Shrimp Muscle Research Articles

Mercury accumulation and metallothionein expression from aquafeeds by Litopenaeus vannamei Boone, 1931 under intensive aquaculture conditions

This study describes the accumulation of Hg and metallothionein gene expression in Litopenaeus vannamei Boone, 1931 with aquafeeds as the major source of Hg. Trials were conducted under controlled conditions in experimental tank facilities with high (indoor tanks) and low (outdoor tanks) Hg aquafeeds concentrations. Aquafeeds were the sole source of Hg for the shrimps and concentrations varied from 5.4 to 124 ng.g-1 d.w.. In the three animal fractions analysed; muscle (6,3 - 15,9 ng.g-1); hepatopancreas (5,1 - 22,0 ng.g-1) and exoskeleton (3,0 - 16,2 ng.g-1), Hg concentrations were significantly lower in the outdoor trials submitted to Hg-poor aquafeeds. Maximum shrimp muscle Hg concentrations were low (36.4 ng.g-1 w.w.) relative to maximum permissible concentrations for human consumption and Hg content in muscle and hepatopancreas were significantly correlated with Hg content in aquafeeds. Highest Hg concentrations in the exoskeleton of animals exposed to Hg-richer aquafeed, suggested that a detoxification mechanism is taking place. On the other hand the metallothionein suffered no variation in its relative expression in any of the experiments, meaning that the contact with feed containing the observed Hg concentrations were not sufficient to activate gene transcription. It was not possible, under the experimental design used, to infer Hg effects on the biological performance of the animals.

Shrimp tissue quality in the lower Gangetic delta at the apex of Bay of Bengal

Seafood containing elevated levels of metals as a result of environmental contamination may entail health risks to human beings. To evaluate such contaminations, our study targeted on the analyses of lead and cadmium, in addition to the toxicologically less-relevant metals, zinc and copper, in the muscle tissue of five commonly edible shrimp species collected from the lower stretch of the River Ganga (in the Sundarbans delta complex). The metals in the tissue samples were determined using an inductively coupled plasma-mass spectrometer. The concentrations in the shrimps ranged as follows: Pb: from below detection limit to 15.9 ± 0.8 ppm dry weight; Cd: from below detection limit to 7.3 ± 0.8 ppm dry weight; Zn: 13.0 ± 0.7–428 ± 3 ppm dry weight; Cu: 8.5 ± 0.3–146 ± 3 ppm dry weight. The concentrations of the metals in the tissues varied significantly depending upon the locations from where the shrimp species were collected. The levels of Pb, Cd, and Cu in shrimp muscle at station 1 were far above the permissible levels for human consumption, while the concentration of Zn was within the normal range at all stations. Station 1 is subjected to maximum stress in terms of pollution, being situated downstream of the River Ganga.

Growth, survival, and fatty acid composition of Indian white shrimp Fenneropenaeus indicus (Milne Edwards) fed diets containing different levels of vitamin E and lipid

The objective of this experiment was to determine the effects of two levels of vitamin E (100 and 300 mg/kg diet) along with two levels of lipid (9 and 14%) and their interaction on growth performance of Indian white shrimp and consequently to evaluate the fatty acid composition and lipid stability of its muscle tissue during frozen storage. Growth of juvenile Indian white shrimp was not significantly affected by dietary vitamin E and lipid levels. Muscle lipid content of shrimp fed diets with 14% lipid was significantly higher than that of with 9% lipid. Obvious effects of the increase in dietary lipid level on muscle fatty acid composition were significant decrease in proportion of 16:0 and increase in proportion of 20:5n-3. The content of vitamin E concentration in shrimp muscle reflected dietary vitamin E concentration and ranged from 6.68 to 14.8 mg/kg muscle corresponding to two (100 and 300 mg/kg) levels of vitamin E in fish diet, respectively. Subsequently, results showed that by increasing the concentration of vitamin E from 100 to 300 mg/kg in diet, the rate of lipid oxidation in the muscle tissue during frozen storage was reduced and, as a result, caused higher HUFA retention in muscle of shrimp fed diet with high lipid level.

Otimização e validação de método multirresíduo para determinação de sulfonamidas em camarão cultivado por cromatografia líquida de alta eficiência com detecção por UV

This work reports the optimization and method validation for sulfonamides (sulfamethazine, sulfaquinoxaline, sulfadimethoxine and sulfathiazole) in shrimp muscle using HPLC-UV. The sulfonamides were extracted with acetonitrile and acetic acid, and the extract cleaned up with a Strata SCX SPE cartridge prior to analysis. The method presented linearity in the range of 20-120 µg kg-1, good linear correlation (r > 0.99), and limits of quantification in the range of 4.7-20.2 µg kg-1. The recovery for shrimp muscles spiked with 50-150 µg kg-1 ranged from 63.2-108.0%. Precision and accuracy analysis showed acceptable relative standard deviation. Commercial shrimps were analyzed and sulfonamides don't were found above of the method limit of quantification.

Lead and cadmium in organisms of commercial importance in the coastal zone of Sinaloa, Mexico: 20 years of studies.

Lead and cadmium may affect the health of ecosystems and of the consumers of food itemscontaminated with these metals, which are widely used in several human activities and are, in addition, among the most important products of Mexican mining activities. Agriculture is the main activity in the State of Sinaloa, and for this reason pesticides and fertilizers are important sources of contamination with both metals. In this article we revise the possible natural sources and the main human activities which may contribute these metalsto the coastal waters of the Mexican State of Sinaloa, and summarize the results of the investigations of the latest two decades on the lead and cadmium contents of the edible tissues of several aquatic organisms of its coastal water bodies. The highest cadmium concentrations were those found in the Altata-Ensenada del Pabellónlagoonal system, possibly as a result of the agricultural activities of the Culiacán plains. Lead concentrations in bivalves seem to indicate a tendency to lower values, possibly due to the use of unleaded gasoline, although this does not coincide with the values found in shrimp muscles. In addition, all lead values are above the level indicating the presence of environmental impact. The cadmium content of some bivalve exceed the level of risk for human health, whereas the lead and cadmium contents of fish and crustacean edible tissues are below the level of concern.

흰반점바이러스(WSSV)의 중화를 위한 재조합단백질 rVP466의 항혈청 생산

본 연구는 WSSV의 재조합단백질 rVP466에 대하여 생산된 항혈청을 사용하여 WSSV에 대한 neutralization (중화) 효과를 확인하고자 수행하였다. 먼저 재조합단백질 rVP466의 생산을 위해 WSSV의 구성단백질 VP466을 암호화하는 유전자인 VP466을 포함하는 재조합 플라스미드 pCold-VP466을 제작한 다음 이것을 발현용 숙주인 E. coli RIPL에서 발현하였다. 발현된 rVP466에 대한 항혈청은 토끼를 사용하여 생산하였으며, 항원 rVP466에 대한 특이면역반응은 Western blot을 통해 확인하였다. WSSV에 대한 항혈청의 중화효과를 확인하기 위해 항혈청과 반응시킨 바이러스액(<TEX>$1{\times}10^4$</TEX> 배로 희석된 WSSV)을 이용하여 실험용 새우(Penaeus chinensis)에게 주사 감염을 통해 공격실험(challenge test)을 수행하였다. 실험 결과, WSSV로 공격실험한 감염대조구(positive control)의 새우들은 감염 후 17일째에 100% 누적폐사율을 보였으며, preimmune serum과 WSSV의 혼합액을 challenge한 preimmune control의 새우들은 감염 후 25일째에 83%의 누적폐사율을 보였다. WSSV와 rVP466 항혈청을 1:0.01, 1:0.1, 1:1로 혼합한 액으로 challenge한 새우들은 감염 후 25일째에 각각 73%, 53%, 46%의 누적폐사율을 보였다. 이상의 결과를 통해 WSSV가 rVP466 항혈청에 의해 농도의존적으로 neutralization됨을 확인하였으며, 이는 WSSV 감염과정에 VP466이 관여함을 나타내는 것이다. This study was carried out to evaluate neutralization effects against WSSV using antiserum produced from recombinant envelop protein, rVP466 of WSSV. The VP466 gene of WSSV was cloned into pCold I expression vector and rVP466 was expressed in E. coli RIPL. The antiserum against rVP466 was produced in white rabbits (New Zealand white rabbit). The specific immunoreactivity to the antigen, rVP466, was confirmed by Western blot. The constant amounts of WSSV at <TEX>$1{\times}10^4$</TEX> diluted stocks were mixed with various antiserum concentrations and then injected to the muscle of shrimp, Penaeus chinensis, for the neutralization challenge. The shrimps challenged with WSSV as a positive control and those with the mixture of WSSV and preimmune serum as a preimmune control showed 100% cumulative mortality at 17 days post challenge and 83% at 25 days post challenge, respectively. The shrimps challenged with 3 different mixtures of WSSV and rVP466 antiserum at ratios of 1:0.01, 1:0.1 and 1:1 showed 73%, 53% and 46% cumulative mortalities at 25 days post challenge, respectively. These results indicated that WSSV could be neutralized by the rVP466 antiserum. These results suggest that envelop protein VP466 is involved in the initial step of WSSV infection in shrimp.

A multi-biomarker approach to assess the impact of farming systems on black tiger shrimp ( Penaeus monodon)

This study examined the advantages of the use of biomarkers as an early warning system by applying it to different shrimp farming systems in Soctrang and Camau provinces, main shrimp producers in Mekong River Delta, Vietnam. Shrimp were collected at 15 different farms divided into four different farming systems: three farms were converted from originally rice paddies into intensive shrimp farming systems (IS1, IS2, IS3); three farms were rice–shrimp integrated farming systems (RS4, RS5, RS6); three farms were intensive farming systems (IS7, IS8, IS9); six farms were extensive shrimp farming systems (From ES1 to ES6). Lipid peroxidation (LPO) and total glutathione (GSH) were measured as well as catalase (CAT), glutathione peroxidase (GPX), glutathione S-transferase (GST) and acetylcholinesterase activities (ACHE). Organ specificity was observed between gills and hepatopancreas with generally higher activity of GST in gills (GSTG) whereas the contrary was observed for LPO level in gills (LPOG). Hierarchical clustering and principal component analysis clearly indicated that shrimp reared in extensive culture system formed a distinct group from those reared in intensive or rice–shrimp integrated systems. CAT in gills (CATG), GPX in gills (GPXG) and hepatopancreas (GPXHP) and ACHE in muscle (ACHEM) of shrimp collected in extensive farms showed a general higher level than those in intensively farmed shrimp. On the contrary, we observed clear high levels of GSTG and GST in hepatopancreas (GSTHP) and LPOG and hepatopancreas (LPOHP) of shrimp sampled in intensive and rice–shrimp integrated systems. Thus, we propose that LPO and CAT, GPX, GST and ACHE can be used as a set of biomarkers for the assessment of health condition and can discriminate between shrimp cultivated in different farming systems. These findings provide the usefulness of integrating a set of biomarkers to define the health status of shrimp in different shrimp culture systems.

SPE–LC Multi-Residue Analysis of Mebendazole and its Metabolites in Grass Carp and Shrimp

An analytical method was developed to detect the residue of mebendazole and its metabolites (hydroxymebendazole and aminomebendazole) in the muscle of grass carp and shrimp by LC–UV detection. Mebendazole and its metabolites were extracted with water and ethyl acetate, defatted with hexane, and purified with MCX solid phase extraction column. The intra- and inter-batch precision (measured by CV%) was <9.0%. The accuracy (measured by relative error, %) was <12%. The LODs were 2.5 μg kg−1 for mebendazole and hydroxymebendazole, 5 μg kg−1 for aminomebendazole; the LOQs were 5 μg kg−1 for mebendazole and hydroxymebendazole, 10 μg kg−1 for aminomebendazole. The mean recoveries of mebendazole and its metabolites from grass carp and shrimp muscle at a concentration range of 5.0–500.0 μg kg−1 were 90.7–97.0% with relative standard deviations below 10%.

Heavy metal (Pb, Co, Cd, Cr, Cu, Fe, Mn and Zn) concentrations in harvest-size white shrimp Litopenaeus vannamei tissues from aquaculture and wild source

The present research project was undertaken to determine heavy metal (Pb, Co, Cd, Cr, Cu, Fe, Mn and Zn) concentrations in intensive aquaculture and wild white shrimp tissues (muscle, shell and liver), and to inform aquatic shrimp food safety. The aquaculture samples were obtained from ten intensive farms and wild animals from one station in Zhanjiang Harbour Bay. Heavy metal concentrations (mg kg −1, dry weight; means ± S.D.) in aquaculture white shrimp muscle, shell and liver tissues were, respectively: Cr – 20.86 ± 5.27, 28.70 ± 8.34 and 18.91 ± 5.50, Cu – 24.26 ± 8.36, 30.86 ± 9.84 and 126.42 ± 90.9, Fe – 61.35 ± 30.76, 55.07 ± 17.72 and 124.04 ± 56.15, Mn – 5.33 ± 2.5, 10.72 ± 4.34 and 8.79 ± 5.29, Zn – 171.56 ± 118.74, 51.84 ± 10.94 and 111.74 ± 84.67. Cadmium was observed only in liver tissue at 3.30 ± 1.29 mg kg −1. Lead could not be detected, and cobalt was detected only in wild shrimp liver tissue. Chromium and manganese mean concentrations in shell tissue were significantly higher than levels in muscle tissue ( P < 0.05), but the zinc mean concentration was significantly lower than noted in muscle tissue ( P < 0.05). Copper and iron mean concentrations in liver tissue were significantly higher than those in muscle or shell tissues ( P < 0.05). Overall, only chromium in white shrimp tissue was the primary risk for human health.

An improved method of cell culture system from eye stalk, hepatopancreas, muscle, ovary, and hemocytes of Penaeus vannamei

Improved methods of cell culture from eye stalk, hepatopancreas, muscle, ovary, and hemocytes of shrimp (Penaeus vannamei) were established using synthetic media and shrimp muscle extract (SME). For hemocytes and ovarian cell cultures, Grace's insect medium supplemented with 10% (v/v) fetal bovine serum and 10% SME (v/v) showed enhanced attachment and proliferation of the cells. The hemocyte and ovarian cell cultures could be maintained for 48 and 66 days, respectively, and have been sub-cultured four and six times, respectively. Both ovary and hemocyte cell cultures contained primarily epithelial-like cells. Cells derived from ovary tissue grew preferably between 26°C and 28°C with 5% CO(2). Although the temperature preference of hemocyte cells was the same as ovarian cells, CO(2) supplementation did not show any difference in the growth of hemocyte cells. When the shrimp were injected with lipopolysaccharide (8 μg/g of shrimp) and hemolymph was drawn 24 h post-injection, the in vitro multiplicity of hemocytes dramatically improved. The growth of eye stalk, hepatopancreas, and muscle-derived cells was much less compared to ovarian cells and hemocytes under the conditions described above. The optimal culture conditions for ovarian cells and hemocytes were also different from that for eye stalk, hepatopancreas, and muscle cell culture. The proliferation efficiencies of primary cultures of hepatopancreas, eyestalk, and muscle cells were about 30, 12, and <7 d, respectively. The improved culture conditions described here, particularly for hemocytes and ovary, will be very useful for in vitro studies involving viruses infecting shrimp and in shrimp genomic studies.

GROWTH RESPONSE AND MUSCLE LIPID QUALITY OF INDIAN WHITE SHRIMP FED DIFFERENT OILS AT TWO DIETARY LIPID LEVELS

ABSTRACT A feeding trial was conducted to evaluate the effects of partial fish oil replacement with vegetable oils at two levels of dietary lipid on growth performance and muscle lipid quality of Indian white shrimp Fenneropenaeus indicus. Final weight of shrimp fed control diet (100% fish oil) was higher than that of shrimp fed the other diets. Partial replacement of fish oil with vegetable oils resulted in reduced levels of eicosapentaenoic acid and docosahexaenoic acid in shrimp muscle tissue. Increasing the dietary lipid level had an effect on lipid content of shrimp, by increasing lipid deposition in muscle tissue, but without a significant effect on growth. Concentrations of cholesterol in shrimp muscle were not significantly affected by dietary lipid source and level (P &gt; 0.05). Shrimp fed diets with 14% lipid had significantly higher concentration of malonaldehyde (thiobarbituric acid reactive substances) in muscle tissue than shrimp fed diets with 9.5% lipid. Vitamin E concentration in muscle of shrimp fed 100% fish oil was significantly (P &lt; 0.05) affected by dietary lipid level.PRACTICAL APPLICATIONSThe high market value of and great demand for penaeid shrimp in the world market provide a strong stimulus for its intensive culture. The Indian white shrimp Fenneropenaeus indicus (Milen Edwards) was the most important shrimp species cultivated in Iran (due to market demand, high market value and local shrimp). It is a species with relatively slow growth potential compared to Penaeus monodon and its production is relatively low in intensive farms. It is mainly reared with commercial feed. Fish oil is an important ingredient used in commercial marine fish and shrimp feeds. The demand for fish oils in the aquafeed industry has increased steadily despite the static global production levels and may not be able to cater to the demand in the near future. Therefore, finding alternative to fish oils is one of the critical research areas in marine fish and shrimp nutrition.

Studies on the immunoreactivity of the major allergen tropomyosin in mantis shrimp (&lt;I&gt;Squilla oratoria&lt;/I&gt;)

Crustacean is one of the eight kinds of allergen sources in coastal areas,which causes the IgE-mediated hypersensitive reactions with clinical manifestations including urticaria,angioedema,asthma,and even fatal anaphylaxis. Tropomyosin (TM) is the major allergen of decapod crustaceans with highly conserved amino acid sequences. The purpose of this study is to confirm whether TM is a major cross-reactive allergen in mantis shrimp (Squilla oratoria) which is taxonomically distinct from decapods and largely consumed as a delicacy in China. Quantification of TM in different species of crustaceans was also conducted. Muscle sample from mantis shrimp was homogenized with phosphate buffer to prepare heated extracts and separated by 12 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). A protein band with molecular mass of about 36 ku was detected by IgE-immunoblotting by all of the sera with crustacean allergy,suggesting it is the major allergen of mantis shrimp. This protein was further purified to homogeneity by acetone powder preparation,isoelectric point precipitation,ammonium sulfate fractionation (40%-60% saturation),and heat treatment. Purified protein was demonstrated to be TM by Western blot using polyclonal antibody against TM from Chinese mitten crab (Eriocheir sinensis). Heated extracts from other crustaceans including mud crab (Scylla serrata),Pacific white shrimp (Penaeus vannamei) and short necked clam (Venerupis variegata) were also prepared,and the cross-reactivity of TM from mantis shrimp with TMs from these crustaceans was confirmed by inhibition immunoblotting using polyclonal antibody against TM and inhibition ELISA using sera with crustacean allergy. Quantification by ELISA using polyclonal antibody against TM revealed that TM content in mantis shrimp muscle is much lower than that in Pacific white shrimp and mud crab muscle,which was about 45 times lower in mantis shrimp muscle than that in Pacific white shrimp muscle. However,its allergenicity seems equivalent to other decapod crustaceans as showed by inhibition ELISA using sera with crustacean allergy. This may be due to the degradation of TM in mantis shrimp by its endogenous serine proteinases and cathepsins which destroyed the IgG-binding epitope but not the IgE-binding epitopes. In conclusion,this study demonstrated that allergenicity of mantis shrimp is almost equivalent to decapods and TM is the major allergen in mantis shrimp with high cross-reactivity to decapod crustaceans.

Genetic heterogeneity of non-O1 and non-O139 Vibrio cholerae isolates from shrimp aquaculture system: a comparison of RS-, REP- and ERIC-PCR fingerprinting approaches

The genetic diversity of Vibrio cholerae isolated from black tiger shrimp (Penaeus monodon) aquaculture farms was determined using three PCR typing methods based on enterobacterial repetitive intergenic consensus (ERIC) sequences, ribosomal gene spacer (RS) sequence and repetitive extragenic palindromic (REP) sequences. Non-O1 and non-O139 V. cholerae isolates were obtained from shrimp pond water, pond sediment, shrimp head and shrimp muscle. RS-PCR yielded fewer bands than REP-PCR and ERIC-PCR. Higher similarity was observed in RS-PCR (75-100%) than in REP-PCR (60-95%) and ERIC-PCR (40-95%). A 100% similarity between V. cholerae isolates was only noticed in RS-PCR. The choleratoxigenic V. cholerae (non-O1 and non-O139) showed greater genetic similarity with ctx-negative V. cholerae than among ctx-positive V. cholerae. The greater similarity of ctx-positive V. cholerae with ctx-negative V. cholerae isolates indicates that the ctx-positive strains (non-O1 and non-O139) might have originated from autochthonous V. cholerae in the aquatic niche.

The Isolation and Identification of Vibrio Species in Marine Shrimps of Sri Lanka

Vibrios are the most common genera associated with crustaceans often causing significant economic losses. Many Vibrio species are pathogenic to human and have been implicated in food borne diseases. Shrimp samples were collected from Mutwal and Chilaw landing sites over a 3 months period (March - May 2007) in order to investigate Vibrio species in the marine environment. Vibrio species from shrimp muscle and brain fluid were isolated and identified at species level using thiosulfate-citrate-bile saltsucrose agar and the designed biochemical key. A total of 159 were isolated and undergone seven biochemical tests shown in the biochemical key. Accordingly, in family Vibrionaceae, twelve species such as, A.hydrophila, V.cholerae, V. metschnikovii, V.parahaemolyticus, V.carchariae or V.harveyi, P.shigelloides, V.vulnificus, V.damsela, V.mimicus, V.furnissii or V.fluvialis were isolated and identified. Among them A.hydrophila, V.parahaemolyticus, V.carchariae or V.harveyi and P.shigelloides species were predominantly present in both locations. V.cholerae and V.metschnikovii most probably present in the two landing sites Chilaw and Mutwal in west and north west coast of Sri Lanka. Keywords: Biochemical key, Isolation; Marineshrimps; Vibrio spp. DOI: 10.4038/jfa.v1i1.1838 Journal of Food and Agriculture Vol.1(1) 2008 pp.36-44

Improved compatibility of liquid chromatography with electrospray tandem mass spectrometry for tracing occurrence of barbital homologous residues in animal tissues

This work describes a liquid chromatography–tandem mass spectrometry (LC–MS/MS) procedure for multiplex screening, ultratrace quantification and reliable confirmation of barbital series residues in animal-derived food matrices. The method is developed based on a distinct dependency of the electrospray ionization (ESI) response of nine structural homologues on LC eluent properties and gas-phase ion chemistry during the ESI process. The “wrong-way-round” negative ionization aspect has been explored to optimize the compatibility of the hyphenated LC–MS/MS technique, which facilitates detection limits at 30–100-fold lower than 0.01 ppm without derivatization or post-column basification step. A mobile phase using methanol modified with 0.01% acetic acid is adopted to achieve an approximately 2–9-fold increase in signal-to-noise ratio over the results under suboptimal conditions. There is no significant differential matrix effects or deuterium isotope effects on chromatographic retention and ESI responsiveness at all levels across the different analyte–matrix pairs. Mean recoveries ranged from 79.6% (barbital) to 108% (secobarbital) at fortified levels of 0.5–20 ng/g within relative standard deviations less than 11%. Between-run repeatability and within-laboratory reproducibility were 3–11% and 5–13%, respectively. An ion ratio criterion for valid detection limit data for simultaneous screening of homologous multiresidues in complex sample matrices is proposed. The satisfactory applicability of the newly described procedure to 43 real samples including pork, poultry meat, swine liver, fish tissue and shrimp muscle demonstrated the LC–MS/MS technique with facile sample handling can serve as an attractive alternative analytical method accepted for regulatory purpose.

Comparative analysis of the fatty acid and sterol profiles of widely consumed Mediterranean crustacean species

Abstract Comparative analysis of FA, NL and sterol profiles in the Nephrops norvegicus (langoustine), Palinurus vulgaris (lobster) and Penaeus kerathurus (shrimp) muscle and cephalothorax showed that C16:0, C16:1ω-7, C18:0, C18:1ω-9, C20:4ω-6, EPA and DHA were found to be their major FA. Highest EPA occurred in langoustine muscle TL, DHA in both shrimp tissues TL while C20:4ω-6 in lobster muscle and cephalothorax TL. Muscle and cephalothorax NL consisted mainly of sterols (42.5–54.4% and 13.7–43.1%) and triacylglycerols (35.4–45.8% and 44.6–59.4%). Cholesterol was the major sterol (70.90–98.58% and 97.10–98.31% of total sterols in muscle and cephalothorax respectively) followed by avenasterol (0.18–20.32% and 0.07–0.70%) and β-sitosterol (0.29–7.30% and 0.23–0.75%). Lower concentrations of brassicasterol, stigmasterol, Δ7-stigmasterol, campesterol and campestanol were also found. The examined crustaceans muscle (edible part) was found to be a good ω-3 PUFA source for the consumers, while the cephalothorax (which is usually discarded) could be used effectively as a source for ω-3 PUFA production.

Effects of Ergothioneine from Mushrooms (Flammulina velutipes) on Melanosis and Lipid Oxidation of Kuruma Shrimp (Marsupenaeus japonicus)

The antimelanosic and antioxidative properties of a hot water extract prepared from the fruiting body of the edible mushroom (Flammulina velutipes) were evaluated by dietary supplementation in Kuruma shrimp (Marsupenaeus japonicus) for possible aquaculture application. The extract contained ergothioneine (ERT) at a level of 2.05 mg/mL. A commercial standard of l-ergothioneine (l-ERT) and the mushroom extract showed inhibitory activity against mushroom polyphenoloxidase (PPO). Feeding of the extract had no adverse effects on the immune systems of the shrimp under the present experimental conditions. Supplementation of the extract in the diet significantly suppressed PPO activities in the hemolymphs of the shrimp. Expression of the prophenoloxidase (proPO) gene decreased in the hemocyte of the Kuruma shrimp fed with the mushroom extract. Consequently, development of melanosis was significantly suppressed in the supplement fed shrimp during ice storage. Lipid oxidation was also effectively controlled in the supplement fed group throughout the storage period. In vitro experiments showed that l-ERT effectively inhibited the activation of proPO in the hemocyte lysate supernatant (HLS). The transcript of the proPO gene in the hemocyte showed lower expression in the l-ERT-treated HLS. It was concluded that dietary supplementation of the mushroom extract in shrimp could be a promising approach to control post mortem development of melanosis and lipid oxidation in shrimp muscles.

Metal and metalloid bioaccumulation in the Pacific blue shrimp Litopenaeus stylirostris (Stimpson) from New Caledonia: Laboratory and field studies

The present work aimed at better understanding metal and metalloid bioaccumulation in the edible Pacific blue shrimp Litopenaeus stylirostris, using both laboratory and field approaches. In the laboratory, the bioaccumulation kinetics of Ag, Cd, Co, Cr, and Zn have been investigated in shrimp exposed via seawater and food, using the corresponding γ-emitting radiotracers (110mAg, 109Cd, 57Co, 51Cr, and 65Zn) and highly sensitive nuclear detection techniques. Results showed that hepatopancreas and intestine concentrated the metals to the highest extent among the blue shrimp organs and tissues. Moulting was found to play a non negligible detoxification role for Co, Cr and, to a lesser extent, Zn. Metal retention by L. stylirostris widely varied (from a few days to several months), according to the element and exposure pathway considered (a given metal was usually less strongly retained when ingested with food than when it was taken up from the dissolved phase). In the field study, Ag, As, Cd, Co, Cr, Cu, Mn, Ni, and Zn were analysed in shrimp collected from a New Caledonian aquaculture pond. Metal concentrations in the shrimp muscles were generally relatively low and results confirmed the role played by the digestive organs and tissues in the bioaccumulation/storage/detoxification of metals in the Pacific blue shrimp. Preliminary risk considerations indicate that consumption of the shrimp farmed in New Caledonia is not of particular concern for human health.

A study on biochemical changes in the penaeid shrimp, Metapenaeus monoceros (Fabricius) following exposure to sublethal doses of organochlorine pesticide (endosulfan)

Endosulfan, a broad-spectrum non-systemic organochlorine (OC) pesticide is extensively used to control a wide variety of pests in agriculture, horticulture and public health programmes. Biochemical changes occurring in the metabolically active tissues of gills (GL), hepatopancreas (HP) and muscle (MU) of the penaeid shrimp, Metapenaeus monoceros (Fabricius) on exposure to two sublethal doses (40 and 60 ng L −1) of endosulfan were studied for 23 days of exposure (DoE). Sublethal doses of endosulfan significantly ( P < 0.05) altered the levels of the total protein (TP), the total carbohydrates (TC), the glycogen (GLY), the total free sugars (TFS) and the total lipids (TL) in test shrimps. Concentrations of biochemical components significantly varied with the DoE but were dose-independent ( P < 0.05). Percent decrease in all biochemical components increased with the progress of the DoE irrespective of the exposure concentrations. The order of percent decrease in the concentrations of the TP, TC, GLY, TL and TFS in different tissues at the end of 23 DoE was found to be MU > GL > HP, HP > GL > MU, MU > HP > GL, HP > MU > GL and MU > GL > HP, respectively. The results of the study revealed that sublethal doses of endosulfan significantly alters the proximate composition of major tissues, particularly the TP levels in the MU tissues thereby reducing the nutritive value of this economically important penaeid shrimp. Since M. monoceros exhibits significant biochemical changes on exposure to endosulfan, this species could possibly be used as biosensor of coastal marine and estuarine pollution by OCs.

Molecular cloning and transcriptional analysis of a newly identified anti-lipopolysaccharide factor gene in kuruma shrimp, Marsupenaeus japonicus

In the present study, we have cloned a new family of anti-lipopolysaccharide factor (ALF) from haemocytes of kuruma shrimp Marsupenaeus japonicus (MjALF2) using RACE method. Transcriptional analysis of MjALF2 gene in the organs of healthy shrimp revealed prominent expression in gills and muscle. In vitro LPS stimulation in the lymphoid organ cells resulted in significant increase in expression at 48, 8 and 12 h poststimulation, compared to the nonstimulated cells. In vivo injection of V. penaeicida does not show any high expression in time course assay. Phylogenetic analysis showed MjALF2 is placed in the group closer to P. monodon isoform 1 and 2 than to MjALF1. The full-length MjALF2 gene consists of 558 bp with a 363 -bp open reading frame, encoding 121 amino acids. The deduced peptide contains a putative signal peptide of 22 amino acids with molecular mass of about 13.8 kDa molecular mass. The deduced amino acid sequence of MjALF2 showed 83.3 and 56.7% identity with ALF sequences of P. monodon. The present work revealed the presence of MjALF2 gene, which is highly expressed in gills and muscle of healthy kuruma shrimp. Further studies are required to clarify the involvement of MjALF2 in immune responses for using as a therapeutic agent. Antimicrobial peptides are promising potential therapeutic agents for disease control in aquaculture. Understanding the relation of MjALF2 with innate immunity mechanism will lead to develop better health management strategies for long-term sustainability of the shrimp industry.