Hemolymph Glucose Research Articles

Effects of water immersion on immune, intestinal flora and metabolome of Chinese mitten crab (Eriocheir sinensis) after air exposure

Air exposure stress can induce stress response of Eriocheir sinensis and affect its normal life activities. The goal of this study was to investigate the effects of water immersion on the recovery of hepatopancreas immune-related enzyme activity, intestinal microbial diversity and metabolic level of Chinese mitten crabs after exposure to air. The results show that immersion can effectively alleviate the adverse effects of air exposure on the antioxidant capacity and immune capacity of Chinese mitten crabs, and the longer the time of immersion, the more obvious the recovery effect. Among them, the levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase and acid phosphatase significantly increased after exposure to air (P < 0.05), reached a peak at 3 h, began to decline after immersion, and returned to a level close to the initial value at 24 h (P < 0.05). In addition, after exposure to air, the glucose and total cholesterol in haemolymph of Eriocheir sinensis were significantly different from the initial values (P < 0.05), gradually recovered to the initial level after re-immersion. However, changes in intestinal flora and hepatopancreas metabolism caused by air exposure did not fully recover after water exposure, and its negative effects did not completely disappear. The sequencing results showed that the species composition and diversity of intestinal microorganisms of Chinese mitten crab changed after air exposure and immersion treatment. The relative abundance of Actinomycetes increased significantly, while that of Proteobacteria and Firmicutes decreased significantly. Metabolomics analysis showed that air exposure and immersion destroyed the metabolic balance of amino acids and carnitine, reduced the level of carnitine metabolism, hindered the absorption of nutrients, and led to the accumulation of harmful substances.

Effect of stocking density on survival and growth of spiny sand lobsters Panulirus homarus in cages at 7‐m depth

AbstractObjectiveThe spiny sand lobster Panulirus homarus is one of the lobster species that has a high price in local and international markets and is economically valuable. The objective of this study was to determine the effect of stocking density on the survival and growth of spiny sand lobsters kept in cylindrical cages (1.00 m high × 1.15 m in diameter) at a depth of 7 m below the water surface, with an average ambient temperature of 29–32°C.MethodsThe 3‐month study was performed in 2022 and had a completely randomized design. There were three treatment groups, each of which had three replications. Stocking densities were 10 lobsters/m2 (10 lobsters/cage) for treatment A, 15 lobsters/m2 (15 lobsters/cage) for treatment B, and 20 lobsters/m2 (20 lobsters/cage) for treatment C. The initial average body weight was 75 g/lobster.ResultThe survival rates for treatment A (mean ± standard error = 91.50 ± 0.25%) and treatment B (93.30 ± 0.51%) were higher than the survival rate for treatment C (86.05 ± 0.45%), but the values were not significantly different. Daily weight gain and the specific daily growth rate for treatments A and C were significantly higher than those of treatment B. The feed conversion ratio was not significantly different among the groups. Total hemocyte count in the hemolymph between treatments A and B as well as initial (day‐0) values were not significantly different, but values for treatments A and B were significantly lower than that for treatment C. The hemolymph glucose value for treatment A was significantly lower than that of treatment C but was not different from that of treatment B.ConclusionThe grow‐out study of spiny sand lobsters at 7‐m depth indicated an optimal stocking density of 15 lobsters/m2. This was supported by several physiological response and production performance parameters, which generally had no significant difference with those of the lower stocking density.

Assessment of Amphibola crenata as a bioindicator of estuarine trace element pollution using biochemical and physiological endpoints

To assess its utility as a bioindicator of estuarine contamination, Amphibola crenata, a pulmonate mud snail, was collected from 17 estuaries in New Zealand. Whole-body soft tissue trace element concentrations were measured via quadrupole inductively coupled plasma mass spectrophotometer (Q-ICP-MS) and were found to be significantly positively correlated with sediment trace element profiles for arsenic, copper and lead. Snails from polluted estuaries generally displayed higher ammonia excretion rates, elevated whole-body soft tissue catalase activity and lipid peroxidation compared to snails from reference sites. Across all sites haemolymph glucose was positively correlated with the tissue burdens of arsenic, copper, nickel and zinc, while haemolymph protein was negatively correlated with arsenic, cadmium and zinc soft tissue concentrations, indicative of altered energy metabolism associated with trace element contamination. Overall, sites were distinguishable by application of the array of measures employed. Our findings suggest that gastropods have significant value as bioindicators of estuarine health.

Palliative effect of abalone insulin-related peptide 2 on emersion stress-induced hyperglycemia in the Pacific abalone (Haliotis discus hannai)

The Pacific abalone (Haliotis discus hannai), an economically important marine mollusk in East Asia, is often exposed to extreme environments, such as high water temperatures during summer and air exposure during handling or transportation. These stressful conditions lead to hyperglycemia and the generation of reactive oxygen species (ROS), resulting in oxidative stress and subsequent cell damage in the organism. To understand the role of insulin-related peptides in the regulation of elevated hemolymph glucose, hyperglycemia was confirmed in abalone under high water temperature and air (emersion) exposure (p < 0.05). To prove the existence of a glucose regulatory system in abalone, changes in hemolymph glucose were measured after bovine insulin injection, and hemolymph glucose was lowered by bovine insulin injection in hyperglycemia caused by both stressors (p < 0.05), supporting the existence of a glucose regulatory system functionally similar to that of vertebrates. Among the three abalone insulin-related peptides (AIPs), which were previously reported to be upregulated along with hemolymph glucose levels upon feeding, the relative mRNA expression of AIP2 in the cerebral ganglion showed a close correlation with high water temperature and emersion stress, which peaked 12 h after treatment (p < 0.05). Based on these results, we focused on the function of AIP2 in glucose homeostasis and characterized the localization of AIP2 mRNA-expressing cells in the cerebral ganglion using in situ hybridization. Positive signals from RNA probes were detected in neural cortex cells in the cerebral ganglion. Furthermore, suppressing AIP2 mRNA expression using RNA interference with double-stranded RNA (dsRNA) revealed that dsRNA-AIP2 injected abalone maintained high hemolymph glucose levels after 24 h of recovery following 1 h of emersion stress (p < 0.01). In contrast, abalones that received dsRNA-GFP and molluscan saline showed relatively rapid clearance of hemolymph glucose. This study demonstrates that AIP2 gene expression affects the recovery rate from emersion stress-induced hyperglycemia by regulating hemolymph glucose.

Comparative transcriptome analysis provides a glance into the regulatory of insulin-like peptide 1 gene in the Pacific white shrimp Litopenaeus vannamei

Insulin-like peptides (ILPs) have been reported to exert significant and diverse regulatory effects through the insulin signaling pathway in invertebrates. However, there is a large research gap in the functional analysis of ILP genes in crustaceans. In this study, we combined RNA interference (RNAi) technology with transcriptome sequencing to investigate the regulatory effects of the ILP1 gene in the Pacific white shrimp Litopenaeus vannamei. After two weeks of continuous RNAi of LvILP1, the shrimp in the dsLvILP1 group exhibited reduced body length gain, weight gain, molting rate, and hemolymph glucose concentration compared to the control dsEGFP group, indicating that knockdown of LvILP1 slowed shrimp growth, molting, and metabolism. Comparative transcriptome analysis of three tissues revealed that knockdown of LvILP1 inhibited muscle growth and development by modulating the mTOR and MAPK signaling pathways, and prevented the renewal of epidermal tissue through the Hippo signaling pathway. Meanwhile, it also suppressed certain growth signals in the hepatopancreas, such as juvenile hormone (JH) synthesis and 20-hydroxyecdysteroid (20E) signal activation. Additionally, it promoted the conversion pathway of some sugars into amino acid metabolism instead of energy metabolism after glycolysis in the hepatopancreas. Interestingly, knockdown of LvILP1 also affected carbohydrate metabolism by regulating the intracellular and extracellular transport of glucose and trehalose, as well as regulating the expression levels of key metabolic genes in three shrimp tissues. This disruption led to a decrease in blood glucose levels and suppressed energy production, ultimately providing less energy for shrimp growth and molting. This study provides valuable insights into the underlying genetic mechanism through which ILP1 regulates glucose metabolism and growth in shrimp. It enhances our understanding of the structure and function of the insulin pathway in crustaceans and serves as an important reference for the identification of candidate genes in molecular genetic breeding of shrimp.

The impact of Roundup® Original DI on the hemolymph metabolism and gill and hepatopancreas oxidative balance of Parastacus promatensis (Crustacea, Decapoda, Parastacidae).

In Brazil, glyphosate is present in more than 130 commercial formulations, and its toxic effects have already been tested in different species to understand its impact on biota Decapod crustaceans are widely used as experimental models due to their biology, sensitivity to pollutants, ease of collection, and maintenance under laboratory conditions. We evaluated the changes in metabolism (hemolymph) and oxidative balance markers (gill and hepatopancreas) of a crayfish (Parastacus promatensis) after exposure to Roundup® (active ingredient: glyphosate). The crayfish were captured in the Garapiá stream within the Center for Research and Conservation of Nature Pró-Mata, Brazil. We collected adult animals outside (fall) and during (spring) the breeding season. The animals were transported in buckets with cooled and aerated water from the collection site to the aquatic animal maintenance room at the university. After acclimatization, the animals were exposed to different concentrations of glyphosate (0, 65, 260, 520, and 780 µg/L). The results showed a significant variation in the hemolymph glucose, lactate, and protein levels. We observed variations in the tissue antioxidant enzymatic activity after exposure to glyphosate. Finally, the increase in oxidative damage required a high energy demand from the animals to maintain their fitness, which makes them more vulnerable to stress factors added to the habitat.

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.

Dietary protein-to-carbohydrate ratio effects development and metabolism in Drosophila larvae and imago

Background. Nutrition during growth and development affects various traits not only in larvae but also imago including lifespan, reproduction, feeding, metabolism, and stress resistance. In this study, we have tested the hypothesis of whether the dietary protein-to-carbohydrate (P:C) ratio in the developmental diet could be related to subsequent changes in metabolic profile and physiological parameters in Drosophila larvae and imago. Materials and Methods. Drosophila melanogaster Canton-S strain were used in this study. Larvae were fed diets with different P:C ratios. Experimental media were composed of either 2 % or 5 % dry yeast and 0 %, 1 %, or 10 % of sucrose. We tested developmental rate, wet or dry body weight and the levels of certain metabolites inclu­ding glucose, glycogen, triacylglycerides and total lipids. The developmental rate was assessed by counting the number of generated pupae every 6/6/12 hours. For wet or dry weight measurement, 20 larvae or flies were weighed and transferred to plastic vial with a cut bottom. The flies were dried at 60 °C with the subsequent weighing after two days. Another two-day flies cohort were separated by sex and frozen in liquid nitrogen for further biochemical assays. Hemolymph glucose, total lipid concentration, triacyl­glycerides (TAG), body glucose and glycogen contents were determined spectrophotometrically. Results. We found that a low 0.08 P:C ratio in the diet slowed down pupation by ~20 % and decreased body weight in larvae. Hemolymph glucose levels in both larvae and imago were inversely associated with dietary P:C. Larvae developing on a diet with a low P:C ratio displayed a lower level of glycogen pool, but a higher level of lipids. Developmental dietary P:C ratio also influences metabolic traits such as hemolymph glucose, glycogen, TAG and total lipids in male and female imago. A higher total protein intake combined with restriction of sucrose consumption had glucose-lowering and lipids-lowering effects. Conclusions. Our study demonstrated that nutritional conditions during larval development trigger adaptive changes that provide a level of regulation necessary to surpass dietary stress in Drosophila imago.

Effect of paprika extracts on growth performance, haemolymph chemistry, intestinal microbiota and antioxidant enzyme activities of white-leg shrimp (Litopenaeus vannamei).

This study aims to evaluate the effects of paprika extract on the survival rate, growth performance and stimulation of the innate immune system of Litopenaeus vannamei. In this experiment, 240 healthy shrimp (3.22 ± 0.12 g) were randomly divided into four groups. The shrimp were fed diets with different concentrations of paprika oil extracts (0%, 0.5%, 1% and 2%) for 8 weeks. The results showed that growth performance, urea, uric acid, creatinine, cholesterol levels, aspartate aminotransferase and alkaline phosphatase activities were not significantly affected by adding paprika extract to the shrimp diet (p > 0.05). Diets containing 1% and 0.5% paprika extract showed the highest levels of total protein and triglyceride, respectively (p < 0.05). There was a significant decrease in haemolymph glucose concentration in shrimp-fed diets containing 1% and 2% paprika extract (p < 0.05). Moreover, a diet containing 0.5% paprika extract resulted in the highest levels of total heamocyte count, hyaline cells and large-granular cells in shrimp (p < 0.05). Higher catalase and superoxide dismutase activities were also exhibited in the paprika groups (p < 0.05). Vibrio sp. bacteria were not significantly reduced by paprika extract in the intestines of L. vannamei (p > 0.05). A significant decrease in heterotrophic bacteria was observed with increasing extract concentrations (p < 0.05). The shrimp culture industry can utilize paprika extract as a cost-effective, efficient and environmentally friendly immune stimulant at a concentration of 0.5%.

Atractylodes macrocephala Koidz. and Cuscuta chinensis Lam. extract relieves insulin resistance via PI3K/Akt signalling in diabetic Drosophila

Background and aimType-2 diabetes mellitus (T2DM) is mainly characterized by insulin resistance (IR) induced by hyperglycaemia and insufficient insulin secretion. We employed a diabetic fly model to examine the effect and molecular mechanism of Atractylodes macrocephala Koidz. and Cuscuta chinensis Lam. (AMK–CCL) extract as traditional Chinese medicine in treating IR and T2DM. Experimental procedureThe contents of the active ingredients (rhamnose, xylose, mannose, and hyperoside) in AMK–CCL extract were determined by high-performance liquid chromatography. Wild-type (Cg-GAL4/+) or diabetic (Cg > InRK1409A) Drosophila flies were divided into the control group or metformin group and AMK–CCL (0.0125, 0.025, 0.05, 0.1 g/ml) groups. Food intake, haemolymph glucose and trehalose, protein, weight, triglycerides (TAG), and glycogen were measured to assess glycolipid metabolism. Phosphatidylinositol-3-kinase (PI3K)/Akt signalling was detected using fluorescent reporters [tGPH, Drosophila forkhead box O (dFoxO)–green fluorescent protein (GFP), Glut1–GFP, 2-NBDG] in vivo. Glut1/3 mRNA levels and Akt phosphorylation levels were detected by quantitative polymerase chain reaction and western blotting, respectively, in vitro. ResultsAMK–CCL extract contained 0.038 % rhamnose, 0.017 % xylose, 0.69 % mannose, and 0.039 % hyperoside. AMK–CCL at 0.0125 g/mL significantly suppressed the increase in circulating glucose, and the decrease in body weight, TAG, and glycogen contents of diabetic flies. AMK–CCL improved PI3K activity, Akt phosphorylation, Glut1/3 expression, and glucose uptake in diabetic flies, and also rescued diabetes-induced dFoxO nuclear localisation. ConclusionsThese findings indicate that AMK–CCL extract ameliorates IR-induced diabetes via the PI3K/Akt signalling pathway, providing an experimental basis for clinical treatment.

Adaptation of Glucose Metabolism to Limb Autotomy and Regeneration in the Chinese Mitten Crab.

Limb autotomy and regeneration represent distinctive responses of crustaceans to environmental stress. Glucose metabolism plays a pivotal role in energy generation for tissue development and regeneration across various species. However, the relationship between glucose metabolism and tissue regeneration in crustaceans remains elusive. Therefore, this study is aimed at analyzing the alterations of glucose metabolic profile during limb autotomy and regeneration in Eriocheir sinensis, while also evaluating the effects of carbohydrate supplementation on limb regeneration. The results demonstrated that limb autotomy triggered a metabolic profile adaption at the early stage of regeneration. Hemolymph glucose levels were elevated, and multiple glucose catabolic pathways were enhanced in the hepatopancreas. Additionally, glucose and ATP levels in the regenerative limb were upregulated, along with increased expression of glucose transporters. Furthermore, the gene expression and activity of enzymes involved in gluconeogenesis were repressed in the hepatopancreas. These findings indicate that limb regeneration triggers metabolic profile adaptations to meet the elevated energy requirements. Moreover, the study observed that supplementation with corn starch enhanced limb regeneration capacity by promoting wound healing and blastema growth. Interestingly, dietary carbohydrate addition influenced limb regeneration by stimulating gluconeogenesis rather than glycolysis in the regenerative limb. Thus, these results underscore the adaptation of glucose metabolism during limb autotomy and regeneration, highlighting its essential role in the limb regeneration process of E. sinensis.

Effects Density and Salinity of Artificial Brackish Water on the Growth and Physiological Perfomance of Whiteleg Shrimp (Litopenaeus vannamei Boone, 1931)

The increase in shrimp culture causes problems with the quality of seawater which is a source of water for whiteleg shrimp culture it self. The use of artificial brackish water as a culture medium is an effort to increase the availability of water sources for whiteleg shrimp culture. This study aims to (1) determine the optimum stocking density of whiteleg shrimp culture using artificial brackish water salinity based on carrying capacity, (2) determine mineral content (Mg, Ca, Na and K) in artificial brackish water, (3) study the effect of salinity on the growth and blood glucose concentration of whiteleg shrimp cultivated in artificial brackish water, (4) examine the effect of salinity on hemolymph osmolarity of whiteleg shrimp cultivated in artificial brackish water. This study used an experimental method consisting of two stages. Each stage was arranged using a completely randomized design (CRD). Obtained stocking densities of SD-50% (20 shrimps/50L), SD (40 shrimps/50L) and SD+50% (60 shrimps/50L). The best growth and performance were obtained in the treatment of 20 individuals, namely survival (SR) 87%, specific growth rate (SGR) 3.26%/day and hemolymph glucose concentration of 30.73 mg/dl. In the next stage of the research, the salinity treatments tested were seawater, 10 ppt, 15 ppt, 20 ppt and 25 ppt. The best shrimp performance was obtained in treatment 20 shrimps/50 L, namely survival rate 70.00%, specific growth rate (SGR) 1.55%/day, feed conversion ratio (FCR) 1.72, glucose haemolymp 73.44 mg/dl. The optimal osmotic work level (OWL) value is obtained at a salinity of 15-20 ppt.

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.

Dietary Cholesterol Requirements of Large Red Swamp Crayfish (Procambarus clarkii).

To investigate the dietary cholesterol requirements of large red swamp crayfish (Procambarus clarkii), crayfish (initial body weight: 13.49 ± 0.22 g) were hand-fed six diets containing 2.47 (C0), 4.27 (C1), 6.80 (C2), 8.77 (C3), 11.74 (C4), and 14.24 (C5) g/kg cholesterol. After 8 weeks of feeding, the maximum weight gain rate and specific growth rate occurred in group C4. The lowest feed conversion ratio was observed in group C3. Total flesh percentage increased significantly by 15.33% in group C2 compared to group C0. The increase in dietary cholesterol resulted in significant quadratic trends in concentrations of crude protein and lipid in muscle and whole body; cholesterol and free fatty acid in hemolymph, hepatopancreas, and muscle; activities of lipase and amylase in hepatopancreas and intestine; and total antioxidant capacity and catalase activity in hepatopancreas. Group C3 experienced a noteworthy increase in hemolymph glucose and total protein content compared to group C0. Additionally, malondialdehyde content and superoxide dismutase activity in hepatopancreas displayed significant linear and quadratic trends. The optimal dietary cholesterol level for large P. clarkii is between 7.42 and 10.93 g/kg, as revealed by the quadratic regression analysis.

Tyramine mediates growth performance, immune response, and physiological regulation of Litopenaeus vannamei through dietary administration

In this study, the effects of feeding diets containing tyramine (TA) to Litopenaeus vannamei at levels of 10 (TA10), 50 (TA50), and 100 (TA100) mg/kg on growth performance, immune parameters, hemolymph glucose and lactate level, biogenic amine synthesis, and resistance against Vibrio alginolyticus were investigated. Results showed that shrimp fed with TA-containing diets had significantly higher feed efficiency and percent weight gain for a period of 56 days. The total hemocyte count increased in TA10, TA50 and TA100 after 7 and 28 days, and increased in TA100 after 14 days. The phenoloxidase activity increased in TA100 after 7 days, and respiratory bursts increased in TA100 after 7 and 14 days. In terms of resistance against V. aliginolyticus infection, the phagocytic activity increased in TA10, TA50 and TA100 after 7–28 days. The clearance efficiency increased in TA10, TA50 and TA100 after 7 and 14 days, and increased in TA 100 after 28 days. Shrimp fed with TA50 and TA100 for 28 days then challenged with V. aliginolyticus had a higher survival rate compared to those fed the basal diet at 48 h–168 h post challenges. The hemolymph glucose level was significantly decreased after being fed with TA10 at 28 days and TA100 for 56 days, while the octopamine synthesis in shrimp was improved after 14 days of being fed with TA50 and TA100, and after 28 days of being fed with TA100. These findings suggest that administering a TA diet regulated the growth performance, immune response and V. aliginolyticus resistance in L. vannamei.

Molecular characterization of four genes encoding abalone insulin-related peptides and their roles in regulation of hemolymph glucose in the Pacific abalone Haliotis discus hannai

Members of the insulin superfamily play key roles in metabolism, growth, reproduction, and longevity via insulin/IGF signaling pathway. The presence of insulin and insulin-related peptides are found among animals. In Mollusca, molluscan insulin-related peptides (MIPs) are involved in their body and shell growth. Hence, this study aimed to understand if there was difference in these peptide levels of fast and slow growing Pacific abalones, Haliotis discus hannai. To this end, four abalone insulin-related peptides (AIPs) are mined from cerebral ganglia transcriptomes of fast-growing and slow-growing abalones: AIP1, −2, −3 and −6. Notably, these AIPs possess 8 cysteine residues in their predicted insulin A and B chains, potentially supporting four disulfide bridges. Further sequence analysis based on the phylogenetic tree and multiple alignment methods indicated that AIPs have sequence features that are conserved among MIPs in many gastropods. The tissue distributions of four AIPs showed ubiquitous and similar expression in the following tissues including the cerebral ganglion, hepatopancreas, gonad, and mantle of both genders. However, there is some difference in other tissue distribution. Since the food consumption is intimately associated with the hemolymph glucose and animal growth, relative AIP1, −2, −3 and − 6 expression levels in the cerebral ganglion were measured following seaweed supplementation. The animals fed with brown seaweed showed increased these AIPs by 1.6–4.7 folds from 3 h after feeding, which lasting to 12 h, coinciding with the peak of hemolymph glucose. Based on these results, we propose that a physiological glucose regulatory mechanism involving AIPs exists in the cerebral ganglion of the H. discus hannai neuroendocrine system.

A mitotoxic fungicide alters post-ingestive glucose signals necessary for associative learning in honey bees

The Proboscis Extension Reflex (PER) paradigm trains honey bees to associate an odor with a sugar reward and is commonly used to assess impacts on associative learning after exposure to pesticides. While the effects of some types of pesticides have been well-investigated, relatively little attention has been focused on fungicides that are applied to flowering crops. We have previously shown that consumption of field-relevant concentrations of the fungicide Pristine® (active ingredients: 25.2% boscalid, 12.8% pyraclostrobin) impairs honey bee performance in an associative learning assay, but the mechanism of its action has not been investigated. We hypothesized that Pristine® interferes with carbohydrate absorption and/or regulation, thereby disrupting the post-ingestive feedback mechanisms necessary for robust learning. To test this hypothesis, we measured hemolymph glucose and trehalose levels at five time points during the ten minutes after bees consumed a sucrose solution. Pristine®-exposed bees had elevated baseline glucose concentrations in the hemolymph relative to control bees. Hemolymph glucose levels rose significantly within five minutes of feeding in control bees, but not in Pristine®-fed bees. These data suggest that the post-ingestive feedback mechanisms necessary for robust learning are disrupted in bees that have consumed this fungicide, providing a plausible mechanistic explanation for its effects on learning performance in the PER assay. Pristine®-exposed bees may have elevated hemolymph glucose levels because the fungicide elicits an inflammatory response. These results provide additional mechanistic understanding of the negative physiological effects of mitotoxic fungicides on this important pollinator.

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.

Metabolic and osmoionic effects of the recombinant crustacean hyperglycemic hormone (rCHH-A) of the Pacific white shrimp Penaeus vannamei on specimens acutely exposed to extreme salinities

The crustacean hyperglycemic hormone (CHH) is a multifunctional neuropeptide that plays a central role in crustacean metabolism and physiology. Experiments were conducted to examine the metabolic and osmoionic capabilities of the white shrimp Penaeus vannamei injected with its recombinant CHH-A (rCHH-A) peptide and acutely transferred from iso- (26 ppt) to hypo- (10 ppt) and hyper-osmotic (40 ppt) conditions. Hemolymph glucose, protein, osmoregulatory capacity (OC), and ionoregulatory capacity (IC) for sodium, chloride, potassium, and calcium were evaluated at four post-injection times (0.5, 1, 2, and 24 h). The rCHH-A peptide had hyperglycemic activity in all salinity conditions, obtaining maximum values at 1 h post-injection. However, in shrimp transferred to hyper-osmotic condition, rCHH-A caused the most significant reductions in OC (2 h), chloride IC (2 h), and total proteins (0.5-2 h) compared to the phosphate-buffered saline (PBS) control. Contrastingly, in shrimp transferred to hypo-osmotic conditions, rCHH-A decreased OC significantly from 2-24 h, strongly decreased chloride and potassium IC at 1 h post-injection, and increased total protein concentration in the hemolymph (1-2 h) when compared to PBS control. Concerning calcium, the rCHH-A injection decreased calcium IC at 10 ppt (1 h) and 26 ppt (2 h), providing insight into a potential role for CHH-A in calcium regulation. The results suggest that glucose and protein mobilization could enhance energy for osmo-ionic regulation under extreme osmotic conditions. This research study contributed to understanding crustacean endocrinology in P. vannamei and related euryhaline crustaceans. Further research should be performed to understand the osmo- and ionoregulatory mechanisms of the different CHH variants in crustaceans exposed to other stress conditions and the relationship with intermediary energy metabolism regulation.

STARVATION DURING DEVELOPMENT AFFECTS METABOLISM IN DROSOPHILA

Aim. To investigate how starvation during early stage of fly development affects carbohydrate metabolism in imago flies and their progeny of F1 generation. Methods. Wild-type Canton-S strain Drosophila melanogaster flies were used in all experiments. Flies of parental and offspring generations were used for the determination of glycogen and glucose content using the diagnostic kit Glucose-Mono-400-P according to the manufacturer's instructions. Results represent as the mean ± SEM of 3-4 replicates per group. According Student's t-test significant difference between groups was P&lt;0.05. Graphing and statistical analysis were performed by using GraphPad Prism. Results. Starvation during development significantly influenced the level of hemolymph and body glucose in imago flies of parental generation. Hemolymph glucose concentration was lower by 34% (P=0.008) and 32% (P=0.033) in experimental females and males, respectively, as compared to control groups. Starvation during development led to lower level of body glucose in adult parental flies of both sexes. Adult males F1, generated by parents that were starved during development, showed 3-fold lower glycogen content, as compared to control. Conclusions. Starvation at early stage of development led to lower hemolymph glucose and body glucose level in imago flies. Moreover, parental starvation decreased glycogen pool in F1 males.