Hepatopancreas Of White Shrimp Research Articles

The in silico and toxicity study of fraction red betel leaf (Piper crocatum) extract on histology lymphoid organ and hepatopancreas in white shrimp (Litopanaeus vannamei)

The shrimp farming industry has experienced a high mortality caused by diseases in recent decades. One of the most common diseases is vibriosis. This research aims is to obtain information on compounds found in fraction one of Piper crocatum as a therapeutic agent using in silico prediction and evaluate the toxicity test with an LC50. The potential of active ingredient content from P. crocatum extract was analyzed by in silico methods. The LC50 test was conducted by injections of the fraction one extract of P. crocatum for 96 hours at doses of 0 (control), 2 μg/g, 4 μg/g, 6 μg/g, 8 μg/g, and 10 μg/g. In silico toxicity (LD50) analysis showed that citral had the highest toxicity value (LD50) at 500 mg/kg and was classified as toxicity class 4, with an abundance value of 3.71%. The 10 μg/g concentration showed an LC50 mortality value of 53.3±5.8%. The percentage of heavy damage was observed at the concentration of 10 μg/g (53.33). These results indicate that the higher the concentration of fraction one extract of P. crocatum, the higher the percentage of damage to white shrimp's hepatopancreas and lymphoid organs.

Effects of a Single Cell Protein (Methylococcus capsulatus, Bath) in Pacific White Shrimp (Penaeus vannamei) Diet on Growth Performance, Survival Rate and Resistance to Vibrio parahaemolyticus, the Causative Agent of Acute Hepatopancreatic Necrosis Disease

The efficacy of a single cell protein (SCP) methanotroph (Methylococcus capsulatus, Bath) bacteria meal (FeedKind®, Calysta, Menlo Park, CA, United States), in Pacific white shrimp (Penaeus vannamei) diets was studied to determine growth performance, survival rate and disease resistance against Vibrio parahaemolyticus causing Acute Hepatopancreatic Necrosis Disease (AHPND). The growth trial was assigned in a completely randomized design (CRD) with four treatments and 5 replicates of each, T1: a fishmeal-based control containing 15% fish meal and 3 diets with graded levels of methanotroph bacteria meal, namely T2: 5% methanotroph bacteria meal, T3: 10% methanotroph bacteria meal, and T4: 15% methanotroph bacteria meal. Shrimp were fed ad libitum for 6 weeks on trial diets to assess growth. Subsequent to the growth trial, three replicates of the same groups were exposed to V. parahaemolyticus by a single bath challenge and held for a further 15 days on the same diets as the growth study to assess survival and resistance. No significant differences (p > 0.05) in survival or in growth performance, including final weight, weight gain, specific growth rate, feed consumption or feed conversion ratio of white shrimp fed feeds containing methanotroph bacteria meal or control diets for 6 weeks. Immune markers such as hemocyte counts, phenoloxidase, superoxide dismutase and lysozyme activity were similar across all groups after the 6-week feeding trial. In a V. parahaemolyticus challenge, methanotroph bacteria meal in the diet significantly promoted the survival rate, and the reduction of Vibrio sp. in the hepatopancreas of white shrimp. Hemocyte count and phenoloxidase activity showed no significant differences (p > 0.05) between diet treatment groups, but hemolymph protein was significantly higher (p < 0.05) in shrimp fed diets containing 15% methanotroph bacteria meal after challenge. The Vibrio colony counts from hepatopancreas in the treatment groups were all significantly lower than the control (p < 0.05). The findings show that methanotroph bacteria meal can entirely replace fishmeal in white shrimp diets and the 15% inclusion of methanotroph bacteria meal in shrimp diet shows no adverse effects on growth performance, feed utilization and survival rate. In addition, shrimp fed methanotroph bacteria meal diets exhibited improved survival rates to an AHPND challenge.

Localization of trypsin-like protease in postmortem tissue of white shrimp (Litopenaeus vannamei) and its effect in muscle softening

Fluorescein-isothiocyanate (FITC) labeled trypsin-like protease was prepared and injected into the hepatopancreas of white shrimp. Different segments of the injected shrimp were analyzed with a fluorescence microscope during storage. FITC-trypsin-like protease can be detected in the first segment of shrimp muscle at day 4, while it cannot be observed in the second segment until day 6. The results showed that trypsin-like protease can migrate from hepatopancreas to the tail portion. Texture profile analysis showed that soybean trypsin inhibitor retarded the softening of the shrimp muscle. The rheological results revealed that the content of myosin heavy chain (MHC) in shrimp muscle was decreased with the extended storage time. Proteomics analysis displayed that trypsin-like protease accelerated the metabolism of postmortem muscle. It can be concluded that trypsin-like protease migrated from the hepatopancreas to the muscle tissue, degraded myofibrillar protein, deteriorated the muscle texture, and eventually leaded to the softening of white shrimp.

Histopathological in Gills, Hepatopancreas and Gut of White Shrimp (Litopenaeus Vannamei) Infected White Feces Disease (WFD)

White Feces Disease which is the presence of white feces floating on the surface of the water. Physical disruption of White Feces Disease attacked by exocytes becoming soft. the cause of white defect disease (WFD) is a decrease in environmental quality such as salinity, DO and ammonia. white feces disease is associated with gregarine protozoa in the Apicomplexa phylum. Gregarine parasitic infection from the type of nematopsis that was interacted with Vibrio and Vibrio type bacteria found in white shrimp (Litopenaeus vannamei) which was attacked by white impurities was V. harveyi. This study aims to analyze the presence of gregarine protozoa in the gills, intestines and hepatopancreas of white shrimp infected with White Feces Disease. The method used in this research is descriptive method. Histopathological results showed that there were protozoa (Gregarine) in the intestine, hepatopancreas and gills of white shrimp infected with WFS in 3 different locations and found Aggregated Transformed Microvilli (ATM) that resembled gregarine .

Use of TPP and ATPS for partitioning and recovery of lipase from Pacific white shrimp (Litopenaeus vannamei) hepatopancreas.

Lipase recovery from Pacific white shrimp hepatopancreas using a three-phase partitioning (TPP) system in combination with an aqueous two-phase system (ATPS) was studied. TPP system was formed with a simultaneous addition of salt directly to crude extract (CE) followed by an organic solvent addition. The various process parameters required for efficient purification of lipase were optimized. The best lipase yield (87.41%) and purification fold (PF) (3.49-fold) were obtained in the interphase of TPP system, which consisted of the CE to t-butanol ratio of 1:1 (v/v) in the presence of 50% (w/v) (NH4)2SO4. Subsequently, TPP fraction was subjected to ATPS. Effects of phase compositions including PEG molecular weight and concentration, types and concentration of salts, NaCl addition and system pH on lipase partitioning were investigated. With the application of 25% (w/w) PEG1000 and 15% (w/w) MgSO4, at pH 5.0 was found most appropriate since high lipase PF (5.19-fold) and yield (78.46%) in top phase were obtained. The partitioned enzyme exhibited optimal activity at pH 8.0 and 55°C and was stable at a temperature range of 0-40°C and a pH range of 7-10. The partitioned lipase showed high tolerance in the presence of ethanol and methanol. Hence, the combined partitioning systems, TPP-ATPS, were found to be an attractive technique for the recovery and partial purification of lipase from Pacific white shrimp hepatopancreas.

Effect of pure microcystin-LR on activity and transcript level of immune-related enzymes in the white shrimp (Litopenaeus vannamei).

Microcystins (MCs) in freshwater and marine waters released by toxin-producing cyanobacteria have negative impacts to the aquatic environment. This study aimed to investigate the effect of pure microcystin-LR on activity and transcript level of immune-related enzymes in the white shrimp Litopenaeus vannamei. After exposed to varying concentrations of pure microcystin-LR (MC-LR) for 30 days, the activity of superoxide dismutase (SOD), lysozyme (LZM), glutathione peroxidase (GPx), peroxidase (POD), acid phosphatase (ACP), alkaline phosphatase (AKP) and transcript level of cMn-sod, lzm, gpx were investigated in the hepatopancreas of white shrimp (L. vannamei). Immune-related enzyme activities responded differently to MC-LR exposure. SOD, GPx, and POD activity in the hepatopancreas were activated in a concentration-dependent manner while LZM activity was significantly inhibited in the treatment groups. ACP and AKP activity showed an increase, followed by a decrease. The transcript levels of cMn-sod, lzm, and gpx were consistent with changes in their encoding enzyme activity. These results demonstrated that sub-chronical exposure to MC-LR induced the alteration of immune-related enzymes and corresponding genes in the hepatopancreas, which may help explain the presence of detoxification mechanisms in crustaceans and how they were protected from MC-LR stress for a long period of time.

Laundry detergent-stable lipase from Pacific white shrimp (Litopenaeus vannamei) hepatopancreas: Effect of extraction media and biochemical characterization

ABSTRACTExtraction and biochemical properties of a new lipase from the hepatopancreas of Pacific white shrimp were studied. Recovery of the hepatopancreas powder with 50 mM Tris-HCl, pH 7.0 containing 0.2% (v/v) Brij35 gave a higher recovery of lipase activity than other extractants tested (p < 0.05). The optimal pH and temperature for lipase activity were 8.5 and 60°C, respectively, when p-nitrophenyl palmitate was used as a substrate. The enzyme was stable to heat treatment up to 40°C and over a pH range of 7.0–10.0 for 30–120 min. Lipase activities continuously decreased as the sodium deoxycholate (NaDC) concentration increased, but activities increased as NaCl concentration increased up to 3.0 M. Hydrolytic activity was enhanced by NaN3, but strongly inhibited by Hg2+, Cu2+, Al3+, and phenylmethanesulfonyl fluoride. The lipase was evaluated as highly stable against surfactants (Tween 20, Tween 80, Triton X-100, and gum arabic). However, the enzyme was unstable against sodium dodecyl sulphate. Stability of the lipase with commercial liquid and solid detergents (Attack®, Bres®, Omo®, and Pao®) was also investigated. The lipase exhibited substantial stability and compatibility with tested commercial liquid and solid laundry detergents for 30–60 min. The overall properties of the lipase from Pacific white shrimp hepatopancreas, thus leading us to propose that it is an excellent candidate for use as biocatalysts for better detergent formulation.

Effects of T-2 toxin on growth, immune function and hepatopancreas microstructure of shrimp (Litopenaeus vannamei)

T-2 toxin (T-2) is a trichothecene-A-type mycotoxin produced by the fungus Fusarium spp., which can contaminate animal feed. It is toxic to many living cells especially the hepatocytes and bone marrow cells. The effects of dietary T-2 on growth, biochemical and immunity parameters, and histopathological changes in the hepatopancreas of white shrimp (Litopenaeus vannamei) were studied. Shrimp with an initial body weight of 3.5±0.5g were fed five diets (n=50/group) containing 0, 0.5, 1.2, 2.4, or 4.8mgkg−1 T-2. Dietary exposure to T-2 significantly decreased shrimp growth rate and specific growth rate (P<0.5), but not the fat content, compared with the control group. Exposure to T-2 damaged the hepatopancreas microstructure in a dose-dependent manner and this may have contributed to a lower hemolymph albumin concentration and lower phenoloxidase activity. Total hemocyte counts and activity of hemolymph glutamic-pyruvic transaminase>glutamic-oxaloacetic transaminase and alkaline phosphatase enzymes increased initially in a dose-dependent manner but declined thereafter. This study showed that T-2 is a multi-organ toxin with effects on the hemolymph, immune system, and hepatopancreas of shrimp. Collectively, these effects may have contributed to the inhibition of growth and decreased survival rate leading to a lower yield and quality of shrimp meat with resultant economic loss. Statement of relevanceIt is proposed that hemolymph phenoloxidase enzyme activity could be used as a cheap non-specific biomarker for initial screening of shrimp exposed to even low T-2 toxin concentrations, instead of more expensive LC-MS/MS T-2 analysis — although a dose response was not evident at high exposure concentrations.

Comparative Study on Virgin Coconut Oil Extraction Using Protease from Hepatopancreas of Pacific White Shrimp and Alcalase

Virgin coconut oil (VCO) was extracted from coconut milk with the aid of crude protease extract (CPE) from hepatopancreas of Pacific white shrimp at different levels (5–15 unit/g protein) at 60C for various hydrolysis times (0–180 min). Yield of VCO increased within the first 90 min (P < 0.05). However, Alcalase showed higher efficacy in VCO extraction, compared with CPE and the control (without enzyme) (P < 0.05). No differences in lipid oxidation of all VCOs extracted by different proteases were observed (P > 0.05). Alcalase and CPE increased the creaming index and induced the collapse of oil droplets in coconut milk, as determined by the confocal laser scanning microscopy and the phase contrast microscopy. VCOs contained medium chain fatty acid, especially lauric acid (C12:0), as the most abundant fatty acid, followed by myristic acid (C14:0). Therefore, VCO could be extracted using CPE or Alcalase at the optimal temperature (60C) for 90 min. Practical Applications Pacific white shrimp hepatopancreas has been known as a potential source of trypsin that can be used to hydrolyze proteins stabilizing emulsion in coconut milk. Trypsin from Pacific white shrimp hepatopancreas could therefore serve as a potential aid for virgin coconut oil extraction with the shorter processing time, particularly when applied under the optimal condition. As a consequence, the cost associated with commercial enzymes could be reduced and the prime quality of virgin coconut oil was still obtained.

Chemical compositions and properties of virgin coconut oil extracted using protease from hepatopancreas of Pacific white shrimp

Virgin coconut oil (VCO) was extracted from coconut milk with the aid of crude protease extract (CPE) from hepatopancreas of Pacific white shrimp at different levels (5–15 U/g protein) for various hydrolysis times (0–24 h). Yield of VCO increased within the first 6 h. The use of CPE exhibited similar extraction efficiency to the typical fermentation process. However, lipids extracted with fermentation process underwent oxidation to a higher extent, compared with those extracted using CPE. CPE increased the creaming index and induced the collapse of oil droplets in coconut milk, as determined by the phase contrast microscope. VCO contained medium chain fatty acid (MCFA), especially lauric acid (C12:0), as the most abundant fatty acid, followed by myristic acid (C14:0). Therefore, VCO could be extracted using CPE at room temperature for 6 h, in which the extraction yield was increased and lipid oxidation was negligible.Practical applications: Pacific white shrimp hepatopancreas can be a major source of proteases, especially trypsin, that can be used for hydrolysis of lipoprotein or to destabilize emulsion. Proteases from shrimp hepatopancreas could be used as a promising processing aid for virgin coconut oil extraction, in which the cost associated with commercial enzymes could be reduced and the proteases from shrimp processing byproducts were better exploited. The proteases from shrimp hepatopancreas can therefore be of benefit for the virgin coconut oil processing enterprise or industry by increasing extraction efficacy via destabilzation of coconut milk emulsion.Proteases from Pacific white shrimp hepatopancreas can be used as the aid for VCO extraction via the accelerated emulsion destabilization without any negative effect on quality.

Impact of enzymatic method using crude protease from Pacific white shrimp hepatopancreas on the extraction efficiency and compositions of lipids

Lipids from striped catfish muscle were extracted with the aid of crude protease extract (CPE) from hepatopancrease of Pacific white shrimp at different levels (5–15unit/g protein) for various hydrolysis times (0–180min) at 60°C. Yield of lipids increased within the first 30min and was in agreement with increasing degree of hydrolysis. The increases in phospholipid and free fatty acid content were observed with hydrolysis times up to 30min (p<0.05). Yield of lipid increased when 50mM CaCl2 was combined with CPE. CPE exhibited similar extraction efficiency to Alcalase. However, the lower yield was obtained in comparison with the solvent extraction. Higher oxidation and hydrolysis were found in lipids extracted with the aid of CPE and Alcalase, compared with those found in lipid extracted using solvents (p<0.05). All lipid samples had triglyceride as the major component and were rich in palmitic acid and oleic acid.

Antioxidative activities of hydrolysates from seabass skin prepared using protease from hepatopancreas of Pacific white shrimp

Antioxidative activities of hydrolysates from skin of seabass (Lates calcarifer) with different degrees of hydrolysis (DH: 10–40%) prepared using an ammonium sulphate precipitated fraction (ASPF) from Pacific white shrimp hepatopancreas and commercial Alcalase were compared. The hydrolysate prepared using ASPF or Alcalase had increases in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging activities and ferric reducing antioxidative power (FRAP) as DH increased (P<0.05). When the hydrolysate prepared using ASPF with 40% DH was subjected to a gastrointestinal model system (GIMs), ABTS radical scavenging activity and chelating activity increased, especially in the duodenal condition. The hydrolysate (500–2000mg/L) could inhibit lipid oxidation in a lecithin liposome system in a dose dependent manner. Based on gel filtration using a Sephadex™ G-15 column, peptide with a molecular weight of 364Da showed the strongest ABTS radical scavenging activity. Therefore, the extract from hepatopancreas could be used to increase DH of hydrolysate from seabass skin.

Profiling of differentially expressed genes in hepatopancreas of white shrimp (Litopenaeus vannamei) exposed to long-term low salinity stress

The Pacific white shrimp, Litopenaeus vannamei, is a euryhaline crustacean capable of tolerating a wide range of ambient salinity (0.5–40psu). To investigate the effect of long-term low salinity stress on gene expression in the hepatopancreas in shrimp, we performed suppression subtractive hybridization (SSH) in juvenile L. vannamei exposed to long-term low salinity. The shrimp (initial body weight, 0.27±0.01g) were cultured at salinity 2 psu and salinity 30 psu for 56days. We then constructed forward and reverse subtractive cDNA libraries. We used bioinformatics tools and vector screening to select a total of 200 (80 from forward, 120 from reverse) randomly selected clones over 100 nucleotides in length for further analysis. Nineteen contigs and 54 singletons were generated from a total of 73 consensuses. The consensuses, upon a sequence homology search using BLASTX (NCBI), revealed that 24.66% (18/73) of them had no significant match to reported sequences in the database, suggesting that they had not previously been found and that they were probably associated with stress-regulated functions. The remaining 75.34% (55/73) of the consensuses encoded proteins were matched to a wide range of functions including immune-related functions, metabolism, ribosomal activity, transfer activity, and apoptosis. The most common group in these SSH libraries was immune-related proteins and enzymes (11/17). Quantitative RT-PCR results confirmed that the relative expression of 5 differentially expressed genes encoding hemocyanin, chitinase, ecdysteroid-regulated protein, trypsin and chymotrypsin 1 was decreased 2-, 1.45-, 11.11-, 1.33- and 1.54-fold, respectively, in the reverse library. This subtractive cDNA library provides a basis for the study of the genetic response of shrimp to environmental stress.

Gene expression of ferritin in tissue of the Pacific white shrimp, Litopenaeus vannamei after exposure to pH stress

Ferritin is a storage protein that plays a key role in iron metabolism. In this study, the gene expression of ferritin in hepatopancreas, muscle and eye of white shrimp Litopenaeus vannamei were analysed by quantitative real-time PCR after exposure to pH stress. The results showed the expression level in the hepatopancreas and eye were about 5 times to those of beta-actin, whereas lower expression levels in muscle were about 1/2 to those of beta actin. The gene expressions of ferritin in hepatopancreas and muscle were up regulated at 1.5 h after exposure to pH stress, but down-regulated in eye slightly. The ferritin transcript in hepatopancreas of white shrimp L. vannamei at pH 9.3 and pH 5.6 treatments increased significantly in 1.5 h and 12 h post-challenge respectively, but both returned to the original values in 24h post-challenge. The present result showed that ferritin expression in tissue of L.vannamei was induced by acute pH stress and ferritin induction varied in response to duration of induction and acidic or alkaline pH treatment, and was more sensitive to high pH than low pH.

In vitro studies of the effects of aflatoxin B 1 and fumonisin B 1 on trypsin-like and collagenase-like activity from the hepatopancreas of white shrimp ( Litopenaeus vannamei)

The in vitro effects of aflatoxin B 1 (AFB 1) and fumonisin B 1 (FB 1) on the trypsin-like and collagenase-like activity from hepatopancreas (HP) of cultured white shrimp ( Litopenaeus vannamei) have been studied. The presence of trypsin-like and collagenase-like enzymes in the shrimp HP was confirmed by inhibition and electrophoresis analysis. Both enzymes were strongly inhibited by phenylmethylsulfonyl fluoride (PMSF) which suggests that they belong to serine proteases. Collagenase-like protease digested native porcine type I collagen. Trypsin-like activity was inhibited by AFB 1 and FB 1. The collagenase-like activity was stimulated by FB 1. Lineweaver–Burk reciprocal plots indicated uncompetitive inhibition of trypsin-like enzyme by AFB 1. Competitive inhibition in collagenase-like enzyme, and increased K m and V max values of collagenase-like enzyme were observed when exposed to FB 1. These findings may suggest that AFB 1 and FB 1 toxicity to shrimp might be due to digestive deficiency and consequential histological changes in its hepatopancreas, as a result of alterations of trypsin and collagenase activity due to the presence of mycotoxins in the shrimp diet.