Acute tolerance and histopathological effects of ammonia on juvenile maroon clownfish Premnas biaculeatus (Block 1790)

This study evaluated the median lethal concentration values (LC₅₀) and the histopathological effects of un‐ionized ammonia (NH₃^‐‐N) on juvenile maroon clownfish Premnas biaculeatus. After 96 h of exposure to different concentrations of ammonia, juveniles were sampled for histopathological evaluation. The 24 and 96 h LC50 values of NH₃^?‐N determined were 1.68 and 0.89 mg L^?1 respectively. Maroon clownfish exposed to different ammonia concentrations displayed histopathological alterations in the gills, kidney, liver and brain. Gill tissue damage included lamellar hyperplasia, lamellar shorting and hyperplasia and hypertrophy of mucous cells. The kidney showed hyperanaemia, enlarged sinusoids within an apparently decreased amount of haematopoietic tissue, oedema on tubular cells and tubular necrosis, and an enlarged Bowman's capsule. The liver presented dilatation of hepatic sinusoids, fatty deposition in hepatocytes and Mallory bodies. Examination of the brain revealed a proliferation of glial cells, and the Virchow‐Robin space indicated a severe perivascular oedema and signs of neuronal suffering with satellitosis. The results of this study indicate that juvenile maroon clownfish are relatively sensitive to ammonia and particular attention must be given to this toxic compound in culture systems.     

Chitosan-Based Films with 2-Aminothiophene Derivative: Formulation,Characterization and Potential Antifungal Activity

In this study, films of chitosan and 2-amino-4,5,6,7-tetrahydrobenzo[b]thiophene-3-carbonitrile (6CN), a 2-aminothiophene derivative with great pharmacological potential, were prepared as a system for a topical formulation. 6CN-chitosan films were characterized by physicochemical analyses, such as Fourier-transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), thermogravimetric analysis (TGA), X-ray diffraction (XRD), and scanning electronic microscopy (SEM). Additionally, the antifungal potential of the films was evaluated in vitro against three species of Candida (C. albicans, C. tropicalis, and C. parapsilosis). The results of the FTIR and thermal analysis showed the incorporation of 6CN in the polymer matrix. In the diffractogram, the 6CN-chitosan films exhibited diffraction halos that were characteristic of amorphous structures, while the micrographs showed that 6CN particles were dispersed in the chitosan matrix, exhibiting pores and cracks on the film surface. In addition, the results of antifungal investigation demonstrated that 6CN-chitosan films were effective against Candida species showing potential for application as a new antifungal drug.     

The larval rearing of the marine ornamental crab, Mithraculus forceps (A. Milne Edwards, 1875) (Decapoda: Brachyura: Majidae)

The goal of this study is to develop a larviculture protocol for Mithraculus forceps, a popular marine aquarium species. Different temperatures (25±0.5°C and 28±0.5°C), stocking densities (10, 20, 40 and 80 larvae L^?1), prey densities (newly hatched Artemia of 1, 4, 7 and 12 nauplii mL^?1) and metamorphosis to crab conditions (Systems A and B) were tested. The best survivorship and faster development were obtained when the larvae were reared at a density of 40 larvae L^?1 for 7 days post hatching (DPH) in System A, at 28°C and fed with 7 mL^?1 of newly hatched Artemia nauplii. After 7 DPH all the megalopa were moved to System B and the same temperature and prey density were maintained. At the end of the experiment, 12 DPH, survivorship of 74.1±4.8% was obtained.     

Effect of dietary carbohydrate-to-lipid ratios on growth, lipid deposition and metabolic hepatic enzymes in juvenile Senegalese sole (Solea senegalensis, Kaup)

A study was undertaken to determine the effect of various dietary carbohydrate‐to‐lipid ratios on growth performance, whole‐body composition and tissue lipid content in Senegalese sole (Solea senegalensis) juveniles. Data on the dietary regulation of key hepatic enzymes of the lipogenic and glycolytic pathways (glucose‐6‐phosphate dehydrogenase, G6PD; malic enzyme, ME; fatty acid synthetase, FAS; pyruvate kinase, PK and glucokinase, GK) were also generated. Four isonitrogenous (crude protein: 52% dry matter (DM)) diets were formulated to contain one of two lipid levels (11% and 21% DM). Within each dietary lipid level, the nature of the carbohydrate fraction (raw or extruded peas) was varied. Triplicate groups of 54 sole (initial body weight: 23.6±1.2 g) were grown in recirculated seawater over 67 days. Fish were fed using automated feeders. At the end of the study, whole‐body, liver, viscera and muscle samples were withdrawn for analyses. During the experimental period, the mean fish weight about doubled in all treatments. No significant differences were found in growth performance (ranging from 1.1% to 1.4% body weight day^?1) among dietary treatments. High‐fat diets increased whole‐body fat content. Similarly, daily fat gain ranged from 0.54 to 0.78 g kg^?1 day^?1 and highest values were found in fish fed high‐lipid diets. Dietary treatments also affected tissue lipid content (liver, viscera and muscle), with highest values in fish fed high‐fat diets. The nature of dietary carbohydrates had little influence on performance criteria, but affected tissue lipid deposition. The activities of G6PD, ME and FAS were depressed by elevated levels of dietary lipid, confirming the inhibitory effect of dietary fats on lipid biosynthesis. At both dietary lipid levels, ME and FAS activities were little affected by dietary carbohydrate. Activities of PK and GK were not affected by the starch level of the diets. In Senegalese sole juveniles, the lipogenic pathway is more susceptible to modulation by dietary means (particularly through lipid intake) than the glycolytic pathway.     

Effects of beneficial bacteria on biomass,photosynthetic parameters and cell composition of the microalga Muriellopsis sp. adapted to grow in seawater

The present study evaluated the effects of five bacterial isolates from the microalga Muriellopsis sp. on the microalgal biomass concentration (g/L), specific growth rate (K), photosynthetic quantum yield (QY), non‐photochemical quenching (NPQ), maximum relative electron transport rates (rETR max) and levels of nutritional components such as carbohydrates, lipids, proteins and carotenoids in Muriellopsis sp. cultivated in seawater under indoor and outdoor conditions. The bacterial strains were Microbacterium sp. Dom 1; Pseudomonas sp. Dom2; Microbacterium sp. Dom 3; Rhizobium sp. Dom 4; and Dietzia sp. Dom 5, as well as the Pseudoalteromonas sp. (SLP1), a bacterium known to promote the growth of microalgae. At the laboratory level, inoculation of the strains SLP1, Dom 1, Dom 3 and Dom 5 resulted in higher specific growth rates and biomass of the microalga. Subsequently, the best results were obtained with a combination of the selected bacterial strains (Dom1, Dom 3, Dom5 and SLP1) under outdoor culture conditions improved the biomass, proteins, carbohydrates and total carotenoids in 22.12%, 48.28%, 19,25% and 48.27%, respectively, compared to cultures without the incorporation of the selected bacteria (control), while no effects were observed on the photosynthetic parameters. Thus, it was demonstrated that positive associations of the selected bacterial strains played an important role in the production of Muriellopsis sp. acclimatized to grow in seawater. This is of crucial importance, especially in desert areas, where solar radiation is high and freshwater is a limited resource.     

Influence of light intensity on biofilm formation and the performance of pink shrimp Farfantepenaeus paulensis juveniles reared in cages

A reduction in light intensity may increase shrimp activity, although it may also negatively affect the development of photoautotrophic organisms present in biofilm, an important natural food source for shrimp. This experiment evaluated the influence of light intensity on biofilm development and on the growth of cultured Farfantepenaeus paulensis juveniles. Six cages were deployed in a shrimp culture pond and stocked with 60 juveniles (0.72 ± 0.03 g). Three cages were randomly chosen and covered with five layers of polyamide net to reduce light incidence (shaded treatment), while the other three cages were not covered (control treatment). Biofilm chlorophyll a and microorganism abundance, as well as shrimp growth, were monitored during the experiment. Chlorophyll a concentration and the abundance of bacteria and flagellates were significantly lower in the shaded treatment (P<0.05). After 75 rearing days, shrimp stocked in control treatment achieved significantly higher (5.98 g) (P<0.05) mean weight than shrimp reared in shaded treatment cages (5.13 g). Similarly, the biomass produced was also significantly higher (P<0.05) in the control treatment (322.92 g) than in the shaded treatment (287.31 g). The results of this study demonstrate that light intensity has a huge influence on shrimp performance mainly due to the increase in natural food availability.     

Phenotypical parameters as a tool to evaluate the immunostimulatory effects of laminarin in Oncorhynchus mykiss

Laminarin is a β‐glucan from the brown algae Laminaria digitata (J.V. Lamour), which can activate innate immune responses. With the aim of developing a strategy to evaluate specific phenotypical parameters of the effects of laminarin in trout innate immunity, we (i) fed fish with laminarin‐supplemented diet (0.2 g kg^?1 day^?1) for 21 days and (ii) treated fish with a single dose of intraperitoneal injected laminarin (0.08 g kg^?1 fish). The evaluation of cellular and humoral immune parameters was established at phenotypic level by the phagocytic activity of headkidney macrophages and detection of inflammatory cytokines in head kidney and gill tissue by indirect ELISA. Results showed that both delivery methods of laminarin produce an increase in the phagocytic activity in head kidney macrophages and a significant increase in the production of TNFα and IL‐8 in gill tissue at day 21. Additionally, some of these parameters were significantly correlated (P < 0.025), which places them as new potential combined markers to detect activation of trout defense mechanisms by laminarin. These results highlight the importance of developing new protocols to quantitate immune parameters, in order to evaluate immunostimulants in fish farming.     

Chemical composition of the Lippia origanoides essential oils and their antigenotoxicity against bleomycin-induced DNA damage

The present work evaluated the chemical composition of the essential oils (EO) obtained from Lippia origanoides and their DNA protective effect against bleomycin-induced genotoxicity. L. origanoides EO chemical composition was determined by gas chromatography–mass spectrometry (GC–MS). The major compounds of the L. origanoides EOs were thymol (34–58%) and carvacrol (26%). The antigenotoxic effects of the EOs, major compounds and standard compound (epigallocatechin gallate) were assayed in co-incubation procedures using the SOS chromotest in Escherichia coli. Both EOs and their major compounds protected bacterial cells against bleomycin-induced genotoxicity indicating that these two compounds were principally responsible for the antigenotoxicity detected in the oils. Thymol and carvacrol antigenotoxicity was lower than those observed with epigallocatechin gallate. The results were discussed in relation to the chemopreventive potential of L. origanoides EOs and their major components, carvacrol and thymol.     

Characterization of symptoms of senescence and chilling injury on inflorescences of Heliconia bihai (L.) cv. Lobster Claw and cv. Halloween

Inadequate temperatures during the shipping and commercialization of cut tropical flowers may accelerate the senescence process and cause chilling injury, leading to symptoms that have not yet been described for Heliconia bihai. The aim of the present study was to evaluate physiological responses in cut inflorescences of H. bihai cv. Lobster Claw (LC) and cv. Halloween (HW) as well as symptoms of senescence and chilling injury. For such, changes in fresh weight, bract color (L*, a* and b*), percentage of absolute integrity (PAI) of cell membranes and leakage of potassium ions (LPI) were determined. The flowering stems were evaluated at five different intervals after harvest (0, 2, 4, 6 and 8 d). A refrigerated treatment (RT) with a temperature of 6.5 °C and 85% relative humidity was compared to a control treatment (CT) at room temperature of 24 °C and 66% relative humidity. Both cultivars stored at 6.5 °C exhibited dryness of bract tissue (symptom of senescence) and dark stains that became brownish and evolved to necrosis (symptom of chilling injury). The visual quality of inflorescences decreased with time in both cultivars maintained without refrigeration. The severity of chilling injury increased with the length of storage time in both cultivars. There was a significant reduction in the fresh weight of inflorescences in both treatments (RT and CT) and both cultivars (LC and HW). Bract color changed in both cultivars at 6.5 °C. There was no change in PAI throughout the evaluation period in the inflorescences stored at room temperature, whereas those stored at 6.5 °C for 6 and 8 d had lower PAI values. The inflorescences in the control treatment underwent no change in LPI values, whereas those stored under refrigeration had increased LPI values after the sixth day of storage. The physiological responses of cut Heliconia flowers were influenced by storage period and temperature, as demonstrated by visual symptoms of chilling injury and senescence.