Haemolymph Biochemical Parameters Research Articles

The effects of acutely elevated ammonia concentrations on haemolymph parameters in the Mediterranean mussel, Mytilus galloprovincialis

In aquaculture systems, ammonia nitrogen is a major component of waste products arising from the metabolism of cultured organisms and can significantly affect the quality of the rearing environment. The purpose of the current study was to determine modulation of the haemocyte population and selected haemolymph biochemical parameters of the Mediterranean mussel Mytilus galloprovincialis (Lamarck, 1919) exposed to acutely elevated ammonia concentrations.In order to investigate the extent and dynamics of an expected reduction of physiological fitness and survival rate of cultured mussels the stress-on-stress test was conducted combining ammonia and air exposure. The latter test illustrated the extent of reduced mussel physiological fitness and survival rate upon 72 h exposure to 10 and 30 mM NH4Cl in experimental tanks followed by additional exposure to air. Mussels cultured for up to 48 h in experimental tanks comprising 5, 10 and 30 mM NH4Cl, revealed dose and time dependent increase in haemolymph enzyme lactate dehydrogenase levels, urea, and uric acid metabolite levels, as well as the corresponding redistribution in haemocyte population profiles, indicating an inherent response of this organism to ammonia. Finally, a significant negative linear correlation between the mussel hyalinocytes subpopulation and the mean urea levels was observed. The obtained result suggests a possible function of basophilic and acidophilic granulocyte subpopulations in ammonia metabolism leading to urea synthesis by means of surplus ammonium ions are sequestered and extracted from the body. Therefore, defined modulation of the mussel haemocyte population structure may serve as an accurate bioindicator of mussels exposure to acutely elevated ammonia concentrations in environmental and aquaculture biomonitoring studies.

Evaluation of Yeast Hydrolysate in a Low-Fishmeal Diet for Whiteleg Shrimp (Litopenaeus vannamei).

An eight-week feeding trial was performed to evaluate the effects of yeast hydrolysate (YH) supplementation in a low-fishmeal diet on the growth, immune responses, intestinal histology and disease resistance of whiteleg shrimp (Litopenaeus vannamei). Five experimental diets were produced by supplementing YH at 0 (CON), 0.5 (YH0.5), 1 (YH1), 2 (YH2) and 4 (YH4) % to a basal diet containing 10% fishmeal and compared with a positive control with 25% fishmeal (FM25). Shrimp with an initial average weight of 0.43 ± 0.005 g (mean ± SD) were stocked in 18 tanks and fed the experimental diets (38% protein and 8% lipid) four times a day. Results showed that shrimp fed the FM25 diet exhibited significantly higher final body weight, weight gain, specific growth rate and protein efficiency ratio than those fed CON, YH0.5, YH1 and YH2 diets (p < 0.05). However, there were no significant differences between shrimp fed the YH4 and FM25 diets (p > 0.05). In addition, there were no significant differences in whole-body proximate composition, hemolymph biochemical parameters and non-specific immune responses among treatments. Intestinal villi length and muscular layer thickness of shrimp fed the YH4 and FM25 diets were significantly higher than the other groups. At the end of the bacterial (Vibrio parahaemolyticus) challenge test, shrimp fed YH4 and FM25 diets showed a significantly higher survival rate than those of shrimp fed CON, YH0.5 and YH1 (p < 0.05). These results suggest that supplementing 4% YH in diet containing 10% fishmeal could beneficially influence growth, intestinal morphology and disease resistance of whiteleg shrimp.

Dietary Riboflavin Requirement of Pacific White Shrimp (Litopenaeus vannamei).

This study was conducted to determine the dietary riboflavin requirement and its effects on growth performance, feed utilization, innate immunity, and diet digestibility of Litopenaeus vannamei. A riboflavin-free basal diet (R0) was formulated as a control, and six other diets were prepared by adding riboflavin of 10, 20, 30, 40, 50, and 60 mg/kg to the basal diet (designated as R10, R20, R30, R40, R50, and R60, respectively). Quadruplicate groups of shrimp (initial average weight 0.17 ± 0.00 g) were fed the diets six times a day for 8 weeks. Weight gain, specific growth rate, and protein efficiency ratio were significantly increased by riboflavin (p < 0.05). The maximum values were observed in shrimp fed R40 diet. The highest activities of phenoloxidase, nitro blue tetrazolium, superoxide dismutase, and glutathione peroxidase were observed in shrimp fed R40 diet. Lysozyme activity was significantly higher in shrimp fed R30 and R40 diets than that of shrimp fed R60 diet (p < 0.05). Intestinal villi were significantly longer in shrimp fed R50 and R60 diets compared to those of all other groups while the shortest villi were observed in R0 group (p < 0.05). Intestinal villi were clearly distinguished in shrimp fed higher levels of riboflavin compared to those of shrimp fed R0 and R10 diets. Apparent digestibility coefficients of dry matter and protein in diets were not significantly affected by riboflavin levels (p < 0.05). Whole-body proximate composition and hemolymph biochemical parameters were not significantly altered by dietary riboflavin (p < 0.05). Therefore, the results of this study indicate that riboflavin is essential to enhance growth performance, feed utilization, nonspecific immunity, and intestine morphology of shrimp. An optimal riboflavin requirement for the maximum growth of L. vannamei seems to be approximately 40.9 mg/kg diet.

Cholesterol supplementation improved growth performance, cholesterol metabolism, and intestinal health of Pacific white shrimp (Litopenaeus vannamei) fed a low fishmeal diet

An 8-weeks feeding trial was conducted to evaluate the effects of dietary cholesterol supplementation on growth performance, hemolymph biochemical parameters, cholesterol metabolism, intestinal health, and disease resistance of Pacific white shrimp (Litopenaeus vannamei) fed a low fishmeal diet. Four experimental diets including HFM (25% fishmeal), LFM (15% fishmeal+10% soy protein concentrate), LC1 (LFM+0.6 g kg−1 cholesterol), and LC2 (LFM+1.2 g kg−1 cholesterol) were formulated, each diet was fed to quadruplicate tanks with 40 shrimp (0.33 ± 0.03 g) per tank for 56 days. The results shown that cholesterol supplementation in the diet did not affect the growth performance of Litopenaeus vannamei (P > 0.05), but the crude lipid content significantly reduced in the LC2 group than that in HFM group (P < 0.05). In the hemolymph biochemical indices, the content of total cholesterol in LFM and LC1 groups were significantly lower than that in HFM group; the HDL-C content was significantly lower in the shrimp fed low fishmeal based-diets (LFM, LC1, and LC2) than those fed in HFM diet; the TG content of two cholesterol supplemented groups were significantly lower than that of HFM group; the MDA content was significantly higher in LFM group than that in HFM group (P < 0.05). In hepatopancreas lipid metabolism, the shrimp fed LFM diet has higher expression in srebp, fas, and acc than those fed the HFM diet, and two cholesterol groups significant decreased the acc relative expression compare with LFM group; compare with HFM group, the LFM group showed significantly up-regulated hmg-CoA, npc1 relative expression, and LC1 group significantly up-regulated scp relative expression, then LC2 group significantly up-regulated scp, npc2 relative expression of hepatopancreas in cholesterol metabolism; LFM, LC1 group significantly up-regulated beclin1, atg12 relative expression and LC2 group up-regulated beclin1 relative expression than those in HFM group of hepatopancreas in cell autophagy (P < 0.05). LC1 group up-regulated imd, relish, toll, myd88, pelle, tube, and dorsal relative expression and LC2 group up-regulated ikkβ, myd88, tube, and dorsal relative expression in intestinal immunity than those in HFM group (P < 0.05). Cholesterol supplementation increased the height of the intestinal folds of shrimp fed the low fishmeal based-diets (P < 0.05), as well as improved the integrity of the mitochondria and the endoplasmic reticulum. Dietary cholesterol supplementation decreased the abundance of harmful bacteria phylum (Chlamydiae) and increased the abundance of beneficial bacteria genus (Pseudoalteromona, Rhodopirellula, and Ruegeria) compared with LFM group (P < 0.05). Shrimp fed the LC1 diet showed higher resistance to V. parahaemolyticus challenge than those fed the LFM diet (P < 0.05). In conclusion, cholesterol supplementation in a low fishmeal diet significantly enhanced the lipolytic capacity of shrimp, promoted the cellular autophagy, improved the intestinal health, and enhanced disease resistance of Litopenaeus vannamei.

Effects of dietary different canthaxanthin levels on growth performance, antioxidant capacity, biochemical and immune-physiological parameters of white shrimp (Litopenaeus Vannamei)

An 8-week feeding trial was conducted to evaluate the effects of different dietary canthaxanthin (CX) levels on growth performance, pigmentation, antioxidant capacity, hemolymph biochemical parameters, immune response, and resistance to hypoxia stress of white shrimp (Litopenaeus vannamei). Juveniles (initial weight 1.15 ± 0.12 g) were fed with five iso‑nitrogenous and isolipidic diets supplemented with/without CX: 0 (control), 50, 100, 200, and 400 mg kg−1 diet. Results showed that the growth performance, survival, and feed conversion ratio improved significantly in CX supplemented treatments compared to the control. The redness of cooked shrimp tended to increase with increasing CX level; however, no significant difference was observed among the treatments that received CX of more than 100 mg kg−1. Further, the activities of digestive enzymes, total antioxidant capacity, and peroxidase increased significantly (P < 0.05), while the activities of glutathione peroxidase, catalase, and malondialdehyde decreased significantly (P < 0.05) in the hepatopancreas with increasing dietary CX levels. The immune-related enzymes (alkaline phosphatase, lysozyme, alanine aminotransferase, and aspartate aminotransferase) and hemolymph biochemical parameters (cholesterol, high-density, and low-density lipoprotein) are significantly affected by different dietary CX levels. Total carotenoids and astaxanthin contents in shrimp muscle, shell, head, and whole-body showed an upward trend with the increment of dietary CX. After exposure to hypoxia stress, juveniles in 200 mg kg−1 supplementation treatment exhibited the highest LT50 value among all the treatments. Moreover, broken-line regression analysis indicated that a dose of 173.73 to 202.13 mg kg−1 of CX was suitable in the diet of L. vannamei as a potential carotenoid source for substituting dietary astaxanthin in the shrimp feed industry.

A Comparison of the Effect of Blast Chilling and Electrical Stunning on Some Hemolymph Parameters in Red Swamp Crayfish (Procambarus Clarkii)

Abstract The aim of the present investigation is to compare the effect of standard method and electrical stunning on some hemolymph biochemical parameters in mid-sized Red Swamp Crayfish. The results showed that electrical stunning time of induction was significantly shorter and recovery was longer than chilling stun. Parameters of an electrical field that stuns crayfish beyond the point of recovery without causing damage have been identified. Based on this experiment, it seems that electrical stunning would be preferable to the other method.

Dietary turmeric, curcumin and nanoencapsulated curcumin can differently fight against salinity stress in Pacific white shrimp Penaeus vannamei Boone, 1931

The influence of diet supplementation with turmeric (TUR), curcumin (CUR) and nanoencapsulated curcumin (NMC) was evaluated on haemolymph biochemical parameters and hepatopancreas antioxidant enzymes of Penaeus vannamei (2.96 ± 0.10 g) exposed to low and high salinity stress at 5 and 55 g L−1 respectively. Nine dietary groups containing different dosages (g kg−1 diet) of TUR (TUR 2.5, TUR 5 and TUR 10), CUR (CUR 0.075, CUR 0.150 and CUR 0.300) and NMC (NMC 0.075, 0.150 and 0.300) along with the control group made the ten treatments. After nine weeks of feeding, CUR additives decreased the haemolymph triglycerides, cholesterol, glucose and cortisol in shrimps under normal and stress conditions compared with the control group; the lowest and highest values were observed in NMC 0.150 and control group respectively (p < 0.05). Adding TUR, CUR and NMC supplements to the diet increased total protein, total haemocytes count and differential haemocytes count in stressed and non-stressed shrimps; the highest was observed in NMC 0.150. The lowest and highest activities of antioxidant enzymes including CAT, SOD and GSH-Px before and after the salinity stress were detected in control group and NMC 0.150 respectively (p < 0.05). After the salinity stress, the experimental treatments increased the survival rate, where NMC 0.150 caused the highest survival (p < 0.05). In conclusion, different forms of dietary CUR improved antioxidant capacity and resistance of P. vannamei to acute salinity stress. Nanomicellar carriers in shrimp nutrition can promote immunity and antioxidant responses under stressful conditions through more efficient delivery of dietary immunostimulants.

Effects of long-term exposure of Mytilus galloprovincialis to thiacloprid: A multibiomarker approach

Thiacloprid is a neonicotinoid insecticide widely exploited in agriculture and easily mobilized towards aquatic environments by atmospheric agents. However, little information about its toxicological effects on aquatic invertebrate bioindicators is available. In this study, specimens of the mussel Mytilus galloprovincialis were exposed to thiacloprid at environmental (4.5 μg L−1) and 100 times higher than environmental (450 μg L−1) concentrations for 20 days. Thiacloprid affected haemolymph biochemical parameters, cell viability in the digestive gland, antioxidant biomarkers and lipid peroxidation in the digestive gland and gills at environmentally relevant concentrations (4.5 μg L−1). In addition, thiacloprid exposure caused histological damage to the digestive gland and gills. Interestingly, the pesticide was detected at levels equal to 0.14 ng g−1 in the soft tissues of sentinels exposed for 20 days to 450 μg L−1 thiacloprid in seawaterμ. Due to its harmful potential and cumulative effects after long-term exposure of M. galloprovincialis, thiacloprid may pose a potential risk to nontarget aquatic organisms, as well as to human health. This aspect requires further in-depth investigation.

How does sublethal permethrin effect non-target aquatic organisms?

Permethrin is belonged to pyrethroids that are one of the substances developed as an alternative to pesticides. Permethrin, which is used especially in agriculture, can bioaccumulate in the water and sediment when mixed into aquatic ecosystems. For this reason, it is necessary to investigate the effect of this substance on aquatic organisms other than the target organism. The aim of this study was the determination of acute and sublethal effects as antioxidant enzyme levels on different organs and hemolymph biochemistry of the non-target aquatic organism, narrow-clawed crayfish (Astacus leptodactylus), after exposure to permethrin, one of the synthetic pyrethroid pesticides, contaminating aquatic ecosystems due to its increase usage. The invertebrate model organism, the narrow-clawed crayfish, was selected for its bioindicator role in food webs as planktivorous grazers epibenthic scavengers and good alternative models in ecotoxicology studies with the importance in conservation of freshwater ecosystems. The 96-h LC50 value of permethrin to experimental species was estimated as 0.903 μg/L (95% CI = 0.5042-2.2734 μg/L) with probit analysis method. The sublethal concentration of the permethrin was determined by 1/10 of 96-h LC50 values as 0.09 μg/L. There were two control (negative and acetone) groups in the experiment. The sampling of hemolymph and the tissues (gills, hepatopancreas, and muscle) were done 48 h and 96 h after exposure of the permethrin. The total hemocyte counts significantly increased in the 96-h exposed group of permethrin (p<0.05). Among the hemolymph biochemical parameters, the hemolymph potassium and chloride values increased statistically (p<0.05). Malondialdehyde levels (MDA) of gills and muscle were significantly increased, whereas the MDA level of the hepatopancreas was significantly decreased at the end of the experiment (p<0.05). Hyperplasia in the lamella was recorded in gills, while the degenerations of the hepatopancreas tissues were observed. According to obtained results, permethrin was extremely toxic as acutely to narrow-clawed crayfish and also effected at sublethal concentrations.

Effect of common salt (NaCl) on reproductive tract dimension, haemolymph haemocyte count and biochemical parameters of Giant African Land snail (A. marginata) during dry season

A study was conducted on the effect of common salt (NaCl) on reproductive tract dimension, haemolymph haemocyte count and biochemical parameters of Giant African Land snail (GALs), Archachatina marginata during the dry season. Forty (40) snails with a weight range of 150 g–180 g were used for this study. Four (4) treatment with each consisting of ten (10) replicates were assigned as follows: T1 (Control: concentrate +0 g of salt (NaCl)), T2 (concentrate +0.05 g of salt (NaCl)), T3 (concentrate+0.15 g of salt (NaCl) and T4 (concentrate + 0.25 of salt (NaCl)). The experiment lasted 13 weeks. Haemolymph was collected at the end of 13 weeks to assess haemocyte count, total protein, albumin, globulin, alanine transaminase (ALT) and aspartate transaminase (AST). The haemocyte count was determined using a Neubeur haemocytometer, while haemolymph biochemical parameters were determined using spectrophotometric methods with the appropriate kit. The snails were also dissected for complete isolation of the reproductive tract for morphometric measurements. Key organs concerned were: Ovo-testis, albumen, little hermaphrodite duct and hepatopancreas. The weight and length of each of these organs were taken together with complete tract weight. Ovo-testis from each treatment was also harvested for histology. A mortality record was taken as the experiment progresses. The result showed that serum biochemical parameters were not significantly (P > 0.05) affected by a various inclusion level of salt used. But it had a significant effect on ovotesis length, albumen weight, little hermaphrodite weight and reproductive tract weight. However, no significant effect was seen in ovotestis weight, albumen length, little hermaphrodite length and hepatopancreas weight and length. It was also discovered that common salt (NaCl) significantly affected (P < 0.01) haemocyte count and mortality record of GALS. Snails fed 0.25 g NaCl had the highest haemocyte count, followed by those fed 0.15 g, while snails fed 0 g and 0.05 g recorded the least value. It was also discovered that 0 g NaCl inclusion into the diet of snail (A. marginata) recorded the highest mortality (60 %) followed by 0.05 g NaCl inclusion level which gave 30 % mortality. However, the inclusion level of 0.15 g and 0.25 g NaCl into snail feed recorded no mortality. Histological evaluation of the ovotestis revealed that snail given 0.25 g inclusion into the concentrate diet had active acini with spermatogonium and well-formed oocyte, followed by those group with 0.15 g with active acini with several ova. It is obvious from this study that NaCl had an immune-modulatory influence on GAS during the dry season via mitigation of mortality without negatively affecting the haemolymph biochemical parameter. Inclusion used in this study also boost reproductive apparatus functions which are usually not active during this period. It is therefore recommended that both 0.15 g and 0.25 g/kg be included into the diet of A. marginata during the dry season to mitigate challenges that confront snail farming during the period to boost production and reduce economic losses due to mortality during this season.

Spirulina platensispowder is an applicable feed additive for Chinese horseshoe crabTachypleus tridentatus

The development of artificial feeds for Tachypleus tridentatus is important given the requirement of Tachypleus amebocyte lysate and the temporary rearing and industrial aquaculture. The effects of Spirulina platensis powder on growth performance, haemolymph biochemical parameter and non-specific immune enzymes were investigated. Addition of S. platensis powder improved artificial feed quality for growth performance. Haemocyanin concentration, total protein and total cholesterol (TCH) did not differ among the groups after 4 weeks of feeding natural feeds. TCH in Treatment 1 decreased significantly after 2 weeks, but this phenomenon was resolved by the addition of S. platensis powder. Total antioxidant, superoxide dismutase and catalase decreased with varying degrees in the groups of semi-artificial feed within the first 2 weeks, but the parameters converged after 4 weeks. Malonaldehyde increased after 4 weeks with the addition of S. platensis powder to semi-artificial feed. Groups with semi-artificial feed had similar contents of acid phosphatase and alkaline phosphatase to those of the group with natural feed. Thus, no differences were found among groups during the 4 weeks. S. platensis powder not only had physiological stimulative effects but also increased the growth of the adult Chinese horseshoe crab T. tridentatus when the crabs were fed with semi-artificial feed. The powder also stabilized the haemolymph biochemical parameters and non-specific immune enzymes. Therefore, the effects of S. platensis powder as feed additive on the adult T. tridentatus were evaluated. This study provides first-hand data for the development of artificial feeds and generates positive results for T. tridentatus.

Accumulation of paralytic shellfish toxins by Southern Rock lobster Jasus edwardsii causes minimal impact on lobster health

Recurrent dinoflagellate blooms of Alexandrium catenella expose the economically and ecologically important Southern Rock Lobster in Tasmania to paralytic shellfish toxins (PST), and it is unknown if PST accumulation adversely affects lobster performance, health and catchability. In a controlled aquaculture setting, lobsters were fed highly contaminated mussels to accumulate toxin levels in the hepatopancreas (mean of 6.65 mg STX.2HCl equiv. kg−1), comparable to those observed in nature. Physiological impact of PST accumulation was comprehensively assessed by a range of behavioural (vitality score, righting ability and reflex impairment score), health (haemocyte count, bacteriology, gill necrosis and parasite load), nutritional (hepatopancreas index and haemolymph refractive index) and haemolymph biochemical (21 parameters including electrolytes, metabolites, and enzymes) parameters during a 63 day period of uptake and depuration of toxins. Exposure to PST did not result in mortality nor significant changes in the behavioural, health, or nutritional measures suggesting limited gross impact on lobster performance. Furthermore, most haemolymph biochemical parameters measured exhibited no significant difference between control and exposed animals. However, the concentration of potassium in the haemolymph increased with PST, whilst the concentration of lactate and the sodium:potassium ratio decreased with PST. In addition, exposed lobsters showed a hyperglycaemic response to PST exposure, indicative of stress. These findings suggest that PST accumulation results in some measurable indicators of stress for lobsters. However, these changes are likely within the adaptive range for Jasus edwardsii and do not result in a significant impairment of gross performance. Our findings support previous conclusions that crustaceans are relatively tolerant to PST and the implications for the lobster fishery are discussed.

EFFECTS OF AESTIVATION DURATION ON TESTOSTERONE, HAEMOLYMPH BIOCHEMICAL PARAMETERS AND REPRODUCTIVE TRACT DIMENSIONS AND WEIGHT OF GIANT AFRICAN LAND SNAIL (Archachatina marginata) DURING DRY SEASON

Aestivation is a process of metabolic inactivity under which energy reserve are manipulated for survival. Reproductive apparatus and haemolymph biochemical agents also undergo phase manipulation as the duration continues. To determine the physiological influence on key hormone of reproduction and reproductive apparatus, seventy five A. marginata snails were used for this study. The snails were divided into five treatments with fifteen replicate each. Treatment include: Zero (0) week, Three (3) weeks, Six (6) weeks, nine (9) weeks and six (6) weeks post-aestivation. Parameters measured were: Testosterone concentration, haemolymph biochemical parameters (Total protein, albumin, globulin, Aspartate transaminase (AST) and Alanine transaminase (ALT), dimensions (length) and weight of the organs and tissues of the reproductive tracts (Ovo-testis, penis, vaginal, oviduct, little hermaphrodite duct, common hermaphrodite duct, vas deferens and albumen gland) gonado-somatic index and percentage mortality. Result showed that level of testosterone at three and six weeks of aestivation significantly reduced compared to the control. Also, at nine weeks of aestivation, the reduction was significantly greater than what was recorded at both three and six weeks of aestivation. But the testosterone levels were reversed at nine weeks post aestivation. Total protein and globulin were significantly influenced with both reaching a peak value at 9 weeks of aestivation while ALT, AST and albumin were not significantly affected. So also, of all the reproductive tract parts measured, organ weight, ovo-testis weight, penis weight and length were significantly influenced (P&lt;0.001; P&lt;0.001; P&lt;0.05; P&lt;0.001). Similarly, vaginal weight, oviduct weight, little hermaphrodite duct weight and length were also significantly affected together with vas deferens length and albumen gland length while aestivation duration had no significant influence on reproductive tract weight, ovo-testis length, vaginal length, oviduct length, little hermaphrodite duct length, vas deferens weight and albumen gland weight. Similarly, gonado-somatic index was also not significantly affected by aestivation duration. It was also obvious from this study that the highest mortality was recorded at 6 weeks of aestivation, followed by 9 weeks of aestivation while 3weeks and 6 weeks post-aestivation had the least mortality with the control intact. &#x0D; In conclusion, it is clear from this study that aestivation duration significantly influenced testosterone concentration, haemolymph biochemical parameters and some selected reproductive apparatus of A. marginata.&#x0D; &#x0D; &#x0D; &#x0D;

Effects of copper supplement on the immune function and blood-chemistry in adult Chinese horseshoe crab Tachypleus tridentatus

Oxygen in hemocyanin binds to copper ions and turns the hemolymph of horseshoe crabs blue. Copper ions not only transport oxygen but also enhance the immune function of horseshoe crabs. A 28-day indoor culture experiment was conducted to study the effects of adding different levels of copper amino acid chelate (0, 50, or 100 mg kg−1) into semi-finished feed on the nonspecific immune enzyme activity and hemolymph biochemical parameters of Tachypleus tridentatus. Results revealed that the activities of serum antioxidant enzyme indicators, namely, copper–zinc superoxide dismutase (Cu/Zn-SOD), catalase (CAT), glutathione peroxidase (GPX), total antioxidant capacity (T-AOC), lysozyme (LZM), alkaline phosphatase (AKP), and acid phosphatase (ACP), of copper-fed adult horseshoe crabs increased; on the other hand, malondialdehyde (MDA) activity decreased. Hemolymph biochemical indicators, triglyceride (TG) and cholesterol (CHO) decreased in the copper-fed group, but hemocyanin concentration (HC) levels increased during the experiment. At the end of experiment, all antioxidant enzymes and blood biochemical indicators in the copper-fed group were significantly different from those without copper supplement. Hemolymph T-AOC, MDA, AKP, ACP values in the low-level copper group (50 mg kg−1) were significantly different from those in the high-level copper group (100 mg kg−1).

Acute exposure to chlorpyrifos and glyphosate induces changes in hemolymph biochemical parameters in the crayfish, Astacus leptodactylus (Eschscholtz, 1823)

Acute and subacute toxicity tests for chlorpyrifos and glyphosate were performed on the crayfish, Astacus leptodactylus. The crayfish were divided into a control group and four experimental groups with exposure from 50.18 to 301.11 μg L−1 of chlorpyrifos for 96 h under semi-static test conditions. In the same experiment, the crayfish were exposed to different concentrations of glyphosate (from 0.0, 8.14 to 13.05 mg L−1) for 96 h. Mortality was recorded, and the median lethal concentrations (LC50) were calculated using probit analysis. The 96 h LC50 values of chlorpyrifos and glyphosate to A. leptodactylus were 49.55 ± 4.66 μg L−1 and 7.83 ± 0.50 mg L−1, respectively. The results showed that chlorpyrifos was a few hundred times more toxic than glyphosate for the crayfish. Then, the crayfish were exposed to different subacute concentrations of chlorpyrifos (12.5 and 25 μg L−1) and glyphosate (2 mg L−1 and 4 mg L−1) for seven days, and their toxicity was evaluated through several hemolymph biochemical parameters. Chlorpyrifos and glyphosate exposure caused a significant (P < 0.01) increase in the activity of lactate dehydrogenase (LDH), and gamma-glutamyl transferase (GGT). The results showed a significant (P < 0.01) decrease in acetylcholinesterase (AChE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and phenoloxidase (PO) activities in the hemolymph of the crayfish exposed to chlorpyrifos and glyphosate. The total protein and cholesterol levels showed a significant decrease (P < 0.01) in the hemolymph of the crayfish after seven days of exposure to chlorpyrifos and glyphosate. There was a significant increase in triglycerides and glucose levels in treated crayfish. These results show that chlorpyrifos is highly toxic, while glyphosate is moderately toxic to A. leptodactylus. According to the results, subacute concentrations of chlorpyrifos and glyphosate could lead to a collapse of cellular homeostasis and changes in the hemolymph biochemical parameters in the crayfish. Physiological changes caused by exposure to chlorpyrifos and glyphosate in A. leptodactylus may have direct effects on the survival of these organisms.

The impact of holding stressors on the immune function and haemolymph biochemistry of Southern Rock Lobsters (Jasus edwardsii)

Lobsters are fished world-wide due to their status as a high value, luxury seafood. A large proportion of the product is sold via live export, with lobsters subject to a range of stressors during holding post-capture. Improving the current understanding of the immune response to these stressors assists in improving efficiency and reducing loss in the chain between capture and consumption. In this study, the immune status of four treatment groups of Southern Rock Lobster (Jasus edwardsii) were studied: controls recently landed from a fishing boat, lobsters displaying advanced shell necrosis, lobsters in an unexplained moribund state and lobsters held in a processing facility for 10 weeks in standard conditions (i.e. high density, fasted). A total of 15 immune parameters and 19 haemolymph biochemical parameters were assayed. Phenoloxidase activity was only sporadically observed in haemocyte lysate and was consistently observed at a low level in the plasma with no difference between treatments for either. Haemocyte lysate prophenoloxidase activity was detected in most individuals, with no differences found between treatments. Prophenoloxidase in the plasma showed the highest level of activity, with the shell necrosis treatment demonstrating an elevated activity level relative to the other three treatments. Cell viability was not affected in any treatment. Lobsters with shell necrosis had a reduced capacity for phagocytosis, a significantly higher total haemocyte count, fewer hyalinocytes and more granulocytes and semigranulocytes. Fasted lobsters showed an opposite shift, with significantly more hyalinocytes compared to the other treatments and very few granulocytes and semigranulocytes. The balance of a range electrolytes, minerals metabolites and enzymes were affected in shell necrosis and fasted treatments, raising them as potential markers for immunocompromised lobsters. Multivariate analysis of all assayed parameters showed that all individuals in the necrosis treatment showed a similar, distinct immune response and that the fasted treatment, along with one control and one moribund individual, showed a separate intermediate response. The remainder of the control and moribund lobsters demonstrated a distinct “non-response” in comparison. These results offer a characterisation of the physiological response to common challenges during post-capture holding of rock lobsters, demonstrating the differential response to pathogenic bacterial infection, long term fasting, non-specific moribundity and the stress of capture and transport.

Utilization of date seed meal in the diet of Pacific white shrimp (Penaeus vannamei): growth performance, body and fatty acid composition, biochemical parameters, and tolerance of salinity stress

Current study aimed to examine the effectiveness of date seed meal (DSM) in the diet of Pacific white shrimp (Penaeus vannamei). Shrimp were fed on diets containing different amounts of DSM including 0, 50, 100, 150, and 230 g/kg for a period of 8 weeks. Growth parameters and body proximate composition showed no significant differences among treatments (p > 0.05). Similarly, no notable differences were observed in hemolymph biochemical parameters in shrimp fed with different amounts of DSM compared with the control group (p > 0.05). The contents of whole body polyunsaturated fatty acids (PUFAs) including docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA), and total n-3 in P. vannamei fed diet containing 100 g/kg DSM were significantly greater than control treatment (p 0.05). Altogether, the results of this study showed that DSM could be utilized as a cheapest source of carbohydrate in the shrimp diet without depressive effects on its performance.

Evaluation of the Protective Roles of Synthetic Zeolite on Some Physiological and Biochemical Parameters after Cadmium Toxicity of Crayfish (Procambarus Clarkii)

Background: the release of heavy metals into the environment through industrial effluents is a major concern, worldwide and removal of such pollutants has been a great concern during the last decades. Heavy metals are not biodegradable and tend to be accumulated in organisms and cause numerous diseases and disorders. Aim of the Work: this study evaluated the protective role of synthetic zeolite against cadmium toxicity of freshwater crayfish (Procambarus clarkii). Patients and Methods: the crayfish was divided into six groups (36 individual in each group), the first group was used as a control group, the second group was exposed to a dose of (50 μg/L) cadmium chloride for 45 days, the third group exposed to a dose of (50 μg/L) cadmium chloride for 45 days and then added zeolite (1 mg/L) for 45 days. The fourth group was exposed to a dose of (50 μg/L) cadmium chloride for 45 days and then added zeolite (5 mg / L) for 45 days, the fifth group was exposed to a dose of (50 μg/L) cadmium chloride plus (1 mg/L) zeolite for 45 days. The sixth group was exposed to a dose of (50 μg/L) cadmium chloride plus (5 mg/L) zeolite for 45 days. After the experimental periods, the crayfish were weighted and hemolymph was collected to measure the biochemical parameters (Glucose, total protein, albumin, globulin, A/G ratio, cholesterol, triglyceride, LDL, HDL, Na+, K+, Cu+2, Ca+2and Mg+2). Results: in G2 the concentration of total protein, albumin, globulin, HDL, Na+, K+, Cu+2 and Ca+2 were significantly decreased (p< 0.05) compared with the control group, while glucose, A/G ratio, cholesterol, triglyceride, LDL and Mg+2 were significantly increased (p< 0.05) compared with the control group. The addition of the ion-exchanging agent, zeolite (1 mg/L and 5 mg/L) to cadmium exposed group (G2) caused improvement in weight and all hemolymph biochemical parameters in G3, G4, G5 and G6. Conclusion: the synthetic zeolite was able to protect crayfish against cadmium toxicity by reducing the transfer of cadmium from polluted water into crayfish tissue and reducing the chance for metal uptake by interacting in the experimental medium which in evidently improves the physiological and biochemical functions.

Evaluation of the Protective Roles of Synthetic Zeolite on Some Physiological and Biochemical Parameters after Cadmium Toxicity of Crayfish (Procambarus Clarkii)

Background: the release of heavy metals into the environment through industrial effluents is a major concern, worldwide and removal of such pollutants has been a great concern during the last decades. Heavy metals are not biodegradable and tend to be accumulated in organisms and cause numerous diseases and disorders. Aim of the Work: this study evaluated the protective role of synthetic zeolite against cadmium toxicity of freshwater crayfish (Procambarus clarkii). Patients and Methods: the crayfish was divided into six groups (36 individual in each group), the first group was used as a control group, the second group was exposed to a dose of (50 μg/L) cadmium chloride for 45 days, the third group exposed to a dose of (50 μg/L) cadmium chloride for 45 days and then added zeolite (1 mg/L) for 45 days. The fourth group was exposed to a dose of (50 μg/L) cadmium chloride for 45 days and then added zeolite (5 mg / L) for 45 days, the fifth group was exposed to a dose of (50 μg/L) cadmium chloride plus (1 mg/L) zeolite for 45 days. The sixth group was exposed to a dose of (50 μg/L) cadmium chloride plus (5 mg/L) zeolite for 45 days. After the experimental periods, the crayfish were weighted and hemolymph was collected to measure the biochemical parameters (Glucose, total protein, albumin, globulin, A/G ratio, cholesterol, triglyceride, LDL, HDL, Na+, K+, Cu+2, Ca+2and Mg+2). Results: in G2 the concentration of total protein, albumin, globulin, HDL, Na+, K+, Cu+2 and Ca+2 were significantly decreased (p< 0.05) compared with the control group, while glucose, A/G ratio, cholesterol, triglyceride, LDL and Mg+2 were significantly increased (p< 0.05) compared with the control group. The addition of the ion-exchanging agent, zeolite (1 mg/L and 5 mg/L) to cadmium exposed group (G2) caused improvement in weight and all hemolymph biochemical parameters in G3, G4, G5 and G6. Conclusion: the synthetic zeolite was able to protect crayfish against cadmium toxicity by reducing the transfer of cadmium from polluted water into crayfish tissue and reducing the chance for metal uptake by interacting in the experimental medium which in evidently improves the physiological and biochemical functions.

The impact of seismic air gun exposure on the haemolymph physiology and nutritional condition of spiny lobster, Jasus edwardsii

There is a critical knowledge gap regarding the impacts of seismic air gun signals on the physiology of adult crustaceans. We conducted four controlled field experiments to examine the impact of seismic acoustic signals on spiny lobster, Jasus edwardsii. Seismic air gun exposure suppressed total haemocyte count (THC) for up to 120days post-exposure, suggesting a chronic negative impact of immune competency. THC levels after 365days post-exposure, were elevated two fold, potentially indicating an immune response to infection. Haemolymph refractive index was reduced after 120days post exposure in one experiment, suggesting a chronic impairment of nutritional condition. There was no effect of air gun exposure on 24 haemolymph biochemical parameters, hepatopancreas index or survival. Collectively these results indicate that the biochemical haematological homeostasis of J. edwardsii is reasonably resilient to seismic acoustic signals, however, air gun exposure may negatively influence the lobster's nutritional condition and immunological capacity.