Integrated mRNA and miRNA analysis reveals the regulatory network of oxidative stress and inflammation in Coilia nasus brains during air exposure and salinity mitigation.

BACKGROUND: Air exposure is an inevitable source of stress that leads to significant mortality in Coilia nasus. Our previous research demonstrated that adding 10‰ NaCl to aquatic water could enhance survival rates, albeit the molecular mechanisms involved in air exposure and salinity mitigation remained unclear. Conversely, salinity mitigation resulted in decreased plasma glucose levels and improved antioxidative activity. To shed light on this phenomenon, we characterized the transcriptomic changes in the C. nasus brain upon air exposure and salinity mitigation by integrated miRNA-mRNA analysis. RESULTS: The plasma glucose level was elevated during air exposure, whereas it decreased during salinity mitigation. Antioxidant activity was suppressed during air exposure, but was enhanced during salinity mitigation. A total of 629 differentially expressed miRNAs (DEMs) and 791 differentially expressed genes (DEGs) were detected during air exposure, while 429 DEMs and 1016 DEGs were identified during salinity mitigation. GO analysis revealed that the target genes of DEMs and DEGs were enriched in biological process and cellular component during air exposure and salinity mitigation. KEGG analysis revealed that the target genes of DEMs and DEGs were enriched in metabolism. Integrated analysis showed that 24 and 36 predicted miRNA-mRNA regulatory pairs participating in regulating glucose metabolism, Ca2+ transport, inflammation, and oxidative stress. Interestingly, most of these miRNAs were novel miRNAs. CONCLUSION: In this study, substantial miRNA-mRNA regulation pairs were predicted via integrated analysis of small RNA sequencing and RNA-Seq. Based on predicted miRNA-mRNA regulation and potential function of DEGs, miRNA-mRNA regulatory network involved in glucose metabolism and Ca2+ transport, inflammation, and oxidative stress in C. nasus brain during air exposure and salinity mitigation. They regulated the increased/decreased plasma glucose and inhibited/promoted antioxidant activity during air exposure and salinity mitigation. Our findings would propose novel insights to the mechanisms underlying fish responses to air exposure and salinity mitigation.

Cortisol enhances aerobic metabolism and locomotor performance during the transition to land in an amphibious fish.

Amphibious fishes on land encounter higher oxygen (O2) availability and novel energetic demands, which impacts metabolism. Previous work on the amphibious mangrove killifish (Kryptolebias marmoratus) has shown that cortisol becomes elevated in response to air exposure, suggesting a possible role in regulating metabolism as fish move into terrestrial environments. We tested the hypothesis that cortisol is the mechanism by which oxidative processes are upregulated during the transition to land in amphibious fishes. We used two groups of fish, treated fish (+metyrapone, a cortisol synthesis inhibitor) and control (-metyrapone), to determine the impact of cortisol during air exposure (0 and 1 h, 7 days) on O2 consumption, terrestrial locomotion, the phenotype of red skeletal muscle, and muscle lipid concentration. Metyrapone-treated fish had an attenuated elevation in O2 consumption rate during the water to air transition and an immediate reduction in terrestrial exercise performance relative to control fish. In contrast, we found no short- (0 h) or long-term (7 days) differences between treatments in the oxidative phenotype of red muscles, nor in muscle lipid concentrations. Our results suggest that cortisol stimulates the necessary increase in aerobic metabolism needed to fuel the physiological changes that amphibious fishes undergo during the acclimation to air, although further studies are required to determine specific mechanisms of cortisol regulation.

Scaling features in high-concentrations PM2.5 evolution: the Ignored factor affecting scarlet fever incidence.

As an acute respiratory disease, scarlet fever has great harm to public health. Some evidence indicates that the time distribution pattern of heavy PM2.5 pollution occurrence may have an impact on health risks. This study aims to reveal the relation between scaling features in high-concentrations PM2.5 (HC-PM2.5) evolution and scarlet fever incidence (SFI). Based on the data of Hong Kong from 2012 to 2019, fractal box-counting dimension (D) is introduced to capture the scaling features of HC-PM2.5. It has been found that index D can quantify the time distribution of HC-PM2.5, and lower D values indicate more cluster distribution of HC-PM2.5. Moreover, scale-invariance in HC-PM2.5 at different time scales has been discovered, which indicates that HC-PM2.5 occurrence is not random but follows a typical power-law distribution. Next, the exposure-response relationship between SFI and scale-invariance in HC-PM2.5 is explored by Distributed lag non-linear model, in conjunction with meteorological factors. It has been discovered that scale-invariance in HC-PM2.5 has a nonlinear effect on SFI. Low and moderate D values of HC-PM2.5 are identified as risk factors for SFI at small time-scale. Moreover, relative risk shows a decreasing trend with the increase of exposure time. These results suggest that exposure to short-term clustered HC-PM2.5 makes individual more prone to SFI than exposure to long-term uniform HC-PM2.5. This means that individuals in slightly-polluted regions may face a greater risk of SFI, once the PM2.5 concentration keeps rising. In the future, it is expected that the relative risk of scarlet fever for a specific region can be estimated based on the quantitative analysis of scaling features in high-concentrations PM2.5 evolution.

Emersion and recovery alter oxygen consumption, ammonia and urea excretion, and oxidative stress parameters, but not diffusive water exchange or transepithelial potential in the green crab (Carcinus maenas).

The green crab (Carcinus maenas) is an inshore species affected by intertidal zonation patterns, facing periods of emersion during low tide and submersion during high tide. During these periods of air and subsequent water exposure, these species can face physiological challenges. We examined changes in O2 consumption rate (MO2), and ammonia and urea excretion rates over sequential 14 h periods in seawater (32 ppt, control), in air and during recovery in seawater after air exposure (13°C throughout). At the end of each exposure, the anterior (5th) and posterior (8th) gills and the hepatopancreas were removed for measurements of oxidative stress parameters (TBARs and catalase in the gills and hepatopancreas, and protein carbonyls in the gills). MO2 remained unchanged during air exposure, but increased greatly (3.4-fold above control levels) during the recovery period. Ammonia and urea net fluxes were reduced by 98% during air exposure, but rebounded during recovery to >2-fold the control rates. Exchangeable water pools, rate constants of diffusive water exchange, unidirectional diffusive water flux rates (using tritiated water) and transepithelial potential were also measured during control and recovery treatments, but exhibited no significant changes. Damage to proteins was not observed in either gill. However, lipid damage occurred in the anterior (respiratory) gill after the air exposure but not in the posterior (ionoregulatory) gill or hepatopancreas. Catalase activity also decreased significantly in recovery relative to levels during air exposure in both the anterior gill and hepatopancreas, but not in the posterior gill. The crabs did not modify water metabolism or permeability. We conclude that MO2 was maintained but not enhanced during air exposure, while ammonia and urea-N excretion were impaired. As a result, all of these parameters increase greatly during re-immersion recovery, and oxidative stress also occurs. Clearly, emersion is not without physiological costs.

ROS-induced moderate autophagy of haemocytes confers resistance of Mercenaria mercenaria to air exposure stress.

Air exposure (AE) is a significant environmental stressor that can lead to desiccation, hypoxia, starvation, and disruption of cellular homeostasis in marine bivalves. Autophagy is a highly conserved catabolic pathway that facilitates the degradation of damaged macromolecules and organelles, thereby supporting cellular stress responses. To date, autophagy-mediated resistance mechanisms to AE stress remain largely elusive in bivalves. In this study, we performed a multi-tool approach to investigate the autophagy-related physiological regulation in hard clams (Mercenaria mercenaria) under different duration of AE (T = 0, 1, 5, 10, 20, 30 days). We observed that autophagy of haemocytes was significantly activated on day 5. However, autophagy activity began to significantly decline from day 10 to day 30. Autophagy was significantly inhibited after antioxidant treatment, indicating that reactive oxygen species (ROS) was an endogenous inducer of autophagy. A significant decline in the survival rate of hard clams was observed after injection of ammonium chloride or carbamazepine during AE stress, suggesting that moderate autophagy was conducive for clam survival under AE stress. We also observed DNA breaks and high levels of apoptosis in haemocytes on day 10. Activation of apoptosis lagged behind autophagy, and the relationship between autophagy and apoptosis might shift from antagonism to synergy with the duration of stress. This study provides novel insights into the stress resistance mechanisms in marine bivalves.

The effect of air exposure and re-water on gill microstructure and molecular regulation of Pacific white shrimp Penaeus vannamei.

The Penaeus vannamei is an important shrimp species with enormous commercial and ecological values. In production process, the air exposure resistance is vital for live transportation without water. We tested the air exposure resistant ability of P. vannamei, and carried out gill histological observation and gene expression analysis. The physiology and molecular response to the air exposure stress of P. vannamei was revealed. We found that body weight could affect the air exposure tolerance. Air exposure caused epithelial cell of gill filament shrinking and tissue fluid exudation within half of hour, and triggered oxidative stress response. After retrieved to water, epithelial cell shrinking and tissue fluid exudation recovered gradually, but oxidative and antioxidant response is still going on. Organisms reduced oxidative stress by regulating levels of antioxidants and antioxidant enzymes that remove reactive oxygen species (ROS) and RNA and DNA processing to repair tissue damage, and expression of apoptosis associated-genes altered. Furthermore, the survive shrimps could live steadily more than 5 days, and their gill filament recovered to normal state, proving that the damage of air exposure is reversible. These findings could be considered in the waterless live transportation of P. vannamei.

New insights into the regulation mechanism of red claw crayfish (Cherax quadricarinatus) hepatopancreas under air exposure using transcriptome analysis.

Red claw crayfish (Cherax quadricarinatus) is an important freshwater shrimp species worldwide with enormous economic value. Waterless transportation is an inherent feature of red claw crayfish transportation. However, the high mortality of red claw crayfish is a severe problem in the aquaculture of crayfish after waterless transportation. In this study, we investigated the responses of the hepatopancreas from the red claw crayfish undergoing air exposure stress and normal conditions on transcriptome levels. We used Illumina-based RNA sequencing (RNA-Seq) to perform a transcriptome analysis from the hepatopancreas of red claw crayfish challenged by air exposure. An average of 57,148,800 clean reads per library was obtained, and 33,567 unigenes could be predicted and classified according to their homology with matches in the National Center for Biotechnology Information (NCBI) non-redundant protein sequences (Nr), Gene Ontology (GO), a manually annotated and reviewed protein sequence database (Swiss-Prot), protein families (Pfam), Clusters of Orthologous Groups (COG) of proteins, and Kyoto Encyclopedia of Genes and Genomes (KEGG) databases. 690 and 3407 differentially expressed genes (DEGs) were identified between the two stress stages of the red claw crayfish. More DEGs were identified in 12 h, indicating that gene expressions were largely changed at 12 h. Some immune-related pathways and genes were identified according to KEGG and GO enrichment analysis. A total of 12 DEGs involved in immune response and trehalose mechanism were verified by quantitative real-time-polymerase chain reaction (qRT-PCR). The results indicated that the red claw crayfish might counteract the stress of air exposure at the transcriptomic level by increasing expression levels of antioxidant-, immune-, and trehalose metabolism-related genes. These transcriptome results from the hepatopancreas provide significant insights into the influence mechanism of air exposure to the trehalose mechanism and immune response in the red claw crayfish.

Untargeted GC-MS metabolomics reveals the metabolic responses in the gills of Chinese mitten crab (Eriocheir sinensis) subjected to air-exposure stress.

Desiccation is a common stress experienced by crabs during aquaculture and transportation. In China, the crustacean, Chinese mitten crab (Eriocheir sinensis), is economically important. However, little is known about the molecular pathways underlying physiological stress. Here, by using untargeted gas chromatography-mass spectrometry metabolomics, we investigated the metabolic responses of the gills of E. sinensis subjected to air-exposure stress by six biological replicates of the control group (CG) and the air-exposure stress group (AG). Metabolomic analysis identified 43 differential metabolites in the AG versus the CG that could be potential biomarkers of desiccation stress. In addition, integrated analysis of key metabolic pathways revealed the involvement of histidine metabolism; glycine, serine and threonine metabolism; the pentose phosphate pathway; the citrate cycle (TCA cycle); and nicotinate and nicotinamide metabolism. These findings indicated the special physiological responses to air-exposure stresses in this species.

RNA-Seq analysis and WGCNA reveal dynamic molecular responses to air exposure in the hard clam Mercenaria mercenaria.

Intertidal bivalves are constantly exposed to air due to daily and seasonal tidal cycles. The hard clam Mercenaria mercenaria is an economically important bivalve species and often subjected to air exposure for more than 10 days during long-distance transportation. Hard clam exhibits remarkable tolerance to air exposure. In this study, we performed RNA sequencing on hemocytes of M. mercenaria exposed to air for 0, 1, 5, 10, 20 and 30 days. The overall and dynamic molecular responses of hard clams to air exposure were revealed by different transcriptomic analysis strategies. As a result, most cytochrome P450 1A and 3A, and monocarboxylate transporter family members were up-regulated during air exposure. Additionally, the dominant molecular process in response to 5-d, 10-d, 20-d and 30-d air exposure was refolding of misfolded proteins in endoplasmic reticulum, lysosome-mediated degradation of phospholipids, protein metabolism and reorganization of cytoskeleton, and activation of anti-apoptotic process, respectively. Our results facilitated comprehensive understanding of the tolerance mechanisms of intertidal bivalves to air exposure.

An accidental discovery of mannan-oligosaccharide's protection effect against air exposure and its potential mechanism in hybrid grouper (Epinephelus lanceolatus ♂ × Epinephelus fuscoguttatus ♀).

The original objective was to explore the potential benefiting effects of three prebiotics in hybrid grouper (Epinephelus lanceolatus♂ × Epinephelus fuscoguttatus♀). Therefore, three experimental diets (basal diet + 1% fructooligosaccharide, Diet F; basal diet + 1% inulin, Diet I; basal diet + 0.3% mannan-oligosaccharide, Diet M) and one basal diet (Diet C) were prepared and a feeding trial was conducted. However, at the end of the fourth week into the feeding experiment, a water-leaking accident occurred and fishes of all groups went through an unexpected air exposure event. Surprisingly, different prebiotic-supplemented groups showed significantly different air exposure tolerance: the mortality of M group was significantly lower (P ≤ 0.05) than all the other groups. Examination of antioxidant, non-specific immunity, and stress parameters revealed that comparing to control group, M group showed significantly increased catalase (CAT), acid phosphatase (ACP), and alkaline phosphatase (AKP) activities, decreased superoxide dismutase (SOD) activity, and similar cortisol level (P ≤ 0.05). Real-time PCR experiment revealed that M group significantly increased the expression of CAT, glutathione peroxidase (GPx), and manganese superoxide dismutase (MnSOD) genes in head kidney (P ≤ 0.05). Overall, M exhibited the best anti-air exposure/antioxidative stress effects among the three prebiotics and could be considered a promising feed additive to relieve air exposure/oxidative stress in hybrid grouper culture.

Acclimation to tidal conditions alters the physiological responses of the green shore crab, Carcinus maenas, to subsequent emersion.

Animals inhabiting the intertidal zone are exposed to abrupt changes in environmental conditions associated with the rise and fall of the tide. For convenience, the majority of laboratory studies on intertidal organisms have acclimated individuals to permanently submerged conditions in seawater tanks. In this study, green shore crabs, Carcinus maenas, were acclimated to either a simulated tidal regime of continuous emersion-immersion ('tidal') or to permanently submerged conditions ('non-tidal') to assess their physiological responses to subsequent emersion. Tidal crabs exhibited an endogenous rhythm of oxygen consumption during continuous submersion with lower oxygen consumption during periods of anticipated emersion, which was not detected in non-tidal crabs. During emersion, tidal crabs were able to buffer apparent changes in acid-base balance and exhibited no change in venous pH, whereas non-tidal crabs developed an acidosis associated with a rise in lactate levels. These results indicate that tidal crabs were better able to sustain aerobic metabolism and had lower metabolic costs during emersion than non-tidal crabs. It is likely that the elevated levels of haemocyanin exhibited by tidal crabs allowed them to maintain oxygen transport and buffer pH changes during emersion. This suggests that acclimation of C. maenas to submerged conditions results in a loss of important physiological mechanisms that enable it to tolerate emersion. The results of this study show that caution must be taken when acclimating intertidal organisms to submerged conditions in the laboratory, as it may abolish important physiological responses and adaptations that are critical to their performance when exposed to air.

Tambaqui (Colossoma macropomum) acclimated to different tropical waters from the Amazon basin shows specific acute-stress responses.

Stress responses in teleosts include the release of hormones into the bloodstream. Their effects depend on the species and on the environmental conditions. The Amazon basin collects waters of diverse chemical composition, and some fish are able to inhabit several of them. However, the effects of these waters on the stress axis are still unknown. Here we show how acute air-exposure differently affects stress biomarkers in tambaqui (Colossoma macropomum), a tropical model species, when acclimated to two Amazonian waters (Rio Negro -RN- water rich in humic acids and poor in ions, and groundwater -IG- with no humic acids and higher concentration of ions). This study described primary and secondary stress responses after air exposure including plasma cortisol, energy metabolites, pH and ions, skin mucus energy metabolites, as well as gills and kidney Na+/K+-ATPase and Na+/H+-exchanger (NHE) activities. Several differences were described in these stress biomarkers due to the acclimation water. The most remarkable ones include increased mucus glucose only in RN-fish, and mucus lactate only in IG-fish after air exposure. Moreover, an inverse relationship between plasma cortisol and Na+ concentrations as well as a direct relationship between plasma ammonia and branchial NHE activity were observed only in RN-fish. Our results demonstrate how important is to study stress responses in fish acclimated to different environments, as physiological differences can be magnified during episodes of high energy expenditure. In addition to having a direct application in aquaculture, this study will improve the management of critical ecosystems such as the Amazon.

High temperature acclimation alters the emersion behavior in the crab Neohelice granulata.

An increase in environmental temperature can deleteriously affect organisms. This study investigated whether the semiterrestrial estuarine crab Neohelice granulata uses emersion behavior as a resource to avoid thermal stress and survive higher aquatic temperatures. We also examined whether this behavior is modulated by exposure to high temperature; whether, during the period of emersion, the animal loses heat from the carapace to the medium; and whether this behavior is altered by the temperature at which the animal has been acclimated. The lethal temperature for 50% of the population (LT50) was determined through 96-h mortality curves in animals acclimated at 20 °C and 30 °C. The behavioral profile of N. granulata during thermal stress was based on monitoring crab movement in aerial, intermediary, and aquatic zones. Acclimation at a higher temperature and the possibility of emersion increased the thermotolerance of the crabs and the synergistic effect of acclimation temperature. The possibility of leaving the hot water further increased the resistance of these animals to thermal stress. We observed that when the crab was subjected to thermal stress conditions, it spent more time in the aerial environment, unlike under control conditions. Under the experimental conditions, it made small incursions into the aquatic environment and stayed in the aerial environment for a longer time in order to cool its body temperature. The animals acclimated at 20 °C and placed into water at 35 °C remained in the aerial zone. The animals acclimated and maintained at 30 °C (control) that were placed in water at 35 °C with the possibility of emerging into hot air transited more frequently between the aquatic and aerial zones than did the animals that were put in water at 35 °C with the possibility of emerging into a cooler air environment. We conclude that emergence behavior allows N. granulata to survive high temperatures and that this behavior is influenced by acclimation temperature.

The role of autophagy in the pathogenesis of exposure keratitis.

Incomplete tear film spreading and eyelid closure can cause defective renewal of the ocular surface and air exposure-induced epithelial keratopathy (EK). In this study, we characterized the role of autophagy in mediating the ocular surface changes leading to EK. Human corneal epithelial cells (HCECs) and C57BL/6 mice were employed as EK models, respectively. Transmission electron microscopy (TEM) evaluated changes in HCECs after air exposure. Each of these models was treated with either an autophagy inhibitor [chloroquine (CQ) or 3-methyladenine (3-MA)] or activator [Rapamycin (Rapa)]. Immunohistochemistry assessed autophagy-related proteins, LC3 and p62 expression levels. Western blotting confirmed the expression levels of the autophagy-related proteins [Beclin1 and mammalian target of rapamycin (mTOR)], the endoplasmic reticulum (ER) stress-related proteins (PERK, eIF2α and CHOP) and the PI3K/Akt/mTOR signalling pathway-related proteins. Real-time quantitative PCR (qRT-PCR) determined IL-1ß, IL-6 and MMP9 gene expression levels. The TUNEL assay detected apoptotic cells. TEM identified autophagic vacuoles in both EK models. Increased LC3 puncta formation and decreased p62 immunofluorescent staining and Western blotting confirmed autophagy induction. CQ treatment increased TUNEL positive staining in HCECs, while Rapa had an opposite effect. Similarly, CQ injection enhanced air exposure-induced apoptosis and inflammation in the mouse corneal epithelium, which was inhibited by Rapa treatment. Furthermore, the phosphorylation status of PERK and eIF2α and CHOP expression increased in both EK models indicating that ER stress-induced autophagy promoted cell survival. Taken together, air exposure-induced autophagy is indispensable for the maintenance of corneal epithelial physiology and cell survival.

Identification of immune-related genes in gills of Chinese mitten crabs (Eriocheir sinensis) during adaptation to air exposure stress.

The Chinese mitten crab, Eriocheir sinensis, is the most important crab in China. Air exposure is regarded as one of the crucial restriction factors in the crab cultivation and transportation process. Numerous studies have shown that air exposure stress can cause many negative effects on aquatic farming animals. However, the molecular mechanisms of drying on Chinese mitten crabs are still poorly studied. In this study, gill reference transcriptome was assembled and differentially expressed gene (DGE) analysis was conducted between air exposure 16 h and normal dissolved oxygen of Chinese mitten crab. A total of 76075 transcripts were generated and 50800 unigenes with a mean length of 1090 bp and N50 length of 1584 bp were observed. Transcriptomic comparison revealed 352 DEGs between air exposure 16 h group and control group, including 122 up-regulated genes and 230 down-regulated genes. Gene ontology (GO) analysis revealed that these DEGs involved in 16 biological process subcategories, 8 cellular component subcategories and 6 molecular function subcategories. Further Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis annotated 352 DEGs to 85 pathways, and some pathways were regarded as related with immune system and diseases, such as phagosome, systemic lupus erythematosus, and alcoholism. Eight genes involved in multiple KEGG signaling pathways were validated by qRT-PCR. This study demonstrates the first gill transcriptomic analysis challenged with air exposure stress in Chinese mitten crab and provides valuable gene resources for understanding the crab gill immunity, which can provides insight into the immune response of crab against air exposure stress.

Cortisol responses of goldfish (Carassius auratus) to air exposure, chasing, and increased water temperature.

Fish can respond to stimuli from the internal or external environment with activation of the hypothalamo-pituitary-interrenal (HPI) axis and the secretion of cortisol. Stimuli that activate the HPI axis of fish include short term air exposure and increases in water temperature. The present study was conducted to determine how quickly cortisol concentrations increase in goldfish subjected to an increase in water temperature, and to compare the response to an increase in water temperature with responses to other stimuli. Plasma cortisol concentrations varied widely between individual goldfish, with concentrations ranging from 9.1 to 516.0 ng/mL in goldfish on the day of arrival from the supplier. Mean cortisol concentrations in undisturbed goldfish were low (4.5 ±â€¯1.0 ng/mL). Mean cortisol concentrations in fish exposed to air for 3 min and in fish that experienced chasing for 10 min were markedly elevated 15 min after the beginning of the stimuli (132.6 ±â€¯31.0 and 121.1 ±â€¯23.9 ng/mL respectively). Mean cortisol concentrations in fish that experienced an increase in water temperature rose to 22.2 ±â€¯7.6 ng/mL after 15 min, declined to <10 ng/mL at 30 and 60 min then increased and were elevated (79.0 ±â€¯10.8 ng/mL) at 240 min. Cortisol measurements can be used to indicate the responsiveness of fish to changes in water temperature and goldfish will be a convenient study species for the development of studies of plasticity in responses of fish to increases in water temperature that are happening due to climate change.

The oxidative stress and antioxidant responses of Litopenaeus vannamei to low temperature and air exposure.

Low temperature and air exposure were the key attributes for waterless transportation of fish and shrimp. In order to investigate the oxidative stress and antioxidant responses of the live shrimp Litopenaeus vannamei in the mimic waterless transportation, live shrimp were cooled at 13 °C for 3 min, stored in oxygen at 15 °C for 12 h, and then revived in water at 25 °C. The survival rate of shrimp under this waterless transportation system was over 86.67%. The ultrastructure of hepatopancreas cells were observed while activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), glutathione peroxidase (GSH-Px), antisuperoxide anion free radicals (ASAFR), total antioxidant capacity (TAOC), reactive oxygen species (ROS) production, content of malondialdehyde (MDA) and relative mRNA expressions of CAT and GSH-Px in the hemolymph and hepatopancreas were determined. Slight distortions of some organelles in hepatopancreas cells was reversible upon the shrimp revived from the cold shock. The activities of SOD, POD, CAT, GSH-Px, TAOC, ROS production and relative mRNA expressions of CAT and GSH-Px increased following the cold shock and reached peak levels after 3 or 6 h of storage, and then decreased gradually. There was no significant difference between the fresh and the revived shrimp in SOD, POD, GSH-Px, TAOC, ROS, MDA and relative mRNA expressions of CAT and GSH-Px. The oxidative stress and antioxidant responses were tissue-specific because hepatopancreas seemed to have a greater ability to defend against organelle damage and was more sensitive to stress than hemolymph based on the results of SOD activity, MDA content and GSH-Px mRNA expression. These results revealed that low temperature and air exposure caused significant oxidative and antioxidant responses, but did not lead to irreversible damages in this waterless system.

Neglected exposure route: cobalt on skin and its associations with urinary cobalt levels.

OBJECTIVES: Cobalt (Co) exposure is associated with adverse health effects including skin sensitisation, asthma and interstitial lung fibrosis. Exposure to Co in industrial settings is often assessed using air samples or biomonitoring in urine. Skin exposure is rarely measured. Aim of this study was to quantify and compare the importance of Co skin exposure and respiratory exposure in determining urinary Co concentrations. METHODS: Co skin exposure was measured in 76 hard metal workers by acid wipe sampling before and at the end of work shifts. Spot urine was collected during a 24-hour period from the start of a shift. Respiratory exposure was measured by personal inhalable dust sampling during a shift in 30 workers. Co was analysed by inductively coupled plasma mass spectrometry. RESULTS: Quantile regression modelling showed that a doubling of Co on skin before or at the end of shift increased the median urinary concentration of Co by 70% (p<0.001) or 32% (p<0.001), respectively. A doubling of Co in air increased median urinary Co by 38% (p<0.001). Co skin exposures were still significantly associated with urinary Co after excluding a group of workers with high respiratory exposure (33%, p=0.021 and 17%, p=0.002). CONCLUSIONS: The results indicate an association between Co skin exposure and urinary Co concentrations. This should be considered when using urinary Co as a biomarker of exposure.

The modulation role of serotonin in Pacific oyster Crassostrea gigas in response to air exposure.

Serotonin, also known as 5-hydroxytryptamine (5-HT), is a critical neurotransmitter in the neuroendocrine-immune regulatory network and involved in regulation of the stress response in vertebrates and invertebrates. In the present study, serotonin was found to be widely distributed in the tissues of Pacific oyster Crassostrea gigas, including haemolymph, gonad, visceral ganglion, mantle, gill, labial palps and hepatopancreas, and its concentration increased significantly in haemolymph and mantle after the oysters were exposed to air for 1 d. The apoptosis rate of haemocytes was significantly declined after the oysters received an injection of extra serotonin, while the activity of superoxide dismutase (SOD) in haemolymph increased significantly. After the stimulation of serotonin during air exposure, the apoptosis rate of oyster haemocytes and the concentration of H2O2 in haemolymph were significantly decreased, while the SOD activity was significantly elevated. Furthermore, the survival rate of oysters from 4th to 6th d after injection of serotonin was higher than that of FSSW group and air exposure group. The results clearly indicated that serotonin could modulate apoptotic effect and redox during air exposure to protect oysters from stress.

Acute exposure to high environmental ammonia (HEA) triggers the emersion response in the green shore crab.

The physiological effects of high environmental ammonia (HEA) exposure have been well documented in many aquatic species. In particular, it has recently been demonstrated that exposure to ammonia in fish leads to a similar hyperventilatory response as observed during exposure to hypoxia. In littoral crabs, such as the green crab (Carcinus maenas), exposure to severe hypoxia triggers an emersion response whereby crabs escape hypoxia to breathe air. We hypothesized that exposure to HEA in green crabs would lead to a similar behavioural response which is specific to ammonia. Using an experimental arena containing a rock bed onto which crabs could emerse, we established that exposure to HEA (4mmol/l NH4HCO3) for 15min triggers emersion in crabs. In experiments utilizing NaHCO3 controls and NH4HCO3 injections, we further determined that emersion was triggered specifically by external ammonia and was independent of secondary acid-base or respiratory disturbances caused by HEA. We then hypothesized that emersion from HEA provides a physiological benefit, similar to emersion from hypoxia. Exposure to 15min of HEA without emersion (no rock bed present) caused significant increases in arterial haemolymph total ammonia (Tamm), pH, and [HCO3-]. When emersion was allowed, arterial haemolymph Tamm and [HCO3-] increased, but no alkalosis developed. Moreover, emersion decreased haemolymph partial pressure of NH3 relative to crabs which could not emerse. Overall, we demonstrate a novel behavioural response to HEA exposure in crabs which we propose may share similar mechanistic pathways with the emersion response triggered by hypoxia.