Naringin-loaded Poly(ε-caprolactone)/Gelatin Electrospun Mat as a Potential Wound Dressing: In vitro and In vivo Evaluation

In the present study, naringin, a flavonoid isolated from the grape and citrus fruit species, was incorporated with poly(ε-caprolactone)/gelatin composite mats in order to develop a potential wound dressing. The composite mats were prepared by electrospinning of poly(ε-caprolactone)/gelatin (1:1 (w/w)) solution incorporated with 1.50 %, 3 % and 6 % (w/w) of naringin. The electrospun mats were evaluated regarding their morphology, contact angle, water-uptake capacity, water vapor transmission rate, tensile properties, drug release, cellular response and in vivo wound healing activity. The study showed that after 2 weeks, the full-thickness excisional wounds of Wistar rats treated with the naringin-loaded dressings achieved a wound closure of higher than 94 % and the dressing containing 6 % (w/w) naringin had almost 100 % wound closure. The sterile gauze, as the control group, showed nearly 86 % of wound closure after this period of time. Our results provided evidence that supports the possible applicability of naringin-loaded wound dressing for successful wound treatment.     

Swim bladder inflation as a possible cause of saddleback-like syndrome malformation in hatchery-reared red spotted grouper <Emphasis Type="Italic">Epinephelus akaara</Emphasis>

Saddleback-like syndrome is one of the most frequently observed malformations in hatchery-reared red spotted grouper Epinephelus akaara. Saddleback-like syndrome is characterized by a V-shaped dorsal-profile depression behind the head region accompanied by deformities or misalignments of the first to fourth neural spines on the vertebral column and the first to fourth pterygiophores of the dorsal fin. Abnormal swim bladder inflation in several finfish species leads to malformations, particularly of the vertebral column. The present study examined the influence of swim bladder inflation on saddleback-like syndrome in hatchery-reared red spotted grouper. The incidence of saddleback-like syndrome increased as swim bladder inflation rate increased. Additionally, the incidence of saddleback-like syndrome among juveniles with inflated swim bladders was significantly higher than that among juveniles with uninflated swim bladders. The larval swim bladder is located just below the first to fifth vertebrae and the first to fifth dorsal spines, where saddleback-like syndrome occurs. Our results suggest that an inflated swim bladder increases the risk of saddleback-like syndrome in hatchery-reared red spotted grouper.     

Cylindrospermopsin effects on cell viability and redox milieu of Neotropical fish <Emphasis Type="Italic">Hoplias malabaricus</Emphasis> hepatocytes

Cylindrospermopsin (CYN) is a cyanotoxin that is cytotoxic to a wide variety of cells, particularly to the hepatocytes. In this study, the toxic effects of purified CYN were investigated in primary cultured hepatocytes of Neotropical fish Hoplias malabaricus. After isolation, attachment, and recovery for 72 h, the cells were exposed for 72 h to 0, 0.1, 1.0, 10, and 100 μg l^?1 of CYN. Then, cell viability and a set of oxidative stress biomarker responses were determined. Catalase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione S-transferase activities were not affected by exposure to CYN. Concentration-dependent decrease of glutathione reductase activity occurred for most CYN-exposed groups, whereas non-protein thiol content increased only for the highest CYN concentration. Lipid peroxidation, protein carbonylation, and DNA damage levels were not altered, but reactive oxygen species levels increased in the cells exposed to the highest concentration of CYN. Cell viability decreased in all the groups exposed to CYN. Thus, CYN may cause a slight change in redox balance, but it is not the main cause of cell death in H. malabaricus hepatocytes.     

Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants,antioxidant, membrane stabilizer,equilibration time and dilution ratio on sperm motility and fertility

Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL^?1) and ascorbic acid (0, 2.5, 5 and 10 U mL^?1), on the post‐thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL^?1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post‐thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post‐thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post‐thaw motility (P < 0.05). The motility of frozen–thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL^?1 BSA significantly improved post‐thaw motility (P < 0.05), but ascorbic acid did not improve post‐thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 ± 5.75) and hatching rates (28.2 ± 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL^?1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.     

Androgen-induced pseudo-hermaphroditic phenotypes in female <Emphasis Type="Italic">Brevimyrus niger</Emphasis> Günther 1866 (Teleostei,Mormyridae)

This paper explores the plasticity of sexually dimorphic characters in subadult female Brevimyrus niger, an African weakly electric mormyrid species. Thirty-five fish were exposed in a staggered fashion (five fish a week) to aromatizable 17α-methyltestosterone over a period of 7 weeks; 18 fish served as untreated controls. 17α-MT induced precocious vitellogenesis that mirrored the natural maturational process during seasonal ovarian recrudescence. At the same time, 17α-MT exposure resulted in complete masculinization of the females’ anal fin support structure normally observed during rainy season in adult males. We discuss possible hormonal mechanisms acting along the brain-pituitary-gonad axis that would explain the occurrence of precocious vitellogenesis and the male-typical transformation of the female’s anal fin ray bases. Our findings are relevant to commercial aquaculture as the use of 17α-MT in fish hatcheries can pose serious environmental issues.     

Free-Living Plant-Parasitic Nematodes do not Affect the Efficiency of Seed Tuber Fungicide Treatment against <Emphasis Type="Italic">Rhizoctonia solani</Emphasis>

Stem canker on germinating potato sprouts is often caused by seed-borne inoculum of the fungus Rhizoctonia solani. However, high amounts of free-living plant-parasitic nematodes have been found in field patches of potato plants with stem canker. Fungicide treatment of the seed tubers can be used to avoid stem canker caused by seed-borne inoculum but it is unknown if nematodes can affect this. To investigate whether free-living plant-parasitic nematodes, the root-lesion nematode Pratylenchus penetrans or a combination of several plant-parasitic nematode genera in a full nematode community, may have a negative effect on the fungicide seed treatment, a pot experiment with seed tubers inoculated with R. solani, half of which were treated with fungicides, was performed. The seed-borne inoculum caused severe damage to the plants, while no fungal damages were observed on the fungicide treated plants. This shows that the nematodes did not affect the fungicide treatment. The probability of black scurf decreased in treatments with a full nematode community, which may be due to the action of fungal-feeding nematodes.     

Electrical properties of graphene/waterborne polyurethane composite films

Graphene is classified as a carbon-based material. Structurally, graphene is made up of carbon-based two-dimensional atomic crystals and a one atom thick planar sheet of sp²-bonded carbon atoms. This sort of arrangement in graphene makes it a unique material with exceptional mechanical, physicochemical, thermal, electrical, optical, and biomedical properties. Methods for graphene-based fabric production mainly use graphene-based materials such as graphene (G), graphene oxide (GO), and reduced graphene oxide (rGO) coated on fabric or yarn. Waterborne polyurethane (WPU) is one of the most rapidly developing and active branches of polyurethane chemistry. More and more attention is being paid to graphene-coated fabrics owing to their low temperature flexibility, the presence of zero or very few VOCs (volatile organic compounds), water resistance, pH stability, superior solvent resistance, excellent weathering resistance, and desirable chemical and mechanical properties. It is used as a coating agent or adhesive for fibers, textiles, and leather. Also, graphene-containing materials have been used to enhance the properties of WPU. In this study, graphene/WPU composite solution and film was prepared to conduct basic research for developing electrical heating textiles which is not harmful to the human body, flexible and excellent in electrical properties. Graphene/WPU composite solutions were prepared with a graphene content of 0, 2, 4, 8, and 16 wt%, and graphene/WPU film was prepared with solution casting method. The graphene contents were analyzed for their surface morphology, electrical properties, and electrical heating properties.     

A fluorescence‐based assay for in vitro screening of Saprolegnia inhibitors

The incidence of fish pathogenic oomycetes, Saprolegnia, has increased significantly in aquaculture since the ban of malachite green. For the efficient characterization of anti‐Saprolegnia therapeutics, simple accurate methods are required. However, the current screening methods are limited by time, and none of them are confirming the viability of treated spores or hyphae. In this study, a modified fluorescence‐based assay for the in vitro screening of Saprolegnia inhibitors has been developed. This method involves the use of FUN‐1 viability dye combined with calcofluor white M2R, and is based on the formation of orange‐red cylindrical intravacuolar structures (CIVS) in metabolically active spores, hyphae and biofilms. Heat‐killed and bronopol‐treated Saprolegnia spores, hyphae and biofilms exhibited diffuse bright green fluorescence which confirms complete loss of viability. For boric acid‐treated spores, no germination was observed. However, tiny CIVS were observed in 50% of treated spores which indicated reduction in their viability. Our results proved that FUN‐1 dye is an efficient tool to distinguish between live and dead Saprolegnia spores, hyphae and biofilms and to monitor the change in Saprolegnia viability during qualitative evaluation of potential anti‐Saprolegnia compounds.