The use of chitosan as an antioxidant in the feed of cultivated P. vannamei shrimp against oxidative stress induced by exposure to microplastics.

Microplastics (MP) are omnipresent in aquaculture and can induce several toxic effects, mainly oxidative stress. Therefore, alternatives to minimize these effects are welcome. In this study, chitosan (1 and 3 g/kg) was supplemented through the feed of farmed shrimp P. vannamei for 30 days. After this period, the shrimp were exposed to MP (0.5 mg/L) for 7 days. The results showed the presence of MP in hepatopancreas, gills and muscle. Hepatopancreas morphological alterations, as well as lipid peroxidation, a decrease in GSH level, and an increase in SOD activity indicated an oxidative stress that was reversed by chitosan. The muscle was also affected by MP, showing decreased CAT activity and increased SOD activity, though no lipid peroxidation was observed. In muscle, chitosan reversed the SOD increase to basal activity. The results obtained showed that chitosan was more effective against oxidative stress than in preventing accumulation and histological damage.

A native strain of Bacillus subtilis increases lipid accumulation and modulates expression of genes related to digestion and amino acid metabolism in Litopenaeus vannamei.

In the field of shrimp aquaculture, the utilization of probiotics represents a promising avenue, due to the well-documented benefits conferred by these microorganisms. In the current study, a Bacillus subtilis strain, referred to as strain E, was isolated from the gastrointestinal tract of the shrimp Litopenaeus vannamei and subsequently identified via molecular methods and phylogeny. The probiotic potential of strain E was characterized, and its application as a feed shrimp additive was evaluated in a 45-day experiment. Several parameters were assessed, including zootechnical performance, muscle tissue proximate composition, hepatopancreas lipid concentration, and the expression of genes associated with digestion, amino acid metabolism, and antioxidant defense mechanisms in various shrimp tissues. Although no significant impact on zootechnical performance was observed, supplementation with strain E led to an increase in lipid concentration within both muscle and hepatopancreas tissues. Furthermore, a marked decrease in the expression of genes linked to digestion and amino acid metabolism was noted. These findings suggest that the addition of the B. subtilis strain E to shrimp feed may enhance nutrient absorption and modulate the expression of genes related to digestion and amino acid metabolism.

Effects of including of Japanese Pumpkin Seeds and Pomace in the Diets of Pacific White Shrimp (Penaeus vannamei).

A 60-day feeding trial was conducted to evaluate the effects of including pumpkin seeds and pomace in the diets of Pacific white shrimp Penaeus vannamei, and the effects of these supplements on growth performance, body composition, and total polyphenol, flavonoid and carotenoid contents, as well as on total antioxidant activity, and body color parameters. Five diets were evaluated: pumpkin seeds (PS) at 50 and 100 g·kg-1, pumpkin pomace (PP) at 50 and 100 g·kg-1, and a control treatment. Pacific white shrimp (P. vannamei) juveniles (0.60 ± 0.01 g) were stocked in 15 tanks (310 L), containing 30 shrimps per tank, and the treatments were randomly distributed in triplicate. At the end of the experiment, shrimps were euthanized, weighed, and dissected for further analyses. The inclusion of PS in the diets impaired growth performance, reduced the total flavonoid content and had a pro-oxidative effect on muscle. The inclusion of PP in the diets did not affect growth performance, improved the feed conversion ratio, increased the total flavonoid content in the diets and hepatopancreas, and improved the antioxidant activity of the feeds and shrimp muscle. The total carotenoid content of the feeds increased with the inclusion of PS or PP in the diets; however, the total carotenoid content of shrimp increased only in those fed PP diets. Shrimp fed with PS diets showed a yellowish color and higher saturation when fresh and a reddish color and yellow hue angle after cooking. Shrimp fed PP diets turned reddish and yellowish, both when fresh and after cooking. The inclusion of PS in P. vannamei diets is not recommended; however, PP can be included at 100 g·kg-1 without affecting the growth parameters. Further studies evaluating the inclusion of higher PP levels in shrimp diets are recommended.

Evaluation of Interactions of Added Soybean Peroxidase with Other Nutrients Present in Fish Feeds Using an In Vitro Digestive Simulation.

Peroxidase (PO) has been applied in different areas of industrial biotechnology, including the control of contaminants like aflatoxin B1 in fish feeds. However, its potential negative interactions with the macro and micro components of feeds have not been evaluated. The aim of this study was to evaluate the impact of PO's addition to a feed on compounds like fatty acids and polyphenols using an in vitro simulation of the digestive tract of the tilapia. The influence on fatty acids was determined by changes in the peroxide index, with the feed including PO presenting values four times higher than those of the control feed. On the other hand, the in vitro digestive simulation also evidenced an effect of PO on the bioaccessibility of polyphenols significantly influenced by the total digestion time and temperature. The bioaccessibility of polyphenol ranged from 2.09 to 16.23 µmol of the total Trolox equivalent antioxidant capacity for the combinations evaluated in the study. The greatest bioaccessibility was observed at the central point under the following conditions of digestive hydrolysis: pH of 7, 30 °C, 4.5 h of digestive hydrolysis and an absence of PO.

Mitigation of aflatoxin B1 in fish feed by peroxidase from soybean meal.

Potential of the enzyme peroxidase (PO) from soybean meal to mitigate aflatoxin B1 (AFB1) in fish feed was evaluated. Reaction parameters studied in the wet stage of the feed production process were enzyme activity (0.01-0.1 U/g), temperature (20-36 °C), time (0-8 h) and humidity content (40-70%). Feed was produced in conformity with the National Research Council and spiked with AFB1 at 10 ng/g. Any residual concentration of AFB1 in the diet was extracted by the QuEChERS method and quantified by a liquid chromatograph with a fluorescence detector. AFB1 mitigation of 90% was reached when feed production conditions were 0.035 U/g, 32 °C, 6 h and 70% humidity. Therefore, application of PO to the feed industry may be considered a promising tool for mitigation of AFB1, considering its toxicity and frequent occurrence. In addition, it guarantees safe food for consumers of fish farming products, as AFB1 can bioaccumulate in the food chain. It also provides an alternative use for soybean meal that would previously be discarded.

Shellfish industrial waste reuse.

The global production of aquatic organisms has grown steadily in recent decades. This increase in production results in high volumes of by-products and waste, generally considered to be of low commercial value and part of them are consequently discarded in landfills or in the sea, causing serious environmental problems when not used. Currently, a large part of the reused aquaculture waste is destined for the feed industry. This generally undervalued waste presents an important source of bioactive compounds in its composition, such as: amino acids, carotenoids, chitin and its derivatives, fatty acids and minerals. These compounds are capable of offering numerous benefits due to their bioactive properties. However, the applicability of these compounds may be opportune in several other sectors. This review describes studies that seek to obtain and apply bioactive compounds from different sources of aquaculture waste, thus adding commercial value to these underutilized biomasses.HIGHLIGHTSVolume of aquaculture industrial waste from crustaceans and mollusks.Quantity and quality of bioactive components in aquaculture waste.Applications of recovered proteins, lipids, chitin, carotenoids and minerals.Future prospects for the destination of aquaculture waste.

Occurrence and bioacessibility of mycotoxins in fish feed.

In this study Aflatoxin B1 (AFB1), ochratoxin A (OTA) and zearalenone (ZEN) occurrence in fish feed, regarding its chemical composition, were investigated. Besides, AFB1 bioaccessibility to fish was evaluated by in vitro digestion. Mycotoxins were extracted by QuEChERS and quantified by HPLC-FLD. Results showed that 93.3% of the samples were contaminated at maximum levels of 16.5, 31.6, and 322 µg/kg in the cases of AFB1, OTA, and ZEN, respectively. A positive correlation between OTA, ZEN contamination, and lipid content was observed. Risk estimation of feed consumption by fish at the highest levels of AFB1, OTA, and ZEN shows that the younger the fish, the higher the risk of exposure to mycotoxins. The AFB1 bioaccessibility assay showed that 85% of this mycotoxin may be absorbed by fish. Therefore, establishing maximum levels in the fishing sector is fundamental to contribute to feed quality and nutritional safety of fish species.

Modulation of nodularin toxicity in shrimp Litopenaeus vannamei (BOONE, 1931) fed with dietary açai (Euterpe oleracea) inclusion.

This study evaluated the effect of dietary inclusion of lyophilized açaí Euterpe oleracea (LEO) on redox status of shrimp Litopenaeus vannamei (initial weight 1.5 ± 0.39 g) upon exposure to cyanotoxin nodularin (NOD) in bioflocs system. Three hundred juvenile shrimps were randomly divided into two groups and fed twice a day with two diets: one containing 0.00 (control diet) and the other 10.0% LEO (w/w) for 30-days. After the feeding period, both shrimp groups were submitted to three treatments (14 L; 7 shrimp/tank) with different concentrations of cyanotoxin NOD (0.00; 0.25; and 1.00 µg/L) dissolved in water with 96 h of exposure. Then, the shrimps were sampled (n = 15/treatment) for the determination of reduced glutathione (GSH), the activity of glutathione-S-transferase (GST), sulfhydryl groups associated to proteins (P-SH), and lipid peroxidation (TBARS) in the hepatopancreas, gills and muscle. The NOD accumulation was measured in the muscle. The results revealed that dietary LEO significantly increased GSH levels in the hepatopancreas and gills of the shrimps exposed to NOD. Toxin exposure did not modify GST activity in all organs. Muscle TBARS levels were lower in the shrimp fed with the LEO diet and exposed to NOD. The NOD toxin did not accumulate in the muscle but notably was detected in the control groups fed or not with dietary LEO. Açaí was able to induce the antioxidant system of L. vannamei, as well as lowered the oxidative damage in shrimps exposed to NOD, suggesting its use as a chemoprotectant against cyanotoxins.

Sources, quantification techniques, associated hazards, and control measures of mycotoxin contamination of aquafeed.

With the productive intensification of fish farming, the partial or total replacement of fishmeal by ingredients of plant origin became a reality within the feed industry, with the aim of reducing costs. However, this practice increased the impact of mycotoxin contamination. Studies have shown that mycotoxins can induce various disorders in fish, such as cellular and organic alterations, as well as impair functional and morphological development, and, in more severe cases, mortality. Thus, studies have been conducted to evaluate and develop strategies to prevent the formation of mycotoxins, as well as to induce their elimination, inactivation or reduction of their availability in feed.

Probiotic expressing heterologous phytase improves the immune system and attenuates inflammatory response in zebrafish fed with a diet rich in soybean meal.

Although aquaculture is among the fastest growing food production sectors in the world, one of the bottlenecks for the continuity of its expansion is the dependence of animal protein on commercial feed formulations. Vegetable proteins are an alternative due to the low cost and high availability. However, this protein source is accompanied by a series of antinutritional and pro-inflammatory compounds, including phytate. Phytases can be added in feed for phytate degradation and increase nutrient availability. However, the use of purified phytases significantly increases the production costs. An interesting alternative is to use probiotics genetically modified as bioreactors for phytase production. In the present study, a strain of Bacillus subtilis secreting a fungal phytase was used to evaluate the effect of a feed with high content of soybean meal on zebrafish (Danio rerio). We analysed the condition factor (K) of fish, and the expression of genes related to the immune system, inflammatory response and oxidative. stress. The results obtained demonstrate that the transgenic probiotic was efficient in improving the fish condition factor, stimulating the immune system, reducing the inflammatory response and oxidative stress. Thus, probiotics acting as phytase bioreactors can be considered an interesting tool for the adaptation of commercial species to feed of lower cost.

Fishmeal substitution for Arthrospira platensis in juvenile mullet ( Mugil liza) and its effects on growth and non-specific immune parameters / Substitución de harina de pescado por Arthrospira platensis en juveniles de lisa (Mugil liza) y sus efectos en el crecimiento y parámetros del sistema inmune no-específico / Substituição da farinha de peixe por Arthrospira platensis em juvenis de tainha (Mugil liza) e seus efeitos no crescimento e nos parâmetros do sistema imune não-especifico

Abstract Background: Cyanobacterium Athrospira platensis (Spirulina) is a potential fishmeal (FM) substitute in fish diets because of its high protein content, antioxidant and immunostimulant properties. Objective: To evaluate the effects of total and partial substitution of FM with A. platensis (0, 30, 50, 70 and 100% substitution) in juvenile mullet (Mugil liza). Methods: Juvenile mullets (n=210) were maintained in a recirculation system under optimal water parameters for the species. Mullets were fed five experimental diets for 80 days. Each diet was tested in triplicate tanks. At the end of the experimental period growth parameters were measured and samples of blood, liver and spleen were taken to evaluate the immune system. Results: Full replacement (100%) of FM resulted in growth deficits and low survival. The FM replacement induced changes in the proportion of macrophages and lymphocytes. Up to 50% FM replacement increased the expression of CD3 receptors in spleen T lymphocytes (T-Cells), whereas >50% FM replacement decreased the expression of CD3 receptors. We also found that partial FM substitution diminished the apoptotic process. Conclusions: Up to 50% FM substitution with A. platensis can improve performance of non-specific immune system of mullets.

Resumen Antecedentes: La cianobacteria Arthrospira platensis (Spirulina) puede usarse como substituto potencial de la harina de pescado (HP) por su alto contenido de proteína, sus antioxidantes y sus propiedades inmunoestimulantes. Objetivo: Analizar el efecto de la substitución parcial y total de HP por A. platensis (0, 30, 50, 70 y 100% de substitución) en juveniles de lisa (Mugil liza). Métodos: Juveniles de lisa (n=210) se mantuvieron en un sistema de recirculación con parámetros de calidad de agua en niveles óptimos para la especie. Las lisas se alimentaron con las dietas experimentales durante 80 días. Cada dieta fue evaluada en triplicado. Al final del periodo experimental se midieron los parámetros de crecimiento y se colectaron muestras de sangre, hígado y bazo para evaluación del sistema inmune. Resultados: La substitución total (100%) resultó en deficiente crecimiento y baja sobrevivencia. El remplazo de HP produjo cambios en las proporciones de macrófagos y linfocitos. La substitución de hasta un 50% HP aumentó la expresión de receptores CD3 en linfocitos T del bazo. Por otro lado, la substitución mayor a 50% HP disminuyó la expresión de receptores CD3. La substitución parcial de HP disminuyó el proceso de apoptosis. Conclusiones: Proponemos una substitución de HP del 50% por A. platensis, lo cual mejora el desempeño del sistema inmune no especifico de las lisas.

Resumo Antecedentes: A cianobactéria Athrospira platensis (Spirulina) é um potencial substituto da farinha de peixe (FP) pelo seu alto conteúdo de proteína, antioxidantes e características imune estimulantes. Objetivo: Foram avaliados os efeitos da substituição parcial e total da FP por A. platensis (0, 30, 50, 70 e 100% substituição) em juvenis de tainha (Mugil liza). Métodos: Juvenis de tainha (n=210) foram mantidos em um sistema de recirculação com os parâmetros da água sendo mantidos em níveis ótimos para a espécie. As tainhas foram alimentadas com as dietas experimentais por 80 dias, cada dieta foi testada em triplicata, ao final do período experimental foram avaliados os parâmetros de crescimento e amostras de sangue, fígado e baço foram coletadas para a avaliação do sistema imune. Resultados: A substituição total de FP resultou em redução do crescimento e baixa sobrevivência. A avaliação do sistema imune demostrou que a substituição da FP produz alterações nas proporções de macrófagos e linfócitos. Provou-se que até 50% de substituição da FP incrementa a expressão de receptores CD3. Além disso, a substituição parcial da FP diminui o processo de apoptose. Conclusão: Baseado em nossos descobrimentos, se propõe a substituição de até 50% da FP por A. platensis que melhorará o desempenho do sistema imunológico não específico das tainhas.

Fish oil and meal replacement in mullet (Mugil liza) diet with Spirulina (Arthrospira platensis) and linseed oil.

This research evaluated the effect of the simultaneous substitution of fish meal and fish oil at 0, 30, 50, 70 and 100% (termed SS0, SS30, SS50, SS70 and SS100, respectively) with both Spirulina (Arthrospira platensis) and linseed oil. The effect on growth, colour, fatty acid (FA) profile, and liver and muscle antioxidant response in mullets (Mugil liza) were evaluated. A full substitution was not possible due to a final body weight reduction (p < 0.05). The colorimetric analyses showed an improvement in fillet colouration (p < 0.05). Also, we found a saturation of carotenoids in muscle at 30% substitution. The content of highly unsaturated fatty acids - HUFA (DHA, EPA and ARA) in muscle diminished as the substitution level increased and, at the same time, the level of these FAs in the diet diminished. We found interesting evidence of elongation from 18:3n-3 to 20:3n-3 when substitution with linseed oil levels was above 50% but no desaturation to 20:4 or 20:5n-3 was found, which could insinuate a certain n-3 HUFA requirement under the experimental conditions. Moreover, we noted an increase in the antioxidant capacity up to the treatment SS70; the SS100 treatment showed a diminution in the antioxidant capacity. It is concluded that a partial inclusion of Spirulina and linseed oil (50%) is possible, and that this replacement can increase antioxidant responses, improve its growth performance and can modulate the mullets fillet quality without affecting negatively fillet fatty acid content.

Biochemical responses over time in common carp Cyprinus carpio (Teleostei, Cyprinidae) during fed supplementation with α-lipoic acid.

The current study aimed to evaluate the influence of lipoic acid (LA) supplementation (439.84±6.71 mg LA/kg feed) on antioxidants responses throughout the time in intestine, liver and muscle of juvenile common carp Cyprinus carpio. Two experimental groups were fed during four weeks with a diet with or without LA. Glutathione-S-transferase (GST) activity, glutathione (GSH) content, antioxidant capacity against peroxyl radicals (ACAP) and lipid peroxidation (TBARS) were evaluated in these organs. Also, a technique to measure protein disulfide bonds and sulfhydryl groups was optimized for intestine samples. GST activity was significantly higher (p<0.05) in intestine after two weeks of supplementation. GSH content was also significantly higher (p<0.05) in intestine, liver and muscle of fish fed with LA after two and three weeks, respectively. Total capacity antioxidant against peroxyl radicals was significantly increased (p<0.05) in the muscle of animals fed with LA after the fourth week. Concentration of disulfide bonds was higher in the intestine of fish fed with LA but this group also showed higher concentration of sulfhydryl groups (p<0.05). It is concluded that supplementation with LA is a safe strategy to induce antioxidant responses and improves the antioxidant status in different organs of common carp. Two week of supplementation are required to induce antioxidant responses in intestine and liver and three week for muscle.

Benzocaína e eugenol como anestésicos para juvenis do pampo Trachinotus marginatus / Benzocaine and eugenol as anaesthetics for pompano juvenile Trachinotus marginatus

O objetivo deste trabalho foi avaliar a eficácia da benzocaína e do eugenol como anestésicos para juvenis de Trachinotus marginatus. Foram testadas concentrações de 25, 50, 75, 100 e 150ppm de benzocaína e de 25, 37,5, 50, 62,5 e 75ppm de eugenol. Os testes foram realizados em aquários de vidro contendo 30l de água (salinidade 30‰ e temperatura 19°C). Os peixes (51,4±13,5g) foram expostos individualmente às diferentes concentrações de cada anestésico (n=5 para cada concentração). Enquanto ainda anestesiados, eles foram medidos e pesados para posteriormente serem colocados em um aquário sem anestésico para recuperação. Posteriormente, os peixes foram colocados em um aquário sem anestésico para observação da recuperação. O tratamento estatístico dos resultados foi feito pela Análise de Variância (One-Way - ANOVA) com 95 por cento de significância e, quando encontradas diferenças significativas, o teste de Duncan foi aplicado. Os períodos para atingir a latência para os juvenis de pampos variaram entre 11min para a concentração de 25ppm e 2min na concentração de 150ppm para a benzocaína e de 10min (25ppm) a 2min (75ppm) para o eugenol. Os tempos de recuperação foram entre 3min para 25ppm e 14min para a concentração de 150ppm para a benzocaína e de 2min (25ppm) a 8min (75ppm) para o eugenol. A dose de 50ppm de benzocaína e eugenol proporcionou um período de latência e recuperação dentro da faixa considerada adequada (3mim para a latência e 5mim para a recuperação). Entretanto, é importante salientar que o custo da benzocaína para anestesiar juvenis de pampo é 1/3 inferior ao custo do eugenol.

The aim of this experiment was to evaluate the efficacy of benzocaine and eugenol as anesthetics for Trachinotus marginatus juveniles. Five concentrations of benzocaine (25, 50, 75, 100, and 150ppm) and eugenol (25, 37.5, 50, 62.5, and 75ppm) were tested. The trials were conducted in glass aquariums filled with 30l seawater (salinity 30‰ and temperature 19°C). The fish (51.4±13.5g) were individually exposed to different concentrations of each anesthetic (n=5 for each concentration). They were measured and weighed under anesthesia, and then returned to another aquarium without anesthetic for recovery. The results were evaluated using Analyses of Variance (One-Way - ANOVA) followed by the test of Duncan (95 percent confidence level) when necessary. Anesthesia time varied from approximately 11min (25ppm) to 2min (150ppm) for benzocaine, and from 10min (25ppm) to 2min (75ppm) for eugenol. Recovery time ranged from 3min (25ppm) to 14min (150ppm) for benzocaine and from 2min (25ppm) to 8min (75ppm) for eugenol. Benzocaine and eugenol, both at 50ppm, induce quick anesthesia and recovery (3mim for anesthesia and 5mim for recovery) for juvenile Trachinotus marginatus, and therefore are considered efficient anesthetics for this species. However, it is important to emphasize that the cost of benzocaine is 1/3 lower than eugenol to anesthetize juvenile pompano.

Criação de juvenis de peixe-rei (Odontesthes argentinensis) em diferentes taxas de arraçoamento / Rearing of pejerrey (Odontesthes argentinensis) juveniles at different feeding rates

Este estudo teve como objetivos avaliar os efeitos de diferentes taxas de arraçoamento sobre a sobrevivência e o crescimento de juvenis do peixe-rei (Odontesthes argentinensis). Juvenis de peixe-rei (100mg) foram estocados na densidade 10 peixes L-1, em tanques circulares de 50L. Foram testadas cinco taxas de alimentação (4, 8 12, 16 e 20 por cento do peso vivo diário) em duplicata durante 30 dias, mais um controle negativo no qual os peixes não foram alimentados. Durante o experimento, a temperatura foi de 23°C, a salinidade 26ë e a água foi constantemente aerada. A mortalidade total dos peixes mantidos em jejum foi observada após 20 dias de criação; entretanto, não foi verificada diferença significativa de sobrevivência entre os peixes submetidos aos demais tratamentos até o final do experimento, pois a mortalidade foi inferior a 10 por cento, independentemente da taxa de arraçoamento testada. Pelos resultados da regressão polinomial quadrática dos dados de ganho em peso e eficiência alimentar, sugere-se que a taxa de arraçoamento ótima para juvenis de peixe-rei entre 100 e 500mg é igual a 9,0 por cento do peso vivo ao dia.

Tolerância de juvenis do pampo Trachinotus marginatus (Teleostei, Carangidae) ao choque agudo de salinidade em laboratório / Acute salinity tolerance of juvenile pompano Trachinotus marginatus (Teleostei, Carangidae) under laboratory conditions

O pampo Trachinotus marginatus é uma espécie com potencial para piscicultura, mas a sua utilizaçäo pode ser limitada pelas variaçöes de salinidade comuns em estuários ou em ambientes super-salinos. Este trabalho foi realizado com o objetivo de determinar a tolerância à salinidade de juvenis de T. marginatus em laboratório. Foram testadas 12 salinidades em duplicata (0, 6, 8, 10, 15, 25, 45, 55, 58, 61, 65 e 75ë) e um tratamento controle (35ë). Dez peixes (comprimento total: 20,7±2,3mm e peso úmido: 427±113mg) foram colocados em tanques plásticos de 3L. A temperatura da água foi mantida em 24ºC com um banho termostatisado, aeraçäo constante e, a cada 24 horas, os meios experimentais foram completamente renovados. Após 96 horas de exposiçäo, as salinidades letais médias inferior e superior, com os respectivos intervalos de confiança (IC 95 por cento), foram estimadas em 6,99ë (IC 95 por cento = 6,86-7,13ë) e 58,50ë (IC 95 por cento = 56,81-60,24ë), respectivamente. Estes resultados permitem caracterizar esta espécie como eurialina. O estudo da influência da salinidade sobre T. marginatus deve ser aprofundado, buscando avaliar principalmente os efeitos sobre o crescimento, de modo que seja possível determinar o potencial do seu cultivo em ambientes com diferentes salinidades