Environmental Effects on Seafood Availability,Safety, and Quality (Chemical & Functional Properties of Food Components). Edited by E. Grażyna Daczkowska-Kozon and Bonnie Sun Pan

Trypsin, with molecular weight of 28 kDa from the intestine of genetically improved Nile tilapia (Oreochromis niloticus), was purified by ammonium sulfate precipitation, gel filtration, and anion-exchange chromatography. Purified trypsin had maximal activity at pH 8.0 and 60°C for hydrolysis of N _α-p-tosyl-L-arginine methyl ester. The enzyme was stable at temperatures up to 50°C and pH range of 6.0–11.0. Its activity was strongly inhibited by metal ions such as Pb²⁺ and Fe³⁺ and protease inhibitors including soybean trypsin inhibitor and phenylmethylsulfonyl fluoride. Also, the ion Ca²⁺ slightly inhibited this activity. The Michaelis-Menten constant (K _m) and catalytic constant (K _cat) of purified trypsin were 0.036 mM and 152 s^?1, respectively. Furthermore, trypsin contained low amounts of hydrophobic and aromatic amino acids as well as β-sheet (20.2%) and β-turn (25.0%).     

Purification and Characterization of an Antioxidant Protein from Pearl Oyster (Pinctada fucata martensii)

Protein from pearl oyster (Pinctada fucata martensii) was purified and characterized. Results showed that the pearl muscle protein (PMP) was composed of a single polypeptide with a molecular weight of approximate 11.6 kDa and contained 95.1% of the protein and 4.92% of the carbohydrate of the oyster. The polypeptide appears to be a glycoprotein, since the Fourier transform infrared (FTIR) spectroscopy of the PMP shows the typical characteristics of protein and polysaccharides. The denaturation temperature of the PMP was 81.3°C by differential scanning calorimetry. Circular dichroism spectroscopy showed that the PMP has a highly ordered structure. The PMP (200 μg/mL) showed a high oxygen radical absorbance activity (ORAC) of 6.57 μmol Trolox/μmol protein. Its antioxidant activity was stable at temperatures from 30–80°C and pH 2–10. The antioxidant activity was significantly inhibited by Zn²⁺, Ca²⁺, and EDTA and enhanced by Mn²⁺. The results suggest that the PMP could be used as a potential natural antioxidant.     

Effects of Fumonisin B1-Containing Feed on the Muscle Proteins and Ice-Storage Life of White Shrimp (Litopenaeus vannamei)

Juvenile (5.5–5.7 g) white leg shrimp were exposed to mycotoxin in one indoor trial by feeding fumonisin B₁ (FB₁) at levels from 0.2 to 2.0 μg g^?1 FB₁ for 30 days. Shrimp growth was affected after exposure to more than 0.6 μg g^?1 FB₁. Soluble muscle protein concentration decreased, and changes in myosin thermodynamic properties were observed in shrimp after 30 days of exposure to FB₁. Marked histological changes in tissue of shrimp fed a diet containing FB₁ at 2.0 μg g^?1 were also observed. Shrimp fed diets containing more than 0.6 μg g^?1 FB₁ showed greater decrease in shear forces after 12 days of ice storage.     

Determination of Trace Lead in Fish Sauces by ID-ICP-MS After Mg(OH)2 Coprecipitation

A method of isotope dilution coupled with inductively coupled plasma mass spectrometry (ID-ICP-MS) after Mg(OH)₂ coprecipitation was employed to determine trace lead (Pb) in fish sauces. The pH values and the amounts of Mg²⁺ in the coprecipitation system and the amounts of isotopically enriched ²⁰⁶Pb reagent added in the measurement were optimized. Results showed that a recovery by adding standard was 92.0% with a relative standard deviation (RSD) of 0.39%. The detection limit estimated with the calculated method used in the study was 7.65 ng g^?1. Through the Mg(OH)₂ coprecipitation, more than 80% of matrix ions such as K⁺, Na⁺, and Ca²⁺ in the samples were effectively removed, and the trace Pb in fish sauces was largely enriched. The concentrations of Pb measured in 12 fish sauces ranged from 0.068 to 0.299 μg g^?1, which was completely within the Pb limit (0.50 μg g^?1), a hygienic standard for aquatic flavorings set in China (GB 10133-2005). It has been well demonstrated that the method of ID-ICP-MS coupled with Mg(OH)₂ coprecipitation is suitable for determining trace Pb in fish sauces containing high levels of salt matrices.     

Changes of Fatty Acid Composition in Frog Legs (Rana esculenta) During Cold Storage Period: Irradiation Effect

This study investigates the changes in fatty acid profile of gamma irradiated frog legs (Rana esculenta) during cold (2°C) storage period. Instead of freezing the frog legs for a prolonged shelf life, gamma irradiated fresh frog legs can be used for consumer satisfaction, because consumers prefer fresh frog legs and are willing to pay more than for frozen in the European market. Frog legs were irradiated at doses of 0, 4, and 5 kGy using ⁶⁰Co sources. The main fatty acids of nonirradiated and irradiated frog legs (4 and 5 kGy) were palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1ω9), linoleic acid (C18:2ω6), and eicosatrienoic acid (C20: 3ω3, ETE). Low doses (4 and 5 kGy) of irradiation treatment had no effect on fatty acid components of frog legs compared to the nonirradiated ones (p > 0.05). The fatty acid composition of frog meat was characterized by its high linoleic acid content (17.1–21.4%). At the initial stage of the storage, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) levels of frog legs remained unchanged with low doses of irradiation (p < 0.05). These fatty acids decreased significantly in the irradiated group (p < 0.05) at the end of the storage period. However, there were no significant changes in nonirradiated frog legs after 10 days of storage.     

A study of the damage of the intestinal mucosa barrier structure and function of <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis> with <Emphasis Type="Italic">Aeromonas hydrophila</Emphasis>

The aim of this study is to explore the effect of Aeromonas hydrophila on the intestinal mucosal barrier structure and intestinal permeability in grass carp (Ctenopharyngodon idella). Histopathological examinations showed that A. hydrophila induced severe intestinal lesions, including inflammatory cell infiltration and intestinal villus fusion and swelling. Messenger RNA (mRNA) expression of the inflammatory cytokines TNF-α, IL-1β, IL-8, IL-10 and MyD88 was significantly increased after infection with A. hydrophila. The permeability of intestinal mucosa was determined using Evans blue (EB) and d-lactic acid. The results indicated that the levels of EB and serum d-lactic acid were significantly increased after infection with A. hydrophila (p < 0.05). Our results also indicated that the intestinal mucosal barrier injury induced by A. hydrophila infection was closely associated with the expression of the tight junction (TJ) protein zonula occludens-1 (ZO-1), occludin, claudin b and claudin c as well as the activity of Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase. Lower mRNA levels of occludin and lower Na⁺, K⁺-ATPase and Ca²⁺, Mg²⁺-ATPase activity in the intestines were observed after challenge. ZO-1 and claudin c were significantly increased 24 h after infection with A. hydrophila. The most interesting finding was that claudin b also significantly increased 24 h after challenge and then decreased to lower levels at 72, 120 and 168 h post-infection compared to the PBS-treated control group. The results demonstrated that grass carp infection with A. hydrophila induced intestinal inflammation and impaired the structure and function of the intestinal mucosal barrier.     

FRESHWATER PRAWN FARMING IN GHER SYSTEMS IN SOUTHWEST BANGLADESH

The livelihoods of a large number of farmers are associated with freshwater prawn (Macrobrachium rosenbergii) farming in gher (modified rice fields with high, broad peripheral dikes) systems in southwest Bangladesh. Most farmers integrated prawn with fish and rice in their gher and followed extensive methods using low inputs. Although farmers have improved their social and economic conditions through prawn farming, constraints include high production costs, low supply of wild fry and snail meat as prawn feed, lack of technical knowledge of farmers, and negative environmental impacts.     

Purification and characterization of trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides

Trypsin from the pyloric ceca of orange-spotted grouper, Epinephelus coioides, was purified by fractionation with ammonium sulfate, ionic exchange, and affinity chromatography. The protein was purified 161.85-fold with a yield of 4%. Purified trypsin had an apparent molecular weight of 24 kDa according to an SDS-PAGE analysis. Optimal profiles of temperature and pH of the enzyme were 50°C and 8–10, respectively, using Nα-benzoyl-l-arginine ethyl ester as the substrate. The results of thermal and pH stability assays showed that the enzyme was stable at temperatures of up to 50°C and in the pH range of 6–8. Trypsin activity decreased with an increasing NaCl concentration (0–0.6 M). The activity of purified trypsin was effectively inhibited by a soybean trypsin inhibitor and N-p-tosyl-l-lysine chloromethyl ketone, and was slightly inhibited by iodoacetic acid, ethylenediaminetetraacetic acid, 1-(l-trans-epoxysuccinyl-leucylamino)-4-guanidinobutane, and pepstatin A. Protein identification of the purified protease showed that the sequences of two peptides, LGEHNI and NLDNDIML, were highly homologous to other fish trypsins. The measurement of trypsin activity in different tissues showed that the highest activity was detected in pyloric ceca, followed by anterior intestine, middle intestine, hind intestine and spleen, but very low activities were found in other tissues. An inverse relationship between the trypsin activity in four tissues of pyloric ceca, anterior intestine, middle intestine and hind intestine and fish body weight as a result of increased pepsin in stomach indicated grouper growth status was increased.     

Stochastic modeling of production risk and technical efficiency of Thai koi (Anabas testudineus) farming in Northern Bangladesh

Thai koi is one of the high commercially valued farm fishes which have expanded tremendously in the northern part of Bangladesh. However, despite its impressive growth in the initial years, output of the industry is highly volatile across different years and farms. This research, following the Just and Pope framework, has made use of Kumbhakar’s (2002 Kumbhakar, S.C. (2002) Specification and estimation of production risk, risk preferences, and technical efficiency. American Journal of Agricultural Economics, 84, 8–22.[Crossref], [Web of Science ®] , [Google Scholar]) extended stochastic frontier model (SFM) to estimate production risk and technical efficiency (TE) in Thai koi farming. Results show that feed is the most important input significantly influencing production. Zeolite and pesticide appear to be risk-reducing, while fingerlings, labor, feed and salt turn out to be risk-increasing inputs. The inefficiency model indicates that labor and pond area significantly reduce the technical inefficiency. Feed, fingerling and lime also act as TE-improving factors. Education of farmers improves TE, while experience, training and frequency of water change do not impact the TE. The mean TE scores are 0.73 and 0.96 for the flexible risk estimate and conventional SFM estimates, respectively. TE values appear to become exaggerated when production risk is excluded from the model. SFM recognizes that risk is involved in Thai koi farming that causes lower TE.     

Use of Aqueous Extracts of Avocado Pear,Pyrus Communis,Leaf as Anaesthetic in Gonadectomy of African Catfish,Clarias gariepinus

This study was carried out to determine the efficacy and effective concentration of avocado pear (Pyrus communis) leaf extract as an anaesthetic in gonadectomy of Clarias gariepinus broodstocks. Fresh avocado leaves were harvested, water extracted, and applied at 0 (control), 100, 130, 160, and 190ml/l to C. gariepinus broodstocks. Onset of individual phases of anaesthesia and recovery rate was determined. Gonadectomy was then carried out on individual anaesthetized fish and the time taken was recorded. Aqueous extracts of avocado pear leaf at concentrations ranging from 130–190 ml/l induced general anaesthesia. The recovery rate was concentration-dependent but non-linear, with a significant threshold in efficacy between 130 and 190 ml/l. There was no mortality recorded during the experimental period. The effective concentration of avocado pear leaf extracts that best sustained the gonadectomy was 190 ml/l with a recovery time of 12 minutes.     

溶藻弧菌感染后凡纳滨对虾鳃组织免疫相关基因的表达

分别给凡纳滨对虾(Litopenaeus vannamei)注射生理盐水、3×10⁶ CFU/mL(低浓度组)和9×10⁶ CFU/mL(高浓度组)溶藻弧菌(Vibrio alginolyticus)菌液, 采用荧光定量PCR技术, 检测不同处理后凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶基因表达量随时间的变化。结果表明, 处理42 h内, 注射生理盐水组对虾的Toll受体和IMD mRNA表达量无显著变化, 溶菌酶mRNA表达量在注射36 h后显著升高。急性感染溶藻弧菌后, 凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶mRNA的表达量峰值分别出现在感染后24、42和36 h; 溶藻弧菌的感染剂量不影响上述基因表达峰值的出现时间, 但显著影响上述基因的表达峰值(P<0.05), 各基因表达量峰值由大到小均依次为高浓度组、低浓度组、生理盐水组。急性感染初期, 对虾鳃组织中Toll受体mRNA表达量呈现显著下调, 而IMD和溶菌酶mRNA表达量在感染初期不存在显著下调现象。与感染前各基因的表达量相比, 高浓度溶藻弧菌感染组Toll受体mRNA表达量在2 h时显著下调, 低浓度溶藻弧菌感染组Toll受体mRNA表达量在3 h显著下调(P<0.05); 高浓度组IMD和溶菌酶mRNA表达量分别在36 h和12 h时开始有显著上调, 而低浓度组IMD和溶菌酶mRNA表达量则分别在42 h和24 h才有显著上调(P<0.05)。表明溶藻弧菌感染对凡纳滨对虾鳃组织中Toll受体、IMD和溶菌酶mRNA表达量有显著影响, 各基因的表达量与感染进程及溶藻弧菌剂量存在一定的相关性。

     

Age, growth, and reproductive characteristics of dolphinfish Coryphaena hippurus in the waters off west Kyushu, northern East China Sea

The growth and reproductive characteristics of dolphinfish Coryphaena hippurus collected in the waters off western Kyushu from May 2008 to April 2011 were determined based on scale and otolith readings and gonad histological examinations, respectively. Based on annual increments in scales and daily increments in sagittal otoliths, the von Bertalanffy growth curves in male and females were determined as $ FL_{t} = 1049[1 - \exp \{ - 0.835(t + 6.975 \times 10^{ - 14} )\} ] $ and $ FL_{t} = 938[1 - \exp \{ - 1.029(t + 6.975 \times 10^{ - 14} )\} ] $ , respectively, where FL _t is the mean fork length (mm) at age t. The spawning period was found to last from June to August for dolphinfish, based on an examination of the monthly changes in the gonadosomatic index and histological observations. Therefore, based on the relationship between the fork length and the developmental stage of the testes or ovaries, male and female dolphinfish were found to reach sexual maturity by the following spawning season after hatching in the northern East China Sea.     

Metabolic and phylogenetic profiles of microbial communities from a mariculture base on the Chinese Guangdong coast

This study examined the phylogenetic and metabolic features of microbial communities in Baisha Bay mariculture areas of Nan’ao Island, a mariculture base on the Chinese Guangdong coast. Large seaweed (Gracilaria lemaneiformis) cultivation zones (LZ), fish culture zones (FZ), and control zones (CZ) were sampled. According to community-level physiological profiling, d-mannitol, d-mannitol/α-d-lactose, and d-mannitol/α-cyclodextrin were the carbon sources with the highest utilization rates in CZ, LZ, and FZ, respectively (R _s  > 1.9). Denaturing gradient gel electrophoresis analysis showed that the dominant bacteria in CZ and LZ were Alphaproteobacteria (39.9 and 34.5%, respectively), whereas in FZ they were Flavobacteria (32.2%). The Shannon diversity in CZ and LZ was significantly higher than in FZ (P < 0.05). Some opportunistic pathogens, such as Vibrio, Pseudoalteromonas and Loktanella, were abundant in FZ. The microbial communities and most of the dominant bacteria in FZ were more affected by environmental factors including salinity, chemical oxygen demand and total phosphorus. Some bacteria, such as Brevundimonas and Polaribater, have the potential to degrade agar in seaweed, and were isolated from samples rich in algae which are dominant in LZ. The cultivation of G. lemaneiformis created the notable structures and functions of macroalgae-associated bacterial communities that were less affected by the environmental factors examined.     

Isolation,Purification, and Biochemical Characterization of Trypsin from Indian Mackerel (Rastralliger kanagurta)

Trypsin from viscera of Indian mackerel (Rastralliger kanagurta) was purified by ammonium sulphate precipitation and chromatographic techniques such as size exclusion, ion exchange, and affinity chromatography, with a 14.4-fold increase in specific activity and 18.7% recovery. The molecular weight of the trypsin was estimated to be approximately 26 kDa using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Purified trypsin showed amidase-specific activity which was determined using benzoyl-dl-arginine-p-nitroanilide (BAPNA). The optimum pH and temperature for isolated trypsin activity were 9.0 and 50°C, respectively. The purified trypsin was strongly inhibited by soybean trypsin inhibitor (SBTI) and N-p-tosyl-1-lysine chloromethyl ketone (TLCK). Purified trypsin showed almost 40% recovery at high NaCl concentration (30%). The N-terminal amino acid sequence of the first 10 amino acids of purified trypsin was IVGGYESQPH. The Michaelis-Menten constant (K_m) and catalytic constant (K_cat) of purified trypsin were 0.430 mM and 0.77 s^?1, respectively, determined using BAPNA as a substrate. Purified trypsin showed digestion of casein similar to bovine trypsin by the fluorometric method.     

Antioxidative Activitiy of Protein Hydrolysate from the Muscle of Common Kilka (Clupeonella cultriventris caspia) Prepared Using the Purified Trypsin from Common Kilka Intestine

Trypsin from the intestine of common kilka (Clupeonella cultriventris caspia) was purified using ammonium sulfate precipitation (30–50% saturation), Sephadex G-75, and DEAE-cellulose chromatography with the purity of 30-fold and the yield of 12%. The molecular weight of trypsin was estimated to be 23.2 kDa based on sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The trypsin had optimal activity at pH 8.0 and 60°C using N-α-benzoyl-DL-arginine-ρ-nitroanilide hydrochloride (BAPNA) as a substrate and showed high stability in the pH range of 7.0–10.0. It was stable up to 50°C. Soybean trypsin inhibitor (SBTI) and N-ρ-tosyl-L-lysine-chloromethylketone (TLCK) significantly inhibited trypsin activity (p < 0.05). Protein hydrolysate from common kilka muscle with different degrees of hydrolysis (DHs; 20, 30, and 40%) was prepared using the purified trypsin, and antioxidative activities were determined. 1,1-Diphenyl-2-picrylhydrazyl (DPPH), 2,2’-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical scavenging activities, ferric reducing antioxidant power, and ferrous chelating activity of hydrolysate increased with increasing DH up to 40% (p < 0.05). Therefore, trypsin from intestine of common kilka could be used as a processing aid for production of fish protein hydrolysate with antioxidative activity.     

Effects of <Emphasis Type="SmallCaps">l</Emphasis>-carnitine against H<Subscript>2</Subscript>O<Subscript>2</Subscript>-induced oxidative stress in grass carp ovary cells (<Emphasis Type="Italic">Ctenopharyngodon idellus</Emphasis>)

This study was designed in vitro to investigate the effects of l-carnitine against H₂O₂-induced oxidative stress in a grass carp (Ctenopharyngodon idellus) ovary cell line (GCO). GCO cells were pre-treated with different concentrations of l-carnitine, followed by incubation with 2.5 mM H₂O₂ for 1 h to induce oxidative damage. The results indicated that adding l-carnitine at concentrations of 0.01–1 mM into the medium for 12 h significantly increased cell viability. Pre-treatment with l-carnitine at concentrations of 0.1–5 mM for 12 h significantly inhibited 2.5 mM H₂O₂-induced cell viability loss. The significant decreases in the level of reactive oxygen species and cell apoptosis were observed in 0.5 mM l-carnitine group compared to the H₂O₂ group. Malondialdehyde values of all of the l-carnitine groups were significantly lower than those of the H₂O₂ group, while total glutathione levels of all of the l-carnitine groups were significantly higher than of the H₂O₂ group. The activity of antioxidant enzymes, such as total superoxide dismutase (0.1 and 0.5 mM l-carnitine), catalase (0.5 mM l-carnitine) and γ-glutamyl cysteine synthetase (0.5 and 1 mM l-carnitine), was significantly increased. In addition, pre-treatment of l-carnitine in GCO cells exposed to 2.5 mM H₂O₂ significantly increased the mRNA expression of copper, zinc superoxide dismutase, catalase (0.5 mM l-carnitine), glutamate cysteine ligase catalytic subunit (0.1–1 mM) and glutathione peroxidase (0.1 mM l-carnitine). In conclusion, l-carnitine promotes GCO cell growth and improves antioxidant function, it plays a protective role against oxidative stress induced by H₂O₂ in GCO cells, and the appropriate supplemental amount of l-carnitine is 0.1–1 mM.     

黄斑蓝子鱼LC-PUFA 合成代谢与渗透压调节的关系研究

为探讨黄斑蓝子鱼(Siganus canaliculatus)长链多不饱和脂肪酸(long-chain polyunsaturated fatty acids,LC-PUFA)合成代谢与渗透压调节的关系,本研究以鱼油(FO)和混合植物油(苏子油与双低菜籽油,VO)为脂肪源配制两种等氮等脂饲料,投喂饲养在3种盐度(10、20和32)下的黄斑蓝子鱼幼鱼8周后,分析了各处理组幼鱼的生长性能和鳃的磷脂脂肪酸组成、Na+/K+-ATPase(NKA)活力及其基因表达。结果显示,相同盐度下,VO组和FO组鱼的生长性能差异不显著(P0.05);FO组鱼鳃磷脂中的n-3 LC-PUFA含量显著高于VO组(P0.05),但VO组鱼的n-6 LC-PUFA水平显著高于FO组(P0.05);VO组鱼鳃的NKA酶活力及其m RNA表达量都显著高于FO组(P0.05)。不同盐度下,无论VO组还是FO组的鱼,盐度10组鱼的生长性能显著低于盐度20和32组(P0.05),而其鳃的LC-PUFA含量、NKA酶活力及其m RNA表达量都显著高于盐度20和32组(P0.05),各指标在后两个盐度组之间差异不显著(P0.05)。由此可见,盐度10对黄斑蓝子鱼具有一定的胁迫性,导致其生长性能较差。摄食鱼油脂肪源饲料,可以提高鱼鳃磷脂的n-3 LC-PUFA水平;而摄食植物脂肪源饲料时,鱼体可能通过自身合成的n-6LC-PUFA调控鳃的NKA基因表达及其酶活力以调节渗透压。     

Carotenoid Pigments Composition of Two Commonly Discarded Decapod Crustaceans in Grand Sole and the Galician-Northern Portugal Coast Fisheries

ABSTRACT

Carotenoid composition of two commonly discarded decapod crustaceans, Liocarcinus depurator and Munida spp., was assessed in different catching months. Total carotenoid content ranged between 7 ± 1 and 22.4 ± 0.7 μg g^?1. Pigment separation was performed by high-performance liquid chromatography coupled with a diode array detector. Astaxanthin accounted for 21 to 50% of total carotenoids, and the remaining percentage corresponded mainly to esters of astaxanthin. The fatty acid profile of carotenoid extracts for each species was evaluated. Overall total unsaturated fatty acids content ranged from 406 to 1,631 μg g^?1. Both species were rich in omega-3 fatty acids, which on average represented 68% of total unsaturated fatty acids. Therefore, the use of these low value by-catch species as a source of carotenoid pigments and fatty acids (e.g., for aquaculture formulations) is a possible alternative to their disposal at sea.