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Sulfated polysaccharides (SPs), especially those with immunostimulant effects on shrimp and fish, are bioactive compounds with potential use in aquaculture. In this study, Pacific white shrimp (Penaeus vannamei) were fed diets containing 0%, 0.5%, 1.0%, 1.5%, or 2.0% crude extracts from the microalga Porphyridium cruentum for 30 days. After dietary supplementation, the shrimp were exposed to Vibrio alginolyticus, allowing the assessment of phenoloxidase activity (PO) and total hemocyte count (THC) pre- and post-challenge, as well as shrimp survival rates. Shrimp that received the 1.0% treatment had greater survival (90%) at 48 h post-challenge, while the 0% and 2.0% treatments had survival rates close to 63% in the same period. Furthermore, at 48 h post-challenge, THC was significantly greater (p < 0.05) in the shrimp that received crude extract supplementation compared to that in shrimp of the control group (0%). PO showed a proportional increase according to crude extract inclusion levels. Such dose-dependency in response to PO increases became significantly greater (p < 0.05) in the 2.0% treatment group compared to that in the 0% treatment group in the pre-challenge period and that in the 0% and 0.5% treatment groups in the post-challenge period. These results showed that crude extracts from P. cruentum could be considered immunostimulants by increasing the resistance of P. vannamei exposed to V. alginolyticus, potentially representing a major breakthrough for marine shrimp farming.

An experiment was conducted for eight weeks, in a BFT system, to test different strategies for using effective microorganisms (AEM) in shrimp rearing. For that, five different treatments were tested. The treatments tested were: Control (commercial feed only), AEM in the feed (addition of AEM in the commercial feed), AEM in the water (addition of AEM directly in the rearing water), AEM in the feed and in the water (addition of AEM in the feed simultaneously with the addition of AEM in the water) and MA in the feed (addition of the microbiological activator-MA in the commercial feed, without adding microorganisms to the feed and water). The use of AEM or MA, although it did not affect the growth of the animals, stimulated the digestive process of the animals, allowing the required absorption of nutrients with a smaller intestinal area. Associated with this, the presence of AME or MA stimulated the immunological parameters of the animals and reduced the presence of Vibrio in the gut of the shrimp. The association of these factors meant that the survival of the animals that received the additives in the diet was substantially higher than the animals that did not receive the additives. The AEM supplementation strategies did not differ among themselves, and the producer could opt for the strategy of lower cost and/or less complexity of use, according to each specific case.

A study was conducted to test the inclusion of Aurantiochytrium sp. meal in the rearing of Penaeus vannamei grown in a clear water system and at a suboptimal temperature of 22 °C. The doses tested were 0 (control), 1, 2, 3, and 4% at a stocking density of 100 shrimp/m3. Rearing was carried out with aeration and individual heaters, and seawater temperature was controlled with a chiller. After nine weeks, shrimp were weighed and hemolymph was collected for hemato-immunological tests, and growth performance were calculated. Shrimp raised at 22 °C and fed a 1% of dietary supplementation of Aurantiochytrium sp. meal were superior in immunological parameters. After the dietary assay, a total of 42 shrimps (10.9 ± 0.06 g) per treatment were infected with White Spot Syndrome Virus (WSSV). They were orally infected with 2.6 ± 106 virus particles per g of the animal, maintained at a suboptimal temperature of 22 °C for 108 h, and acclimated to an optimal temperature of 28 °C for an additional 48 h. At 7 days post infection, surviving shrimp were collected for hemato-immunological analysis. Cumulative mortality results showed that shrimp fed diets containing 3% and 4% Aurantiochytrium sp. meal had higher survival than other treatments when challenged with WSSV.

ABSTRACT Nile tilapia (Oreochromis niloticus) has an opportunistic omnivorous feeding behavior and studies have demonstrated the consumption of microorganisms by the species from bioflocs, especially in the early phases of life. One may thus assume that when reared in biofloc systems, there may be a reduction in the feeding rates in the Nile tilapia nursery system when compared to the ponds. This study evaluated the effects of different feeding management options in Nile tilapia nurseries in a biofloc system. A total of 700 fingerlings (2.37 ± 0.55 g) were stocked in 20 experimental units (100 L) and inoculated with mature biofloc for the evaluation of five different feeding management options. After 49 days, it was observed that the higher feed rates caused an increase in nitrogen compounds and solids in the water, worsening the feed conversion rate. Furthermore, the excess feed supplied to the tilapia in the biofloc caused a reduction in the utilization of protein nitrogen, an increase in the liver and visceral fat, in addition to an increase in the production of solids and consumption of alkalinizer by fish biomass. On the other hand, low feeding rates cause a decrease in the growth and uniformity of the tilapia juveniles. Taking the results into consideration, it is recommended to use the feed management option proposed in treatment TC (26% below the highest feed rate) in the tilapia nursery in the biofloc system.

The objective of this work was to evaluate the use of biofloc technology to reared Amazon River prawn (Macrobrachium amazonicum). One hundred Amazon River prawn juveniles were divided into two experimental units (250 L), 50 animals per each. Prawns were fed two times per day, with 3% of the prawn biomass. Dissolved oxygen and temperature were performed twice a day. Total suspended solids, pH, alkalinity, ammonia nitrogen, nitrite and nitrate were monitored twice a week. Prawns showed 77.67%, survival, 2.98 feed conversion, weekly weight gain of 0.29 g day-1, and yield of 822.0 g m-3 after six weeks of rearing. Dissolved oxygen, temperature and pH were suitable for the species, however, for some weeks, ammonia nitrogen and nitrite were higher than the limits recommended for prawn. In conclusion it is possible to use BFT to maintain and rearing Amazon River Prawn (M. amazonicum), however to improve the performance it is necessary to define nutritional requirements, as well improve the management techniques for this species in BFT.

Abstract The objective of this work was to evaluate the effect of Lactobacillus plantarum on the preservation of fresh mussels and its effect on the attractiveness, consumption, and midgut bacterial microbiota of Pacific white shrimp broodstock. The experiment evaluated mussels stored with L. plantarum at 4ºC. The controls were: mussels stored at -18ºC without L. plantarum; mussels stored at -18ºC with L. plantarum; and mussels stored at 4ºC without L. plantarum. Microbiological analyses on mussels were performed on days 1, 7, 15, 30, 45, and 60 after processing. Additionally, mussels preserved with L. plantarum and stored at 4ºC, and mussels stored at -18ºC without L. plantarum were evaluated after 15 days for attractiveness, consumption, and midgut bacterial microbiota of shrimps. Mussels preserved with L. plantarum showed higher lactic acid bacteria counts and lower counts of Vibrio spp., as well as of total heterotrophic bacteria, after 60 days of storage. No differences were observed for attractiveness or consumption between treatments. The bacterial microbiota of midgut in shrimp fed mussels preserved with L. plantarum showed higher lactic acid bacteria count and lower Vibrio spp. The use of L. plantarum inhibits Vibrio spp. and preserves feed without changing attractiveness or consumption for shrimp.

ABSTRACT The objective of this study was to evaluate the use of a diet supplemented with Lactobacillus plantarum on performance, immunological parameters, and bacterial microbiota of the digestive tract of white leg shrimp (Litopenaeus vannamei). We used six earthen ponds (1.2 ha) stocked with 14 shrimp m−2 (3 g mean weight). Three ponds received a diet supplemented with L. plantarum and three were maintained as control. The survival and feed efficiency of shrimp fed the supplemented diet were greater than that observed in shrimp fed the control diet, with survival values of 83.02±6.12% and 74.65±9.07% and feed efficiencies of 117.97±4.45% and 104.46±7.30%, respectively. However, we observed no differences in weight gain. The intestines of shrimp fed probiotics had lower counts of Vibrio spp. and higher counts of lactic acid bacteria, compared with those of control shrimp. Diets supplemented with L. plantarum alter the intestinal bacterial microbiota of shrimp, resulting in increased survival and feed efficiency.

This study investigated bacterial strains with probiotic potential isolated from the middle portion of healthy hybrid surubim catfish foregut (Pseudoplatystoma reticulatum female × P. corruscans male). Twenty surubims weighing 1.5 ± 0.3 kg were used for bacterial isolation. In total, 41 strains of bacteria were selected in vitro. Ten strains had inhibition zones >10 mm against Aeromonas hydrophila. Five of those strains presented inhibition zones > 9 mm against other pathogenic bacteria and reached concentrations greater than 105 CFU mL−1 in tubes containing de Man, Rogosa and Sharpe (MRS) medium. In particular, Weissella cibaria (P36) reached 106 CFU mL−1 in MRS and was able to reduce the pH of the medium to 3.85. In the in vivo intestinal colonization studies, 72 healthy hybrid surubims were fed with a commercial diet supplemented with probiotic W. cibaria for 15 days. Changes in gut community composition were then analyzed, and probiotic profile of W. cibaria was determined molecularly by amplification of rRNA 16S gene was performed using PCR. Compared to control fish, W. cibaria-supplemented fish showed an increase in RBC. These results show the efficacy of our haematological approach to probiotic screening in hybrid sorubim. Keywords: Fish nutrition, Sorubim, Lactic acid bacteria, Weissela cibaria

ABSTRACT: A supplementary diet with the probiotic bacteria Weissella cibaria on the efficacy of surubim hybrid immunization against a specific hemorrhagic septicemia caused by Aeromonas hydrophila was evaluated on the following treatments: fish fed a supplemented probiotic diet, vaccinated fish and vaccinated fish fed a supplemented probiotic diet, and untreated fish (control). Fish from the probiotic treatments were fed a diet containing W. cibaria for 41 days. On the 15th day of the experiment, fish from vaccine treatments were intraperitoneally vaccinated, with posterior oral booster for four days. One week after the oral booster, three fish from each experimental unit were sampled. The probiotic supplementation increased the number of thrombocytes and lysozyme concentration compared with surubim that did not receive W. cibaria in the diet. On the other hand, the vaccination increased agglutination titer, lysozyme concentration, and antimicrobial activity compared with surubim that were not vaccinated. However, there was no interaction between diet with probiotics and vaccination in the surubim hybrid in the analysed parameters.

This work aimed to evaluate Nannochloropsis spp. as feed additive in the diet of Pacific white shrimp for their effect on midgut microbiology, thermal shock resistance and immunological parameters. Initially, the digestibility of the microalgae meal was assessed, and the apparent digestibility coefficient (ADC) was determined. The ADC was, in general, high in lipids (78.88%) and eicosapentaenoic fatty acid (73.86%). Then, Nannochloropsis spp. were included in diets at four levels (0, 0.5, 1 and 2% inclusion). The shrimp were reared in 500 L clear water tanks containing 20 shrimp per tank with an initial weight of 6.05 ± 0.06 g and fed four times a day. Shrimp fed with supplemented diets containing Nannochloropsis spp. (0.5 and 2%) presented higher resistance to thermal shock when compared to the non-supplemented group (control). Shrimp fed with 1 and 2% of algae inclusion had a higher production of reactive oxygen species (ROS) when compared to other treatments. No statistical difference was observed in the immunological parameters and microbiology of the intestinal tract. Thus, the inclusion of Nannochloropsis spp. in shrimp diets at 0.5 and 2% levels increases resistance to thermal shock and ROS production in shrimp.

ABSTRACT This study aimed to evaluate 1) the in vitro effect of organic salts on the growth of the probiotic Lactobacillus plantarum and then 2) the combined use of a probiotic with organic salts on the in vitro inhibition of V. alginolyticus, A. hydrophila, E. coli, P. aeruginosa, and S. agalactiae. In vitro tests were performed with eight different organic salts, including butyrate, propionate, succinate, citrate, formate, fumarate, glutamate, and acetate, at two pH values (6.2 and 7.1) to determine their effect on the growth kinetics of L. plantarum. In addition, each organic salt was tested alone and in combination with L. plantarum to evaluate the inhibitory effect against the pathogenic bacteria noted above in either condition. Sodium citrate and formate inhibited the growth of L. plantarum, but sodium glutamate, succinate and fumarate stimulated it. Sodium propionate, butyrate, and acetate did not affect probiotic growth at all. Inhibition against all pathogens was significantly higher in the presence of the probiotic and lower pH. Comparing all organic salts at the two pH values, butyrate, acetate, and propionate exhibited more inhibition against V. alginolyticus than the others, while propionate had higher inhibition against A. hydrophila, E. coli, P. aeruginosa, and fumarate successfully inhibited S. agalactiae. Based on these results, it can be concluded that organic salts showed better in vitro inhibition against the aquaculture pathogenic bacteria tested when combined with the probiotic L. plantarum.

ABSTRACT The objective of this study was to evaluate the effect of different stocking densities and the presence/absence of two artificial substrates on water quality and production rates of marine shrimp in a biofloc shrimp nursery culture. Two experiments were performed: different stocking densities with mosquito netting substrate and the presence/absence of polyester-type substrate. The first experiment lasted 38 days, and shrimp at an initial weight of 0.013±0.010 g were stocked in 24 tanks. The second experiment lasted 35 days, and shrimp at an initial weight of 0.037±0.002 g were stocked in six tanks. Weekly biometric measurements were performed to adjust the amount of feed. Suspended solids were higher at a density of 6000 PL m−3 and mosquito netting substrate. Final weight and specific growth rate were higher in treatments with mosquito netting substrate. However, survival was significantly lower with this substrate. Yield was significantly higher at a density of 6000 PL m−3. Polyester-type substrate had no significant effect on production rates or variables of water quality. However, this substrate could reduce the production of sludge. The results indicate that it is possible to culture shrimp in nursery stage up to 6000 PL m−3 in a biofloc system.

Litopenaeus vannamei shrimp were reared in a bioflocs system and fed different levels of soybean protein concentrate as a replacement for fishmeal, and both immunological parameters of this marine shrimp and its resistance to Vibrio sp. infection (CPQBA 378-12 DRM01) were evaluated. Four different diets were formulated with 0, 33, 66 and 100 % of soybean protein concentrate as a substitute for fishmeal. Shrimp were reared in a biofloc system in twelve 800 L tanks (250 shrimp m-3) maintained at constant aeration and temperature. After 42 days, 36 animals (14.21 ± 0.89 g) per treatment were challenged with Vibrio sp. (1 x 105 CFU mL-1 – LD10), and hemolymph was collected before and after challenge to perform immunological assays (agglutination titer, concentration of protein and phenoloxidase activity). Shrimp fed with the experimental diets showed no difference in their resistance to infection and haemato-immunological parameters. Thus, rearing L. vannamei in a biofloc system on diets containing either partial or total replacement of fishmeal for soybean protein concentrate did not affect either immunocompetence or susceptibility to infection.

ABSTRACT A life-cycle assessment was conducted during semi-intensive and super-intensive commercial cultivation of marine shrimp from December 2011 to June 2012, considering all phases from the preparation of the nursery to harvesting of the shrimp, to determine the carbon footprints of each process. Inputs and outputs associated to the production of 1 kg of shrimp were evaluated using the CML-1A baseline method, V3.01 EU25, with Software SimaPro® 8.0.2, to identify the factors of impact that are most relevant to the category global warming, measured in kg of CO2 eq. The grow-out phase contributed the most to the final results in super-intensive culture, which had a higher carbon footprint, 47.9967 kg of CO2 eq., which was 1.0042 kg of CO2 eq. in the semi-intensive culture. The most important impacting factor is the use of electrical energy, which is required to maintain dissolved oxygen and the biofloc particles in suspension in the super-intensive culture and for movement of large volumes of water in the semi-intensive system.

ABSTRACTThe objective of this work was to evaluate the performance of Litopenaeus vannameifed different levels (0, 25, 50, 75, and 100%) of soybean protein concentrate (63.07% crude protein, CP) to replace fish meal-by product (61.24% CP). The study was conducted in clear water in fifteen 800 L tanks equipped with aeration systems, constant heating (29 ºC), and daily water exchange (30%). Each tank was stocked with 37.5 shrimp/m3 (3.03±0.14 g). Feed was supplied four times a day, at 6% of the initial biomass, adjusted daily. After 42 days, the weight gain of shrimp fed diets with 0 and 25% protein replacement was higher than that observed in shrimp fed 100% replacement, and there were no differences among those fed the other diets. Feed efficiency and survival did not differ among shrimp fed different protein replacements. There was a negative linear trend for growth parameters and feed intake as protein replacement with soybean protein concentrate increased. Fish meal by-product can be replaced by up to 75% of soybean protein concentrate, with no harm to the growth of Pacific white shrimp.

Aiming at assessing the cryopreservation potential of Litopenaeus vannamei sperm cells, 74 spermatophores were manually extracted from the sexually mature individuals. After the toxicity test to define the cryoprotectant concentration, suspensions of spermatic cells were cryopreserved in the groups in freezing solutions comprising different cryoprotectants such as dimethyl sulfoxide (DMSO) and ethylene glycol (EG) at 10% concentration. Each treatment was divided in subgroups for storage in liquid nitrogen during 0, 30, 60 and 90 days, in triplicate. After thawing at 25ºC for 40 seconds, cell viability in the suspensions was analyzed under the microscope in eosin-nigrosin stain and flow cytometry. There were no significant differences between the cryoprotectants used. For all the treatments, lower and higher mortalities occurred in the 0 and 90 days, respectively. By applying the eosin-nigrosin technique, lower and higher mortalities were 23.17 and 82.11% for DMSO and 29.94 and 83.72% for EG, while the flow cytometry registered mortalities of 2.42 and 55.13% for DMSO and 0.90 and 55.56% for EG. The Spearman correlation coefficient indicated a positive correlation (R=0.91) between the two techniques used. It was concluded that there was a decrease in cell viability within a longer cryopreservation time.

The objective of this work was to isolate strains of lactic acid bacteria with probiotic potential from the digestive tract of marine shrimp (Litopenaeus vannamei), and to carry out in vitro selection based on multiple characters. The ideotype (ideal proposed strain) was defined by the highest averages for the traits maximum growth velocity, final count of viable cells, and inhibition halo against nine freshwater and marine pathogens, and by the lowest averages for the traits duplication time and resistance of strains to NaCl (1.5 and 3%), pH (6, 8, and 9), and biliary salts (5%). Mahalanobis distance (D²) was estimated among the evaluated strains, and the best ones were those with the shortest distances to the ideotype. Ten bacterial strains were isolated and biochemically identified as Lactobacillus plantarum (3), L. brevis (3), Weissella confusa (2), Lactococcus lactis (1), and L. delbrueckii (1). Lactobacillus plantarum strains showed a wide spectrum of action and the largest inhibition halos against pathogens, both Gram-positive and negative, high growth rate, and tolerance to all evaluated parameters. In relation to ideotype, L. plantarum showed the lowest Mahalanobis (D²) distance, followed by the strains of W. confusa, L. brevis, L. lactis, and L. delbrueckii. Among the analyzed bacterial strains, those of Lactobacillus plantarum have the greatest potential for use as a probiotic for marine shrimp.

O objetivo deste trabalho foi avaliar o uso de suplemento prebiótico, probiótico e simbiótico, na dieta de camarões marinhos (Litopenaeus vannamei) e seus efeitos sobre o crescimento, a microbiota intestinal, a resposta imune e a resistência ao desafio experimental com Vibrio alginolyticus. Foram utilizados quatro tratamentos: prebiótico inulina; probiótico Lactobacillus plantarum; simbiótico Lactobacillus plantarum + inulina; e controle. Os camarões foram distribuídos em 16 tanques de dez mil litros de água, povoados com 200 camarões cada, cultivados por seis semanas. Avaliaram-se a microbiologia do trato intestinal dos camarões e a reposta imune, antes e após o desafio com V. alginolyticus. A concentração de Vibrio spp. no trato digestório foi menor em camarões alimentados com dieta suplementada com prebiótico, probiótico e simbiótico, enquanto a concentração de bactérias acidoláticas foi superior somente nos camarões alimentados com probiótico e simbiótico. O título aglutinante do soro contra V. alginolyticus aumentou no grupo probiótico e simbiótico, antes da infecção, e foi maior em todos os tratamentos após infecção com V. alginolyticus, em comparação ao controle. Não foi observada diferença entre os tratamentos quanto aos demais parâmetros avaliados. As dietas probióticas, prebióticas e simbióticas alteram a microbiota intestinal e aumentam o título aglutinante do soro contra V. alginolyticus; contudo, não alteram a resistência ao desafio nem o crescimento dos camarões.

O objetivo deste trabalho foi determinar o método de coleta de fezes mais adequado para camarões marinhos (Litopenaeus vannamei) na determinação do coeficiente de digestibilidade aparente da proteína bruta e da matéria seca de dietas comerciais, e avaliar o efeito da suplementação de Lactobacillus plantarum na dieta sobre o coeficiente de digestibilidade aparente da proteína, da energia e da matéria seca. Foram testados três sistemas de coletas de fezes: decantação, sifonamento e dissecação dos animais. A coleta por decantação foi o sistema mais adequado para a determinação in vivo dos coeficientes de digestibilidade aparente da proteína bruta e da matéria seca das dietas, por apresentar baixo grau de lixiviação das fezes e por ser de fácil execução. Nos camarões alimentados com a dieta suplementada com L. plantarum, foi observado maior coeficiente de digestibilidade aparente da proteína (84,18±0,35%) e da energia (81,28±0,16%), quando comparado ao grupo controle (82,73±1,01 e 78,63±0,93%, respectivamente). O uso de L. plantarum como aditivo probiótico na ração pode trazer benefícios nutricionais a L. vannamei, com melhor aproveitamento dos nutrientes.

The aim of this study was to evaluate the immune response of hybrid surubim vaccinated by intraperitoneal injection and by immersion against Aeromonas hydrophila. Vaccine was prepared with formalin-inactivated A. hydrophila. Forty eight fishes (47±9g) were distributed in 12 tanks (4 fish / tank) in the following treatments (4 tanks / treatment): unvaccinated fish, vaccinated intraperitoneally and vaccinated by immersion. After 21 days, intraperitoneally vaccinated fishes showed the highest values of total protein and immunoglobulins, agglutination titer and serum antimicrobial activity. Lysozyme concentration in the serum was higher in the fishes vaccinated by immersion than unvaccinated fishes. Intraperitoneal vaccination induced the highest immune response and could be used to improve the fish resistance against motile Aeromonas.

The aim of this study was to isolate, identify and determine the ability of potential pathogenic bacteria hemolytic mite Disco (Symphysodon discus) and Neon Rainbow (Melanotaenia praecox), and define which of the antibiotic Trimethoprim, Florfenicol, Chloramphenicol, Norfloxacin, Erythromycin, Bacitracin, Enrofloxacin, Tetracycline, and Furazolidone Clindamycin has better inhibitory effect in vitro, and determine the minimum inhibitory concentration (MIC) for Chloramphenicol, Norfloxacin, Erythromycin and Enrofloxacin. In an outbreak of ornamental fish deaths were isolated nine strains of Gram negative, hemolytic where 3 were identified, two as Vibrio cholerae and one as Citrobacter braakii. With these strains were performed antibiogram and was determined the minimum inhibitory concentration at two different temperatures (22 and 30 ° C). Antibiotics Enrofloxacin, Norfloxacin, florfenicol, chloramphenicol and trimethoprim showed the best results in vitro inhibition against the Vibrio cholerae, and Citrobacter braakii. The MIC increased in 30 ° C for erythromycin and enrofloxacin against pathogens, while for Florfenicol and norfloxacin the MIC was not influenced by temperature.O objetivo deste estudo foi isolar, identificar e determinar a capacidade hemolítica de possíveis bactérias patogênicas do Acará Disco (Symphysodon discus) e Neon Arco-íris (Melanotaenia praecox), e definir qual o antibiótico entre Trimetoprim, Florfenicol, Cloranfenicol, Norfloxacina, Eritromicina, Bacitracina, Enrofloxacina, Tetraciclina, Furazolidona e Clindamicina possui melhor efeito inibitório in vitro, e determinar a concentração inibitória mínima (MIC) para Cloranfenicol, Norfloxacina, Eritromicina e Enrofloxacina. Em um surto de mortalidade de peixes ornamentais foram isoladas nove cepas de bactérias Gram negativas, onde três hemolíticas foram identificadas, duas como Vibrio cholerae e uma como Citrobacter braakii. Com estas foram realizados antibiogramas e determinada a concentração inibitória mínima em duas diferentes temperaturas (22 e 30°C). Os antibióticos Enrofloxacina, Norfloxacina, Florfenicol, Cloranfenicol e Trimetoprim apresentaram os melhores resultados de inibição in vitro frente ao Vibrio cholerae e Citrobacter braakii. O MIC da Eritromicina e Enrofloxacina frente aos patógenos aumentou a 30°C, enquanto que para o Florfenicol e a Norfloxacina o MIC não foi influenciado pela temperatura.

The objective of this study was to investigate the growth of juvenile fat snook (Centropomus parallelus) in laboratory conditions when fed a diet supplemented with the probiotic Lactobacillus plantarum. Changes in the intestinal flora, haematological parameters and growth performance were assessed using 180 fishes (54.2 ± 13.4 g each). The fishes were subjected to two treatments divided into six cages: 1) feed supplemented with probiotic, and 2) feed without probiotic (control). The temperature, dissolved oxygen and salinity were maintained at 25 ± 1 ° C, 4.0 mg.L-1 and 33 ‰, respectively. After 10 weeks of culture, the fishes fed probiotic had reduced viable culturable heterotrophic bacteria and Vibrio spp. and increased lactic acid bacteria in the intestinal tract, as well as a higher number of thrombocytes, leukocytes and lymphocytes in the blood. No significant difference was observed in the growth, survival or body composition, but the hepatosomatic index was significantly higher in the fishes fed with probiotic and control.

O objetivo deste trabalho foi avaliar a substituição de dois níveis de inclusão de farinha de peixe por outros ingredientes, nas dietas de Litopenaeus vannamei cultivados em sistema bioflocos. Foram avaliadas dietas sem uso de farinha de peixe (dieta A, 100% de substituição), com inclusão de 12,5% de farinha de peixe (dieta B, 40% de substituição) e direta controle com inclusão de 21% (dieta C, 0% de substituição). Nas dietas A e B, o farelo de soja e as farinhas de carne e vísceras foram os principais substitutos proteicos. Foram analisados os índices de desempenho dos camarões e os parâmetros físicos e químicos da água de cultivo. Os camarões alimentados com a dieta B apresentaram maior peso final (11,63±1,38 g), em comparação aos camarões alimentados com a dieta A (peso final, 9,39±0,31 g) e com a dieta C (peso final, 10,20±1,10 g). Os demais parâmetros de desempenho como produtividade, conversão alimentar e sobrevivência não apresentaram diferenças entre os tratamentos. A redução de até 40,0% da farinha de peixe pode ser feita em cultivos superintensivos de L. vannamei com bioflocos, sem interferir em seu desempenho zootécnico e na qualidade de água do cultivo.

This study evaluated the pathogenic characteristics of bacteria isolated from Litopenaeus vannamei during an outbreak at the Laboratory of Marine Shrimp, UFSC, Santa Catarina State, Brazil. Their virulence potential in larvae and juvenile shrimp and theeffects on the total haemocyte count, phenoloxidase activity and serum agglutinate titre were examined after experimental infection. Bacterial strains were isolated from larvae and adult shrimps, identified by the AP120E biochemical system as: two strains of Vibrioalginolyticus, three of Aeromonas salmonicida and one of Pasteurella multocida sp. and Pasteurella sp. All the bacterial strains isolated in this study caused mortality in shrimp. One strain of V. alginolyticus was responsible for 97.3 and 88.7% mortality in larvae and juvenil shrimps, respectively. The shrimp immunological system was influenced by experimental infection with V. alginolyticus. Decrease in the total haemocyte count and increase in the phenoloxidase activity and the serum agglutinate titre (p < 0.05) were observed. The results showed the high pathogenicity of V. alginolyticus isolated from larvae and juvenile reared marine shrimp.Este estudo avaliou as características patogênicas de cepas de bactérias isoladas de Litopenaeus vannamei durante surto de mortalidade no Laboratório de Camarões Marinhos, UFSC, Estado de Santa Catarina, Brasil. Seu potencial de virulência em larvas e juvenis de camarão marinho e os efeitos sobre a contagem total de hemócito, atividade de fenoloxidase e título aglutinante do soro foramavaliados após infecção experimental. As cepas bacterianas foram isoladas de larvas e de camarões adultos e identificadas bioquimicamente pelo sistema API20E como: duas cepas de Vibrio alginolyticus, três de Aeromonas salmonicida e uma de Pasteurella sp. e P. multocida. Todas as cepas isoladas provocaram mortalidade em L. vannamei, e uma de V. alginolyticus resultou em mortalidade de 97,3 e 88,7% para larvas e juvenis de camarões, respectivamente. O sistema imunológico dos camarões juvenis sofreu influência da infecção experimental com V. alginolyticus, representado por redução significativa no número total de hemócitos circulantes e aumento na atividade da fenoloxidase e do título aglutinante do soro (p < 0,05). Os resultados deste ensaio demonstraram que a cepa de V. alginolyticus isolada apresentou alto poder de virulência para larvas e juvenis de camarão marinho cultivado.

O objetivo deste trabalho foi isolar bactérias ácido-lácticas do intestino de tilápias-do-nilo, e avaliar seu potencial probiótico. Foram isoladas cepas de bactérias ácido-lácticas, e foi avaliada a inibição aos patógenos in vitro. As cepas com os melhores resultados foram identificadas e utilizadas no experimento de colonização do trato intestinal de tilápias-do-nilo, via suplementação na dieta, em delineamento inteiramente ao acaso, com três tratamentos e quatro repetições. Foram avaliados: o total de bactérias, as bactérias ácido-lácticas, Vibrio ssp. e Pseudomonas ssp. A cepa com melhor resultado foi utilizada na infecção experimental, em delineamento inteiramente ao acaso, em esquema fatorial 2x3: dieta suplementada com a cepa e dieta-controle; e os peixes não submetidos à injeção, peixes submetidos à injeção de solução salina e à injeção de Enterococcus durans, com três repetições. Foram avaliados os parâmetros hematológicos. As duas cepas identificadas foram: Lactobacillus plantarum e Lactobacillus brevis, que colonizaram o trato intestinal de tilápias, contudo L. plantarum teve menor número total de bactérias e de Pseudomonas ssp. Foi observado maior número total de eritrócitos, trombócitos, leucócitos, linfócitos, neutrófilos e monócitos, em peixes alimentados com L. plantarum e submetidos à injeção de E. durans. O L. plantarum tem efeito probiótico e melhora o sistema imune das tilápias.The objective of this work was to isolate lactic-acid bacteria from the intestines of Nile tilapia, and to assess their potential as probiotic. Strains of lactic-acid bacteria were isolated, and inhibition against pathogens was evaluated in vitro. Strains with best results were identified and used in tilapia intestinal tract colonization experiment through supplementation in the diet, in a completely randomized design, with three treatments and four replicates. Total number bacteria, lactic-acid bacteria, Vibrio ssp. and Pseudomonas ssp. were evaluated. The strain with best result was used for experimental infection, in a completely randomized design, in factorial arrangement 2x3: diet supplemented with strain and control diet; and fish not submitted to injection, fish submitted to injection of saline solution, and fish submitted to injection of Enterococcus durans, with three replicates. Hematological parameters were evaluated. Two strains were identified as Lactobacillus plantarum and Lactobacillus brevis, which colonized fishes' intestinal tract. However, L. plantarum had the lowest number of total bacteria and of Pseudomonas ssp. Greater number of red blood cells, thrombocytes, leukocytes, lymphocytes, neutrophils and monocytes were observed in fish fed with L. plantarum and submitted to injection of E. durans. L. plantarum has probiotic effect on Nile tilapias and improve their immune system.

The objective of this work was to assess the time-related action of probiotic Lactobacillus plantarum in the bacterial microbiota of the digestive tract of Litopenaeus vannamei, and the relation of total haemocyte count and serum phenol oxidase activity of shrimp challenged with Vibrio harveyi. Shrimps were fed with a probiotic-supplemented diet, for eight days, then shifted to a commercial diet. Shrimps fed only with the commercial diet served as control. Evaluations were made on the 8th day of experiment and repeated two, four, six and eight days later. Total lactic bacteria in the digestive tract was higher until the 4th day of evaluation in the probiotic-supplemented group. Vibrio spp. counts were higher in the control at days zero and two. Until the 4th day of evaluation, the total haemocyte counts in shrimps after challenge with V. harveyi were higher in probiotic-supplemented group than in control group. Significant difference was not observed in phenol oxidase activity. On the 6th day after shifting from supplemented to control diet, all parameters were equal in both groups, suggesting that the time-related action of L. plantarum in shrimp is short.O objetivo deste trabalho foi avaliar o tempo de atuação de Lactobacillus plantarum, na microbiota bacteriana intestinal de Litopenaeus vannamei, e sua relação com a contagem total de hemócitos e a atividade da fenoloxidase após a exposição a Vibrio harveyi. Os camarões foram alimentados com dieta suplementada com probióticos por oito dias e, depois, substituída por dieta comercial. Os camarões alimentados com dieta comercial constituíram o controle. As avaliações foram realizadas no oitavo dia de experimento e repetidas dois, quatro, seis e oito dias depois. A contagem total de bactérias lácticas no intestino foi maior até o quarto dia de avaliação, no grupo alimentado com probióticos. A contagem de Vibrio spp. foi superior no grupo controle nos dias zero e dois. Até o quarto dia, a contagem total de hemócitos dos camarões após a inoculação de V. harveyi foi superior no grupo alimentado com probiótico em relação ao grupo controle. Não foi observada diferença na atividade da fenoloxidase. No sexto dia, após a substituição da dieta suplementada com probióticos por dieta controle, todos os parâmetros avaliados foram iguais nos dois grupos, o que indica que o tempo de atuação de L. plantarum em camarões é curto.

This study evaluated the survival, post-larvae quality, and the population of bacteria in Litopenaeus vannamei after the addition of two strains of lactic-acid bacteria (2 and B6) experimentally infected by Vibrio harveyi. Fifteen hundred nauplii were distributed in 20 L capacity tanks with four replicates. The survival of control animals was lower (21%) than that of animals fed with the strains B6 (50%) and 2 (44%). Total bacterial population in the water and larvae, as well as of the Vibrio ssp. in water was not different among the treatments. No difference was observed in the population of Vibrio ssp. between the control larvae (5.5±0.5 log UFC/mL) and that fed with strain 2 (5.4±0.1 log UFC/mL). Shrimp from control and fed with strain 2 showed significantly higher bacterial population than those fed with strain B6 (1.2±0.2 log UFC/mL). It was detected the lower load of Vibrio ssp. bacteria with potential of pathogenicity after feeding with strain B6.Moreover, these larvae showed more active behavior and low number of necrosis in relation to the control group and to that fed with strain 2.Este trabalho avaliou a adição de duas cepas de bactérias lácticas (2 e B6) na sobrevivência, qualidade de pós-larva e na população de bactérias na larvicultura de Litopenaeus vannamei experimentalmente infectado por Vibrio harveyi. Mil e quinhentos náuplios foram distribuídos em tanques de 20 L com quatro repetições. A sobrevivência dos animais controle foi menor (21%) do que a dos alimentados com as cepas B6 (50%) e 2 (44%). Sobrevivência de misis após desafio com V. harveyi foi maior em B6 do que nos outros tratamentos. A população total de bactérias na água e nas larvas, bem como de Vibrio ssp. na água não foi diferente entre os tratamentos. Não houve diferença, também, entre a população de Vibrio ssp. em larvas do grupo controle (5,5±0,5 log UFC/mL) e larvas alimentadas com a cepa 2 (5,4±0,1 log UFC/mL). Camarões do grupo controle e alimentados com cepa 2 apresentaram maior população de bactérias do que os alimentados com cepa B6 (1,2±0,2 log UFC/mL). Foi comprovada a menor presença de bactérias entéricas com potencial de patogenicidade nos animais alimentados com a cepa B6, apresentando também comportamento mais ativo e menor número de necroses em relação ao controle e cepa 2.

The aim of the present work was to establish in vitro conditions for the induction, stabilization and proliferation of embryogenic cultures of A. angustifolia. Pre-cotyledonary staged zygotic embryos inoculated BM medium supplemented with 5 µM 2,4- D, 2 µM BAP and Kin, and 3% maltose or sucrose resulted in 66.7% induction rate. The rate of induction of embryogenic cultures was affected by the carbon source, as well the multiplication and morphology of the embryogenic cultures. Embryogenic cultures maintained in BM medium with maltose presented bipolar morphology. Globular somatic embryos were obtained BM medium with 9% (PEG) and (9%) maltose. These results could establish an in vitro regenerative protocol towards the conservation and improvement of this important species.No presente trabalho foram investigadas as condições para a indução, estabilização e proliferação de culturas embriogênicas de A. angustifolia. Foram também descritos os padrões de morfogênese e histodiferenciação de culturas embriogênicas e embriões somáticos desta espécie em resposta a diferentes fontes e concentrações de carboidratos. Embriões zigóticos no estágio pré-cotiledonar, inoculados em meio de cultura BM, suplementados com 5 µM de 2,4-D, 2 µM de BAP e Kin e 3% de maltose ou sacarose resultaram em uma taxa de indução de 66,7%. A fonte de carbono afetou a taxa de indução, a multiplicação e a morfologia das culturas. Culturas embriogênicas mantidas em meio de cultura BM suplementado com maltose apresentaram morfologia bipolar. Embriões somáticos globulares foram obtidos em meio de cultura BM suplementado com PEG e maltose. Os resultados obtidos permitirão avançar na direção de um protocolo regenerativo in vitro visando à conservação e o melhoramento genético desta espécie.

This study evaluated the pathogenic characteristics of bacteria isolated from Litopenaeus vannamei during an outbreak at the Laboratory of Marine Shrimp, UFSC, Santa Catarina State, Brazil. Their virulence potential in larvae and juvenile shrimp and the effects on the total haemocyte count, phenoloxidase activity and serum agglutinate titre were examined after experimental infection. Bacterial strains were isolated from larvae and adult shrimps, identified by the AP120E biochemical system as: two strains of Vibrio alginolyticus, three of Aeromonas salmonicida and one of Pasteurella multocida sp. and Pasteurella sp. All the bacterial strains isolated in this study caused mortality in shrimp. One strain of V. alginolyticus was responsible for 97.3 and 88.7% mortality in larvae and juvenil shrimps, respectively. The shrimp immunological system was influenced by experimental infection with V. alginolyticus. Decrease in the total haemocyte count and increase in the phenoloxidase activity and the serum agglutinate titre (p < 0.05) were observed. The results showed the high pathogenicity of V. alginolyticus isolated from larvae and juvenile reared marine shrimp.

Objetivou-se averiguar o efeito da bactéria Lactobacillus plantarum e células inativas das bactérias Vibrio alginolyticus, Aeromonas salmonicida e Pasteurella multocida na sobrevivência larval de Litoenaeus vannamei, no teste de estresse e infecção experimental com Vibrio harveyi. Foram utilizados tanques cônicos de 30 L, povoados com 400 larvas cada, no estádio de pós-larva cinco. Tratamentos em triplicatas foram consistidos de: 1: ração comercial (controle), 2: ração comercial + bacterina via oral na artemia, 3: ração comercial + bacterina via imersão e 4: ração comercial com inóculo de Lactobacillus plantarum. A aplicação da bacterina ocorreu seis horas antes da infecção e do teste de estresse; enquanto o Lactobacillus plantarum foi administrado por 15 dias antes dos desafios. As pós-larvas do tratamento 4 (ração suplementada com L. plantarum) obtiveram maior índice de sobrevivência no teste de estresse (87,86 ± 2,35%), seguido dos tratamentos 2 e 3 (bacterina via imersão e oral) com 81,54±1,50% e 80,16 ± 2,15% respectivamente, superiores ao índice do controle (72,63 ± 3,34%). Já no desafio com V. harveyi, os animais do grupo tratado com a adição de bacterina via imersão apresentaram maior sobrevivência (79,60 ± 7,12%). As pós-larvas dos tratamentos com bacterina via oral na artêmia e alimentadas com o probiótico L. plantarum, apresentaram sobrevivências de 65,60 ± 5,18% e 69,60 ± 10,43 %, respectivamente, sendo superiores ao controle (56,4 ± 5,58%), quando desafiados com V. harveyi. Os resultados demonstram que o uso de ração com L. plantarum e bacterina aumentam a sobrevivência das pós-larvas de L. vannamei frente aos testes de estresse e infecções experimentais com V. harveyi.

ABSTRACT This study aimed to assess the final body weight, weekly weight gain, yield, apparent feed efficiency, protein efficiency ratio, and feed intake of the Pacific white shrimp (Litopenaeus vannamei) fed four diets containing different levels of soy protein concentrate (SPC) as a replacement for fishmeal, reared in a super-intensive biofloc system. Diets consisted of replacing 209 g kg-1 fishmeal at 0, 33, 66, and 100% SPC. Shrimp were raised in a biofloc system using twelve experimental units stocked with 250 shrimp m-3 under constant aeration (O2>5mg L-1) and temperature (29±0.5 °C). No significant differences among treatments were observed based on water quality parameters. Shrimp fed diets with 0 and 33% substitution exhibited the highest weekly growth (1.88 and 1.79 g per week) and final weights (15.2 and 14.7 g) compared with shrimp fed the 66 and 100% replacement. A lower feed intake was observed for shrimp fed the 33% SPC diet (3.18 kg per experimental unit) compared with 0% replacement (3.62 kg). Shrimp fed the 33% replacement achieved a similar performance and lower feed intake than animals fed diet without replacement.

ABSTRACT This study evaluated the performance, immunology, and survival of the Pacific white shrimp Litopenaeus vannamei to experimental challenge to Vibrio alginolyticus based on the use of the probiotic Lactobacillus plantarum and the combined use of probiotic and butyrate. Four different diets resulted from the addition of additives: butyrate, probiotic, butyrate + probiotic, and control (no additives). The attractiveness of the diets was assessed by the percentage of positive choices and rejections, using a dual-choice Y-maze format aquarium. The shrimps were fed during four weeks and performance parameters, intestinal microbiota, and immunological parameters were all evaluated. Subsequently, the shrimps were challenged with V. alginolyticus and after 48 h, survival and immunological parameters were evaluated. The results showed increased attractiveness and intake, but only with diets supplemented with sodium butyrate. However, other diets were not rejected. No difference in performance or immunological parameters was observed among the different diets. Also, among the treatments, no difference in Vibrio spp., or total heterotrophic bacteria counts, was found in the intestinal tract. However, the lactic acid bacteria count was higher in the intestinal tract of shrimps fed diets supplemented with probiotic. After bacterial challenge, shrimp fed all diets had a greater survival when compared with the control group. Lactobacillus plantarum and sodium butyrate increase the resistance of shrimp to infection with V. alginolyticus, but do so without affecting performance, immunological parameters, or Vibrio spp., and total heterotrophic bacteria counts in the intestinal tract.