Identification of tropomyosin as a major allergen in the octopus Octopus vulgaris and elucidation of its IgE-binding epitopes

ABSTRACT: The major allergen (named Oct v 1) in the muscle of the octopus Octopus vulgaris was purified by gel filtration on Sephacryl S-300, anion-exchange fast protein liquid chromatography on Mono Q and reverse-phase high-pressure liquid chromatography on TSKgel Octadecyl-4PW. In addition to the molecular mass, amino acid composition and cross-reactivity with Tur c 1 (turban shell Turbo cornutus allergen), the determined partial amino acid sequence clearly demonstrated that Oct v 1 is tropomyosin, similar to the known molluscan and crustacean allergens. Using peptide fragments isolated from the lysylendopeptidase digest of Oct v 1, competitive enzyme-linked immunosorbant assay inhibition experiments showed that IgE-binding epitopes of Oct v 1 are contained in two peptides (77–112 and 148–160) in the central region and one peptide (269–281) in the C-terminal region. In the peptide 77–112, the same sequence as the IgE-binding epitope proposed for Cra g 1 (oyster Crassostrea gigas allergen) is recognized at 92-105. Moreover, the peptide 148-160 partly overlaps with the IgE-binding epitopes suggested for Pen i 1 (shrimp Penaeus indicus allergen) and Pen a 1 (shrimp Penaeus aztecus allergen), and the peptide 269–281 with those for Tur c 1 and Pen a 1.     

Growth and mortality of common octopus Octopus vulgaris reared at different stocking densities in Mediterranean offshore cages

Four ongrowing cycles of common octopus ( Octopus vulgaris ), each with two stocking densities, were carried out in an offshore area in order to evaluate the effect of the following variables on growth ( G , % body weight day⁻¹) and mortality ( M , % day⁻¹): weight ( W , 0.5–3.0 kg), temperature ( T , 14–26 °C), stocking density (SD, 6–46 kg m⁻³), size dispersion, expressed as coefficient of variation (CV, 13–42%) and significant wave height (SWH: 0.4–1.2 m). The assays were performed in an 8 m³ stainless-steel floating cage divided into two compartments of 4 m³ each. For the range of temperatures considered (14–26 °C), G depended significantly on T and SWH, with maximum G values being obtained at 18.5 °C and with an important negative effect of SWH. M depended significantly on T, W and CV, mortality being minimal at 18 °C. The two other variables had an antagonistic effect, mortality increasing with greater size dispersion – suggesting that animals should be graded throughout the process – although the effect diminished as the sizes increased. The results point towards two alternatives for the commercial ongrowing of octopus under the particular conditions of the present study: (A) two ongrowing cycles of 3.5 months to reach a final weight of 2.5 kg or (B) one 5-month cycle to reach a final weight of 3.5 kg.     

Development of an in vitro model to assess protein bioavailability in diets for common octopus (Octopus vulgaris)

An in vitro model designed to assess protein bioavailability in diets for growing Octopus was developed. The model required a previous assessment of some functional features of protein digestion in this species like the main producing organs, optimum pH for activity and total production per g tissue. The main producing organs identified were the salivary glands and the hepatopancreas (HP), being optimum pH for protease activity quite different in both organs (mostly alkaline in the posterior salivary glands and acid in the HP). In spite of the high‐specific protease activity measured in the salivary glands, a major role of the HP in protein hydrolysis is suggested due to the much bigger size of this viscera. All this information was used as a basis to develop an in vitro two‐step hydrolysis process, which simulated protein hydrolysis performed by these two organs using the Octopus enzymes. The assay was used to evaluate differences in amino acid bioavailability from several protein sources (casein, gelatin, fish meal, squid meal and krill meal) that could be used as feed ingredients for this species. As significant differences were detected both in total amount and in rate of release of the amino acids from such proteins, the model is proposed as a complementary tool in the selection and nutritional evaluation of protein ingredients to be used in diets for Octopus.     

Evaluation of relationship between Chilean octopus (Octopus mimus Gould, 1852) egg health condition and the egg bacterial community

The objective of this study was to evaluate how bacterial community associated with Chilean octopus (Octopus mimus) egg was related to egg health condition using a culture dependent method and PCR‐DGGE fingerprinting technique. Total heterotrophic bacterial number of fresh egg was much lower than infected egg. However, biodiversity of culturable bacterial community associated with the fresh egg exhibited a higher diversity than the infected egg. Result of a culture dependent method showed that Roseobacter clade was predominant in the fresh egg, while predominant species in the infected egg was γ‐proteobacteria. DGGE fingerprinting technique showed that fresh egg associated unculturable bacterial community was constituted of Roseobacter clade and Bacteroidetes, whereas Bacteroidetes was predominant bacteria in the infected egg. These results suggest that there might be some sort of relationship between octopus eggs associated bacterial community and egg health condition. Moreover, Roseobacter clade and Bacteroidetes might be potential symbiotic bacteria associated with the octopus egg, and some γ‐proteobacteria might be involved in octopus egg disease. In particular, Roseobacter clade may play an important role in octopus egg health and it raises the possibility that this clade can be utilized as potential probiotics for octopus aquaculture.     

Live transportation of Macrobrachium rosenbergii (De Man) in chilled sawdust

Three cooling rates of 1.26±0.09°C h^?1 within 8 h (slow, T₁), 2.52±0.18°C h^?1 within 4 h (moderate, T₂) and 5.04±0.36°C h^?1 within 2 h (fast, T₃) were tested to cold‐anaesthetize farm raised Macrobrachium rosenbergii (De Man) (45–52 g) in each case from 25°C down to 15±1°C in a refrigerated chilling tank, provided with aeration. The cold‐anaesthetized prawns subjected to each chilling rate were packed in an insulated cardboard box (triplicate) between two layers of moist and chilled (2–3°C) sawdust, and kept inside a chilled storage cabinet at 15±1°C, for set durations of 6, 9, 12, 15 and 18 h. Survival was determined by revitalizing the prawns in aerated water with an initial temperature of 20°C, which was raised to 29±1°C within 3 h. The experiment was repeated using berried females acclimated to brackishwater of 12 g L^?1 salinity and the percentage survival recorded after live storage for durations ranging from 6 to 24 h at intervals of 3 h. Statistically valid safe durations for obtaining 100% survival of the cold anaesthetized and live stored prawns were determined using probit analysis at the three chilling rates tested, and were found to be 7.39, 6.98 and 4.54 h in the case of adult prawns, and 7.87, 8.17 and 6.43 h for berried females for T₁, T₂ and T₃ respectively. For practical purposes, the durations that yielded 95% survival rates were computed to be 16.47, 12.14 and 8.35 h in the case of adult prawns and 18.49, 19.02 and 11.11 h for berried females for T₁, T₂, and T₃ respectively. The berried prawns revitalized after live storage were incubated in tanks and the zoea larvae reared up to postlarvae (PL‐5), and compared against a control. No significant difference was found in larval hatch fecundity, survival rate and the production of PL L^?1 between the treatment and control, indicating that the method of cold anaesthetization and live storage of berried prawns could be used for successful transportation of broodstock.     

Effects of fish hydrolysate (CPSP®) on growth and digestive gland lipid composition of Octopus vulgaris (Cuvier, 1797) juveniles

Effects of artificial diets were tested on growth and digestive gland (DG) lipid composition of juvenile Octopus vulgaris. For Experiment I, three diets were used: (i) SQUID (Loligo gahi) as a control diet; (ii) Squid paste and fish hydrolysate CPSP^®, agglutinated with gelatine (GEL20); and (iii) Squid paste, fish hydrolysate CPSP^® and gelatine (GEL40). For Experiment II, three diets were used: (i) SQUID, control diet; (ii) Squid paste, CPSP^® and alginate (ALG10); and (iii) Squid paste, CPSP^® and alginate (ALG20). For both experiments, growth rates for octopuses fed control were higher (P < 0.05), while artificial diets were not different (P > 0.05) between them. All diets promoted similar growth, regardless of the different CPSP^® concentrations and binders. Growth rates obtained were among the highest obtained for O. vulgaris with artificial diets until now. The lower growth obtained with the artificial diets was greatly reflected in the DG fatty acid composition. The most important fatty acid groups, such as n‐3 highly unsaturated fatty acids, decreased in the DG of animals fed artificial diets. Lipid content, particularly neutral lipids, was higher in the DG of octopuses fed squid, indicating higher nutritional condition of these animals compared to those fed the artificial diets.     

Enrichment of Artemia metanauplii in phospholipids and essential fatty acids as a diet for common octopus (Octopus vulgaris) paralarvae

Highly unsaturated fatty acids (HUFA), like the eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and polar lipids (essentially phospholipids, PL) have been identified as essential nutrients for common octopus (Octopus vulgaris) paralarvae. However, they are not available in sufficient amounts in live preys as Artemia, making necessary a supplementation of these nutrients previous use. A commercial emulsion, soya liposomes, and marine and soya lecithins were used to supply HUFA and PL to Artemia metanauplii, those being regarded as suitable size preys for octopus paralarvae. Our results prove that a simultaneous enrichment in HUFA and PL is possible using enrichment diets combining HUFA‐ and PL‐rich products in short‐term (4 h) incubations. Particularly interesting was the enrichment efficiency shown by the marine lecithin, which enabled the enhancement of the PL fraction of Artemia metanauplii and, importantly, also their HUFA with a remarkable 13% DHA of total fatty acids. Marine lecithin arises as a novel enrichment diet for Artemia and more effective than some commercial products currently used in hatcheries worldwide.     

Effect of different chilling rates for cold anaesthetization of Penaeus monodon (Fabricius) on the survival, duration and sensory quality under live storage in chilled sawdust

The present work was carried out with a view of determining the chilling rates for cold anaesthetization and live storage of Penaeus monodon (Fabricius) in chilled sawdust, for live transportation. Three chilling rates of 1.38 ± 0.16 °C h^?1 within 8 h (slow), 2.76 ± 0.32 °C h^?1 within 4 h (moderate) and 5.52 ± 0.64 °C h^?1 within 2 h (fast) were tested to cold anaesthetize 144 farm‐raised P. monodon (22–25 g) in each chilling rate at 15 g L^?1 salinity, from 25 °C down to 14 ± 1 °C in plastic net boxes kept in a refrigerated chilling tank provided with aeration. Eight cold‐anaesthetized shrimps subjected to each chilling rate were packed in an insulated cardboard box (triplicate) between two layers of moist and chilled (2–3 °C) sawdust, and kept inside a chilled storage cabinet at 14 ± 1 °C for durations of 16, 20, 24, 28, 32 and 36 h. Survival was determined by revitalizing the shrimps in aerated water with an initial temperature of 20 °C, which was raised to 28 ± 1 °C at 2.7 °C h^?1 within 3 h. Statistically valid, safe durations for obtaining 100% survival were 22.9 ± 1.1, 19.1 ± 0.4 and 14.6 ± 1.1 h, using probit analysis at the slow, moderate and fast chilling rates respectively. The weight loss (1.5–8.8%) due to cold anaesthetization and subsequent revitalization of the packed shrimp was insignificant. Sensory evaluation showed significant improvement in general appearance, and also in colour as well as in flavour of the meat when compared with that of the freshly harvested dead shrimps. The texture as well as odour of the raw and cooked meat between treated and untreated shrimps was unaffected. The effects of chilling rates and shipping durations on sensory quality were insignificant. One hundred per cent survival was obtained for 24, 20 and 16 h for the slow, moderate and fast chilling rates respectively. Percentage survival of the shrimps at different durations was significantly different among the chilling rates, but pair‐wise comparison revealed that the slow and moderate chilling rates were identical. Hence, the moderate chilling rate, which took only 4 h, can be considered the optimum, though the choice of different chilling rates depends on the duration of live storage desired.     

Growth of juvenile common octopus (Octopus vulgaris) in Korean waters is affected by shelter type

This study evaluated the effects of shelter pipes (PS), tube shelters (TS), and no shelter (NS) on the growth of juvenile common octopus (Octopus vulgaris). The grow-out was studied in situ by conducting a passive follow-up survey using integrated transponder tags. The growth and survival rates were evaluated in two experiments. Each experiment included two replicates. Both experiments ran for 60 days, for which each trial included 24 octopuses that were fed once a day a diet of frozen sardines (Sardinops melanostictus) and swimming crabs (Portunus trituberculatus), at 3–8% body weight. We investigated the relationship between the beak microstructure of the common octopus and possible stress events caused by culture conditions at 15 °C for 15 days. The nine juveniles were separated into three groups (NC: natural conditions, PS: culture conditions as a group, and TS: culture conditions with individuals kept separately) to analyze the amount of stress caused by rearing conditions. Stress marks were observed using a high-resolution stereo fluorescence microscope. In Experiment 1, the specific growth rates (SGRs) were 0.20 ± 0.15% per day in the PS group, 0.24 ± 0.10% per day in the TS group, and 0.68 ± 0.48% per day in the NS group. The survival rate was 62.5 ± 6.25%, 37.5 ± 18.75%, and 12.5% in the TS, PS, and NS groups, respectively. The experimental conditions of Experiment 2 were the same as those in Experiment 1, except that Experiment 2 involved different individuals and occurred during a different season. In Experiment 2, the SGRs were 0.54 ± 0.20% per day in the PS group, 0.59 ± 0.22% per day in the TS group, and 0.44 ± 0.10% per day in the NS group. Survival rate was 87.5 ± 6.25%, 75 ± 6.25%, and 75% in the TS, PS, and NS groups, respectively. Regression analysis showed that the water temperature (from 5.4–23.1 °C) affected the growth of octopuses in Experiments 1 and 2. Capture stress was detected in six of the nine individuals (NC, PS, and TS combined). Statistically significant differences in the stress levels were detected between the PS and TS groups (P ≤ 0.1), with the NC group exhibiting no stress lines after initial adaptation, unlike PS. In conclusion, octopuses kept in TS were not affected by stress after initial adaptation.     

Effective use of glucose rather than starch in formulated semimoist diets of common octopus (Octopus vulgaris)

The aim of the present work was to test the capacity of O. vulgaris to use carbohydrates supplied in three diets: a diet without added carbohydrates (diet C0: 500 g kg^?1 water, 200 g kg^?1 gelatine, 100 g kg^?1 egg yolk powder, 50 g kg^?1 freeze‐dried Sardinella aurita and 150 g kg^?1 freeze‐dried Todarodes sagittatus), and two obtained by substituting 50 g kg^?1 of T. sagittatus by glucose (diet GLU50) or by starch (diet STA50). The most stable and best accepted diet was STA50 (SFR 1.26%BW day^?1), although there were no significant differences in the growth rates obtained with the three diets: 10.1 g day^?1, 9.4 g day^?1 and 11.2 g day^?1 for C0, GLU50 and STA50, respectively (P > 0.05). The feed efficiency indices were better for GLU50, of particular note being the protein productive value of 72% and a feed conversion ratio lower than 1. Protein and lipid digestibility were similar in all three diets (96–98% for proteins and 85–94% for lipids), whereas carbohydrate digestibility was higher in GLU50 (98%) than in C0 (84%) and STA50 (0.33%). The content of carbohydrates increased in muscle and the digestive gland as a consequence of the increased carbohydrates intake.     

Effective use of glucose rather than starch in formulated semimoist diets of common octopus (Octopus vulgaris)

The aim of the present work was to test the capacity of Octopus vulgaris to use carbohydrates supplied in three diets: a diet without added carbohydrates (diet C0: 500 g kg^?1 water, 200 g kg^?1 gelatine, 100 g kg^?1 egg yolk powder, 50 g kg^?1 freeze‐dried Sardinella aurita and 150 g kg^?1 freeze‐dried Todarodes sagittatus) and two obtained by substituting 50 g kg^?1 of T. sagittatus by glucose (diet GLU50) or by starch (diet STA50). The most stable and best‐accepted diet was STA50 (SFR 1.26%BW day^?1) although there were no significant differences in the growth rates obtained with the three diets: 10.12 g day^?1, 9.37 g day^?1 and 11.22 g day^?1 for C0, GLU50 and STA50, respectively (P > 0.05). The feed efficiency indices were better for GLU50, of particular note being the protein productive value of 71.88% and a feed conversion ratio lower than 1. Protein and lipid digestibility were similar in all the three diets (96–98% for proteins and 85–94% for lipids), whereas carbohydrate digestibility was higher in GLU50 (98%) than in C0 (84%) and STA50 (0.33%). The content of carbohydrates increased in muscle and the digestive gland as a consequence of the increased carbohydrates intake.     

Changes in lipid composition of different tissues of common octopus (Octopus vulgaris) during short‐term starvation

In this work, we study the variations in concentration (mg 100 g^?1 dry weight) and total content (mg individual^?1) of different lipid classes in muscle and the digestive gland of Octopus vulgaris during an 8‐day starvation period. In all the samples analysed, polar lipids (PL) predominated in muscle (267.3–337.2 mg 100 g^?1) compared with neutral lipids (66.9–104.7 mg 100 g^?1). A significant positive tendency was observed in muscle for the content and concentration of monoglycerides as a consequence of starvation (P < 0.05). Furthermore, a higher cholesterol (CHO) content was detected in this tissue after the fourth day of starvation compared with the fed animals (P < 0.05). Neutral lipids predominated in the digestive gland (12 958–14 151 mg 100 g^?1) compared with PL (3157–6517 mg 100 g^?1), with triglycerides, free fatty acids and monoglycerides being the major lipid classes. The concentration of PL and monoglycerides showed a positive trend with starvation, while the triacylglycerol showed a negative tendency (P < 0.05). The results suggest that triglycerides and CHO are transported simultaneously from the digestive gland to the muscular tissues during starvation and the preferential use of PL and CHO during growth phases. It was estimated that lipids contributed 26% of the energy costs of the animals during starvation, mainly in the form of triglycerides from the digestive gland.     

Current status of the giant freshwater prawn ( Macrobrachium rosenbergii) industry in China,with special reference to live transportation

The giant freshwater prawn Macrobrachium rosenbergii was first introduced into mainland China by the Chinese Academy of Agricultural Sciences from Japan in 1976. It was cultured at the Guangdong Province Fishery Institute (now as Pearl River Fishery Institute, Chinese Academy of Fishery Sciences). Following these early efforts, giant freshwater prawn culture has gradually spread to northern and inland areas. At this time (2011), China has become the largest producer of giant freshwater prawn in the world. Due to a preference of cooking with fresh aquatic products, Chinese retail markets have developed which have special live‐holding counters. From small rural shops to urban supermarkets and specialty restaurants, live seafoods are commonly available whereby customers pick out the products they wish to purchase and consume. Giant freshwater prawns are produced in over 35 countries throughout the world. They are usually sold either fresh, on‐ice, or frozen. What makes the Chinese prawn industry unique is its focus on a live transportation and marketing model. As a result, numerous innovative live‐transport technologies have been developed which have been key to the success of this expanding domestic industry. In this article, we would like to give a general overview of the giant freshwater prawn industry, its status in China, and the special role live‐transportation has for expanding production of giant freshwater prawns.     

Efficient utilization of dietary lipids in Octopus vulgaris (Cuvier 1797) fed fresh and agglutinated moist diets based on aquaculture by‐products and low price trash species

The aim of this study was to evaluate growth, biochemical composition and dietary nutrients utilization in Octopus vulgaris fed on four diets based on bogue Boops boops, from different origin and in two presentations: fresh discarded bogue (aquaculture by‐product) (DB‐f), fresh wild bogue (low price trash species) (WB‐f), discarded bogue agglutinated moist diet (DB‐m) and wild bogue agglutinated moist diet (WB‐m). Diets based on DB showed higher lipid content (19–26% dw) than those based on WB (5–6% dw). Octopuses fed on DB‐based diets showed higher growth (1.5–1.9% day^?1) and higher protein efficiency ratio (0.64–0.69) than those fed on WB‐based diet (1.1–1.5% day^?1 and 0.36–0.37 respectively), which suggests good utilization of dietary lipids and also a possible protein sparing effect by lipids in O. vulgaris. Octopuses fed on diets presented fresh showed a higher growth (1.9–1.5% day^?1) and a higher feed efficiency (62–65%) than those fed on agglutinated diets (1.1–1.5% and 52–60% day^?1 respectively). Regarding fatty acids, the digestive gland clearly reflected dietary lipid and fatty acid profile, while muscle showed a more stable composition. Low dietary ARA content reflected in octopus tissues, especially in specimens fed on DB‐based diets, which did not seem to affect growth during the experimental period.     

热应激对银鲫生化指标和HSP70 mRNA表达的影响

本研究旨在探讨热应激对银鲫(Carassius auratus gibelio)生化指标及肝脏HSP70 mRNA表达的影响,选取平均体重为(72.8±10.1) g的银鲫,分别在热应激前(25℃,对照组)、热应激后(32℃,0 h、2 h、6 h、10 h)和恢复适温后6 h (25℃,恢复组)的条件下,测定机体血浆乳酸(LD)、血浆葡萄糖(Glu)、肝糖原(Gly)、己糖激酶(HK)、丙酮酸激酶(PK)和肝脏HSP70 mRNA表达量变化。研究表明,实验组乳酸含量从热应激开始至恢复期,差异始终不显著(P>0.05);实验组血浆葡萄糖水平在热应激后0 h、2 h、6 h,极显著高于对照组(P<0.01);实验组肝糖原水平在热应激0 h时,极显著高于对照组(P<0.01),其余时间点差异不显著(P>0.05);己糖激酶活力在热应激后2 h,显著高于对照组(P>0.05);丙酮酸激酶活力在热应激后6 h,极显著高于对照组(P<0.01);热应激后2 h,肝脏中HSP70 mRNA表达量达到最高(P<0.01),恢复至适温后,表达量与应激前无差异(P>0.05)。本研究为进一步探究银鲫的高温应激作用机制及调控机理提供参考数据,同时,也为银鲫养殖和育种等生产实践提供相应参考依据。     

Supplementary oxygen and temperature management during live transportation of greenlip abalone, Haliotis laevigata (Donovan, 1808)

Live greenlip abalone, Haliotis laevigata , are highly valued in Australian export markets with demand increasingly being met with cultured stock. Live transportation of abalone requires the maintenance of favourable conditions within transport containers for periods exceeding 35 h. We examined the combined effects of temperature regulation (ice provision) and of supplemental oxygen (60% and 100% concentrations) on mortality rates of abalone over 7 days following a 35-h simulated live-transport experiment. We also examined the physiological condition of greenlip abalone (oxygen consumption rate, haemolymph pH and weight) during the simulation experiment. The provision of ice and supplementary oxygen reduced abalone mortalities. Omission of ice and supplementary oxygen during the transport simulation resulted in mortality rates ranging from 70% to 100%. The addition of ice to containers with ambient oxygen concentrations decreased average mortality rates by 50%. While supplementary oxygen further reduced these rates, the provision of both ice and 100% oxygen was by far the most effective combination, reducing mortalities to between 2% and 6%. Supplementary oxygen increased oxygen consumption rates of abalone above those transported at ambient oxygen concentrations. Live-transport decreased haemolymph pH in all treatments but was most pronounced in treatments without ice or supplementary oxygen. On average, abalone lost 7–13% of their weight during the simulation but this loss was independent of transport treatment.     

Utilization of diets with different fish oil content in common octopus (Octopus vulgaris Cuvier, 1797) and resulting changes in its biochemical composition

The aim of the present work was to obtain the lipid utilization of Octopus vulgaris supplying formulated semi‐moist diets with different contents in cod oil (reduced from water content): 0 g kg^?1 (A0, 138 g kg^?1 lipids DW; N = 4), 100 g kg^?1 (A100, 286 g kg^?1 lipids DW; N = 6) and 200 g kg^?1 (A200, 388 g kg^?1 lipids DW; N = 6). The rest of the ingredients were constant in the three diets: 200 g kg^?1 gelatin, 100 g kg^?1 egg yolk powder, 150 g kg^?1 freeze‐dried Todarodes sagittatus and 50 g kg^?1 freeze‐dried Sardinella aurita). Survival was 100% with the three diets. The highest absolute feeding (15.8 ± 1.2 g day^?1), growth (9.6 ± 1.4 g day^?1; 0.91% BW day^?1) and feed efficiency rates (60.3%) were obtained with diet A0. This diet also showed greater retention of lipid and protein than A100 and A200. Protein digestibility was above 95% in all of the diets. Only diet A0 led to a high lipid digestibility coefficient (81.25%), which fell drastically to 12.3% in A200. It was notable the high polar lipid digestibility rates (83–89%) respect to neutral lipids (2–87%) in all diets. The best results were obtained with lipid feeding rates of around 1 g day^?1 and a suitable lipid content on 130–140 g kg^?1 DW in formulated diets for O. vulgaris.