Berberine attenuates oxidative stress and hepatocytes apoptosis via protecting mitochondria in blunt snout bream <Emphasis Type="Italic">Megalobrama amblycephala</Emphasis> fed high-fat diets

High-fat diets may have favorable effects on growth and cost, but high-fat diets often induce excessive fat deposition, resulting in liver damage. This study aimed to identify the hepatoprotective of a Chinese herb (berberine) for blunt snout bream (Megalobrama amblycephala). Fish were fed with a normal diet (LFD, 5 % fat), high-fat diet (HFD, 15 % fat) or berberine-supplemented diets (BSD, 15 % fat with berberine 50 or 100 mg/kg level) for 8 weeks. After the feeding, histology, oxidative status and mitochondrial function of liver were assessed. The results showed that HFD caused fat accumulation, oxidative stress and apoptosis in hepatocytes of fish. Hepatocytes in HFD group appeared to be hypertrophied, with larger liver cells diameter than these of LFD group. Berberine-supplemented diets could attenuate oxidative stress and hepatocytes apoptosis. HFD induced the decreasing mitochondrial complexes activities and bulk density and surface area density. Berberine improved function of mitochondrial respiratory chain via increasing the complex activities. Moreover, the histological results showed that berberine has the potential to repair mitochondrial ultrastructural damage and elevate the density in cells. In conclusion, our study demonstrated that berberine has attenuated liver damage induced by the high fat mainly via the protection for mitochondria.     

Molecular analysis of the major capsid genes (<Emphasis Type="Italic">g23</Emphasis>) of T4-type bacteriophages in an upland black soil in Northeast China

Bacteriophages (phages) are the most abundant biological entities on the planet and are important as the greatest genomic reservoirs in both marine and terrestrial environments. In this study, we analysed T4-type phage communities in an upland black soil by monitoring g23 clones in DNA extracted from seasonal soil samples with no fertilizer, chemical fertilizers, chemical fertilizers plus manure, and natural restoration treatments. PCR products with degenerate primers MZIA1bis and MZIA6 were subjected to denaturing gradient gel electrophoresis. In total, 46 clones with different g23 sequences were obtained. Phylogenetic analyses indicated that T4-type phage communities in the upland black soil were distinctly different from those in marine environments and in an Antarctic lake, which strongly suggested that T4-type phage communities in soil differed from those in aquatic environments. Among 46 clones, 18 clones formed clusters with the clones from rice field soils, 14 clones formed three new clusters, and 13 clones were left as ungrouped, which indicated that T4-type phage communities in the upland black soil were relatively similar to those in rice field soils but that specific communities also inhabit in the upland black soil exclusively.     

Synthetic seed production by encapsulating nodal segment of <Emphasis Type="Italic">Capparis decidua</Emphasis> (Forsk.), in vitro regrowth of plantlets and their physio biochemical studies

Synthetic seed technology is an emerging and broadly used technique in the field of plant biotechnology to conserve economically important plants. In the present study, nodal segments of Capparis decidua were entrapped in calcium alginate gel matrix to produce firm and uniform synthetic seeds. 3% sodium alginate and 75 mM calcium chloride were found best for encapsulation. Among all the concentrations and combinations of thidiazuron (TDZ) either singly or with indole -3- acetic acid (IAA) augmented in Murashige and Skoog medium used, TDZ (5.0 µM) + IAA (0.5 µM) was found most effective in conversion of synthetic seeds into plantlets as 79% plantlets were developed on this combination with 13.2 ± 0.87 shoots and 5.5 ± 0.40 cm shoot length after 8 weeks of culture. Further, synthetic seeds stored at low temperature (4 °C) can retain their viability up to 4 weeks and showed maximum conversion rate (93%) into plantlets, when placed back to regeneration medium. Root formation was also occurred in the same regeneration medium and roots were healthy. Plantlets were successfully hardened in culture room in plastic cups filled with sterile vermiculite and after 4 weeks, they were transferred to greenhouse where they exhibited normal growth with 80% survival. Growth parameters were evaluated in micropropagated plants and compared with the seedlings of same age. Effect of different days of acclimatization were also recorded on various physio-biochemical activities and showed a positive response that can be interpreted as better protection mechanism of micropropagated plants against the stress possibly generated due to reactive oxygen species when transferred to ex vitro environment.     

Effect of Humic Acid on the Nitrate Removal by Strong Base Anion Exchanger Supported Nanoscale Zero-valent Iron Composite

To probe the effect of common coexist substances on the nitrate removal by polymeric resin supported nanoscale zero-valent iron composite (D201-nZVI), humic acid (HA) was added into the nitrate removal system to elaborate the different interactions between each two and among all in the system including HA, nitrate, and D201-nZVI. The results showed that the effect of HA on the reduction of nitrate by D201-nZVI was concentration-dependent. At low HA concentration (<?5 mg/L), HA coating formed by the HA adsorption on the surface of the nZVI particles enhanced the dispersion of the particles, which led to a more evenly distribution of nZVI particles in the solution, and thus a higher nitrate reduction activity. When HA concentration was increased to 5 mg/L or more, the competitive adsorption of HA and NO₃^? on the surfaces of the D201-nZVI dominated, and the nitrate removal rate and ammonia nitrogen production were decreased. When the HA concentration reached to a further high level (>?20 mg/L), HA acted as an electron shuttle to accelerate the reduction of Fe(III) to Fe(II) in the D201-nZVI, and thus the nitrate reduction rate was accordingly enhanced. The ammonia production increased by 24.8% at HA concentration of 40 mg/L as compared with that of the control (without addition of HA). This research elucidated the interaction of HA within different HA concentration in the complicate system of anions removal by organic support-nanoscale metal particle composite, which may shade some new light on the potential application of nanoscale zero-valent materials in the practical remediation of natural water.     

An attempt to improve the reproductive efficiency of Nile tilapia brood stock fish

A field study was conducted on brood stock Nile tilapia to increase the propagation. Both sexes were individually stocked into Habas (enclosures) in an earthen pond and fed for 19 days on a basal diet supplemented with different additives at graded levels of each (0.5, 1.0 and 2.0 g Therigon^®; 1.0, 2.0, and 3.0 g Nuvisol Hatch P^®; 20, 40 and 60 mg Gibberellic acid and 700, 900 and 1,100 mg L-carnitine/Kg diet). The obtained results were evaluated, and the best treatment for each sex was chosen for mating. Results indicated that all pretreatments for male and female brood stocks of Nile tilapia positively affected the total count of the offspring produced. Yet, the Haba, in which the females were pretreated with 0.5 g Therigon^®/Kg diet and the males pretreated with 700 mg L-carnitine/Kg diet, gave the highest total count of the offspring comparing with the other Habas. But, because of the high feed cost due to the additives cost, 0.5 g Therigon^®/kg diet as pretreatment for ♀ only (3rd Haba), 2 g Nuvisol Hatch P^®/Kg diet as pretreatment for ♀ only (5th Haba), followed by 0.5 g Therigon^® and 700 mg L-carnitine/Kg diet for ♀ and ♂, respectively (4th Haba), respectively were the best economically.     

Effect of sublethal α-cypermethrin exposure on main macromolecules concentration,energy content,and malondialdehyde concentration in free-feeding <Emphasis Type="Italic">Danio rerio</Emphasis> larvae

α-Cypermethrin (Cyp) is a synthetic insecticide used to control pests in agricultural crops and to protect human health against noxious insects; this toxic can reach aquatic systems through ground infiltration or by runoff and could affect the aquatic biota. The present study was aimed at evaluating the acute toxicity of Cyp on zebrafish (Danio rerio) exogenous feeding larvae of 10 and 20 days post-fertilization (dpf), and of sublethal concentrations on only 10-dpf larvae. Proteins, lipids, carbohydrates, glycogen concentration, and total energy contents, as well as malondialdehyde (MDA) quantification, through thiobarbituric acid reactive substances, as a lipid peroxidation biomarker, were assessed in free-feeding larvae exposed to sublethal Cyp concentrations. The LC₅₀ for 10-dpf larvae was 1.94 µg L^?1, and these were more sensitive than 20-dpf larvae (3.56 µg L^?1). The amount of protein, carbohydrates, and glycogen were not significantly affected (p > 0.05), but sublethal Cyp concentrations exposure caused decrement in lipids from 9.05 to 3.74 µg larva^?1, as well as a reduction in MDA and in the total energy content, which affected significantly the development of this fish. Although Cyp is considered an insecticide of reduced residual effect in the environment, the present study revealed that relatively low Cyp concentrations produced significant toxic effects on exogenous feeding fish larvae, a situation that could contribute to increase deaths during this already critical developmental stage in which high mortality is observed frequently.     

Gastrointestinal and hepatic enzyme activities in juvenile silvery-black porgy (<Emphasis Type="Italic">Sparidentex hasta</Emphasis>) fed essential amino acid-deficient diets

As amino acids (AAs) are vital molecules in the metabolism of all living organisms and are the building blocks of enzymes, a 6-week feeding trial was conducted for determining the influence of dietary essential amino acid (EAA) deficiencies on pancreatic, plasma, and hepatic enzyme activities in silvery-black porgy (initial weight 4.7?±?0.01 g) juveniles. Eleven isoproteic (ca. 47%) and isoenergetic (ca. 20.5 MJ kg^?1) diets were formulated including a control diet, in which 60% of dietary nitrogen were provided by intact protein (fish meal, gelatin, and wheat meal) and 40% by crystalline AA. The other 10 diets were formulated by 40% reduction in each EAA from the control diet. At the end of the experiment, fish fed with threonine-deficient diet showed the lowest survival rate (P?<?0.05), whereas growth performance decreased in fish fed all EAA-deficient diets, although the reduction in body growth varied depending on the EAA considered. Pancreatic enzymes (trypsin, lipase, α-amylase, and carboxypeptidase A) activities significantly decreased in fish fed the EAA-deficient diets in comparison with fish fed the control diet (P?<?0.05). Fish fed with the arginine-deficient diet had the highest plasma and liver alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels (P?<?0.05). Plasma and liver lactate dehydrogenase and superoxide dismutase showed the highest and lowest values, respectively, in fish fed the arginine and lysine-deficient diets (P?<?0.05). Plasma metabolites were significantly affected by dietary EAA deficiencies (P?<?0.05). The results of this study suggesting dietary EAA deficiencies led to reduction in growth performance as well as pancreatic and liver malfunction. Furthermore, arginine and lysine are the most limited EAA for digestive enzyme activities and liver health in silvery-black porgy.     

Evaluation of interferon gamma (IFN-γ) of <Emphasis Type="Italic">Labeo rohita</Emphasis> as an immunomodulator: in vitro expression model

Interferon gamma (IFN-γ) or type II interferon is a cytokine that is critical for innate and adaptive immunity against viral and some bacterial and protozoal infections. The importance of IFN-γ in the immune system lies in its ability to inhibit viral replication directly and most importantly from its immunomodulatory effects. Previously, we successfully co-administered IFN-γ along with GAPDH gene of Edwardsiella tarda as bicistronic DNA vaccine in Labeo rohita. In order to ascertain the individual role of IFN-γ, the present study involves cloning and expression of 552-bp IFN-γ open-reading frame (ORF) of L. rohita in striped snakehead (SSN-1) cell line using eukaryotic expression vector system (pQE-TriSystem) followed by transfection in peripheral blood lymphocytes (PBMCs) to evaluate its immunomodulatory ability in comparison to polyinosinic-polycytidylic acid (Poly I:C)-treated PBMCs. The 18.7-kDa protein, expressed in the pQE-IFNγ-transfected SSN-1 cells, reacted with anti-His antibody in Western blot confirming it to be recombinant IFN-γ, whereas the relative expression of IFN-γ, iNOS, Mx, and IL-1β genes in PBMCs was quantified at 24 h and 48 h post treatment by qPCR. The comparative kinetics of all four genes showed significantly (p?<?0.05) high upregulation pattern in both pQE-IFNγ-transfected cell group and Poly I:C-treated cell group demonstrating recombinant IFN-γ as an equally efficient inducer like Poly I:C. Thus, our in vitro experiment results highlight the immunomodulatory potential of recombinant IFN-γ as an analogue to synthetic Poly I:C which warranted future studies to further explore the potential of recombinant IFN-γ as an effective vaccine adjuvant against different microbial invasion.     

Rapid de-adhesion of northern pike <Emphasis Type="Italic">Esox lucius</Emphasis> eggs using sodium hypochlorite

The elimination of egg stickiness is required for effective artificial reproduction of northern pike, but until now, available methods have required at least 40 min. Sodium hypochlorite was tested under laboratory conditions, and exposure to aqueous concentrations of 0.025–0.05% for 40 s effectively eliminated stickiness without adverse effects. Fertilization and hatching rates in laboratory trials were similar to those observed in eggs treated with traditional methods using clay or milk for 40 or 60 min, respectively, as well as those without treatment. Testing using conventional hatchery incubation techniques did not reveal differences in fertilization rates, while the number of hatched larva was significantly higher in eggs treated with sodium hypochlorite vs. clay. Eggs treated with sodium hypochlorite retained transparency, which facilitated monitoring of embryo development.