Dietary simultaneous replacement of fish meal and fish oil with blends of plant proteins and vegetable oils in yellowfin seabream (Acanthopagrus latus) fry: Growth,digestive enzymes,antioxidant status and skin mucosal immunity

A 56‐day nutritional research was performed to examine the influence of alternative vegetal protein and lipid sources on performance of yellowfin seabream fry (Acanthopagrus latus) (0.5 ± 0.0 g). In this regard, five isoproteic (Ca. 500 g/kg) and isolipidic (Ca. 150 g/kg) diets were formulated in which fish meal (FM) and fish oil (FO) were simultaneously replaced with blends of plant proteins (PP, soybean meal and corn gluten) and vegetal oils (VO, canola and soybean oils) at 20% (SR20), 40% (SR40), 60% (SR40) and 80% (SR80) levels, respectively; meanwhile, a control diet (SR0) was formulated based on FM and FO. Growth and feed utilization were not influenced by experimental diets. The fatty acid profile of fillet drastically altered by dietary treatments. Fish fed with the SR60 and SR80 feeds had higher total protease, trypsin and α‐amylase activities than other treatments. The antioxidant enzyme activities and glutathione content in liver were enhanced in fish fed with the SR40, SR60 and SR80 diets. Skin mucosal immune parameters including total protein content, alkaline phosphatase and alternative complement pathway activities in the control group were relatively lower than the vegetal treatments. According to these results, it is recommended that 410 g/kg of FM and 45 g of FO/kg can be replaced with alternative vegetal sources in diet for A. latus fry.     

Extrapituitary gonadotropin-releasing hormone (GnRH) binding sites in goldfish

In teleosts, as in other vertebrates, the secretion of pituitary gonadotropin (GTH) is mediated by the hypothalamic decapeptide, gonadotropin-releasing hormone (GnRH). Recent findings in teleosts indicate that GnRH receptors are not restricted to the pituitary gonadotropes and are also associated with somatotropes as well as being present in a number of other tissues. In the present study, we provide novel information on GnRH binding in a number of extrapituitary tissues in goldfish. However, we do not intend to provide full characterization of GnRH binding sites in various extrapituitary tissues in goldfish as this would clearly be outside the scope of this paper. In this study we examined GnRH binding in a number of extrapituitary tissues in goldfish and observed specific binding in ovary, testis, brain, liver and kidney. No specific GnRH binding was observed in muscle, skin, gut, gill and heart. In general, the present findings together with the results of other studies carried out in our laboratory demonstrate that mature goldfish ovary and testis contain two classes of GnRH binding sites, high affinity/low capacity and low affinity/high capacity sites with binding characteristics similar to those of the pituitary GnRH receptors. The brain of goldfish was also found to contain two classes of GnRH binding sites, a super-high affinity/low capacity and a low affinity/high capacity sites. Furthermore, study of goldfish liver and kidney demonstrated the presence of a single class of GnRH binding sites with characteristics different from those of pituitary, ovary, testis and brain. Overall, it is evident that goldfish contains a family of GnRH binding sites which can be classified into four groups based on binding affinities: 1) A class of high affinity binding sites present in the pituitary, ovary and testis, 2) a class of super high affinity sites so far only detected in the brain, 3) a class of intermediate-affinity GnRH binding sites in the liver and kidney, and 4) a class of low affinity binding sites present in all the tissues containing specific GnRH binding sites except for liver and kidney.     

A study of goldfish oocyte meiosisin vitro: effects of 2,4-dinitrophenol and adenosine-5-triphosphate

Germinal vesicle migration (GVM) and/or dissolution (GVD) were measured in goldfish oocytes, treated with 17α, 20β dihydroxyprogesterone (DHP) and other compounds considered to effect the cytoskeleton and oxidative phosphorylation,in vitro. Administration of DHP reinitiated meiotic maturation, increasing GVM and GVD in goldfish oocytes. Addition of 2,4-dinitrophenol (DNP) to the incubation medium significantly inhibited DHP-induced GVM and GVD. The DNP effect was found to be partially reversible after 24 h and could be reversed fully after a further delay of approximately 24h. Treatment of goldfish oocytes with demecolcine (DE; a colchicine derivative also known as colcemid) induces GVM to the micropyle without effecting GVD; while Cytochalasin-B which inhibits microfilament polymerization impairs both GVM and GVD. Administration of DNP, significantly inhibited DE-induced GVM, suggesting that GVM as well as GVD are dependent upon the process of oxidative phosphorylation. Addition of adenosine-5′ -triphosphate (ATP) at low concentrations (0.01–0.1 mM) did not effect DHP-induced or DNP-inhibited GVD in goldfish oocytes. The present results are consistent with the idea that migration of the oocyte nucleus during meiosis reinitiation has an energy requirement and involves participation by the cytoskeleton.     

Quercetin attenuates H2O2‐induced toxicity of rooster semen during liquid storage at 4°C

Current study was carried out to examine the protective effects of quercetin against toxicity induced by hydrogen peroxide in rooster semen in vitro. Semen samples were collected from ten roosters (Ross 308 broiler breeder males, 32 weeks old) twice a week by abdominal massage method. Samples with ≥70% progressive motility were selected, pooled, diluted and used for the study. Experimental groups consisted of negative control, control that received solvent of quercetin, H₂O₂ (40 μM) and combination groups which incubated with constant dose of H₂O₂ (40 μM) plus various levels of quercetin (20, 40 and 80 μM). Measurement of total hydroperoxide (HPO), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (TAC) and superoxide dismutase activity as well as routine sperm tests were done at 0, 24 and 48 hr of storage at 4°C. Results revealed that exposure to hydrogen peroxide significantly increased HPO (138.43 ± 7.32 vs. 66.08 ± 3.97 μmol/g protein), MDA (7.21 ± 0.08 vs. 5.71 ± 2.16 μmol/g protein) and NO (0.367 ± 0.013 vs. 0.215 ± 0.011 μmol/g protein) levels and decreased sperm progressive motility (27.28 ± 1.21 vs. 47.49 ± 1.29%), and amounts of TAC (11.49 ± 0.39 vs. 15.70 ± 0.79 mmol/g protein) compared to control at 24 hr (p < 0.05). Changes at mentioned variables were repeated at 48 hr of storage. Also, co‐administration of quercetin (especially at 40 and 80 μM) with hydrogen peroxide restored the toxic effects of hydrogen peroxide on rooster semen parameters such as primary and secondary lipid peroxidative indicators and other evaluated variables. The study concluded that rooster semen enrichment with quercetin would protect lipid peroxidative and nitrosative hydrogen peroxide‐mediated damage during cold liquid storage of rooster semen.     

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods:In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results:Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC₅₀ for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion:Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). Key Words: Fed Batch, recombinant protein, Pseudomonas aeruginosa, scFv     

Sperm motility and seminal plasma characteristics in Barbus sharpeyi (Günther, 1874)

Spermatozoa concentration, ionic composition, osmolality, glucose and total protein contents of seminal plasma and sperm motility were determined in Barbus sharpeyi (Cyprinidae, Teleosotei). Spermatozoa concentration ranged from 9.77 to 20.20 × 10⁹ spermatozoa mL^?1. Osmolality (mOsmol kg^?1) and ionic contents (mM L^?1) of the seminal plasma were 274.5±9.0, 70.0±3.4 Na⁺, 28.8±0.9 K⁺, 101.7±3.1 Cl^?, 0.9±0.1 Mg²⁺ and 2.1±0.1 Ca²⁺ respectively. Total protein and glucose were 5.3±0.2 g L^?1 and 76.7±4.3 mM L^?1 respectively. Sperm motility was initiated in a hypo‐osmotic condition, composed of either an ionic (KCl or NaCl) or a non‐ionic (sucrose) activation medium. Duration of sperm motility was very short: <2 min after activation in distilled water. Percentage of motile spermatozoa was significantly higher in an activation medium containing NaCl compared with that of distilled water. An activating medium containing NaCl or KCl higher than 150 mM or sucrose higher than 275 mM totally inhibited the activation of sperm motility. Immediately after sperm activation, wave(s) propagated along the flagellum, but waves were restricted to the proximal part of the flagellum (close to the head) at 1 min post activation. Studied characteristics in the present study were compared with those of other cyprinids for understanding inter‐species differences.     

Mucoadhesive Chitosan Electrospun Nanofibers Containing Tetracycline and Triamcinolone as a Drug Delivery System

The mucoadhesive Chitosan (CS) nanofibers as a drug delivery system were developed. Chitosan was modified via the immobilization of thiol groups from L-cysteine as a mucoadhesive reagent. The mucoadhesive properties of the chitosan nanofibers were evaluated by tensiometer set and via tensile studies. Drug and mucoadhesive agent loading lead to decrease diameters and increased porous of nanofibers. The release of Tetracycline (Tet) and Triamcinolone (Tri) were increased with increasing immersion time and it became constant at long immersion times. Mucoadhesion studies were done at pH 2–7 and in pH 6 maximum mucoadhesive properties observed. Release studies demonstrated a sustained release of both drug continued up to 48 hours. Microbial studies were performed on the nanofibers. The drug delivery system represented a novel tool for improve the therapeutic efficacy of various drugs that are poorly absorbed from the gastrointestinal tract. Also it is an efficient system for treatment of oral ulceration.     

Response of European seabass (<Emphasis Type="Italic">Dicentrarchus labrax</Emphasis>) to canola oil diets: effect on growth performance,fish health and liver and intestine histomorphology

This study was undertaken to assess the effects of feeding European seabass (Dicentrarchus labrax) canola oil-added diets on growth, health status and liver and intestine histomorphology. Seabass (56.18 ± 0.16 g initial body weight) were fed one of three fish meal-based diets with ~48 % crude protein and ~16.0 % lipid, combining fish oil (FO) and canola oil (CO) for 12 weeks. The diets contained: zero (control, CTRL), 45 (CO50) or 63 (CO70) g CO kg^?1 assigned in triplicates to three dietary groups. The results indicated that neither dietary oil type (FO or CO) nor CO level adversely affected (P > 0.05) the growth, feed utilization or major blood constituents’ composition as an indicator of the overall health status of fish. Despite the CO diets influence on head kidney macrophage activity being unappreciable (P > 0.05), there was a reducing trend with an increase in CO level incorporation. The CO70 diet induced a minor fat infiltration in hepatocytes and leucocytes infiltration, hyperplasia of the basal nuclei and supranuclear vacuolization of the enterocytes of the distal intestine. The present observations suggest that it is possible to incorporate up to 63 g CO Kg^?1 in the feed for European seabass juveniles without major negative effects on growth, health status or liver and intestinal histomorphology.     

The Effects of Estrogen Receptors' Antagonist on Brain Edema,Intracranial Pressure and Neurological Outcomes after Traumatic Brain Injury in Rat

Background:

In previous studies, the neuroprotective effect of 17β-estradiol in diffuse traumatic brain injury has been shown. This study used ICI 182,780, a non-selective estrogen receptor antagonist, to test the hypothesis that the neuroprotective effect of 17β-estradiol in traumatic brain injury is mediated by the estrogen receptors.

Methods:

The ovariectomized rats were divided into eight groups. Brain injury was induced by Marmarou’s method. Estrogen was injected 30 minutes after traumatic brain injury, and ICI 182,780 was injected before traumatic brain injury and also before estrogen treatment. In one group only ICI 182,780 was injected. The brain water content and Evans blue dye content were measured 24 and 5 hours after traumatic brain injury, respectively. The neurologic outcomes and intracranial pressure were assessed before, 4, and 24 hours after traumatic brain injury.

Results:

Brain water content and Evans blue content were less in estrogen-treated group comparison to vehicle group. ICI 182,780 eliminated the effects of estrogen on brain edema and brain blood barrier permeability. Intracranial pressure was increased significantly after trauma, and estrogen decreased intracranial pressure at 4 and 24 hours after traumatic brain injury in comparison to vehicle. This inhibitory effect was also eliminated by treatment with ICI182,780. ICI 182,780 also inhibited the estrogen induced increase in neurologic outcomes following traumatic brain injury. However, the use of ICI 182,780 alone had no neuroprotective effect after traumatic brain injury.

Conclusion:

The results suggest that classical estrogen receptors have probably a role in the neuroprotective function of estrogen following traumatic brain injury.Key Words: Estrogens, Intracranial pressure (ICP), Brain edema