedling stage, we conducted potting experiments using the Huayouza 62 variety. Five different concentrations of IBA-K for seed soaking (0, 10, 20, 40, 80 mg·L- 1) were tested. The promotional impact of IBA-K on rapeseed demonstrated an initial increase followed by a decline, reaching a peak at 20 mg·L- 1. Therefore, 20 mg·L- 1 was determined as the optimal concentration for subsequent experiments. To further understand the mechanism of IBA-K's action on salt-stressed rapeseed seedlings, we utilized the moderately salt-resistant cabbage rapeseed variety Huayouza 158R and the highly salt-resistant Huayouza 62 as specimens. The investigation focused on their response and repair mechanisms under 150 mmol·L- 1 NaCl stress. The findings demonstrated that compared with the sole NaCl stress, the 20 mg·L- 1 IBA-K seed soaking treatment under salt stress significantly enhanced the plant height, stem diameter, and leaf area of both rapeseed varieties. It also led to greater biomass accumulation, increased chlorophyll content, and improved photosynthetic efficiency in rapeseed. Furthermore, this treatment bolstered the activity of antioxidant enzymes such as superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), while significantly reducing the levels of electrolyte leakage (EL) and malondialdehyde (MDA). Consequently, it alleviated the membrane lipid peroxidation damage induced by NaCl stress, enhanced the accumulation of soluble proteins, maintained cellular osmotic pressure, and effectively mitigated the adverse effects of NaCl stress on rapeseed.

UT-1 Transporter Expression in the Spiny Dogfish (Squalus acanthias): UT-1 Protein Shows a Different Localization in Comparison to That of Other Sharks.

The original UT-1 transporter gene was initially identified in the spiny dogfish (Squalus acanthias), but localization of the UT-1 protein was not determined. Subsequent UT-1 expression was shown to localize to the collecting tubule (CT) of the shark nephron in other shark species, with expression in a closely related chimaera species also located additionally at a lower level in the intermediate-I segment (IS-I) of the nephron. In spiny dogfish, two UT-1 splice variants are known (UT-1 long and short), and there was also a second UT-1 gene described (here termed Brain UT). In this study, a second splice variant of the second Brain UT gene was discovered. Expression profiles (mRNA) of UT-1 long and short and Brain UT were determined in a number of spiny dogfish tissues. Quantitative PCR in kidney samples showed that the level of the short variant of UT-1 was around 100 times higher than the long variant, which was itself expressed around 10 times higher than Brain UT cDNA/mRNA (in kidney). For the long variant, there was a significantly higher level of mRNA abundance in fish acclimatized to 75% seawater. Ultimately, three UT-1 antibodies were made that could bind to both the UT-1 short and long variant proteins. The first two of these showed bands of appropriate sizes on Western blots of around 52.5 and 46 kDa. The second antibody had some additional lower molecular weight bands. The third antibody was mainly bound to the 46 kDa band with faint 52.5 kDa staining. Both the 52.5 and 46 kDa bands were absent when the antibodies were pre-blocked with the peptide antigens used to make them. Across the three antibodies, there were many similarities in localization but differences in subcellular localization. Predominantly, antibody staining was greatest in the intermediate segment 1 (IS-I) and proximal (PIb) segments of the first sinus zone loop of the nephron, with reasonably strong expression also found at the start and middle of the late distal tubule (LDT; second sinus zone loop). While some expression in the collecting tubule (CT) could not be ruled out, the level of staining seemed to be low or non-existent in convoluted bundle zone nephron segments such as the CT. Hence, this suggests that spiny dogfish have a fundamentally different mode of urea absorption in comparison to that found in other shark species, potentially focused more on the nephron sinus zone loops than the CT.

Inhibition of WNK Kinases in NK Cells Disrupts Cellular Osmoregulation and Control of Tumor Metastasis.

INTRODUCTION: The serine/threonine with-no-lysine (WNK) kinase family function in blood pressure control, electrolyte homeostasis, and cellular osmoregulation. These kinases and their downstream effectors are considered promising therapeutic targets in hypertension and stroke. However, the role of WNK kinases in immune cells remains poorly understood. METHODS: Using the small-molecule WNK kinase inhibitors WNK463 and WNK-IN-11, we investigated how WNK kinase inhibition affects natural killer (NK) cell physiology. RESULTS: WNK kinase inhibition with WNK463 or WNK-IN-11 significantly decreased IL-2-activated NK cell volume, motility, and cytolytic activity. Treatment of NK cells with these inhibitors induced autophagy by activating AMPK and inhibiting mTOR signaling. Moreover, WNK kinase inhibition increased phosphorylation of Akt and c-Myc by misaligning activity of activating kinases and inhibitory phosphatases. Treatment of tumor-bearing mice with WNK463 impaired tumor metastasis control by adoptively transferred NK cells. CONCLUSION: The catalytic activity of WNK kinases has a critical role of multiple aspects of NK cell physiology and their pharmacologic inhibition negatively impacts NK cell function.

Another fly diuretic hormone: tachykinins increase fluid and ion transport by adult Drosophila melanogaster Malpighian 'renal' tubules.

Insects such as the model organism Drosophila melanogaster must modulate their internal physiology to withstand changes in temperature and availability of water and food. Regulation of the excretory system by peptidergic hormones is one mechanism by which insects maintain their internal homeostasis. Tachykinins are a family of neuropeptides that have been shown to stimulate fluid secretion from the Malpighian 'renal' tubules (MTs) in some insect species, but it is unclear if that is the case in the fruit fly, D. melanogaster. A central objective of the current study was to examine the physiological role of tachykinin signaling in the MTs of adult D. melanogaster. Using the genetic toolbox available in this model organism along with in vitro and whole-animal bioassays, our results indicate that Drosophila tachykinins (DTKs) function as diuretic hormones by binding to the DTK receptor (DTKR) localized in stellate cells of the MTs. Specifically, DTK activates cation and anion transport across the stimulated MTs, which impairs their survival in response to desiccation because of their inability to conserve water. Thus, besides their previously described roles in neuromodulation of pathways controlling locomotion and food search, olfactory processing, aggression, lipid metabolism and metabolic stress, processing of noxious stimuli and hormone release, DTKs also appear to function as bona fide endocrine factors regulating the excretory system and appear essential for the maintenance of hydromineral balance.

The effects of dissolved organic carbon and model compounds (DOC analogues) on diffusive water flux, oxygen consumption, nitrogenous waste excretion rates and gill transepithelial potential in Pacific sanddab (Citharichthys sordidus) at two salinities.

Many flatfish species are partially euryhaline, such as the Pacific sanddab which spawn and feed in highly dynamic estuaries ranging from seawater to near freshwater. With the rapid increase in saltwater invasion of freshwater habitats, it is very likely that in these estuaries, flatfish will be exposed to increasing levels of dissolved organic carbon (DOC) of freshwater origin at a range of salinities. As salinity fluctuations often coincide with changes in DOC concentration, two natural freshwater DOCs [Luther Marsh (LM, allochthonous) and Lake Ontario (LO, autochthonous) were investigated at salinities of 30 and 7.5 ppt. Optical characterization of the two natural DOC sources indicate salinity-dependent differences in their physicochemistry. LO and LM DOCs, as well as three model compounds [tannic acid (TA), sodium dodecyl sulfate (SDS) and bovine serum albumin (BSA)] representing key chemical moieties of DOC, were used to evaluate physiological effects on sanddabs. In the absence of added DOC, an acute decrease in salinity resulted in an increase in diffusive water flux (a proxy for transcellular water permeability), ammonia excretion and a change in TEP from positive (inside) to negative (inside). The effects of DOC (10 mg C L-1) were salinity and source-dependent, with generally more pronounced effects at 30 than 7.5 ppt, and greater potency of LM relative to LO. Both LM DOC and SDS increased diffusive water flux at 30 ppt but only SDS had an effect at 7.5 ppt. TA decreased ammonia excretion at 7.5 ppt. LO DOC decreased urea-N excretion at both salinities whereas the stimulatory effect of BSA occurred only at 30 ppt. Likewise, the effects of LM DOC and BSA to reduce TEP were present at 30 ppt but not 7.5 ppt. None of the treatments affected oxygen consumption rates. Our results demonstrate that DOCs and salinity interact to alter key physiological processes in marine flatfish, reflecting changes in both gill function and the physicochemistry of DOCs between 30 and 7.5 ppt.

Metagenomic insights into the prokaryotic communities of heavy metal-contaminated hypersaline soils.

Saline soils and their microbial communities have recently been studied in response to ongoing desertification of agricultural soils caused by anthropogenic impacts and climate change. Here we describe the prokaryotic microbiota of hypersaline soils in the Odiel Saltmarshes Natural Area of Southwest Spain. This region has been strongly affected by mining and industrial activity and feature high levels of certain heavy metals. We sequenced 18 shotgun metagenomes through Illumina NovaSeq from samples obtained from three different areas in 2020 and 2021. Taxogenomic analyses demonstrate that these soils harbored equal proportions of archaea and bacteria, with Methanobacteriota, Pseudomonadota, Bacteroidota, Gemmatimonadota, and Balneolota as most abundant phyla. Functions related to the transport of heavy metal outside the cytoplasm are among the most relevant features of the community (i.e., ZntA and CopA enzymes). They seem to be indispensable to avoid the increase of zinc and copper concentration inside the cell. Besides, the archaeal phylum Methanobacteriota is the main arsenic detoxifier within the microbiota although arsenic related genes are widely distributed in the community. Regarding the osmoregulation strategies, "salt-out" mechanism was identified in part of the bacterial population, whereas "salt-in" mechanism was present in both domains, Bacteria and Archaea. De novo biosynthesis of two of the most universal compatible solutes was detected, with predominance of glycine betaine biosynthesis (betAB genes) over ectoine (ectABC genes). Furthermore, doeABCD gene cluster related to the use of ectoine as carbon and energy source was solely identified in Pseudomonadota and Methanobacteriota.

Acute and chronic toxicity of rare earth elements-enriched sediments from a prospective mining area: Effects on life history traits, behavioural and physiological responses of Gammarus fossarum (Crustacea Amphipoda).

Rare earth elements (REE) have an essential role and growing importance in the world's economy. They are attracting interest from society, policymakers, and scientists. The rapidly growing global demand for REE in several strategic industrial and agricultural sectors led many countries to consider the (re)-opening of mining activities for REE extraction. Hence, their increasing use led to the disruption of their biogeochemical cycles with anthropic abnormalities already observed in aquatic ecosystems. Nonetheless, REE remain less studied, and their mechanisms of toxicity actions are not fully understood. As amphipods, Gammarus fossarum represent an important part of the aquatic macroinvertebrate assemblage and are generally used in ecotoxicological studies for their high ecological relevance. However, their use for the study of REE effects has been rather limited so far. The current study aims to assess the potential effects of two naturally REE-enriched sediments (N2 and B4) on G. fossarum. Effects on life history traits, behavioural and physiological responses have been evaluated. Exposing G. fossarum males for 72h to sediments N2 and B4 led to a decrease in haemolymph osmolality and locomotion while an increase in ventilatory activity was observed. Exposing G. fossarum pre-copula pairs with females at the same reproductive stage to the naturally REE-enriched sediments, for one moult cycle duration (∼30 days) showed that sediment B4 led to i) a significant uptake of REE, ii) a significant decrease in the proportion of females with oocytes and iii) a significant reduction in the total number of juveniles. The physicochemical analyses of sediments showed that B4 contains the highest amount of REE with a higher proportion of light REE. The present study gives the first insights into the potential toxicity of REE on G. fossarum as they may have deleterious effects on G. fossarum population's dynamics, which may alter the functioning of aquatic ecosystems.

Inter-subspecies diversity of maize to drought stress with physio-biochemical, enzymatic and molecular responses.

Background: Drought is the most significant factor limiting maize production, given that maize is a crop with a high water demand. Therefore, studies investigating the mechanisms underlying the drought tolerance of maize are of great importance. There are no studies comparing drought tolerance among economically important subspecies of maize. This study aimed to reveal the differences between the physio-biochemical, enzymatic, and molecular mechanisms of drought tolerance in dent (Zea mays indentata), popcorn (Zea mays everta), and sugar (Zea mays saccharata) maize under control (no-stress), moderate, and severe drought stress. Methods: Three distinct irrigation regimes were employed to assess the impact of varying levels of drought stress on maize plants at the V14 growth stage. These included normal irrigation (80% field capacity), moderate drought (50% field capacity), and severe drought (30% field capacity). All plants were grown under controlled conditions. The following parameters were analyzed: leaf relative water content (RWC), loss of turgidity (LOT), proline (PRO) and soluble protein (SPR) contents, membrane durability index (MDI), malondialdehyde (MDA), and hydrogen peroxide (H2O2) content, the antioxidant enzyme activities of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT). Additionally, the expression of heat shock proteins (HSPs) was examined at the transcriptional and translational levels. Results: The effects of severe drought were more pronounced in sugar maize, which had a relatively high loss of RWC and turgor, membrane damage, enzyme activities, and HSP90 gene expression. Dent maize, which is capable of maintaining its RWC and turgor in both moderate and severe droughts, and employs its defense mechanism effectively by maintaining antioxidant enzyme activities at a certain level despite less MDA and H2O2 accumulation, exhibited relatively high drought tolerance. Despite the high levels of MDA and H2O2 in popcorn maize, the up-regulation of antioxidant enzyme activities and HSP70 gene and protein expression indicated that the drought coping mechanism is activated. In particular, the positive correlation of HSP70 with PRO and HSP90 with enzyme activities is a significant result for studies examining the relationships between HSPs and other stress response systems. The discrepancies between the transcriptional and translational findings provide an opportunity for more comprehensive investigations into the role of HSPs in stress conditions.

Molecular pathways of osmoregulation in response to salinity stress in the gills of the scalloped spiny lobster (Panulirus homarus) within survival salinity.

Scalloped spiny lobster (Panulirus homarus) aquaculture is the preferred strategy to resolve the conflict between supply and demand for lobster. Environmental conditions, such as salinity, are key to the success of lobster aquaculture. However, physiological responses of P. homarus to salinity stress have not been well studied. This study investigated the gill histology, osmoregulation and gill transcriptome of the early juvenile P. homarus (weight 19.04 ± 3.95 g) cultured at salinity 28 (control), 18, and 38 for 6 weeks. The results showed that the gill filaments of P. homarus exposed to low salinity showed severe separation of the cuticle and epithelial cells due to water absorption and swelling, as well as the dissolution and thinning of the cuticle and the rupture of the septum that separates the afferent and efferent channels. The serum osmolarity of P. homarus varied proportionately with external medium salinity and remained consistently above ambient osmolarity. The serum Na+, Cl-, K+, and Mg2+ concentrations P. homarus exhibited a pattern similar to that of serum osmolality, while the concentration of Ca2+ remained unaffected at salinity 18 but significantly increased at salinity 38. Gill Na+/K+-ATPase activity of P. homarus increased (p < 0.05) under the both salinity stress. Salinity 18 significantly increased Glutamate dehydrogenase (GDH) and Glutamicpyruvic transaminase (GPT) activity in the hepatopancreas of P. homarus (p < 0.05). According to transcriptome analysis, versus control group (salinity 28), 929 and 1095 differentially expressed genes (DEGs) were obtained in the gills of P. homarus at salinity 18 and 38, respectively, with these DEGs were mainly involved in energy metabolism, transmembrane transport and oxidative stress and substance metabolism. In addition, the expression patterns of 8 key DEGs mainly related to amino acid metabolism, transmembrane transport and oxidative stress were verified by quantitative real-time PCR (RT-qPCR). The present study suggests that salinity 18 has a greater impact on P. homarus than salinity 38, and P. homarus demonstrates effective osmoregulation and handle with salinity fluctuations (18 to 38) through physiological and functional adaptations. This study provides an improved understanding of the physiological response strategies of P. homarus facing salinity stress, which is crucial for optimizing aquaculture practices for this species.

Hyposalinity elicits physiological responses and alters intestinal microbiota in Korean rockfish Sebastes schlegelii.

Global warming significantly impacts aquatic ecosystems, with changes in the salt environment negatively affecting the physiological responses of fish. We investigated the impact of hyposalinity on the physiological responses and intestinal microbiota of Sebastes schlegelii under the context of increased freshwater influx due to climate change. We focused on the osmoregulatory capacity, oxidative stress responses, and alterations in the intestinal microbiome of S. schlegelii under low-salinity conditions. Our findings revealed compromised osmoregulatory capacity in S. schlegelii under low-salinity conditions, accompanied by the activation of oxidative stress responses, indicating physiological adaptations to cope with environmental stress. Specifically, changes in Na+/K+-ATPase (NKA) activity in gill tissues were associated with decreased osmoregulatory capacity. Furthermore, the analysis of the intestinal microbiome led to significant changes in microbial diversity. Exposure to low-salinity environments led to dysbiosis, with notable decreases in the relative abundance of Gammaproteobacteria at the class level and specific genera such as Enterovibrio, and Photobacterium. Conversely, Bacilli classes, along with genera like Mycoplasma, exhibited increased proportions in fish exposed to low-salinity conditions. These findings underscore the potential impact of environmental salinity changes on the adaptive capacity of fish species, particularly in the context of aquaculture. Moreover, they highlight the importance of considering both physiological and microbial responses in understanding the resilience of aquatic organisms to environmental stress. Additionally, they highlight the importance of intestinal microbiota analyses in understanding the immune system and disease management in fish.

Physiological responses of the invasive blue crabs Callinectes sapidus to salinity variations: Implications for adaptability and invasive success.

This study provides a comprehensive analysis of the eco-physiological responses of the blue crab (Callinectes sapidus) to variations in salinity, shedding light on its adaptability and invasive success in aquatic environments. Gender-specific differences in osmoregulation and Electron Transport System (ETS) activity highlight the importance of considering sex-specific aspects when understanding the physiological responses of invasive species. Females exhibited increased ETS activity at lower salinities, potentially indicative of metabolic stress, while males displayed constant ETS activity across a range of salinities. Osmoregulatory capacity which depended on gender and salinity, was efficient within meso-polyhaline waters but decreased at higher salinities, particularly in males. These findings provide valuable understandings into how C. sapidus specimens in an invaded area responds to salinity changes, important for considerate its distribution through saline pathways during tidal cycle fluctuations. This study shows the importance of interdisciplinary research for effective management of invasive species and conservation of affected aquatic ecosystems.

Roles and occurrences of microbiota in the osmoregulatory organs, gills and gut, in marine medaka upon hypotonic stress.

Gills and gut are the two primary osmoregulatory organs in fish. Recently, studies have expanded beyond the osmoregulatory mechanisms of these organs to explore the microbiota communities inhabiting them. It is now known that microbial communities in both organs shift in response to osmotic stress. However, there are limited studies identifying the major contributors and co-occurrence among these microbiota in both organs under seawater and freshwater transfer conditions. The current data mining report performed a bioinformatics analysis on two previous published datasets from our group, aiming to provide insights into host-bacteria relationships under osmotic stress. We divided the samples into four groups: control seawater gills (LSW); control seawater gut (TSW); freshwater transfer gills (LFW); and freshwater transfer gut (TFW). Our results showed that LSW had higher diversities, richness, and evenness compared to TSW. However, both the LFW and LSW did not show any significant differences after the freshwater transfer experiment. We further applied co-occurrence network analysis and, for the first time, reported on the interactions of taxa shaping the community structure in these two organs. Moreover, we identified enriched ectoine biosynthesis in seawater samples, suggesting its potential role in seawater environments. Increased mRNA expression levels of Na+/K+-atpase, and cftr, were observed in gills after 6 h of ectoine treatment. These findings provide a foundation for future studies on host-bacteria interactions under osmotic stress.