Effectiveness of l‐ascorbyl‐2‐monophosphate Na/Ca as a vitamin C source for yellowtail Seriola quinqueradiata juveniles

A feeding experiment was conducted to examine the efficacy of a vitamin C (L‐ascorbic acid, AsA) derivative, l ‐ascorbyl‐2‐monophosphate Na/Ca (AMP‐Na/Ca) by comparing to l ‐ascorbyl‐2‐monophosphate‐Mg (AMP‐Mg) in terms of growth, AsA content in tissues, and hematology in yellowtail Seriola quinqueradiata juvenile (1.10 ± 0.01 g). Furthermore, a stress resistance test for the species fed AMP‐Na/Ca was also conducted. In experiment 1, 11 test pellet diets with different levels of AMP‐Na/Ca (0, 12, 43, 88, 440 and 881 mg AsA kg^?1diet) and AMP‐Mg (0, 16, 52, 106, 595 and 1164 mg AsA kg^?1 diet) were formulated and fed to the yellowtail three times per day for 50 days. In both the vitamin C sources, the survival rates of the fish fed diets without supplemental AsA were lower than 50% at day 20, and more than 50% mortality occurred in fish that fed the diets containing 12 or 16 mg AsA kg^?1 after day 30. However, no significant differences were detected in survival and growth among fish that fed the diets containing more than 43 mg AsA kg^?1. Liver and brain AsA concentrations generally increased with increasing dietary AsA level in both sources. In experiment 2, test diets were formulated to contain 43, 88, 440 and 881 mg AsA kg^?1 using AMP‐Na/Ca, and after 60 days of feeding, yellowtail juveniles were subjected to low salinity and air exposure stress. The fish that received diets with 440 mg AsA kg^?1 showed significantly higher tolerance to low salinity stress and higher survival rate in air exposure stress than those of other groups. The present study demonstrated that yellowtail juveniles could utilize AMP‐Na/Ca as an AsA source like AMP‐Mg, and that supplementation of 43–52 mg AsA kg^?1 diet was optimum for normal growth. However, this study showed that dietary inclusion of 440 mg AsA kg^?1 would be necessary to improve stress resistance of this species.     

Evaluating environmental impacts of the Japanese beef cow–calf system by the life cycle assessment method

The objectives of this study were to evaluate the environmental impacts of a beef cow–calf system using a life cycle assessment (LCA) method and to investigate the effects of scenarios to reduce environmental impacts on the LCA results. The functional unit was defined as one marketed beef calf, and the processes associated with the cow–calf life cycle, such as feed production, feed transport, animal management, the biological activity of the animal and the treatment of cattle waste were included in the system boundary. The present results showed that the total contributions of one beef calf throughout its life cycle to global warming, acidification, eutrophication and energy consumption were 4550 kg of CO₂ equivalents, 40.1 kg of SO₂ equivalents, 7.0 kg of phosphate (PO₄) equivalents and 16.1 GJ, respectively. The contribution of each process to the total environmental impact in each environmental impact category showed a similar tendency to the contribution of each process in each environmental category reported in the case of the beef fattening system as a whole. The results from this analysis showed that shortening calving intervals by 1 month reduced environmental impacts by 5.7–5.8% in all the environmental impact categories examined in the current study, and increasing the number of calves per cow also reduced environmental impacts in all the categories, although the effects were smaller.     

Differentiation of canine bone marrow stromal cells into voltage- and glutamate-responsive neuron-like cells by basic fibroblast growth factor

We investigated the in vitro differentiation of canine bone marrow stromal cells (BMSCs) into voltage- and glutamate-responsive neuron-like cells. BMSCs were obtained from the bone marrow of healthy beagle dogs. Canine BMSCs were incubated with the basal medium for neurons containing recombinant human basic fibroblast growth factor (bFGF; 100 ng/ml). The viability of the bFGF-treated cells was assessed by a trypan blue exclusion assay, and the morphology was monitored. Real-time RT-PCR was performed to evaluate mRNA expression of neuronal, neural stem cell and glial markers. Western blotting and immunocytochemical analysis for the neuronal markers were performed to evaluate the protein expression and localization. The Ca²⁺ mobilization of the cells was evaluated using the Ca²⁺ indicator Fluo3 to monitor Ca²⁺ influx. To investigate the mechanism of bFGF-induced neuronal differentiation, the fibroblast growth factor receptor inhibitor, the phosphoinositide 3-kinase inhibitor or the Akt inhibitor was tested. The bFGF treatment resulted in the maintenance of the viability of canine BMSCs for 10 days, in the expression of neuronal marker mRNAs and proteins and in the manifestation of neuron-like morphology. Furthermore, in the bFGF-treated BMSCs, a high concentration of KCl and L-glutamate induced an increase in intracellular Ca²⁺ levels. Each inhibitor significantly attenuated the bFGF-induced increase in neuronal marker mRNA expression. These results suggest that bFGF contributes to the differentiation of canine BMSCs into voltage- and glutamate-responsive neuron-like cells and may lead to the development of new cell-based treatments for neuronal diseases.     

Mitigation of nitrous oxide (N2O) emission from swine wastewater treatment in an aerobic bioreactor packed with carbon fibers

Mitigation of nitrous oxide (N₂O) emission from swine wastewater treatment was demonstrated in an aerobic bioreactor packed with carbon fibers (CF reactor). The CF reactor had a demonstrated advantage in mitigating N₂O emission and avoiding NOx (NO₃ + NO₂) accumulation. The N₂O emission factor was 0.0003 g N₂O‐N/gTN‐load in the CF bioreactor compared to 0.03 gN₂O‐N/gTN‐load in an activated sludge reactor (AS reactor). N₂O and CH₄ emissions from the CF reactor were 42 g‐CO₂ eq/m³/day, while those from the AS reactor were 725 g‐CO₂ eq/m³/day. The dissolved inorganic nitrogen (DIN) in the CF reactor removed an average of 156 mg/L of the NH₄‐N, and accumulated an average of 14 mg/L of the NO₃‐N. In contrast, the DIN in the AS reactor removed an average 144 mg/L of the NH₄‐N and accumulated an average 183 mg/L of the NO₃‐N. NO₂‐N was almost undetectable in both reactors.     

Effects of dietary arginine and lysine levels on growth performance and biochemical parameters of juvenile Japanese flounder Paralichthys olivaceus

ABSTRACT: To investigate the effects of disproportionate levels of dietary arginine and lysine on juvenile Japanese flounder Paralichthys olivaceus , growth performance and biochemical parameters were evaluated by feeding five test diets, comprising different levels of arginine and lysine, to triplicate groups of juveniles (initial bodyweight 1.85 g) for 40 days. Crystalline amino acids were supplemented to test diets to correspond to the amino acid pattern found in the whole body protein of the Japanese flounder, except for arginine and lysine. After the feeding trials, plasma arginine, lysine and urea levels, excreted ammonia-N, and liver arginase activity were analysed. Survival, specific growth rate, feed conversion efficiency, and apparent protein retention were adversely affected ( P  <   0.05) by the deficiency in dietary arginine or lysine concentrations. An excess of either lysine or arginine in the diet did not depress growth when the diets contained adequate levels of either arginine or lysine, proving that there is no evidence for an arginine–lysine antagonism. Results for plasma arginine, lysine and urea levels, excreted ammonia, and liver arginase activity also demonstrated that Japanese flounder juveniles are not sensitive to excess dietary levels of lysine and arginine.     

The addition of calcium ion chelator,EGTA to thawing solution improves fertilizing ability in frozen–thawed boar sperm

In our previous study, seminal plasma effectively suppressed the induction of sperm to capacitation‐like status and acrosome loss during the thawing process. However, because boar seminal plasma is contaminated with various kinds of bacteria and/or viruses, it is necessary to develop a thawing solution without animal‐derived materials. In this study, we focused on the increase of intracellular Ca²⁺ ([Ca²⁺]_i) in sperm after thawing and the negative effects of sperm qualities. After thawing, the fluorescent intensity of [Ca²⁺]_i indicator, Fluo‐3/AM, and the level of phosphorylated tyrosine residue of protein were increased in the sperm. Next, we investigated whether the addition of Ca²⁺ chelators (ethylenediaminetetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA)) improved post‐thawed sperm motility. When the frozen–thawed sperm were treated with 6 mmol/L EDTA + 6 mmol/L EGTA, sperm motility was significantly increased as compared with control (6 mmol/L EDTA alone) at all incubation periods (P < 0.05). The combinational treatment significantly suppressed the elevation of [Ca²⁺]_i and the tyrosine phosphorylation, which improved the acrosomal status and fertilizing ability in vitro. Furthermore, when the thawing method was applied for artificial insemination, the fertilization rate was significantly higher than control (P < 0.05, 33% vs. 82%). Thus, we conclude that the addition of EDTA + EGTA to thawing solution is a beneficial tool for artificial insemination using frozen–thawed boar sperm.     

Seasonal variations in the incidence of pine wilt and infestation by its vector, Monochamus alternatus, near the northern limit of the disease in Japan

During the last few decades, pine wilt disease has spread to cool-climate regions in Japan and, more recently, the potential risk of it spreading into the European midwest has also become a concern. In a coastal pine stand (84.7?ha) in Akita, near the northern limit of pine wilt disease in Japan, we investigated seasonal variations in the incidence of damage caused by the disease to trees and oviposition by the disease’s insect vector, Monochamus alternatus, during a two-year period. Foliage discoloration occurred throughout each year, and its seasonal variation showed a bimodal pattern in Pinus thunbergii (a higher peak in May–June and a smaller peak in October) and a clear peak in June in P. densiflora, which differed from the patterns in seasonal variation seen for warm-climate regions. Oviposition scars by M. alternatus were found in 40–45% of the trees damaged each year. The percentage of trees that had oviposition scars was higher in P. thunbergii than in P. densiflora. This appeared to reflect the difference in seasonal discoloration pattern between the two species. Analysis of the oviposition risk showed that trees that exhibited discoloration starting between July and October had a significantly higher risk or significantly higher oviposition scar densities, particularly for those that became discolored between August and September (2.5–14.6-fold higher risk than during other months). Oviposition scar densities per damaged tree were similar within the period of higher oviposition risk. Considering both oviposition risks and scar densities, we concluded that trees with discoloration that become apparent between July and October are important targets for preventing the spread of pine wilt disease in Akita.     

Interactive effect of two methyl‐group sources on the growth and phospholipids’ content of kuruma shrimp Marsupenaeus japonicus Bate post‐larvae

Six micro‐bound diets were formulated to contain three levels of choline chloride (CC) (0.0, 0.6 and 1.2 g kg⁻¹) and 2 levels of methionine (Met) (0 and 15 g kg⁻¹). Soybean protein isolates (SPI) were used as the main protein source for its limited Met content. A significant (P < 0.05) interaction was determined between CC and Met on the survival (S %), weight gain (WG%), specific growth rate (SGR % day⁻¹), feed efficiency ratio (FER), protein efficiency ratio (PER) and phosphatidylcholine (PC), phoshphatidylethanolamine (PE) and Met contents of the whole body of shrimp. The shrimp group did not receive either supplemental CC or Met showed lower (P < 0.05) values of the above‐mentioned parameters than other shrimp groups fed with 0.6 and 1.2 g kg⁻¹ supplemental CC with or without Met supplementation. The present study showed that supplementation of 1.2 g kg⁻¹ CC in the diets could compensate shrimp post‐larvae with the needed methyl group when received Met‐deficient diets. The study also assumed that the biosynthesis of PC in the shrimp’s body can be achieved by the methylation of PE through the S‐adenosylmethionine (SAM) pathway and/or through the cytosine di‐phosphoryl (CDP) choline pathway directly from dietary choline.     

Classification of the genus Bacillus based on MALDI-TOF MS analysis of ribosomal proteins coded in S10 and spc operons

A rapid bacterial identification method by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) using ribosomal proteins coded in S10 and spc operons as biomarkers, named the S10-GERMS (the S10-spc-alpha operon gene encoded ribosomal protein mass spectrum) method, was applied for the genus Bacillus a Gram-positive bacterium. The S10-GERMS method could successfully distinguish the difference between B. subtilis subsp. subtilis NBRC 13719(T) and B. subtilis subsp. spizizenii NBRC 101239(T) because of the mass difference of 2 ribosomal subunit proteins, despite the difference of only 2 bases in the 16S rRNA gene between them. The 8 selected reliable and reproducible ribosomal subunit proteins without disturbance of S/N level on MALDI-TOF MS analysis, S10, S14, S19, L18, L22, L24, L29, and L30, coded in S10 and spc operons were significantly useful biomarkers for rapid bacterial classification at species and strain levels by the S10-GERMS method of genus Bacillus strains without purification of ribosomal proteins.