Haemolymph Parameters Research Articles

Effect of supplementing heterotrophic and photoautotrophic biofloc, on the production response, physiological condition and post-harvest quality of the whiteleg shrimp, Litopenaeus vannamei

The effect of supplementing biofloc produced under heterotrophic and photoautotrophic conditions, on the production response, physiological condition and postharvest quality of juvenile L. vannamei, intensively farmed under greenhouse conditions was assessed. Heterotrophic bioflocs were produced under a restricted light condition, using an unspecific marine consortium as inoculum and maintaining the C:N ratio around 10-12. The photoautotrophic biofloc were produced under direct light exposition; using Navicula incerta as inoculum and maintaining the C:N ratio around 2-3. No significant differences on the water quality variables were observed among treatments except for TAN which was higher in the control. The production response was affected and a lower survival and higher FCR were recorded with heterotrophic bioflocs. The hemolymph parameters were similar in shrimp between groups, except for cholesterol which increased by more than 4-fold in the control. The postharvest quality of shrimp was qualified as good in general terms, without significant differences between groups, but the mean of the total qualifiers was slightly better in the treatment with photoautotrophic biofloc. The results of the study suggest that supplementation of both types of biofloc has not negative effect on the water quality, on the physiological condition of shrimp and on their postharvest quality.

Assessing the effects of neonicotinoid insecticide on the bivalve mollusc Mytilus galloprovincialis

In the present work, the marine invertebrate Mytilus galloprovincialis was used as model organism to evaluate the toxic effects of the neonicotinoid Calypso 480 SC (CAL) following 20 days of exposure to sub-lethal concentrations of 7.77 mg L−1 (0.1% 96 h-LC50) and 77.70 mg L−1 (1% 96 h-LC50), and a recovery period of 10 days in uncontaminated seawater. Results revealed that exposure to both concentrations of CAL increased significantly mortality rate in the cells of haemolymph and digestive gland, while digestive gland cells were no longer able to regulate cell volume. Exposure significantly reduced haemolymph parameters (Cl−, Na+), affected the enzymatic activities of superoxide dismutase of digestive gland and catalase of gill, and caused also histopathological alterations in digestive gland and gills. Main histological damages detected in mussels were lipofuscin accumulation, focal points of necrosis, mucous overproduction and infiltrative inflammations. Interestingly, alterations persisted after the recovery period in CAL-free water, especially for haemocyte parameters (K+, Na+, Ca2+, lactate dehydrogenase, glucose). A slight recovery of histological conditions was detected. These findings suggested that sub-chronic exposure to the neonicotinoid insecticide caused significant alterations in both cell and tissue parameters of M. galloprovincialis. Considering the ecologically and commercially important role of mussels in coastal waters, a potential risk posed by neonicotinoids to this essential aquatic resource can be highlighted.

Evaluation of some haemolymph biochemical properties and F‐cell prevalence in hepatopancreas of white leg shrimp (Litopenaeus vanammei) after fed diets containing apple cider vinegar and propionic acid

The present study aimed to evaluate the effect of dietary ACV (apple cider vinegar) and PA (propionic acid) supplementation on some biochemical properties of haemolymph and fibrillar cell (F-cell) number in the hepatopancreas of shrimp. Five different diets supplemented with 0%, 1%, 2%, 4% ACV and 0.5% PA diets were fed to Litopenaeus vanammei. Some biochemical parameters of haemolymph (cortisol, alkaline phosphatase, albumin, urea, uric acid, creatinine, phosphorus, total hemocyte count) and prevalence of F-cell in hepatopancreas were examined after 60 days feeding trial. The albumin concentration and total hemocyte count in 2% and 4% ACV and PA groups were significantly higher than those of other groups (p < .05). Moreover, the highest phosphorus concentration was detected in shrimp fed 4% ACV and PA experimental diets (p < .05). No significant differences in haemolymph cortisol, urea, uric acid, creatinine, alkaline phosphatase and tubule area were observed among the experimental groups (p > .05). 2% ACV group showed more F-cell number in hepatopancreas than other groups while the reduction in F-cell number was observed in the PA group. Based on these observations, the tested ACV and PA diets remarkably exhibited positive impacts on nutritional value and they may be a potential for adding to shrimp diet.

Effect of biofloc technology on growth of speckled shrimp, Metapenaeus monoceros (Fabricus) in different feeding regimes

The effects of biofloc technology (BFT) were tested on growth performance, water quality, haemolymph parameters and hepatopancreas histology with and without BFT for the speckled shrimp. In this study, eight different experimental groups were formed (four BFT and four control groups) to measure the effects of different feeding rates on compensatory growth of speckled shrimp. BFT treatments were performed with zero water exchange and planned as BFT1: gradually feed decreasing group 4, 2 and 1% bw, BFT2: 4% bw, BFT3: 2-days feed with 4% bw, 1-day fasted, and BFT4: 1-day feed with 4% bw, 1-day fasted. In control treatment, the same trial groups were constituted for comparing with BFT at a 50% daily water exchange: C1 (gradually feed decreasing group: 4%, 2% and 1% bw), C2 (4% bw), C3 (2-days feed with 4% bw, 1-day fasted), C4 (1-day feed with 4% bw, 1-day fasted). There were significant differences between BFT and C groups in terms of some water quality (p < 0.05). In general, BFT had an effect on haemocyte profile. At the histological examination, there were no pathological findings in both BFT and control groups. The growth parameters of speckled shrimps in BFT groups, as well as the whole body crude protein ratios and crude ash contents, were higher than the control group (p < 0.05). The results confirmed that biofloc utilization increased with decreasing feeding.

Impact of climate change on the American lobster (Homarus americanus): Physiological responses to combined exposure of elevated temperature and pCO2

The physiological consequences of exposing marine organisms to predicted future ocean scenarios, i.e. simultaneous increase in temperature and pCO2, have only recently begun to be investigated. Adult American lobster (Homarus americanus) were exposed to either current (16 °C, 47 Pa pCO2, pH 8.10) or predicted year 2300 (20 °C, 948 Pa pCO2, pH 7.10) ocean parameters for 14–16 days prior to assessing physiological changes in their hemolymph parameters as well as whole animal ammonia excretion and resting metabolic rate. Acclimation of lobster simultaneously to elevated pCO2 and temperature induced a prolonged respiratory acidosis that was only partially compensated for via accumulation of extracellular HCO3– and ammonia. Furthermore, acclimated animals possessed significantly higher ammonia excretion and oxygen consumption rates suggesting that future ocean scenarios may increase basal energetic demands on H. americanus. Enzyme activity related to protein metabolism (glutamine dehydrogenase, alanine aminotransferase, and aspartate aminotransferase) in hepatopancreas and muscle tissue were unaltered in future ocean scenario exposed animals; however, muscular citrate synthase activity was reduced suggesting that, while protein catabolism may be unchanged, the net energetic output of muscle may be compromised in future scenarios. Overall, H. americanus acclimated to ocean conditions predicted for the year 2300 appear to be incapable of fully compensating against climate change-related acid-base challenges and experience an increase in metabolic waste excretion and oxygen consumption. Combining our study with past literature on H. americanus suggests that the whole lifecycle from larvae to adult stages is at risk of severe growth, survival and reproductive consequences due to climate change.

Response of Pacific white leg shrimp (Litopenaeus vannamei) on exposure to aflatoxin in feed

A research was accomplished to find the response of Litopenaeus vannamei to different levels of aflatoxin in feed. Two hundred live L. vannamei were collected from a shrimp breeding station located on the south of Tiab, in Hormozgan province. Aflatoxin contaminated diets with 0.18, 96.21, 184.74, 711.35, 977.11 and 1605.61 µg/kg were prepared and shrimps in six experimental groups with three replicates in each group for 28 days were fed with these diets. The results showed that activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) in haemolymph and histological alternation index (HAI) value of hepatopancreas in treatement groups were significantly higher than control group (p < 0.05). Decreased total protein and fat in haemolymph and increased protein in muscle of treatment shrimps were observed compared with the control group (p < 0.05). The HAI value of 977.11 and 1605.61 μg/kg groups were significantly higher than the control and other groups (p < 0.05). Histological study results showed that calculated HAI (more than 100) in these groups as a result of necrosis lesion observed in hepatopancreas. Moderate to severe alteration was recognized in group fed by 711.35 µg/kg aflatoxin. Minor aflatoxin levels (0.18, 96.21 and 184.74 µg/kg) caused inappreciable tissue alteration. This study demonstrated body composition, haemolymph parameters and histology of hepatopancreas were affected in the aflatoxin‐contaminated feed when the concentration was over 955.14 µg/kg.

Dietary exposure to melamine and cyanuric acid induced growth reduction, oxidative stress and pathological changes of hepatopancreas in Pacific white shrimp

This study examined the effects of dietary melamine (MEL) and cyanuric acid (CYA) singly and in combination on growth, nutrient utilization, immunological responses, oxidative stress, and histological changes in Pacific white shrimp. Seven experimental isonitrogenous (35%) and isolipidic (8%) diets were formulated, namely diet 1 (a control diet without MEL and CYA); diets 2–5 (with MEL and CYA at 2.5 + 2.5, 5 + 5, 7.5 + 7.5 and 10 + 10 g kg−1 diet); diet 6 (with only MEL at 10 g kg−1 diet) and diet 7 (with CYA alone at 10 g kg−1 diet). The shrimp with initial body weight 2.37 ± 0.02 g were fed with these diets for 10 weeks. The results indicate that all the diets with MEL and CYA singly or in combination had adverse effects on growth and nutrient utilization relative to the control diet (p < 0.05). Total protease and trypsin activities were significantly lowered by all diets containing MEL (p < 0.05). Haemolymph parameters, including total hemocyte count, phenoloxidase (PO) activity, respiratory burst, and lysozyme activity, were significantly decreased (p < 0.05) in shrimp receiving MEL alone (10 g kg−1 diet) and at high combination dosages (10 + 10 g kg−1 diet). Moreover, MEL and CYA induced oxidative stress, damaged hepatopancreas, decreased antioxidant responses, increased lipid peroxidation, and caused abnormality of hepatocytes.

The effects of claw ligatures in American lobster (Homarus americanus) storage: a preliminary study of haemolymph parameters

American lobsters are crustaceans that are offered for sale live and are stored in controlled temperature recirculating aquaria. During marketing, they are subjected to stressors that can affect their welfare, such as air exposure, confinement, and handling. European legislation does not provide specific criteria or retention requirements, and so their management depends largely on the common sense of food business operators. Claw ligatures before and during storage are not legally required but are recommended because they prevent lobsters from damaging each other and ensure workers’ safety. The aim of the present study is to evaluate the effect of claw ligatures on the lobsters’ welfare by analysing eight different haemolymph stress indicators, vitality, and weight. The calcium level showed significant differences in the two experimental groups (P&lt; 0.05). Our results suggested that the absence of rubber bands did not offer any significant contribution to the lobsters’ welfare.

Effects of Oxfendazole on Some Biochemical Parameters in Hemolymph of Galleria mellonella L. (Lepidoptera: Pyralidae) Larvae Reared on Artificial Diet

Oxfendazole is used as an antihelminthic drug in parasite infections of animals. Oxfendazole was added at different concentrations (0, 0.0015, 0.015, 0.15, and 1.5%) to artificial diet and the wax moth Galleria mellonella larvae were reared to 7th stage. The changes in metabolic enzymes activity in G. mellonella hemolymph tissue were investigated. It was found that the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) was increased which are cell damage biomarkers. The highest concentration of this antihelmintic was increased the enzme activity (ALT) from 329 ± 4.33 to 529. 5 ± 3.17 U/L. Gamma glutamyl transferase (GGT) enzyme activity was increased from 0.87 ± 0.1 to 4.32 ± 0.22 U/L. Creatine kinase (CK) and amylase (AMYL) activities were significantly increased in hemolymph of G. mellonella larvae fed with the highest oxfendazole concentration (1.5%) when compared to the control group. Our results indicate that oxfendazole can be used as an insecticide with a well-adjusted concentrations because of its low acute toxicity to humans and other non-target organisms.

Dosage and frequency effects of the microalgae Dunaliella sp. on the diet of Litopenaeus vannamei challenged with Vibrio parahaemolyticus.

Carotenoid sources in shrimp diets have shown to be effective for improving survival, growth, reproductive capacity, stress resistance, and also for diminishing disease. Dunaliella sp. is known to have high levels of β-carotenes, which works as pro-vitamin A, enhancing the immune response in shrimp. However, the administration of Dunaliella sp. in shrimp diet needs to be evaluated to determine the appropriate dose and frequency of administration needed to optimize performance in cultured white shrimp. Diets with three different concentrations of Dunaliella sp. flour (1.5, 2 and 3%) were tested, and each one was administered at three different time frequencies: daily, and at 3- and 7-days intervals. Shrimp fed for 20 days were then infected with Vibrio parahaemolyticus (1 × 106 CFU/mL). Hemolymph parameters including protein, glucose, lactate, cholesterol and triglycerides were analyzed to evaluate shrimp stress status. Additionally, L. vannamei innate non-specific immune response was examined by evaluating the activity of prophenoloxidase (proPO), phenoloxidase (PO) and superoxide dismutase (SOD) in hemolymph; shrimp survival was also recorded. Survival after infection with V. parahaemolyticus was higher for shrimp fed with diets consisting of 2% Dunaliella sp. administered every 3 and 7 days. Shrimp fed a diet consisting of 2% or 3% Dunaliella sp. administered every third day showed positive physiological and immune responses to infection. A decrease in lipid oxidation in plasma triglycerides was observed at 48 h post inoculation in shrimp fed at all diets regimes due to Dunaliella sp. antioxidant action. Experimental results suggest the importance of Dunaliella sp. dosage and feeding frequency in L. vannamei diet to improve the survival and immune response.

Dietary supplementation with sulfated polysaccharides from Gracilaria birdiae promotes a delayed immunostimulation in marine shrimp challenged by the white spot syndrome virus

Immune system stimulation by dietary supplementation has been widely explored in aquaculture as an effective practice in increasing shrimp resistance against infectious diseases. Given the recurring impacts of white spot syndrome virus (WSSV) on shrimp farming worldwide, the search for immunostimulant compounds is of urgent importance. This study investigated the effect of a diet supplemented with sulfated polysaccharides (SP) from the red seaweed Gracilaria birdiae on the immune response of Pacific white shrimp (Litopenaeus vannamei) challenged with a non-lethal dose of WSSV. The health status and immunocompetence of infected shrimp were monitored by assessing multiple immune parameters in hemolymph. Dietary supplementation with SP from G. birdiae (32 days) was able to promote a delayed immunostimulation by enhancing the agglutinating capacity and PO activity and an increase in the survival of L. vannamei challenged with a non-lethal dose of WSSV. Our findings support the potential application of this marine seaweed in shrimp farming and contribute to a better understanding of the effects of immunostimulants on shrimp immune and physiological responses.

Effect of dietary supplementation with apple cider vinegar and propionic acid on hemolymph chemistry, intestinal microbiota and histological structure of hepatopancreas in white shrimp, Litopenaeus vannamei

This experiment was conducted to evaluate the effects of dietary supplementation of Apple cider vinegar (ACV) and propionic acid (PA) on biochemical parameters of hemolymph, intestinal microbiota and histology of hepatopancreas in white shrimp (Litopenaeus vannamei). Five experimental diets were evaluated in this study including diets supplemented with 1.0, 2.0 and 4.0% of ACV, 0.5% propionic acid, and a control diet with no supplements. Shrimps (initial weight of 10.2 ± 0.04 g) in triplicate groups with the density of 25 shrimps per tank were fed the diets for 60 days. At the end of the feeding trial, shrimps fed with ACV and PA supplemented diets had significantly higher total protein level than those fed the control diet (P < 0.05). The number of Vibrio spp., R-cells (lipid storage cells) of hepatopancreas and cholesterol level in shrimps fed the diets containing ACV and PA were lower compared to the control group (P < 0.05). However, there was no remarkable variations in glucose concentration, B-cell number and tubule diameter among the experimental diets (P > 0.05). In addition, shrimps fed the ACV diets had significantly lower total heterotrophic marine bacteria compared to the control or PA groups, and the lowest bacterial number was observed in shrimp fed 4% ACV supplemented diet (P < 0.05). Supplementation of 2 and 4% ACV as well as 0.5% PA in the diet led to a significantly higher calcium concentration than the control treatment (P < 0.05). The lowest triglyceride concentration was observed in the shrimps fed diets containing 2.0 and 4.0% ACV, which resulted in 15 and 20% reduction, respectively (P < 0.05). Overall, the findings indicates that ACV and PA possess antimicrobial activity and demonstrate beneficial effects on health status, so they can be potentially used as feed additive in the feeding of L. vannamei.

Responses of Mytilus galloprovincialis to challenge with the emerging marine pathogen Vibrio coralliilyticus

Vibrio coralliilyticus has emerged as a coral pathogen of concern throughout the Indo-Pacific reef. The interest towards understanding its ecology and pathogenic potential has increased since V. coralliilyticus was shown to be strongly virulent also for other species; in particular, it represents a serious threat for bivalve aquaculture, being one of the most important emerging pathogen responsible for oyster larval mortalities worldwide. V. coralliilyticus has a tightly regulated temperature-dependent virulence and it has been related to mass mortalities events of benthic invertebrates also in the temperate northwestern Mediterranean Sea. However, no data are available on the effects of V. coralliilyticus in the mussel Mytilus galloprovincialis, the most abundant aquacultured species in this area. In this work, responses of M. galloprovincialis to challenge with V. coralliilyticus (ATCC BAA-450) were investigated. In vitro, short term responses of mussel hemocytes were evaluated in terms of lysosomal membrane stability, bactericidal activity, lysozyme release, ROS and NO production, and ultrastructural changes, evaluated by TEM. In vivo, hemolymph parameters were measured in mussels challenged with V. coralliilyticus at 24h p.i. Moreover, the effects of V. coralliilyticus on mussel early embryo development (at 48 hpf) were evaluated. The results show that both in vitro and in vivo, mussels were unable to activate immune response towards V. coralliilyticus, and that challenge mainly induced lysosomal stress in the hemocytes. Moreover, V. coralliilyticus showed a strong and concentration-dependent embryotoxicity. Overall, the results indicate that, although M. galloprovincialis is considered a resistant species to vibrio infections, the emerging pathogen V. coralliilyticus can represent a potential threat to mussel aquaculture.

Biochemical and histological effects of sub-chronic exposure to atrazine in crayfish Cherax destructor

Atrazine (ATR) is a triazine herbicide banned in the European Union. It remains one of the most widely used herbicides in other parts of the world. Considering the scarcity of data on its possible harm to the environment and to human health, we assessed sub-chronic effects of a 14-day exposure at the environmentally relevant concentration of 6.86 μg/L and at 10% of the 96hLC50 (1.21 mg/L) in crayfish Cherax destructor and their recovery in a 14-day period in ATR-free water. Indicators assessed were behavior; hemolymph biochemical profile; oxidative and antioxidant parameters in gill, hepatopancreas, and muscle; and histology of gill and hepatopancreas. Crayfish exposed to the environmental concentration showed significant differences (P < 0.01) from controls in biochemical parameters of hemolymph (lactate, alkaline phosphatase) and activity of superoxide dismutase, as well as in histology of gill tissue. The higher concentration led to low motor activity, differences in biochemical profile of hemolymph (lactate, alkaline phosphatase, ammonia, glucose), antioxidant biomarkers (superoxide dismutase, catalase, glutathione reductase, glutathione S-transferase, reduced glutathione), as well as gill and hepatopancreas histology. Some observed effects persisted after 14-days recovery in ATR-free water. The results provide evidence that environmental concentrations of ATR produce negative effects on freshwater crayfish.

Study on establishing normal ranges of chosen biochemical parameters of haemolymph of Cornu aspersum maxima and Cepaea nemoralis gastropods.

The aim of the study was to establish normal ranges for chosen biochemical parameters of haemolymph of snails (Gasropoda: Mollusca), in the light of the use of these animals as experi- mental models in various types of studies. The study was conducted on 100 specimens of Cornu aspersum maxima (CAM) and 100 specimens of Cepaea nemoralis (CN). The haemolymph col- lected from the animals was analysed using colorimetry to assay aspartate transaminase (AST) activity, alanine transaminase (ALT) activity, amylase activity and the concentrations of urea and triglycerides. In the further part of the study, the influence of administering doxycycline with feed on the change of AST and ALT activity in snail haemolymph has been studied. The normal values established for CAM are as follow: AST activity: 26-38 u/l, ALT activity: 0-11 u/l, amylase activity 9-16 u/l, concentration of urea: 3-6 mg/dl, concentration of triglycerides: 16-20 mg/dl. For CN, the following data have been obtained: AST activity: 30-80 u/l, ALT activity: 0-15 u/l, amylase activity 12-15 u/l, concentration of urea: 5-8 mg/dl, concentration of triglycerides: 18-24 mg/dl. It has been shown that doxycycline presents a high workload on the hepatopancreas of snails, which is reflected by a statistically significant (p ⟨ 0.05) increase of AST and ALT activity in the haemolymph of the specimens which obtained doxycycline in feed, as compared to the groups with antibiotic-free feed. The haemolymph activity of both studied parameters increased together with study time and tetracycline administration time.

IMPACT OF NURSERY USING DIFFERENT PHYSICAL FILTRATION SYSTEM ON HEMOLYMPH GLUCOSE LEVEL AND SURVIVAL RATE OF SPINY LOBSTER JUVENILE Panulirus homarus

The problems in culture of early juvenile phase of spiny lobster Panulirus homarus is low survival rate of the seed. One of strategies to improve production is using filtration system during nursery of spiny lobster. The aim of this study to evaluate differentation of physical filtration system on responsse stress and survival rate of juvenile P. homarus. Experimental design of this study consisted of four treatments and two replications. The treatments were using flow through system without protein skimmer and filter (K), filtration system with protein skimmer (SK), filtration system with top filter (F) and filtration system with combinations of protein skimmer and top filter (SKF). P. homarus with initial weight 0.18+0.01 g were cultured for 60 days with density 250 lobster/tank. During maintenance, P. homarus were fed trash fish with feeding rate 20% of body weight. Parameters of glucose hemolymph were evaluated to determine stress response. Stress response was determined at day 0, 6, 20 and 60. Parameters of survival rate were determined at the end of experiment. The result showed that the treatment of filtration system with combination of protein skimmer and top filter (SKF) decreased stress responsse by decreasing glucose hemolymph of juvenile P. homarus during maintenance. Survival rate also higher (P&lt;0.05) in treatment SKF (33.2%) than other treatments. It is concluded that SKF was the best physical filtration system for juvenile period of spiny lobster P. homarus Keywords: filtration system, glucose hemolymph, spiny lobster, survival rate

Gender-related variations in hemolymph parameters of Carabus lefebvrei (Coleoptera: Carabidae): HPLC analysis and phenoloxidase activity

We characterized the enzymatic activity of basal and total phenoloxidase and HLPC and SDS- PAGE profiles in hemolymph of Carabus lefebvrei males and females at different reproductive status. The phenoloxidase activity was activated by trypsin and inhibited by phenoloxidase activity specific inhibitor phenylthiourea. Our results demonstrated that both in males and females, there were no significant differences in the basal phenoloxidase activity between reproductive and virgin beetles, while the total phenoloxidase activity increased significantly in virgin specimens. Thus, resources seem to be invested to increase the humoral response in pre-reproductive phase forming a barrier against pathogens and preserving the fecundity and longevity of both sexes. The hemolymph DOPA-MBTH assay on polyacrilamide gel electrophoresis showed a high activity of monomeric form with an apparent molecular weight of 90 kDa and a dimer of about 170 kDa, also multimeric bands were present in both sexes. In the SDS-PAGE general protein pattern, specific bands were evident for reproductive and virgin males and females as biochemical markers of sexual difference in immunocompetence. Reproducible differences in peaks were recorded in HPLC analysis performed on virgin and reproductive males and females

Evaluation of dietary soybean meal as fish meal replacer for juvenile whiteleg shrimp, Litopenaeus vannamei reared in biofloc system

Different levels of dietary soybean meal (SBM) as a fish meal (FM) replacer, with and without amino acid supplementation, for whiteleg shrimp, Litopenaeus vannamei reared in the biofloc system was examined in eight weeks of feeding trial. Eight experimental diets consisted of a basal diet with 0% FM replacement by SBM provided in clear sea water without biofloc system (S0SW), four diets replacing FM at 0% (S0), 33% (S33), 67% (S67) and 100% (S100) by SBM, and three diets replacing FM at 33% (S33A), 67% (S67A) and 100% (S100A) by SBM supplemented with amino acids (methionine and lysine) in the seawater biofloc system. Results of water quality analyses showed significantly lower total suspended solids and nitrate for S0SW group than all other treatments. Diets S0 and S33A resulted in higher weight gain and specific growth rate among all groups, with no significant differences with S33 group. In addition, whole-body protein and amino acid compositions of shrimp fed S0SW were lower than most biofloc groups. Haemolymph parameters showed significant differences in total protein, cholesterol and triglyceride between groups S0 and S0SW. Also, superoxide dismutase activity showed a decreasing trend with increasing replacement level. In conclusion, based on these results, SBM could replace up to 33% of FM with or without amino acid supplementation in juvenile whiteleg shrimp diets reared in the biofloc system.

New intravital method for hemolymph collection from Cornu aspersum snails and the establishment of standards for selected biochemical parameters of their hemolymph

The aim of the study was to develop an intravital method of collecting hemolymph and to establish standards for the biochemical parameters of the hemolymph of the Cornu aspersum snail species. The research material consisted of 80 Cornu aspersum maxima snails. Hemolymph was collected from the main vessel with a sterile syringe and needle after removing a fragment of the shell. Hemolymph was studied biochemically by the colorimetric method. The parameters evaluated were aspartate transaminase (AST), alanine transaminase (ALT), amylase (AMYL), urea (UREA), and triglycerides (TG). An average of 0.54 ml hemolymph was collected from the animals tested. After 24 hours, a film formed in place of the removed shell fragment and it underwent mineralization within a few days. All animals included in the study survived, and from some of them it was possible to collect hemolymph several times at intervals of several days. The proposed method of hemolymph collection made it possible to obtain material to develop standards for the biochemical parameters of hemolymph and consequently to monitor the health status of these mollusks.

Spatial Variability in Condition of Southern Rock Lobsters (Jasus edwardsii) at the Start of the Tasmanian Fishing Season.

The southern rock lobster (Jasus edwardsii) industry in Australia favours red lobsters, which are usually caught in shallow waters, over paler (brindle) lobsters. This preference is driven partly by the Chinese market, where red is associated with luck and prosperity, and additionally, by the widely held perception within the industry that brindles have greater mortality rates during out of water transport than reds. Limited scientific evidence supports these industry observations; however, these studies did not evaluate the initial condition of lobsters. This study aimed first, to determine which variables better describe condition in J. edwardsii and second, to compare condition among lobsters in several sites around Tasmania at the typical time of high transport mortality. Male lobsters were collected from the South West, South East, East and North coast of Tasmania in late November/December 2014, which correspond to the start of the Tasmanian fishing season. A comprehensive condition assessment was applied by measuring tissue proximal composition, Brix index, Total Haemocyte Count, pH, haemocyanin and another 16 haemolymph parameters of interest. A useful framework to compare condition in J. edwardsii was established by first, using Brix index as a measure of nutritional condition, second, using pH, magnesium, and bicarbonate to evaluate differences in physiological condition and finally, using THC counts as a proxy for lobster health condition. Lobsters from different sites had different nutritional, physiological and health condition, consistent with industry observations, however our results indicate that some red shallow water lobsters exhibited poorer nutritional and health condition, while some deep water brindle lobsters were in good condition. Differences in condition could not be directly associated to catch depth of lobsters and was related to other spatially discrete factors which sometimes vary over distances <3 km.