Effect of diatom supplementation during the nursery rearing of Litopenaeus vannamei (Boone, 1931) in a heterotrophic culture system

Bio-floc shrimp culture systems have been investigated in an attempt to optimize water use and prevent the discharge of effluent into the environment. The importance of microalgae in maintaining water quality and nutrition of the shrimp is well known in conventional systems; however, its maintenance amid bio-flocs and its role in the shrimp performance in this system are still poorly understood. The aim of this study was to evaluate the contribution of diatoms in the performance of Litopenaeus vannamei reared during the nursery phase in intensive system with minimal water exchange. Shrimp (0.31 ± 0.10 g) were reared among diatoms, bio-flocs and the combination of the two forming the mixture medium. The survival of shrimp was high in all treatments (90–97%). However, the shrimp reared among diatoms showed higher weight gain (P\0.05) and feed conversion ratio significantly more efficient, reaching a value of 0.47. The results indicate the importance of diatoms in bio-floc culture systems and points out to future research in an attempt to maintain a constant presence of these microalgae in culture medium without requiring successive inoculations.

Análise do comportamento e crescimento do camarão branco Litopenaeus vannamei (Boone, 1931) em diferentes substratos inconsolidados

The aim of this study was to test the sediment preference of L. vannamei shrimp. It was observed shrimp visit frequency, swimming and burying behaviour at different sediment compositions for 24h. Juvenile (0.93 ± 0.29g) and sub-adult shrimps (10.0 ± 1.18g) were obtained from the aquaculture station at Universidade Federal Rural do Semi-Árido UFERSA, and held in a plastic tank (water volume 500 L) supplied with aerated water and kept at constant temperature, pH, and salinity. Shrimp was fed by commercial shrimp dry food. The experimental substrates were composed by A: medium sand + thick sand + very thick sand + gravel; B: very fine sand + fine sand; and C: silt + clay. Thus, six different substrate combinations were tested: A, B, C, A+B, A+C, B+C. To test preference, it was used a cylindrical tank (40 l) divided into six differently substrate compartments. A single shrimp was introduced each tank and the frequency at which this shrimp visited each compartment was recorded over a 24h study period. It was tested 54 shrimp (18 sub-adult males, 18 subadult females and 18 juveniles). For each trial, sediment and water were changed to avoid pheromones and residues influence. Shrimp were weighted and sub-adults were divided by sex: males present petasma and females present thelycum. Data were collected on the experimental day at 19:30; 20:30; 00:30; 1:30; 05:30; 06:30; 13:30 and 14:30 h. At each time point, shrimp were observed for 20-min periods, in which we noted down which compartment the shrimp was occupying at 2-min intervals. Thus, for each period we had eleven observations (88 observations per day). For observations at night, it was used dim red light that did not affect shrimp behaviour. At each 20-min period, it was observed visit frequency in each substrate, if shrimp was burred or not or if it was swimming. There was not significant difference between light and dark burry activity for females. Swimming activity was significantly higher at night, mainly at 00:30 and 01:30 h. All L. vannamei shrimp showed preference for sediment B. This animal presents cyclic activity, spends the day light period buried and swims at night

Experimental pathogenecity of white spot syndrome virus (WSSV) in Litopenaeus vannamei

The pathogenecity of white spot syndrome virus (WSV) was studied experimentally with challenge exposure of two hundred shrimp with average weight 10 to 12 grams of Litopenaeus vannamei. The shrimp L. vannamei before introducing examined with IQ 2000 detection Kit for WSV. The Fenneropenaeus indicus that showed the clinical sign and PCR positive of white spot disease (WSD) was used the source of WSV. The challenge exposures were accomplished by feeding minced tissue of F. indicus for 24 hours. The result showed L. vannamei after three days revealed the clinical sign of WSV, the PCR examined was positive and all shrimp died after ten days. The shrimp that showed sign of disease were collected for histpathology in Davidson fixator and a part of samples preserved in Ethyl alcohol %75to %90 for PCR. The histopathology showed the effect of virus and cowdly type A inclusion body can see in all tissue except hepatopancreas. The PCR also indicate the virus infected the shrimp Litpeneaus vannamei after 3 days. The SOI and ROI determined the severity of infection and rate of infection in different tissue.

Biodiversidade e abundância da ictiofauna associada ao cultivo orgânico de Litopenaeus vannamei (Boone, 1931)

The expansion of shrimp farming has caused a series of environmental impacts, often as a result of lack of planning and adequate management. Organic aquaculture has emerged as an alternative to conventional shrimp farming, and differently, aims at the economical, ecological and farming potential of other organisms, such as fishes, oysters and seaweeds. The present study aimed at evaluating the biological diversity and abundance of the ichthyofauna associated to Litopenaeus vannamei organic culture at PRIMAR farm (Rio Grande do Norte, Brazil). The sampling period consisted of four culture cycles (1, 2, 3, and 4) accomplished in four grow-out ponds (V1, V7, V2, and V4) in 2005. The first two culture cycles were carried out during the rainy season, and the last two cycles, during the dry season. Environmental (temperature, salinity, transparency, and rainfall), biodiversity (diversity and uniformity and abundance indexes) of the ichthyofauna, and shrimp productivity data were collected throughout these four culture cycles. The results obtained for the environmental variables showed that both salinity and transparency oscillated in a significant way (p < 0.001) between culture cycles. In relation to the abundance of the ichthyofauna, fifty four species associated to the organic culture of Litopenaeus vannamei were collected and identified at PRIMAR. For the biodiversity criterion, larger species richness (S = 46) and uniformity (E = 0.59 ± 0.16) were observed during cycles 3 and 4 than in cycles 1 and 2 (S = 41 and E = 0.15 ± 0.12). A positive and significant correlation was obtained for the transparency with diversity and uniformity of fish species between cycles 1 and 2 (wet season) and cycles 3 and 4 (dry season) (p < 0.017 - Shannon Index; p < 0.008 - Pielou Index, respectively). A correlation could not be established between shrimp productivity and fish biomass. However, fish biomass decreased from cycles 1 and 2 to cycles 3 and 4, concomitant with an increase in shrimp productivity, most probably due to a higher (from 58.0% to 71.0%) shrimp survival. Regarding the culture potential of the ichthyofauna, three species (Mugil curema, Mugil liza and Chaetodipterus faber) were identified as potential farming alternatives, either singly or in consortium with Litopenaeus vannamei. A fourth species (Centropomus undecimalis) was indicated as an alternative for single culture or in consortium with other fish species. In conclusion, the large diversity of estuarine fishes associated to Litopenaeus vannamei farming observed at PRIMAR clearly indicated the ecological feasibility for organic aquaculture in northeastern Brazil

Efeitos bioquímicos da exposição ao nanomaterial óxido de grafeno em diferentes tecidos do camarão branco (Litopenaeus vannamei; Crustacea, Decapoda), através da suplementação na ração

O desenvolvimento da nanotecnologia vem se intensificando nos últimos anos. Sendo que os NM já estão sendo utilizados em vários produtos disponíveis no mercado. Dentre os NM mais utilizados estão os compostos de carbono que embora sejam compostos somente por este elemento podem ter estruturas diferentes que refletem em suas aplicações e possivelmente em seus efeitos. Dentre os NM de carbono, o grafeno e o óxido de grafeno apresentam promissoras características que ampliam sua utilização em diversos segmentos desde eletrônicos até a distribuição de medicamentos. A intensificação da produção e utilização destes NM é acompanhada pela liberação destes nanomateriais no ambiente que pode afetar os organismos vivos, principalmente os animais aquáticos. Entretanto, pouco se sabe sobre os efeitos do óxido de grafeno em crustáceos de importância comercial como é o caso do camarão branco Litopenaeus vannamei. Portanto, a presente dissertação teve como objetivo avaliar os efeitos biológicos da exposição ao óxido de grafeno em diferentes tecidos do camarão.

Potencial terapêutico de um heparinóide isolado do invertebrado marinho Litopenaeus vannamei

The occurrence of bioactive compounds in marine organisms comes awaking the interest of the pharmaceutical industry. Heparin, a sulfated polysaccharide which presence was already identified in several marine invertebrates, is very attractive due its remarkable functional versatility. Besides to intervene in blood coagulation, this molecule has a great anti-inflammatory potential. However, its strong anticoagulant activity difficult the clinical exploitation of its anti-inflammatory properties. Thus, the aims of this work were to evaluate the effect of a heparin-like compound (heparinoid), isolated from the cephalotorax of the Litopenaeus vannamei shrimp, on the inflammatory response, hemostasia and synthesis of antithrombotic heparan sulfate by endothelial cells, besides studying some aspects concerning its structure. The purified heparinoid was structurally characterized following an analytical boarding, involving electrophoresis and chromatography. The structural analysis have shown that this compound possess a high content of glucuronic acid residues and disulfated disaccharide units. In contrast to mammalian heparin, the heparinoid was incapable to stimulate the synthesis of heparan sulfate by endothelial cells in the tested concentrations, beyond to show reduced anticoagulant activity and hemorrhagic effect. In a model of acute inflammation, the compound isolated from the shrimp reduced more than 50% of the cellular infiltration. Besides reduce the activity of MMP-9 and proMMP-2 of the peritoneal lavage of inflamed animals, the heparinoid also reduced the activity of MMP-9 secreted by activated human leukocytes. These results demonstrate the potential of heparinoid from L. vannamei to intervene in the inflammatory response. For possessing reduced anticoagulant activity and hemorrhagic effect, this compound can serve as a structural model to direct the development of more specific therapeutical agents to the treatment of inflammatory diseases

Protection of white leg shrimp (Litopenaeus vannamei) against white spot disease virus using oral recombinant bioencapsulated VP28

Aquaculture, is perceived as having the greatest potential to meet the growing demand for aquatic food. Crustaceans form one of the main value added components in aquaculture and among them, shrimp aquaculture is the predominant one. Industrial shrimp fanning, in combination with poor management in shrimp aquaculture, has quickly led to severe pollution in shrimp ponds, thereby creating a suitable environment for development of bacterial and virus diseases. White spot disease is one of the most deadly diseases that are caused heavy loss in all Penaeid shrimps family. In Iran during 2002 to 2004 in the Kuzestan province and in 2005 in Bushehr province, the most ponds and farms infected with white spot and the entire industry was facing threat of closure. Owing to the impact of WSSV infection to shrimp aquaculture, there is an urgent need to develop suitable strategies to protect cultured shrimps and make aquaculture more sustainable. Therefore, this study aimed to examine the possibility of protecting shrimp against white spot syndrome virus using bioencapsulated Anemia with E. coil containing the recombinant protein VP28, designed. Virus genome was extracted from naturally infected Litopenaeus vannamei in the Choebdch farms and VP28 gene by designed primers was amplified, extracted, purified and cloned in E. coli TGI. Protein expression evaluated and inactivated bacteria containing recombinant protein encapsulated in Artemia nauplii. White shrimp post larvae stage 5 were fed for 5 days with recombinant nauplii and twice on days 7 and 25 after feeding with Artemia nauplii were challenged with white spot virus. The results of the first experiment revealed that cumulative mortality percent in the group receiving the bacteria containing recombinant plasmid (pMal + VP28) was %14.44±1.11 and the relative percent survival %80.30±1.51. In this group the mortality rates in the various repetitions varied from the 13.33% to 16.66% and relative percent survival of 77.27% to 81.81%. in the Non-recombinant plasmid group (pMal) Mean percent mortality was% 33.33±3.84 and the Relative Percent Survival %54.54±5.24 and in the group that received bacteria contained no recombinant plasmid the Mean cumulative mortality percent was%48.88 ± 5.87 and Relative Percent Survival%33.33± 8.01.

Efeito anti-inflamatório do heparinóide isolado do camarão Litopenaeus vannamei sobre a peritonite aguda

In recent years the heparin has been the subject of several studies that aim to expand its use as a therapeutic agent, due to its ability to modulate the activity of various proteins that play important roles in the regulation of pathophysiological processes. In several experiments and preclinical trials, heparin has demonstrated an anti-inflammatory role. However, its clinical use is limited, due to its strong anticoagulant activity and hemorrhagic complications. For this reason, considerable efforts have been employed in discovery of heparin analogous (heparinoid) with reduced side effects, that retain the anti-inflammatory properties of heparin. In this context, a heparinoid obtained from the head of Litopenaeus vannamei shrimp, which presents a structural similarity to heparin, showed, in previous studies, anti-inflammatory activity in a model of acute peritonitis with reduced anticoagulant effect in vitro and low hemorrhagic activity. Thus, the present work had as objective to evaluate the effect the heparinoid of the cephalothorax of gray shrimp on the acute inflammatory response in different times (3 or 6 hours after the induction of inflammatory stimulus), using the model of acute peritonitis induced in mice. It was also analyzed the HL effect over the activity of elastase, an enzyme involved in leukocyte recruitment. Furthermore to check if the different doses of heparin and heparinoid change the hemostatic balance in vivo, was assessed the effect of these compounds on the plasma clotting time in animals submitted to inflammation. The results show that in 3 hours, all doses of heparinoid were able to prevent efficiently in the acute inflammatory process without any anticoagulant effects, unlike the extrapolation dose of heparin, which has induced a large hemorrhage due its high anticoagulant activity. However, 6 hours after induction of inflammation, only the dosages of 0.1 and 1.0 μg/Kg of heparin and 1.0 μg/Kg of heparinoid kept anti-migratory effect, without changing of the hemostatic balance. These results indicate that the anti-migratory effect of theses compounds depends on the dosage and time of inflammatory stimulus. The HL and heparin were also able to inhibit the activity of the enzyme elastase. The discovery of this bioactive compound in the cephalothorax of shrimps can arouse great interest in biotechnology, since this compound could be useful as a structural model interesting for the development of new therapeutic agents for peritonitis

Digestibility of different wheat products in white shrimp Litopenaeus vannamei juveniles

Dry matter, energy, crude protein and amino acid apparent digestibility coefficients (ADCs) were determined in white shrimp juveniles for six wheat products: hard red winter whole grain meal (HWG), Rayon whole grain meal (RWG), Durum whole grain meal (DWG), hard red winter clear flour (HCF), mixed wheat 2nd clear flour (MCF) and semolina (S). The test diets included 30% of the test ingredients and 70% of a ground commercial diet supplemented with 1% chromic oxide and 1% sodium alginate. Amino acid contents in the ingredients, diets and feces were analyzed, and digestibility was determined by difference in order to minimize the impact of endogenous amino acid losses; crude protein and amino acids ADCswere adjusted for dietary preprandial losses in seawater. In general, nutrients digestibility was far higher in the wheat products than in the fish meal-based reference diet. Drymatter and crude proteinADCswere not statistically different amongwheat products (from84 to 96% and from88 to 107% respectively). Energy ADCs were significantly higher for clear flours (96% for HCF and MCF) than forwhole grainmeals and S (from83 to 86%). Total amino acids (TAA) and essentialamino acids (EAA) ADCs, once adjusted for preprandial leaching fromthe experimental diets, ranged from81 to 89% and from 58 to 81% respectively, and were statistically comparable among wheat products. Low Thr ADCs appear as a common feature of the amino acids digestibility profiles for whole grain meals, clear flours, or semolina.

Apparent digestibility of dry matter, crude protein, and amino acids of six rendered by-products in juvenile Litopenaeus vannamei

Apparent digestibility coefficients (ADC) of dry matter, crude protein (CP), and amino acids (AA) were evaluated in diets with six rendered by-products used to feed juvenile Pacific white shrimp: two poultry meals (poultry meal 1, 69% CP; poultry meal 2, 72% CP), two feather meals (89% CP), one blood meal (96% CP), and one pork meal (57% CP). Experimental diets were formulated with 30% of the test ingredient and 70% of a commercial diet supplemented with 1% of chromium oxide as inert marker. AA contents in ingredients, diets, leached diets, and feces were determined by high performance liquid chromatography. Preprandial AA losses attributed to leaching were higher in the blood meal diet (15%) and pork meal diet (10%). Poultry meal diets 1 and 2 showed mean AA losses of 3% and 5%, respectively, while the reference diet had a mean AA leaching of 6%. The AA that had the highest leaching rates were lysine (21%), methionine (15%), and histidine (12%). The ADC of dry matter was higher for poultry meals 1 (70%) and 2 (73%), followed by pork meal (69%), feather meals (61%), and blood meal (57%). The digestibility of CP was higher for poultry meals (78–80%), followed by pork meal (76%), and blood meal and feather meals (65–67%). The digestibility of CP in the reference diet (83%) was higher than that observed for all the animal by-product meals except the poultry meals. The ADC of the sum of AA adjusted for nutrient leaching fluctuated from 65% for blood meal to 80% for poultry meals.

Análise do comportamento e crescimento do camarão branco Litopenaeus vannamei (Boone, 1931) em diferentes substratos inconsolidados

The aim of this study was to test the sediment preference of L. vannamei shrimp. It was observed shrimp visit frequency, swimming and burying behaviour at different sediment compositions for 24h. Juvenile (0.93 ± 0.29g) and sub-adult shrimps (10.0 ± 1.18g) were obtained from the aquaculture station at Universidade Federal Rural do Semi-Árido UFERSA, and held in a plastic tank (water volume 500 L) supplied with aerated water and kept at constant temperature, pH, and salinity. Shrimp was fed by commercial shrimp dry food. The experimental substrates were composed by A: medium sand + thick sand + very thick sand + gravel; B: very fine sand + fine sand; and C: silt + clay. Thus, six different substrate combinations were tested: A, B, C, A+B, A+C, B+C. To test preference, it was used a cylindrical tank (40 l) divided into six differently substrate compartments. A single shrimp was introduced each tank and the frequency at which this shrimp visited each compartment was recorded over a 24h study period. It was tested 54 shrimp (18 sub-adult males, 18 subadult females and 18 juveniles). For each trial, sediment and water were changed to avoid pheromones and residues influence. Shrimp were weighted and sub-adults were divided by sex: males present petasma and females present thelycum. Data were collected on the experimental day at 19:30; 20:30; 00:30; 1:30; 05:30; 06:30; 13:30 and 14:30 h. At each time point, shrimp were observed for 20-min periods, in which we noted down which compartment the shrimp was occupying at 2-min intervals. Thus, for each period we had eleven observations (88 observations per day). For observations at night, it was used dim red light that did not affect shrimp behaviour. At each 20-min period, it was observed visit frequency in each substrate, if shrimp was burred or not or if it was swimming. There was not significant difference between light and dark burry activity for females. Swimming activity was significantly higher at night, mainly at 00:30 and 01:30 h. All L. vannamei shrimp showed preference for sediment B. This animal presents cyclic activity, spends the day light period buried and swims at night

Biodiversidade e abundância da ictiofauna associada ao cultivo orgânico de Litopenaeus vannamei (Boone, 1931)

The expansion of shrimp farming has caused a series of environmental impacts, often as a result of lack of planning and adequate management. Organic aquaculture has emerged as an alternative to conventional shrimp farming, and differently, aims at the economical, ecological and farming potential of other organisms, such as fishes, oysters and seaweeds. The present study aimed at evaluating the biological diversity and abundance of the ichthyofauna associated to Litopenaeus vannamei organic culture at PRIMAR farm (Rio Grande do Norte, Brazil). The sampling period consisted of four culture cycles (1, 2, 3, and 4) accomplished in four grow-out ponds (V1, V7, V2, and V4) in 2005. The first two culture cycles were carried out during the rainy season, and the last two cycles, during the dry season. Environmental (temperature, salinity, transparency, and rainfall), biodiversity (diversity and uniformity and abundance indexes) of the ichthyofauna, and shrimp productivity data were collected throughout these four culture cycles. The results obtained for the environmental variables showed that both salinity and transparency oscillated in a significant way (p < 0.001) between culture cycles. In relation to the abundance of the ichthyofauna, fifty four species associated to the organic culture of Litopenaeus vannamei were collected and identified at PRIMAR. For the biodiversity criterion, larger species richness (S = 46) and uniformity (E = 0.59 ± 0.16) were observed during cycles 3 and 4 than in cycles 1 and 2 (S = 41 and E = 0.15 ± 0.12). A positive and significant correlation was obtained for the transparency with diversity and uniformity of fish species between cycles 1 and 2 (wet season) and cycles 3 and 4 (dry season) (p < 0.017 - Shannon Index; p < 0.008 - Pielou Index, respectively). A correlation could not be established between shrimp productivity and fish biomass. However, fish biomass decreased from cycles 1 and 2 to cycles 3 and 4, concomitant with an increase in shrimp productivity, most probably due to a higher (from 58.0% to 71.0%) shrimp survival. Regarding the culture potential of the ichthyofauna, three species (Mugil curema, Mugil liza and Chaetodipterus faber) were identified as potential farming alternatives, either singly or in consortium with Litopenaeus vannamei. A fourth species (Centropomus undecimalis) was indicated as an alternative for single culture or in consortium with other fish species. In conclusion, the large diversity of estuarine fishes associated to Litopenaeus vannamei farming observed at PRIMAR clearly indicated the ecological feasibility for organic aquaculture in northeastern Brazil

Potencial terapêutico de um heparinóide isolado do invertebrado marinho Litopenaeus vannamei

The occurrence of bioactive compounds in marine organisms comes awaking the interest of the pharmaceutical industry. Heparin, a sulfated polysaccharide which presence was already identified in several marine invertebrates, is very attractive due its remarkable functional versatility. Besides to intervene in blood coagulation, this molecule has a great anti-inflammatory potential. However, its strong anticoagulant activity difficult the clinical exploitation of its anti-inflammatory properties. Thus, the aims of this work were to evaluate the effect of a heparin-like compound (heparinoid), isolated from the cephalotorax of the Litopenaeus vannamei shrimp, on the inflammatory response, hemostasia and synthesis of antithrombotic heparan sulfate by endothelial cells, besides studying some aspects concerning its structure. The purified heparinoid was structurally characterized following an analytical boarding, involving electrophoresis and chromatography. The structural analysis have shown that this compound possess a high content of glucuronic acid residues and disulfated disaccharide units. In contrast to mammalian heparin, the heparinoid was incapable to stimulate the synthesis of heparan sulfate by endothelial cells in the tested concentrations, beyond to show reduced anticoagulant activity and hemorrhagic effect. In a model of acute inflammation, the compound isolated from the shrimp reduced more than 50% of the cellular infiltration. Besides reduce the activity of MMP-9 and proMMP-2 of the peritoneal lavage of inflamed animals, the heparinoid also reduced the activity of MMP-9 secreted by activated human leukocytes. These results demonstrate the potential of heparinoid from L. vannamei to intervene in the inflammatory response. For possessing reduced anticoagulant activity and hemorrhagic effect, this compound can serve as a structural model to direct the development of more specific therapeutical agents to the treatment of inflammatory diseases

Utilização de embriões liofilizados e flocos de Artemia na diversificação nutricional de pós-larvas do camarão marinho Litopenaes vannamei (Boone, 1931)

The objective of the current study was to evaluate the zootechnical performance (survival and growth) of Litopenaeus vannamei post-Iarvae fed an artificial shrimp diet supplemented with Artemia flakes or freeze-dried Artemia embryos. For that purpose, 20 culturing units were individually stocked with 50 shrimp post-Iarvae (average dry weight of 0,3 ± 0,03 mg) at a stocking density of 20 post-larvae per liter, and fed the experimental diets to satiation during 20 days. The experimental design consisted of four diets (T1, T2, T3 and T4) with five repetitions each. For treatments T1, T2 and T3, dietary supplements of 5mg of Artemia flakes (T1), freeze-dried Artemia embryos (T2), and of the commercial shrimp diet (T3) were offered 2 hours after the shrimp were initially fed the commercial shrimp diet. For treatment T4 (control), no additive was offered 2 hours after the initial feeding. Shrimp survival, absolut (GPA) and relative increase in weight (GPR), and specific growth rate (TCR) were used as evaluation criteria. After the experimental period, no significant statistical differences (p>0,05) in survival were observed. Regarding growth, the dietary treatment which used freeze-dried Artemia embryos as an additive (T2) presented the best results for GPA (6,7 ± 0,7 mg). There were no statistical differences within treatments T1, T3 and T4 (p>0,05). AIso, post-larvae fed freeze-dried embryos (T2) showed a relative increase in weight (2241,4%) which differed significantly (p<0,05) from T4(1911,7%) but not from T1 (1801,6%) or T3 (1946,7%). In conclusion, the results of the current study indicate that an artificial shrimp diet supplemented with freeze-dried Artemia embryos fulfils the nutritional requirements of post-larvae L. vannamei and promotes a better growth than diets not supplemented with Artemia flakes