Hemocyanin Levels Research Articles

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.

Cold temperatures as a source of stress: seasonality, sand burrowing and hemocyanin levels in Atlantic Canada sandy beach amphipods

Cold temperatures as a source of stress: seasonality, sand burrowing and hemocyanin levels in Atlantic Canada sandy beach amphipods

Hemocyanin as a biological indicator of artificial light at night stress in sandy beach amphipods

The influence of artificial light at night (ALAN) is becoming evident in marine sandy beaches. These habitats are dominated by species reliant on natural daylight/night regimes, making the identification of biological indicators a priority. We assessed the applicability of hemocyanin, an oxygen-transport protein in the hemolymph of many invertebrates, as an indicator of ALAN-related stress. Unlike total proteins, hemocyanins signal metabolic function and stress, so we expected them to increase in response to ALAN. We adapted spectrophotometry protocols to describe spatial variation in hemocyanins and total proteins in four populations of the talitroid amphipod Americorchestia longicornis. Then, a two-week experiment tested for changes in response to ALAN. Hemocyanin levels increased by 17% and 40% with respect to experimental controls after 7 and 14 d, respectively, and were higher than any measurements conducted in the field. These results suggest good prospects for hemocyanin as an indicator of ALAN effects.

Sublethal behavioral and physiological effects of claw removal on Jonah crabs (Cancer borealis)

Jonah crabs (Cancer borealis) have been caught as incidental bycatch for over 80 years, but in recent years they have become a targeted fishery, especially in Southern New England where the American lobster (Homarus americanus) population has declined, and harvesters have begun seeking alternative fisheries. This shift has prompted the need for more biological data for Jonah crabs so that this expanding fishery can be managed more effectively. For example, in some areas of its range males have both of their claws removed at sea and only the claws are landed; however, little is known about what happens to the crabs that survive this practice. The focus of this study was to evaluate the impacts of claw removal on Jonah crab mating success, feeding ability and locomotion. In mating trials, all males, regardless of claw status, managed to successfully mate with females that molted while they were paired together (clawed n = 4, claws removed n = 8). When Jonah crab activity was measured in the laboratory, crabs with intact claws were significantly more active than crabs with both claws removed (clawed: 25.0 ± 8.0% of each hour active, 1 claw removed: 18.1 ± 7.3% of each hour active, clawless: 13.6 ± 9.04% of each hour active, mean ± SD). Similarly, based on data from a tag/recapture experiment, crabs with both claws missing moved about half the distance per day as control crabs, between the time they were released and first recaptured (clawed: 117 ± 43.1 m/day, clawless: 50 ± 11.3 m/day, mean ± SE), although the difference was not significant. These changes in locomotion could be the result of the blood loss associated with claw removal. We found that hemocyanin levels dropped significantly after the removal of both claws and remained so for up to two weeks. Finally, crabs with no claws were able to feed, but they had difficulty opening mussel shells, which might influence their diet in their natural habitat. These data suggest that those Jonah crabs that do survive the claw removal process might be impaired, but should be able to forage, mate, and help sustain the population.

Effects of the biomedical bleeding process on the behavior and hemocyanin levels of the American horseshoe crab (Limulus polyphemus)

Effects of the biomedical bleeding process on the behavior and hemocyanin levels of the American horseshoe crab (Limulus polyphemus)

The protective effects of melatonin on oxidative damage and the immune system of the Chinese mitten crab (Eriocheir sinensis) exposed to deltamethrin

Deltamethrin (Del), an important broad-spectrum insecticide, is widely used in agricultural activities. However, Del is an effective reactive oxygen species (ROS) inducer that induces oxidative stress damage in cells or tissues. Del is significantly more toxic to aquatic organisms, especially crustaceans, than to mammals and birds. This study was designed to evaluate the protective effect of melatonin (MT) on the toxicity-induced damage of Del after 6 h in Eriocheir sinensis. The results showed that Del exposure significantly induced oxidative damage in the hepatopancreas and mitochondria, with malondialdehyde (MDA) and glutathione (GSH) levels being significantly increased and superoxide dismutase (SOD) activity being significantly decreased. Moreover, Del exposure significantly induced functional damage of the hepatopancreas and mitochondria, with a significant increase in alanine aminotransferase (ALT), aspartate aminotransferase (AST), acid phosphatase (ACP) and alkaline phosphatase (AKP) activities in the hepatopancreas and the ratio of albumin/globulin (ALB/GLB) in serum, which indicated the permeability and integrity of the membranes were damaged and had caused cell damage. In addition, ATP content, Na+-K+-ATPase activity and cytochrome C (Cyt‑C) content in mitochondria decreased significantly, which indicated that Del exposure destroyed the normal respiratory chain of mitochondria. We also evaluated the hematological parameters. Although there were no significant differences in total hemocyte count (THC) levels, hemocyte apoptosis was significantly induced by Del exposure, and the hemocyte phagocytic activity and the hemocyanin levels decreased significantly with Del exposure. However, MT pretreatment not only prevented oxidative damage and functional damage caused by Del exposure to the hepatopancreas and mitochondria, but it also restored the hemocyte apoptotic rate and phagocytic activity to normal levels. In short, Del exposure caused significant oxidative and functional damage to the hepatopancreas, mitochondria and hemocytes of E. sinensis, whereas the use of MT almost completely eliminated the damage caused by Del exposure.

Effect of Probucol in combination with Enalapril-folic acid tablets on endothelial function in elderly patients with H-type hypertension

Objective To investigate the effect of Probucol combined with Enalapril-folic acid tablets on endothelial function in elderly patients with H-type hypertension. Methods A total of 180 elderly patients with H-type hypertension were randomly divided into three groups (n=60, each): Group A (conventional treatment), Group B (conventional treatment + a tablet containing 10 mg enalapril and 0.8 mg Folic acid/day) and Group C (conventional treatment + 500 mg Protocol twice daily + a tablet containing 10 mg Enalapril and 0.8 mg folic acid/day). Plasma hemocyanin (Hcy) and asymmetric dimethylarginine (ADMA) levels and serum nitric oxide (NO) and endothelial NO synthase (eNOS) levels were measured before treatment, and 1 and 6 months after treatment. Results In Group A, plasma Hcy and ADMA levels and serum NO and eNOS levels had no significant differences before treatment versus 1 and 6 months after treatment (P>0.05). Plasma Hcy and ADMA levels decreased and serum NO and eNOS levels increased in Group B 6 months after treatment, compared with those at pretreatment or 1 month after treatment, which were either lower or higher than those in Group A 6 months after treatment (all P<0.05). Plasma Hcy and ADMA levels decreased and serum NO and eNOS levels increased in Group C 6 months after treatment, compared with those at pretreatment or 1 month after treatment, which were either lower or higher than those in Groups A and B 6 months after treatment (all P<0.05). After adjustment for other risk factors, partial correlation analysis showed that plasma ADMA was positively correlated with plasma Hcy, while plasma ADMA and Hcy were negatively correlated with serum eNOS and NO (all P<0.05). Conclusions Probucol in combination with Enalapril-folic acid tablets may be beneficial in improving endothelial function and mitigating the development of atherosclerosis. Key words: Hypertension; Endothelium, vascular

Is Copper Supplementation Required in Formulated Feeds forOctopus vulgaris(Cuvier, 1797)?

The aim of this study was to test the effect of copper supplementation in a formulated feed on the growth, feed efficiency, and nutritional composition of subadults Octopus vulgaris, with particular reference to the differences in the content of copper in the tissues (muscle, digestive gland, and hemolymph). Two formulated feeds were supplied to subadults octopuses kept individually for 56 days: Basal diet [11.6 mgCu/kg dry weight (dw)], and another feed with a similar composition but including a copper supplementation (copper diet: 115.2 mgCu/kg dw). No significant differences (P > 0.05) were detected in the respective growth rates (0.88% body weight/day), FE (79.5%–82.1%), proximate composition, hemolymph hemocyanin (1.6–1.8 mmol/L), hemolymph copper (0.19–0.20 mgCu/mL), or muscle copper levels (22.7–23.2 mgCu/kg dw). No significant changes were detected on proximate composition, hemocyanin and hemolymph, and muscle copper levels between animals fed formulated feeds and an initial group fed natural diet based on crab and fish (P > 0.05). The only parameter which showed a significant difference (P < 0.05) was the copper level in the digestive gland: 1,797, 390, and 1,148 mgCu/kg dw for initial (natural diet), basal and copper diet, respectively (P < 0.05). Considering the experimental conditions of this study, copper supplementation is not effective in improving the performance of a formulated feed for octopus.

Oxidative damage of hepatopancreas induced by pollution depresses humoral immunity response in the freshwater crayfish Procambarus clarkii

Previous studies provide evidences for the possible oxidative damage of toxic environmental pollutants to tissue protein in fish and amphibian, but little information is available about their effects on immunity response in crustacean. In the present study, we evaluated the relationship between oxidative damage and immune response induced by both typical pollutants (viz. copper and beta-cypermethrin), by exposing the freshwater Procambarus clarkii to sub-lethal concentrations (1/40, 1/20, 1/10 and 1/5 of the 96 h LC50) up to 96 h. Five biomarkers of oxidative stress, i.e. reactive oxygen species (ROS), superoxide dismutase (SOD), catalase (CAT), malondialdehyde (MDA) and protein carbonyl in hepatopancreas, and two immune factors, i.e. phenoloxidase (PO) and hemocyanin in haemolymph were determined. The results indicated that there was a significant increase (P < 0.05) in the contents of ROS, MDA and protein carbonyl accompanied by markedly decreased (P < 0.05) PO and hemocyanin levels in a dose and time dependent manner. The significant and positive correlation (P < 0.01) between protein carbonyls induction and MDA formation was observed in crayfish hepatopancreas at 96 h. The production of these protein carbonyls could significantly depress (P < 0.01) the levels of phenoloxidase and hemocyanin in hemolymph. Higher contents of ROS enhanced the risk of lipid peroxidation, protein carbonylation and immunosuppression of crayfish, and hepatopancreas might play an important role in immune system of crustaceans. Protein oxidation may be one of the main mechanisms for pollution-induced immunotoxicity in P. clarkii.

Sublethal Behavioral and Physiological Effects of the Biomedical Bleeding Process on the American Horseshoe Crab, Limulus polyphemus

The hemolymph of the American horseshoe crab, Limulus polyphemus, is harvested from over 500,000 animals annually to produce Limulus amebocyte lysate (LAL), a medically important product used to detect pathogenic bacteria. Declining abundance of spawning Limulus females in heavily harvested regions suggests deleterious effects of this activity, and while mortality rates of the harvest process are known to be 10%-30%, sublethal behavioral and physiological effects are not known. In this study, we determined the impact of the harvest process on locomotion and hemocyanin levels of 28 female horseshoe crabs. While mortality rates after bleeding (18%) were similar to previous studies, we found significant decreases in the linear and angular velocity of freely moving animals, as well as changes in their activity levels and expression of circatidal behavioral rhythms. Further, we found reductions in hemocyanin levels, which may alter immune function and cuticle integrity. These previously unrecognized behavioral and physiological deficits suggest that the harvest of LAL may decrease female fitness, and thus may contribute to the current population decline.

The Paradoxical Effects of Lead in Interferon-Gamma Knockout BALB/c Mice

It has been reported that lead (Pb) exposure enhances interleukin (IL)-4 and inhibits interferon-gamma (IFNgamma) production in wild-type (WT) BALB/c mice. Here, we examined Pb effects on immunity in IFNgamma knockout (KO) mice. Lead significantly enhanced serum IgG1 anti-keyhole limpet hemocyanin (KLH) levels in WT mice compared to the controls; Pb also increased serum IgG2a anti-KLH levels, but the IgG1:IgG2a ratio was greater with Pb. In addition, total serum IgE levels, but not IgE anti-KLH levels, were increased. In the KO mice, the serum IgG1, IgG2a, IgE anti-KLH, and total IgE levels were significantly lower than those of WT mice. Surprisingly, Pb significantly enhanced IgG1 and IgG2a anti-KLH levels in the KO mice. However, for these mice, unlike the WT mice, Pb caused a greater percentage change in IgG2a than in IgG1 anti-KLH, indicating less skewing toward type-2 immunoglobulins. Lead also enhanced the delayed-type hypersensitivity (DTH) response in WT mice. Not surprisingly, very low DTH occurred in the KO mice; however, Pb induced a strong KLH-specific DTH response. The in vivo Pb exposure significantly increased in vitro production of IL-4, IL-5, and IL-10, but not IFNgamma, IL-2 and IL-12, by KLH-induced WT and KO spleen cells. In contrast to KLH, dinitrofluorobenzene contact hypersensitivity (DNFB CHS) was detected in all groups, and Pb did not affect this response, which suggests that Pb has only a slight effect on CD8+ T cell-related responses. As previously reported, Pb enhances Th2 responses in WT mice; however, in the KO mice, Pb enhanced Th1-related anti-KLH production and a Th2-related DTH. The Pb enhancement of DTH in IFNgamma-deficient mice is likely due to promotion of type-2 cytokines and enhancement of major histocompatibility complex (MHC) class II expression.

Effects of molting and environmental factors on trace metal body-burdens and hemocyanin concentrations in the American lobster, Homarus americanus

Hemocyanin concentrations in the hemolymph of marine crustacea are dependent on the molt cycle and on environmental conditions. Studies in our laboratories have found that hemocyanin levels in blue crabs are reduced after ecdysis and under conditions of environmental stress (Engel, Brouwer, & McKenna, 1993. Hemocyanin concentrations in marine crustaceans as a function of environmental conditions. Marine Ecology Progress Series, 93, 233–244). We have extended those studies to include the American lobster, Homarus americanus. Hemolymph and digestive gland tissues from Long Island Sound lobsters were analyzed for hemocyanin, copper, and zinc during different stages of the molt cycle. Hemocyanin, copper and zinc in the hemolymph were highest in premolt stages (D 1–D 4), and lowest in the postecdysal papershell stages (B 1–B 2). Concomitantly, copper in digestive glands decreased significantly following ecdysis, but no significant changes in the metals bound to metallothionein (MT) were observed. Copper-MT was the predominant form throughout the molt cycle, presumably because lobsters were obtained from copper-contaminated areas. To examine the effects of environmental factors, intermolt lobsters were collected from locations of different environmental quality along the Atlantic coast, and were analyzed for hemocyanin and trace metals. In general, animals from areas with a history of contamination showed the highest hemocyanin concentrations.

Hemocyanin re-uptake in the renal and branchial heart appendages of the coleoid cephalopod Sepia officinalis.

The renal and branchial heart appendages of Sepia officinalis L. were investigated in order to elucidate a possible involvement of their excretory epithelia in hemocyanin metabolism. Immunocytochemical findings and tracer experiments indicate that after passing the barrier of ultrafiltration the hemocyanin molecules are taken up by the epithelial cells of the renal and branchial heart appendages and are subsequently carried back to the circulatory system, suggesting a mechanism of hemocyanin recycling. Apart from a function in maintaining constant hemocyanin levels, the present study indicates that the renal and branchial heart appendages are also sites of temporary hemocyanin storage.

Hemocyanin levels in the giant keyhole limpet, Megathura crenulata, from the coast of California

1. 1. The concentration of hemocyanin in the hemolymph of the giant keyhole limpet Megalhura crenulata, has been determined. 2. 2. In 56 specimens of M. crenulata the hemocyanin concentration ranged from 1.53 to 10.81 g/1 with a mean of 5.45 g/1 and a standard deviation of 2.71 g/1. There is no correlation between the hemocyanin concentration and the weight of the animal. The hemolymph volume and the colour of the mantle are not related to the hemocyanin level either. 3. 3. The hemocyanin concentration shows a seasonal variation, the means and the standard deviations for February, April, July, and September being 6.48 ± 2.58, 4.44 ± 1.72, 2.62 ± 1.05, and 7.65 ± 1.54 g/1, respectively. 4. 4. The lability of the hemocyanin concentration in Megathura (7-fold variation) is less pronounced than it is in California Haliotis, but comparable to that of the Australian abalones. 5. 5. It is difficult to reconcile the wide fluctuations observed in the concentration of hemocyanin with an essential respiratory task for this protein.

Hemocyanin levels in Octopus vulgaris and the cuttlefish Sepia officinalis from the aegean sea

1. 1. The concentration of hemocyanin in the hemolymph of Octopus vulgaris and the cuttlefish Sepia officinalis has been determined. 2. 2. In fourteen specimens of 5. officinalis the hemocyanin concentration ranges from 58.4 to 88.9 g/l, the mean value being 71.1 and the standard deviation 10.9 g/l. There is no correlation between the weight of the animal and the hemocyanin concentration; neither does the sex of the animal have any bearing on the abundance of the copper protein. 3. 3. In five specimens of O. vulgaris the hemocyanin concentration ranges from 47.6 to 66.9 g/l, the mean being 55.9 and the standard deviation ± 7.4 g/l. 4. 4. The wide range of hemocyanin concentrations observed in gastropods and crustaceans apparently is not a characteristic of cephalopods. In these highly aerobic molluscs with closed circulatory systems the synthesis and the breakdown of hemocyanin seems to be tightly controlled and, as a result, the concentration of the copper protein in the hemolymph shows little fluctuation among the individual animals.