Hemolymph Levels Research Articles

Positive effects of a mixture of fish oil and soybean oil as a dietary lipid source on the ovarian development and health of female giant river prawn, Macrobrachium rosenbergii broodstock

Lipids are key nutrients that affect the development of the ovary in aquatic animals. This study aimed to investigate the effects of different lipid sources on the ovarian development and health of Macrobrachium rosenbergii broodstock. The M. rosenbergii broodstock, with an initial body weight of 8.07±0.74 g, were fed diets containing 5 % fish oil (FO), soya bean oil (SO), rapeseed oil (RO), coconut oil (CO), and a 1:1 mixture of fish oil and soybean oil (MO) as lipid sources for a period of 56 days. The results indicated that there were no significant differences in survival rate (SR) and hepatopancreatic index (HSI) among the different groups (P > 0.05). However, the specific growth rate (SGR) and gonadosomatic index (GSI) in the MO group were significantly higher than other groups (P < 0.05). Additionally, fatty acid levels in diets directly influenced the fatty acid composition of prawn hepatopancreas, showing a similar trend to that observed in the diet itself. Furthermore, compared to the other trial groups, haemolymph levels of 17-β estradiol (E2) and progesterone (PROG) were significantly increased in the MO group (P < 0.05). The ovarian section also exhibited better histomorphology and larger oocyte diameter in this group. Moreover, total antioxidant capacity was significantly increased in the MO group (P < 0.05). Saturated fatty acids in RO and CO groups could promote the expression of lipid synthesis-related genes fatty acid synthase (fas) and acetyl-CoA carboxylase (acc), concurrently suppressing the expression of lipid catabolism-related genes acyl-CoA oxidase-1 (acox-1) and AMP-activated protein kinase alpha (ampkα) (P < 0.05). Conversely, polyunsaturated fatty acids in the MO group significantly up-regulated the expression levels of fatty acid binding protein-3 (fabp-3) and peroxisome proliferator-activated receptor gamma (pparγ) (P < 0.05). Based on the findings of hepatopancreatic tissue structure, it was determined that the use of a single vegetable oil resulted in some damage to the hepatopancreatic tissue structure of M. rosenbergii broodstock, leading to increased oxidative stress. In conclusion, it is recommended that a 1:1 mixture of fish oil and soybean oil be used as a dietary lipid source to effectively meet the nutritional requirements during the ovarian development period of M. rosenbergii broodstock. This combination also enhances antioxidant properties, promotes the synthesis of haemolymph steroid hormones and lipid metabolism in female prawns, thereby facilitating ovarian development and overall health of M. rosenbergii broodstock.

Environmentally relevant concentrations of the tricyclic antidepressant, amitriptyline, affect feeding and reproduction in a freshwater mollusc

Antidepressant drugs (ADDs) are one of the most extensively used pharmaceuticals globally. They act at particularly low therapeutic concentrations to modulate monoamine neurotransmission, which is one of the most evolutionary conserved pathways in both humans and animal species including invertebrates. As ADDs are widely detected in the aquatic environment at low concentrations (ng/L to low µg/L), their potential to exert drug-target mediated effects in aquatic species has raised serious concerns. Amitriptyline (AMI) is the most widely used tricyclic ADD, while monoamines, the target of ADDs, are major bioregulators of multiple key physiological processes including feeding, reproduction and behaviour in molluscs. However, the effects of AMI on feeding, reproduction and mating behaviour are unknown in molluscs despite their ecological importance, diversity and reported sensitivity to ADDs. To address this knowledge gap, we investigated the effects of environmentally relevant concentrations of AMI (0, 10, 100, 500 and 1000 ng/L) on feeding, reproduction and key locomotor behaviours, including mating, in the freshwater gastropod, Biomphalaria glabrata over a period of 28 days. To further provide insight into the sensitivity of molluscs to ADDs, AMI concentrations (exposure water and hemolymph) were determined using a novel extraction method. The Fish Plasma Model (FPM), a critical tool for prioritization assessment of pharmaceuticals with potential to cause drug target-mediated effects in fish, was then evaluated for its applicability to molluscs for the first time. Disruption of food intake (1000 ng/L) and reproductive output (500 and 1000 ng/L) were observed at particularly low hemolymph levels of AMI, whereas locomotor behaviours were unaffected. Importantly, the predicted hemolymph levels of AMI using the FPM agreed closely with the measured levels. The findings suggest that hemolymph levels of AMI may be a useful indicator of feeding and reproductive disruptions in wild population of freshwater gastropods, and confirm the applicability of the FPM to molluscs for comparative pharmaceutical hazard identification.

Metabolic and immune status of Pacific white shrimp Penaeus vannamei concerning farming conditions

Biomarkers for monitoring shrimp health have been proposed but scarcely evaluated at the farm level. We analyzed several indicators of energy status in shrimp under farming conditions concerning stocking densities (100 m-2 with biofloc, 15 and 7 m-2). The influence of the year's season (temperature) was also analyzed, and, finally, an unfortunate event of White spot syndrome virus (WSSV) infection occurred on the 7 m-2 farms, adding another condition. At shrimp sampling from ponds, the effect of acute handling stress was also analyzed for indicators typically affected by such procedure with a 3- and 10-fold increase in glucose and lactate levels in hemolymph, respectively, regardless of density. This response was partially blunted at lower temperatures and WSSV incidence. Increased levels of protein in the hepatopancreas, adenylic energy charge (AEC) in both hepatopancreas and muscle and phosphagens in muscle were observed in shrimp from the 100 m-2 farms, suggesting a better nutritional and energetic status in shrimp cultured at high density with biofloc technology. Shrimp with WSSV presented lower hemocyanin levels in hemolymph, most likely associated with its role in the immune response. In WSSV-infected shrimp, the stress response regarding glucose increase was blunted, whereas a stress-induced increase in triglycerides (TG) levels in hemolymph was observed only with WSSV. Increased TG levels in those shrimp hepatopancreas could indicate a switch from carbohydrate to lipid-based metabolism associated with the preferential use of carbohydrates (Warburg effect) for virus replication in the early infection state.

Behavioral and physiological responses of nocturnal marine shellfish to predation pressure

Predator-prey interactions are critical in ecological communities. As the wild resources of Pacific abalone are on the verge of exhaustion, efforts to reveal the effects of predation by hostile organisms on the efficiency of its bottom sowing and proliferation are crucial for the success of resource restoration. In this study, abalone and its predator, starfish Asterias amurensis, were placed in a customized monitoring unit. Through continuous infrared video monitoring, predation pressure was found to cause a significant decrease in the distance moved, duration of movement, cumulative feeding duration and orexin levels in hemolymph (P < 0.01). It also caused a significant increase in the average velocity of movement, group cohesion, coordination and cholecystokinin levels in hemolymph (P < 0.01). Surprisingly, when a shelter was set up in the environment, the cumulative feeding duration of abalone was significantly extended in the presence of the predator (P < 0.001), whereas when the predator was removed, the distance moved, duration of movement, and cumulative feeding duration of abalone were found to significantly increase during continuous 24 h monitoring (P < 0.01). These results first identified the anti-predation strategy by nocturnal marine organisms when exposed to predation risk, and supported the pivotal role of shelter in helping abalone make a trade-off between predation pressure and survival needs, which not only provides a new insight to optimizing the strategy for prevention and control of marine ecological disasters and strengthening the protection of marine endangered species, but also results in a productive attempt to restore the marine benthic ecosystem.

Implications of dietary phosphorus sources on zootechnical performance and stress response of the pacific white shrimp Litopenaeus vannamei

The phosphorous (P) is an essential element for several functions, including the synthesis of high energy compounds in cells like adenosine triphosphate (ATP) and arginine phosphate (ArgP). The dietary requirements of P could change according to physiological and environmental conditions, particularly if organisms are exposed to stress conditions. As the P availability could be related to its chemical form in the diet, the present study analyzed the influence of different P sources (KH2PO4, NaH2PO4, NH4H2PO4, Na5P3O10), on the stress response of the Pacific white shrimp Litopenaeus vannamei. Final weight was 8.5% higher (p < 0.01) in shrimp fed on tripolyphosphate (Na5P3O10) diet compared with the monophosphate diets. Weight gain and specific growth also trend to increase, whereas feed conversion ratio trend to decrease with this polyphosphate source, although not significantly. Stress response of shrimp exposed to handling followed by hypoxia was characterized by an overall increase of lactate and glucose levels in hemolymph, as well as a decrease in Adenylate Energy Charge (AEC) and ArgP in the muscle. No effect of dietary P sources was observed on baseline biochemical composition and overall indicators of stress response, except for higher carbohydrate levels in hepatopancreas and RNAm expression of arginine kinase (AK) in muscle with the NaH2PO4 diet, suggesting a potentially better energy status with this diet.

Growth and Health Status of Spiny Lobsters and Snubnose Pompano with Multilevel Floating Net Cage System

The cultivation of multilevel floating net cages (MFNC) systems is believed to be able to increase production capacity without increasing the horizontal area of ​​the cultivation area. Research on the effect of using MFNC on the growth and health performance of Spiny lobsters and Snubnose pompano has never existed, so a study was conducted with the aim of comparing the growth and health performance of spiny lobsters and Snubnose pompano in floating net cage (FNC) and MFNC cultivation systems. The research method used was a field experiment with a comparative analysis of spiny lobsters and Snubnose pompano cultivation with different cultivation systems, namely the MFNC and FNC systems with 3 replications. The rearing lasted for 2 months and was fed trash fish as much as 5% of the biomass every day. Parameters of spiny lobsters and Snubnose pompano growth performance observed included weight growth, carapace length growth, total length growth, and specific growth rate. The health status of spiny lobsters observed quantitatively was the total hemocytes counts and hemolymph glucose levels, and Snubnose pompano by counting erythrocytes and blood leukocytes. The results showed that there was no difference in growth performance, survival and health status of spiny lobsters and Snubnose pompano reared using MFNC and FNC systems based on paired two sample for means, because p value &gt; 0.05 for all growth parameters survival and health status. Thus, the use of a multilevel floating net cage system in lobster cultivation can increase FNC productivity and be efficient in optimal space utilization and does not affect water quality, especially the parameters of temperature, pH, dissolved oxygen and salinity. There were no significant differences in the growth performance, survival, and health status of lobsters and Snubnose pompano when reared using the MFNC and FNC cultivation systems.

Mechanism of hyperproteinemia-induced damage to female reproduction in a genetic silkworm model

Hyperproteinemia is a metabolic disorder characterized by abnormally elevated plasma protein concentrations (PPC) in humans and animals. Here, a genetic silkworm model with high PPC was employed to investigate the effect of elevated PPC on female reproduction. Transcriptomic analysis revealed that high PPC induces downregulation of the ovarian development-related genes and disrupts ovarian sugar metabolism. Biochemical and endocrinal analyses revealed that high PPC increases trehalose and glucose levels in hemolymph and glycogen content in the fat body through activation of the gluconeogenic pathway and inhibition of the Insulin/Insulin-like growth factor signaling pathway-the serine/threonine kinase (IIS-AKT) pathway, thus disrupting characteristic metabolic homeostasis of sugar in the ovary. These resulted in ovarian developmental delay as well as reduced number and poor quality of eggs. Insulin supplementation effectively increased egg numbers by lowering blood sugar. These collective results provide new insights into the mechanisms by which high PPC negatively affects female reproduction and support the potential therapeutic effects of insulin.

Effects of high stocking density on growth performance and expression of MyD88, and its temporal expression under the challenge of Vibrio parahaemolyticus in the noble scallop Chlamys nobilis

High stocking density has been regarded as an adverse factor in bivalve aquaculture. However, its subsequent molecular response to pathogenic bacteria has been little studied. In order to study the question, a novel MyD88 was first cloned using adult noble scallops Chlamys nobilis (CnMyD88), and its tissue distribution was investigated. Then, 1860 juvenile scallops were divided into two groups with two initial densities of high density (200 individuals/layer, HD) and normal density (110 individuals/layer, ND) and in-situ cultured for three months, in which their growth, survival, and the differential expression of CnMyD88 were examined, respectively. Finally, scallops were injected with the Vibrio parahaemolyticus to assess the temporal expression of CnMyD88. As the results show, CnMyD88 cDNA has a full length of 2241 bp and contains an 1107 bp ORF that encodes a 368-derived protein. It was widely expressed in examined tissues with a significantly higher level in hemolymph, intestine, mantle, and gonad than others. Besides, the HD group showed lower growth (0.39 ± 0.05 mm/day) and survival (37.00 ± 8.49%) than the ND group (0.55 ± 0.02 mm/day and 76.82 ± 5.78%). More importantly, the HD group exhibited significantly lower expression levels of CnMyD88 in their examined tissues than the ND group. After V. parahaemolyticus challenging, CnMyD88 had significantly lower expression levels in the scallops from the HD group than that of the scallops from the ND group at 6th, 24th, and 36th. The present results indicated that high stocking density not only made adverse impacts on growth and survival but also may induce immunosuppression in the noble scallop. Therefore, appropriate low stocking density may be worth considering to adopt in scallop aquaculture.

Field Efficacy, Sub-lethal, and Biochemical Effects of Certain Biorational Insecticides Against the New Intruder, Spodoptera frugiperda in Bani-Suef, Upper Egypt

Frequent inspections on sorghum and maize crops during seasons of 2021 and 2022 in some regions in Bani-Suef governorate, Egypt, discovered unprecedented invasions of Spodoptera frugiperda (J. E. Smith). Accordingly, our study on Beauveria bassiana and spinetoram was supporter to the Food and Agriculture Organization’s tendency in adopting biorational insecticides against S. frugiperda in Egypt. Exposure toxicity of LC25 values at 48 h of B. bassiana were 2.7 × 106 and 5.2 × 106 conidia mL−1 and spinetoram were 0.019 and 0.048 mg L−1 against the 2nd and 4th instar larvae laboratory strain of S. frugiperda, respectively. Sub-lethal effects (LC25) were accomplished on biological parameters against both instar larvae. LC25 of B. bassiana reduced adult emergency (89.91 and 91.05%) more than spinetoram (75.99 and 79.49%) against the 2nd and 4th instar larvae, respectively. The 2nd instar larvae exposed to LC25 of B. bassiana suppressed female fecundity (0.00 eggs) more than spinetoram (19.74 eggs). Enzymatic activity of lipase in hemolymph, fat bodies, and mid-gut of the 4th instars at 48 h showed significant drop in B. bassiana more than spinetoram. Glutathione-S-transferase (GST) levels in hemolymph for both insecticides were equal and exceeded the control. Fat bodies and mid-gut possessed the highest GST activity in B. bassiana followed by spinetoram and the control. Residual efficacy of spinetoram exceled B. bassiana at their field rates under semi-field condition in Bani-Suef along the two seasons of maize crop against both instars. Eventually, B. bassiana alongside spinetoram could afford good control especially on early instar larvae of S. frugiperda.

Consumption of some Nigerian local rice varieties and effect on development of insulin resistance and obesity in Drosophila melanogaster

There is evidence to suggest that the increased incidence of insulin resistance and obesity is associated with the consumption of energy-dense and highly processed foods like polished rice. This study, therefore, investigated the effects of consumption of selected local rice varieties on some markers of insulin resistance and obesity using Drosophila melanogaster. Five different local rice varieties (Jamila, Kwandala, Mai-bakin carki, Yar-katabore and Yar-washagi) were subjected to proximate composition analysis, which showed that the unpolished local varieties had a better deposit of nutrients. Thereafter, adult flies were raised on diets containing either polished or unpolished forms of the five different local rice varieties for seven days and thereafter, analysed for weight and locomotor ability, as well as the levels of glucose, trehalose, glycogen and triglycerides in the hemolymph and expression levels of ACC and Pepck genes. Drosophila melanogaster flies on the polished rice groups showed increased weight, hyperglycaemia and hyperlipidaemia, compared to those on the unpolished rice (p&lt;0.05). Changes in gene expression were in line with the biochemical changes. The study, thus, suggested that the substitution of polished with unpolished local rice may reduce the risk of insulin resistance and obesity, as well as their related complications.

Identification and profiling of long non-coding RNAs during molt cycle: An involvement of lnc1182 in the molt of white shrimp, Litopenaeus vannamei

Molt is an essential physiological process for crustacean growth. It is known to be controlled by factors such as hormones and steroids, but the role of long non-coding RNA (lncRNA), a potential regulatory molecule, has not been studied. In this study, we investigated the role of lncRNA in the molt process of white shrimp, Litopenaeus vannamei. The 9437 of 58,142 unigenes of de novo assembled transcriptome were identified as putative lncRNAs, and 768 lncRNAs were differentially expressed during molt cycles. Five differentially expressed lncRNAs were selected to determine their expressions in different tissues and molt stages. Most were expressed more in gills and epidermis than other tissues. Expressions of lnc1182 and lnc336 significantly decreased during premolt stages and increased at postmolt in gills and epidermis, while lnc179 expression increased during premolt stages. RNA interference was used to investigate the role of lnc1182 in the molt process. The molt of lnc1182 knockdown shrimp progressed to early premolt faster than shrimp injected with dsGFP, and also shortened molt duration. In addition, total sugar level in hemolymph was higher in the lnc1182 knockdown shrimp than the control shrimp, whereas gill Na+/K+ ATPase activity, total hemocyte count, and expression of molt-inhibiting hormone (MIH) were not changed. These suggested that lnc1182 is likely involved in the control of sugar metabolism during molting but not in ion exchange activity and MIH expression. Furthermore, lnc1182 and 20-HE probably regulated each other. RNA sequencing was used to investigate the set of genes associated with lnc1182. Fifty-five transcripts were differentially expressed in gills of lnc1182 knockdown shrimp. Expressions of candidate metabolism and immune-related genes were validated for their differential expressions in lnc1182 knockdown shrimp. Our work produced lncRNA profiles and indicated possible functions of lnc1182 in metabolism and immunity during the molt cycle in shrimp.

Physiological responses to honeybee venom poisoning in a model organism, the firebug Pyrrhocoris apterus

In this study, the biochemical and physiological features of the firebug Pyrrhocoris apterus were investigated to understand the impact of the honeybee Apis mellifera venom on them using physiological methods (mortality, total level of metabolism), biochemical methods (ELISA, mass spectrometry, polyacrylamide gel electrophoresis, spectrophotometry) and molecular methods (real-time PCR). Together, the obtained findings suggest that venom injection increased the level of adipokinetic hormone (AKH) in the CNS of P. apterus, indicating that this hormone plays a key role in activating defence responses. Furthermore, histamine levels in the gut increased significantly after envenomation and did not seem to be modulated by AKH. In contrast, histamine levels in the haemolymph increased after treatment with AKH and AKH + venom. In addition, we found that vitellogenin levels in haemolymph decreased in both males and females after venom application. Lipids, which are the main energy metabolites used by Pyrrhocoris, were significantly exhausted from the haemolymph after the administration of venom and the co-application with AKH reversed this effect. However, we did not find much influence on the effect of digestive enzymes after the injection of venom. Our research has highlighted the noticeable effect of bee venom on P. apterus' body and provided new insights into the role of AKH in controlling defensive responses. However, it is also likely that there will be alternative defence mechanisms.

Assessment of metabolic flexibility by RER in the Gromphadorhina portentosa

Metabolism is the series of chemical reactions utilized by organisms to create energy to drive physiological and cellular functions. Understanding the metabolism of ectothermic organisms has been an area of interest that is widely understudied with limited model organisms. Using the Gromphadorhina portentosa (Giant Madagascar Hissing Cockroach) as a model organism, ectothermic metabolism was measured by respirometry using a closed animal chamber and iWorx gas analyzer. The metabolic rate was calculated as the Respiratory Exchange Ratio (RER), which is the ratio of carbon dioxide produced to oxygen consumed. We investigated whether changes in metabolism were allometric functions dependent on the size of cockroaches. Experiments were conducted to study the effects of high temperatures, exercise and diet on the metabolic rate of G. portentosa compared to control. Preliminary data showed significant changes in RER under several of these experimental conditions. We have also investigated whether the environmental conditions produced sex- and age-specific changes in RER. Lastly, we utilized biochemical assays to determine why several experimental groups exhibit RERs &gt;1.0. Preliminary results show glucose and triglyceride levels in hemolymph accommodate metabolic flexibility requirements in the above experimental conditions. This work represents the most comprehensive evaluation of this ectotherm and adds to our understanding of one of the largest insects. APU This is the full abstract presented at the American Physiology Summit 2023 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Metabolomics analysis of dietary restriction results in a longer lifespan due to alters of amino acid levels in larval hemolymph of Bombyx mori

Dietary restriction (DR) has been a very important discovery in modern aging biology research. Its remarkable anti-aging effect has been proved in a variety of organisms, including members of Lepidoptera, but mechanisms by which DR increases longevity are not fully understood. By using the silkworm (Bombyx mori), a model of lepidopteran insect, we established a DR model, isolated hemolymph from fifth instar larvae and employed LC–MS/MS metabolomics to analyze the effect of DR on the endogenous metabolites of silkworm, and tried to clarify the mechanism of DR to prolong lifespan. We identified the potential biomarkers by analyzing the metabolites of the DR and control groups. Then, we constructed relevant metabolic pathways and networks with MetaboAnalyst. DR significantly prolonged the lifespan of silkworm. The differential metabolites between the DR and control groups were mainly organic acids (including amino acid), and amines. These metabolites are involved in metabolic pathways such as amino acid metabolism. Further analysis showed that, the levels of 17 amino acids were significantly changed in the DR group, indicating that the prolonged lifespan was mainly due to changes in amino acid metabolism. Furthermore, we identified 41 and 28 unique differential metabolites in males and females, respectively, demonstrating sex differences in biological responses to DR. The DR group showed higher antioxidant capacity and lower lipid peroxidation and inflammatory precursors, with differences between the sexes. These results provide evidence for various DR anti-aging mechanisms at the metabolic level and novel reference for the future development of DR-simulating drugs or foods.

Dietary compounds activate an insect gustatory receptor on enteroendocrine cells to elicit myosuppressin secretion.

Sensing of midgut internal contents is important for ensuring appropriate hormonal response and digestion following the ingestion of dietary components. Studies in mammals have demonstrated that taste receptors (TRs), a subgroup of G protein-coupled receptors (GPCRs), are expressed in gut enteroendocrine cells (EECs) to sense dietary compounds and regulate the production and/or secretion of peptide hormones. Although progress has been made in identifying expression patterns of gustatory receptors (GRs) in gut EECs, it is currently unknown whether these receptors, which act as ligand-gated ion channels, serve similar functions as mammalian GPCR TRs to elicit hormone production and/or secretion. A Bombyx mori Gr, BmGr6, has been demonstrated to express in cells by oral sensory organs, midgut and nervous system; and to sense isoquercitrin and chlorogenic acid, which are non-nutritional secondary metabolites of host mulberry. Here, we show that BmGr6 co-expresses with Bommo-myosuppressin (BMS) in midgut EECs, responds to dietary compounds and is involved in regulation of BMS secretion. The presence of dietary compounds in midgut lumen after food intake resulted in an increase of BMS secretions in hemolymph of both wild-type and BmGr9 knockout larvae, but BMS secretions in BmGr6 knockout larvae decreased relative to wild-type. In addition, loss of BmGr6 led to a significant decrease in weight gain, excrement, hemolymph carbohydrates levels and hemolymph lipid levels. Interestingly, although BMS is produced in both midgut EECs and brain neurosecretory cells (NSCs), BMS levels in tissue extracts suggested that the increase in hemolymph BMS during feeding conditions is primarily due to secretion from midgut EECs. Our studies indicate that BmGr6 expressed in midgut EECs responds to the presence of dietary compounds in the lumen by eliciting BMS secretion in B. mori larvae.

Nutrient sensing signaling and metabolic responses in shrimp Litopenaeus vannamei under acute ammonia stress

Ammonia is the primary environmental factor affecting the growth and health of crustaceans. It would induce oxidative stress and metabolic disorders. Extra amount of energy was demanded to maintain the physiological functions under ammonia stress. However, limited information was available on its effects on the main nutrient metabolism, as well as the nutrient sensing signaling pathways. In the present study, shrimp Litopenaeus vannamei were exposed to acute ammonia stress and injected with amino acid solution. The results showed that acute ammonia exposure resulted in lower free amino acid levels in hemolymph, incomplete activation of the mechanistic target of rapamycin (mTOR) signaling and cascaded less protein synthesis in muscle. It induced autophagy and activated the AMP-activated protein kinase (AMPK) pathway. Meanwhile, ammonia exposure enhanced glycolysis and lipogenesis, but inhibited lipolysis. The results characterized the integrated metabolic responses and nutrient signaling to ammonia stress. It provides critical clues to understand the growth performance and physiological responses in shrimp under ammonia stress.

Regulatory role of myo-inositol in the ovarian development of female Chinese mitten crab (Eriocheir sinensis)

The ovarian development of female crab can directly affect the nutritional value, broodstock fertility and health status of offspring and are essential for the success of the crab industry. This study evaluates the influences of dietary myo-inositol (MI) on the ovarian development of female Chinese mitten crab (Eriocheir sinensis). Three diets containing different MI levels (0, 1600 and 3200 mg/kg) were fed to 144 crabs (initial weight: 72.38 ± 2.53 g) for 13 weeks. Crabs fed the diets with 1600, and 3200 mg/kg MI supplementation had significantly higher gonadosomatic index and yolk granule deposition in the ovary than those fed diets without MI supplementation. Crabs fed 1600 and 3200 mg/kg MI supplementation significantly enhanced vitellogenin synthesis in the hepatopancreas, vitellogenin contents in the hemolymph and vitellogenin receptor mRNA level in the ovary compared with those fed diets without MI supplementation. Simultaneously, cyclic adenosine monophosphate content and gene expression of retinoid X receptor, protein kinase A and ecdysteroid receptor in the hepatopancreas, and ecdysteroid content in hemolymph significantly increased when 1600 and 3200 mg/kg MI were supplemented in diets. Moreover, crabs fed 1600 and 3200 mg/kg MI supplementation had significantly higher lipid content in the ovary than those fed the diets without MI supplementation. The neutral lipid content in the hepatopancreas tended to decrease as the dietary MI level increased, while the polar lipid content in the hepatopancreas and the neutral and polar lipid contents in the ovary tended to increase. Dietary MI supplementation significantly enhanced the expression of genes related to lipid catabolism, lipid absorption and transport in the hepatopancreas, and gene expression related to lipid absorption in the ovary, suggesting that MI could promote the transport of lipids to the ovary from the hepatopancreas for ovarian development. This study indicates that dietary 1600 mg/kg MI supplementation is sufficient to promote the ovarian development of E. sinensis.

Polystyrene microplastics alleviate adverse effects of benzo[a]pyrene on tissues and cells of the marine mussel, Mytilus galloprovincialis

As two major ubiquitous pollutants, microplastics (MPs) and polycyclic aromatic hydrocarbons (PAHs) coexist in the marine environment. However, the role of MPs in altering the toxicity of PAHs to marine organisms is poorly understood. We therefore investigated the accumulation and toxicity of benzo[a]pyrene (B[a]P, 0.4 nM), in the marine mussel Mytilus galloprovincialis over a 4-day of exposure with or without the presence of 10 μm polystyrene microplastics (PS MPs) (10 particles/mL). The presence of PS MPs significantly decreased B[a]P accumulation in soft tissues of M. galloprovincialis by approximately 6.7%. Single exposure of PS MPs or B[a]P decreased the mean epithelial thickness (MET) of digestive tubules and enhanced reactive oxygen species (ROS) levels in haemolymph, while upon co-exposure the adverse impacts were alleviated. Real-time q-PCR results showed that most selected genes involved in stress response (FKBP, HSP90), immune (MyD88a, NF-κB) and detoxification (CYP4Y1) were induced for both single exposure and co-exposure. The co-presence of PS MPs down-regulated the mRNA expression of NF-κB in gills compared with of B[a]P alone. The uptake and toxicity reductions of B[a]P might result from the decrease of its bioavailable concentrations caused by the adsorption of B[a]P by PS MPs and the strong affinity of B[a]P to PS MPs. Adverse outcomes for the co-existence of marine emerging pollutants under long-term conditions remain to be further validated.

Comparison of maturation and secondary spawning of fleshy shrimp Penaeus chinensis fed different kinds of diets

The study carried out a 15-day feeding trial of the fleshy shrimp broodstock that have just finished their first spawn using five different maturation feed types (three types of manufactured feed, fresh feed, and commercial feed) to induce multiple spawning in the shrimp and then analyzed and compared hemolymph levels of total protein, total lipid, hemocyanin, glucose, and cholesterol as well as ovary histology between the treatments to identify the potential of the experimental diets to be utilized as maturation feed. The study found that the manufactured diets of polychaetes + clam and polychaetes + clam + squid resulted in significantly higher levels of lipid and protein in shrimp hemolymph suggesting an indicator of multiple spawning in the fleshy shrimp. The other manufactured diet of polychaets + squid led to the highest 2nd spawning with significantly lower levels of lipid, protein, and cholesterol in shrimp hemolymph. The fresh feed treatment showed similar hematological as well as histological responses to the manufactured diet of polychaetes + squid, but with a significantly lower spawning volume compared to the other treatments. The study demonstrated that the manufactured diets could replace fresh feed as maturation feed and attain a higher shrimp maturation than the commercial feed.

Exercise and emersion in airand recovery in seawater in the green crab (Carcinus maenas): Effects on nitrogenous wastes and branchial chamber fluid chemistry.

At low tide, the green crab, which is capable of breathing air, may leave the water and walk on the foreshore, carrying branchial chamber fluid (BCF). N-waste metabolism was examined in crabs at rest in seawater (32 ppt, 13°C), and during 18-h recovery in seawater after 1 h of exhaustive exercise (0.25 BL s-1 ) on a treadmill in air (20°C-23°C), or 1 h of quiet emersion in air. Measurements were made in parallel to O2 consumption (ṀO2 ), acid-base, cardio-respiratory, and ion data reported previously. At rest, the ammonia-N excretion rate (ṀAmm = 44 µmol-N kg-1 h-1 ) and ammonia quotient (AQ; ṀAmm /ṀO2 = 0.088) were low for a carnivore. Immediately after exercise and return to seawater, ṀAmm increased by 65-fold above control rates. After emersion alone and return to seawater, ṀAmm increased by 17-fold. These ammonia-N bursts were greater, but transient relative to longer-lasting elevations in ṀO2 , resulting in temporal disturbances of AQ. Intermittent excretion of urea-N and urate-N at rest and during recovery indicated the metabolic importance of these N-wastes. Hemolymph glutamate, glutamine, and PNH3 did not change. Hemolymph ammonia-N, urea-N, and urate-N concentrations increased after exercise and more moderately after emersion, with urate-N exhibiting the largest absolute increments, and urea-N the longest-lasting elevations. All three N-wastes were present in the BCF, with ammonia-N and PNH3 far above hemolymph levels even at rest. BCF volume declined by 34% postemersion and 77% postexercise, with little change in osmolality but large increases in ammonia-N concentrations. Neither rapid flushing of stored BCF nor clearance of hemolymph ammonia-N could explain the surges in ṀAmm after return to seawater.