Depletion study and estimation of the withdrawal period for enrofloxacin in pacu (Piaractus mesopotamicus)

Defining the pharmacokinetic parameters and depletion intervals for antimicrobials used in fish will help in the development of important guidelines for future regulations by Brazilian agencies on the use of these substances in fish farming. This paper presents a depletion study for enrofloxacin (ENR) and its main metabolite, ciprofloxacin (CIP), in pacu (Piaractus mesopotamicus) fillets. The depletion study was carried out under monitored environmental conditions, with the temperature controlled at 27 °C to mimic the fish farming conditions in Brazil. ENR was administered orally via medicated feed for 10 consecutive days at daily dosages of 10 mg/kg body weight (b.w.). The fish were slaughtered at 6, 12, and 24 h and 2, 3, 5, 8, 12, 17, and 24 days after the medication period. Considering a maximum residue limit of 100 μg/kg for the sum of the ENR and CIP residues in the fillet, the results obtained in the depletion study allowed the estimation of a half‐life for ENR of 2.75 days and a withdrawal period of 23 days. The results obtained in this study are important for the farming of pacu in tropical regions.     

Observation of pulsed gamma-rays above 25 GeV from the Crab pulsar with MAGIC

One fundamental question about pulsars concerns the mechanism of their pulsed electromagnetic emission. Measuring the high-end region of a pulsar's spectrum would shed light on this question. By developing a new electronic trigger, we lowered the threshold of the Major Atmospheric gamma-ray Imaging Cherenkov (MAGIC) telescope to 25 giga-electron volts. In this configuration, we detected pulsed gamma-rays from the Crab pulsar that were greater than 25 giga-electron volts, revealing a relatively high cutoff energy in the phase-averaged spectrum. This indicates that the emission occurs far out in the magnetosphere, hence excluding the polar-cap scenario as a possible explanation of our measurement. The high cutoff energy also challenges the slot-gap scenario.     

Debaryomyces hansenii L2‐enriched diet enhances the immunity status,gene expression and intestine functionality in gilthead seabream (Sparus aurata L.)

This study evaluates the effects of the dietary administration of the live yeast Debaryomyces hansenii strain L2 on the immune responses of gilthead seabream for 4 weeks. Cellular immune parameters were measured from serum and head‐kidney leucocytes respectively. The expression levels of immune‐associated genes were quantified by real‐time polymerase chain reaction. In addition, the profile of intestinal microbiota was studied by denaturing gradient gel electrophoresis. The results showed that seabream fed a diet containing D. hansenii had significantly increased cellular immune parameters. The yeast‐supplemented diet up‐regulated the expression of most seabream genes at week 2 and down‐regulated all of them at week 4, except in the head‐kidney. Finally, a reduction in the diversity of the intestinal microbiota was detected in those specimens receiving the yeast‐supplemented diet. These results support the idea that the live yeast D. hansenii strain L2 stimulates the immune system of gilthead seabream.     

Metabolism of 14C uniformly labeled organic material by woodlice [Isopoda: Oniscoidea] and soil microorganisms

Uniformly labeled ¹⁴C-yeast was fed to woodlice and soil microorganisms together and independently. Mineralization was more rapid and extensive in treatments in which both groups were present. Two days after a single feed of the labeled yeast to freshly-collected or wood-reared animals, approximately 12 per cent of the ¹⁴C had been respired, 28 per cent excreted, 44 per cent assimilated, and 15 per cent unaccounted for. Yeast-reared animals were 6.6 per cent less efficient in assimilating the labeled food. After 26 days, maintenance consumption had resulted in dissipation of 65.8 per cent of the assimilated label, with almost 90 per cent of this amount eliminated as CO₂ and 10 per cent excreted. The elimination rate dropped from 6.8 per cent of the assimilated label per day to 0.6 per cent over the 1 month period following the single feeding of ¹⁴C. Three-quarters of the labeled faecal material excreted by the woodlice was mineralized by the soil microogranisms within 1 month; however, the rate of degradation of the faeces was significantly slower than was the rate of degradation of the labeled yeast. The ¹⁴C method appeared to give high recoveries of label and reproducible results.     

Assessment of the relative contribution of dietary nitrogen from fish meal and biofloc meal to the growth of Pacific white shrimp (Litopenaeus vannamei)

The relative contribution of the dietary nitrogen supplied by fish meal and a biofloc meal to the growth of Pacific white shrimp was evaluated using stable isotope analysis. Biofloculated material was obtained from an experimental tilapia culture system. Five formulated diets were supplied. Two of them consisted in isotopic controls having only fish meal or biofloc meal as protein source. Three mixed diets were formulated with varying proportions of these ingredients on a dietary nitrogen basis (75:25, 50:50 and 25:75). At the end of the trial, survival rates were similar (92–100%) but significant differences in mean final weight were observed and a negative correlation between the inclusion of biofloc meal and weight gain was evidenced. Mean final weight in shrimp fed on diet containing only fish meal was 2.8 g, while mean final weight of animals fed on diet containing 50% biofloc was 1.9 g. Isotopic mixing models indicated that all diets contributed higher proportions of dietary nitrogen from fish meal than from biofloc meal. Dietary nitrogen available in diets containing 25%, 50% and 75% of biofloc meal was incorporated in muscle tissue as 5%, 41% and 64% respectively. Diet supplying 25% of nitrogen from biofloc was the only mixed diet eliciting growth comparable to diet containing only fish meal. Lower growth and nitrogen deposition in shrimp fed on diets containing high proportions of biofloc meal were possibly associated to the use of only two protein sources and a restriction of essential amino acids.     

In vivo and in vitro assessment of Lactobacillus acidophilus as probiotic for tilapia (Oreochromis niloticus,Perciformes:Cichlidae) culture improvement

In this study, the use of Lactobacillus acidophilus as a probiotic for Nile tilapia (Oreochromis niloticus) culture was studied. Fish survival and the expression of some genes involved in the immune response were assessed. Diet supplementation with L. acidophilus for 15 days caused a significant increase in fish survival during a challenge with Aeromonas hydrophila and variations in immune response related to IL‐1β and transferrin expression in Nile tilapia spleen and kidney. Moreover, extracellular products (ECPs) of L. acidophilus showed high antibacterial activity against fish pathogens such as A. hydrophila and Streptococcus agalactiae in vitro. It was also observed that viable L. acidophilus was able to disrupt quorum sensing activity in Chromobacterium violaceum.     

Bioactive Ascochlorin Analogues from the Marine-Derived Fungus Stilbella fimetaria

The marine-derived fungus Stilbella fimetaria is a chemically talented fungus producing several classes of bioactive metabolites, including meroterpenoids of the ascochlorin family. The targeted dereplication of fungal extracts by UHPLC-DAD-QTOF-MS revealed the presence of several new along with multiple known ascochlorin analogues (19–22). Their structures and relative configuration were characterized by 1D and 2D NMR. Further targeted dereplication based on a novel 1,4-benzoquinone sesquiterpene derivative, fimetarin A (22), resulted in the identification of three additional fimetarin analogues, fimetarins B–D (23–25), with their tentative structures proposed from detailed MS/HRMS analysis. In total, four new and eight known ascochlorin/fimetarin analogues were tested for their antimicrobial activity, identifying the analogues with a 5-chloroorcylaldehyde moiety to be more active than the benzoquinone analogue. Additionally, the presence of two conjugated double bonds at C-2′/C-3′ and C-4′/C-5′ were found to be essential for the observed antifungal activity, whereas the single, untailored bonds at C-4′/C-5′ and C-8′/C-9′ were suggested to be necessary for the observed antibacterial activity.     

Incorporation of fatty acids from dietary neutral lipid in eye, brain and muscle of postlarval turbot fed diets with different types of phosphatidylcholine

Previous results demonstrated the stimulating effect of soybean phosphatidylcholine (PC) on the utilization of dietary neutral lipid in larval and postlarval fish. The present study further investigated the effect of the degree of saturation of dietary PC on the enhancement of dietary fatty acid incorporation in lipids of turbot. Newly-weaned turbot were fed for 20 days on four isolipidic diets containing the same amount of highly unsaturated fatty acids (HUFA), presented either as neutral lipid, i.e. fish oil ethyl esters, or as polar lipid. Diet FO was a phospholipid-free control diet. Diets HPC, SPC and FPC were supplemented with 3% hydrogenated soybean PC, 3% native soybean PC and 3% marine fish roe PC, respectively.The three PC-supplemented diets resulted in better growth and higher muscle triacylglycerol levels than the PC-free diet FO. The fish fatty acids were determined in 3 lipid classes (neutral lipid, PC, phosphatidylethanolamine) of 3 organs or tissues (eye, brain and muscle). Despite the identical amounts of n-6 and n-3 fatty acids provided by the soybean oil and by the HUFA ethyl esters, the substitution of 3% hydrogenated coconut oil in diet FO by 3% hydrogenated PC in diet HPC caused, averaged over the various tissues and lipid classes, a 7 to 12% higher incorporation of 18:2n-6, 20:4n-6, 20:5n-3 and a 32% higher 22:6n-3 level in turbot lipid. Diet HPC appeared as efficient as diet SPC for enhancing the incorporation of the n-3 HUFA from the ethyl esters. Feeding diet FPC, in which the n-3 HUFA were provided through the marine PC source, resulted in slightly higher levels of these fatty acids in the fish than feeding the ethyl ester HUFA diets, even if supplemented with PC. Present results confirm the positive effect of PC, either hydrogenated or native, on the utilization of fatty acids provided in the diet as neutral lipid. The slightly higher incorporation of HUFA, when esterified on dietary PC instead of neutral lipid, raises the question regarding the form of intestinal absorption of PL in fish.p>     

Chromomycin A2 Induces Autophagy in Melanoma Cells

The present study highlights the biological effects of chromomycin A2 toward metastatic melanoma cells in culture. Besides chromomycin A2, chromomycin A3 and demethylchromomycin A2 were also identified from the extract derived from Streptomyces sp., recovered from Paracuru Beach, located in the northeast region of Brazil. The cytotoxic activity of chromomycin A2 was evaluated across a panel of human tumor cell lines, which found IC₅₀ values in the nM-range for exposures of 48 and 72 h. MALME-3M, a metastatic melanoma cell line, showed the highest sensitivity to chromomycin A2 after 48h incubation, and was chosen as a model to investigate this potent cytotoxic effect. Treatment with chromomycin A2 at 30 nM reduced cell proliferation, but had no significant effect upon cell viability. Additionally, chromomycin A2 induced accumulation of cells in G₀/G₁ phase of the cell cycle, with consequent reduction of S and G₂/M and unbalanced expression of cyclins. Chromomycin A2 treated cells depicted several cellular fragments resembling autophagosomes and increased expression of proteins LC3-A and LC3-B. Moreover, exposure to chromomycin A2 also induced the appearance of acidic vacuolar organelles in treated cells. These features combined are suggestive of the induction of autophagy promoted by chromomycin A2, a feature not previously described for chromomycins.     

Inhibition of Bacterial Quorum Sensing by Extracts from Aquatic Fungi: First Report from Marine Endophytes

In our search for quorum-sensing (QS) disrupting molecules, 75 fungal isolates were recovered from reef organisms (endophytes), saline lakes and mangrove rhizosphere. Their QS inhibitory activity was evaluated in Chromobacterium violaceum CVO26. Four strains of endophytic fungi stood out for their potent activity at concentrations from 500 to 50 μg mL⁻¹. The molecular characterization, based on the internal transcribed spacer (ITS) region sequences (ITS1, 5.8S and ITS2) between the rRNA of 18S and 28S, identified these strains as belonging to four genera: Sarocladium (LAEE06), Fusarium (LAEE13), Epicoccum (LAEE14), and Khuskia (LAEE21). Interestingly, three came from coral species and two of them came from the same organism, the coral Diploria strigosa. Metabolic profiles obtained by Liquid Chromatography-High Resolution Mass Spectrometry (LC-HRMS) suggest that a combination of fungal secondary metabolites and fatty acids could be the responsible for the observed activities. The LC-HRMS analysis also revealed the presence of potentially new secondary metabolites. This is, to the best of our knowledge, the first report of QS inhibition by marine endophytic fungi.