Molecular cloning and expression profiling of procollagen α1 (I) of cultured Pacific bluefin tuna

Type I collagen is widely distributed in most organs in teleosts. It plays a role not only in intercellular adhesion, but also in molecular signaling. In this study, Pacific bluefin tuna (PBT) procollagen α1 (I) cDNA was cloned and characterized. The nine fragments of a procollagen α1 (I) chain cDNA clone were prepared and spliced together to create the complete coding region. The resulting amino acid sequence was homologous with that of other teleosts. The mRNA expression profile of PBT procollagen α1 (I) in various tissues and the phylogenetic analysis with other vertebrate procollagen α1 (I) chains suggest that PBT procollagen α1 (I) could be a precursor form of the PBT type I collagen α1 chain. In addition, its level of expression in PBT larvae and early juveniles gradually increased with somatic growth. This increase was related to the standard length, wet body weight, and protein content of each individual fish. Therefore, the expression profile of procollagen α1 (I) may be a useful indicator for somatic growth in fish larvae and juveniles.     

Soy protein concentrate as an alternative in replacement of fish meal in the feeds of hybrid grouper,brown‐marbled grouper (Epinephelus fuscoguttatus) × giant grouper (E. lanceolatus) juvenile

Hybrid grouper juveniles (body weight, 6.1 ± 0.7 g) (brown‐marbled grouper, Epinephelus fuscoguttatus × giant grouper, E. lanceolatus) were fed with six isoproteic (50% crude protein) and isolipidic (12% crude lipid) feeds containing different levels of soy protein concentrate (SPC) in replacement of fish meal (SPC at 20%, 30%, 40%, 50% and 60% protein) and control feed (SPC0) for 6 weeks. Hybrid grouper juveniles were cultured in 100‐L fibreglass tank equipped with flow‐through water system and fed twice a day to apparent satiation level. The highest and lowest growth was recorded in fish fed SPC20 and SPC60 respectively. However, growth of SPC20 was not significantly higher than those fed SPC0, SPC30, SPC40 and SPC50 (p > .05). A decreasing growth trend was observed with the increasing level of SPC from feed SPC40 to SPC60. A noticeable better feed utilization was also observed in fish fed SPC0, SPC20, SPC30, SPC40 and SPC50 compared to fish fed SPC60 (p < .05). The fish condition factor, hepatosomatic index, viscerosomatic index and whole body proximate content of the fish were not affected by the graded levels of SPC. However, the body lipid content was significantly lower in fish fed SPC40 to SPC60 (p < .05). The apparent digestibility coefficient (ADC) of protein and lipid was significantly higher in fish fed SPC0 and SPC20 compared to other dietary treatments (p < .05). Based on the regression analysis on specific growth rate, the study suggests that the hybrid grouper grow best at 21.4% and can utilize up to 50% inclusion level of SPC in protein without significantly affect their growth and its body condition.     

Chimeric mice with humanized liver as a model for testing organophosphate and carbamate pesticide exposure

BACKGROUND

Diagnosis of acute intoxication with organophosphate (OP) or carbamate (CM) pesticides in humans is achieved by measuring plasma butyrylcholinesterase (BuChE) activity. However, BuChE activity is not an ideal biomarker in experimental animal models. The aim of this study was to establish an experimental mouse model for evaluating exposure to OP and CM pesticides by monitoring BuChE activity using chimeric mice in which the liver was reconstituted with human hepatocytes.

RESULTS

A single oral administration of acephate (300 mg/kg), chlorpyrifos (10 mg/kg), fenobucarb (300 mg/kg) or molinate (250 mg/kg) in chimeric mice led to inhibition of >95%, > 95%, 28% and 60% of plasma BuChE activity after 7, 0.5, 0.5 and 7 h, respectively. Dose‐dependent decreases in plasma BuChE activity were also observed for acephate and chlorpyrifos. A 5‐day repeated‐dose study with 10 or 30 mg/kg acephate found a constitutive reduction in plasma BuChE activity to 80% and 70% of pre‐dose levels, respectively.

CONCLUSION

Changes in plasma BuChE activity in chimeric mice with humanized liver clearly reflected the exposure levels of OP and CM pesticides. These results suggest that the humanized‐liver mouse model may be suitable for estimating levels of exposure to these pesticides in humans. © 2017 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.     

The optimal dietary level of ascorbic acid for juvenile Pacific bluefin tuna, Thunnus orientalis

A feeding experiment was conducted to evaluate the efficacy of, and determine the requirements for dietary AsA (l-ascorbic acid) using its popular derivative l-ascorbyl-2-monophosphate-Mg (APM) for juvenile Pacific bluefin tuna (PBT). The five test diets (with APM-0, -400, -800, -1200 and -1600) were prepared by adding 0, 400, 800, 1200 and 1600 mg of APM kg diet^?1, respectively. These diets, together with chopped sand lance (the control diet), were administered, in duplicate to 0.27 g PBT at 25 days after hatching (300 juveniles per 15 m³ tank). The PBT were fed to apparent satiety 6 times a day for 2 weeks. While final body weight, feed conversion and survival showed no differences among diets APM 400–1600, PBT fed APM-0 indicated lower growth performance together with anorexia, dark pigmentation and ataxia from 5 days and reached 80 % mortality at 12 days after the start of the feeding trial. In addition, liver and brain ascorbic acid (AsA) concentrations increased with increasing dietary APM levels, the values were similar in PBT fed diets with APM-1200 and APM-1600, and liver and brain concentrations of AsA were significantly higher in these fish than in the PBT fed the other diets (P < 0.05). The PBT responded to increasing dietary APM, and the fish fed APM-1200 had the highest weight gain, protein and lipid content. The data indicate that PBT can utilize APM as an AsA source and suggest that dietary supplementation with APM-1200 (454 mg AsA kg^?1 diet) would result in satisfactory growth.     

Quality improvement of the thermal gel of horse mackerel by sodium citrate and its inhibition of the insoluble metalloproteinase

Modori-inducing proteinases (MIPs) in horse mackerel and the effect of sodium citrate on the modori phenomenon were investigated. Cysteine protease caused the modori phenomenon only at 60 ℃, while metalloproteinase caused it at both 60 ℃ and 70 ℃. Moreover, the autolysis of myofibrillar proteins was mainly inhibited by ethylene diamine tetra-acetic acid (EDTA), verifying the existence of insoluble metalloproteinase. From the results comparing thermal gel properties and protease activities, we confirmed that increased activity of insoluble metalloproteinase corresponded to the weakening of the thermal gel. Thus, insoluble metalloproteinase was the major MIP in horse mackerel. Moreover, sodium citrate could markedly inhibit insoluble metalloproteinase activity at 60 ℃. The addition of sodium citrate significantly improved the breaking force of the thermal gel at 60 ℃ and 70 ℃, suggesting that sodium citrate could suppress the modori phenomenon by inhibiting insoluble metalloproteinase. Therefore, it is desirable to use sodium citrate in horse mackerel surimi-based products to prevent the modori phenomenon.

     

Antioxidant activity of some protein hydrolysates and their fractions with different isoelectric points

Antioxidant activities of commercially available enzymatic hydrolysates of milk and plant proteins were examined. Among them, soy protein and wheat gluten hydrolysates showed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity and antioxidation activity against linoleic acid oxidation in emulsion systems. Peptide fractions with higher antioxidant activities than crude enzymatic hydrolysates of gluten and soy protein were prepared without toxic solvents and reagents. Peptides in these plant protein hydrolysates were fractionated on the basis of the amphoteric nature of sample peptides by preparative isoelectric focusing without adding chemically synthesized carrier ampholytes, which is termed autofocusing. The acidic fractions from both protein hydrolysates showed stronger DPPH radical scavenging activities than the basic fractions, while the basic fractions strongly suppressed 2,2'-azobis (2-amidinopropane) dihydrochloride-induced oxidation of linoleic acid in an emulsion system. These acidic and basic peptide fractions would be useful to examine the mechanism underlying the antioxidant activities of peptides in food.     

Pepsinogens and pepsins from mandarin fish (Siniperca chuatsi)

Four pepsinogens (PG-I, PG-II, PG-III(a), and PG-III(b)) were highly purified from the stomach of the freshwater fish mandarin fish (Siniperca chuatsi) by ammonium sulfate fractionation, anion exchange, and gel filtration. The molecular masses of the four purified PGs were 36, 35, 38, and 35 kDa, respectively. All the pepsinogens converted into their active form pepsins within a few minutes under pH 2.0. The optimum pH and temperature of the four enzymes were 3.0-3.5 and 45-50 degrees C, using hemoglobin as a substrate. The N-terminal amino acid sequences of PG-I and PG-II were determined to the 12th and 17th amino acid residues, respectively. Western blot analysis using antisea bream polyclonal antibodies cross reacted with PG-I, PG-II, and PG-III(b) while no cross reaction with PG-III(a) was detected, suggesting the diversity of pepsinogens in fish.     

The tendency of damage of stone walls in terrace fields in the northwestern Kumamoto City by the 2016 Kumamoto Earthquake and heavy rain in 2016 and 2006

Paddy and Water Environment - The topographical features of farmland stone walls that were damaged by two heavy rains and the 2016 Kumamoto Earthquake were studied. Topographical analyses were...     

Development of new natural polymer-based wood adhesives III: effects of glucose addition on properties of chitosan

The effects of a Maillard reaction between glucose and chitosan on the resultant chitosan films and the bonding properties of chitosans with different molecular weights were investigated. In film preparation, chitosan and glucose were dissolved in 1% acetic acid and dried in a Petri dish at 50°C. The bonding properties of the Maillard-reacted chitosan in three-ply plywood were evaluated by a tensile shear test. The weight, color, free amino groups, insoluble fraction, and thermal properties of the film changed significantly as the amount of added glucose increased. However, few differences in these properties due to the use of chitosan of different molecular weights were observed. The effect of glucose addition on the tensile strength of the film differed for chitosans of different molecular weights. Improvement of the strength by 10 wt% glucose addition was observed in low-molecular-weight chitosans. The dry- and wet-bond strengths were significantly enhanced with increasing glucose addition for low-molecular-weight chitosans. In addition, good bond strength was maintained even in 1% acetic acid solution. It was thus clarified that the bonding properties of low-molecular-weight chitosan were improved markedly by the environmentally safe method of glucose addition.