Phenotypic and functional analysis of bovine peripheral blood dendritic cells before parturition by a novel purification method

Dendritic cells (DCs) are specialized antigen presenting cells specializing in antigen uptake and processing, and play an important role in the innate and adaptive immune response. A subset of bovine peripheral blood DCs was identified as CD172a⁺/CD11c⁺/MHC (major histocompatibility complex) class II⁺ cells. Although DCs are identified at 0.1%–0.7% of peripheral blood mononuclear cells (PBMC), the phenotype and function of DCs remain poorly understood with regard to maintaining tolerance during the pregnancy. All cattle used in this study were 1 month before parturition. We have established a novel method for the purification of DCs from PBMC using magnetic‐activated cell sorting, and purified the CD172a⁺/CD11c⁺ DCs, with high expression of MHC class II and CD40, at 84.8% purity. There were individual differences in the expressions of CD205 and co‐stimulatory molecules CD80 and CD86 on DCs. There were positive correlations between expression of cytokine and co‐stimulatory molecules in DCs, and the DCs maintained their immune tolerance, evidenced by their low expressions of the co‐stimulatory molecules and cytokine production. These results suggest that before parturition a half of DCs may be immature and tend to maintain tolerance based on the low cytokine production, and the other DCs with high co‐stimulatory molecules may already have the ability of modulating the T‐cell linage.     

Hair cortisol levels of lactating dairy cows in cold‐ and warm‐temperate regions in Japan

We compared the hair cortisol levels of lactating dairy cows in a cold‐ and a warm‐temperate region out of four climatic zones in Japan. We simultaneously investigated the effects of calving number, lactation period and month of hair sampling. Hair of nine Holstein lactating cows chosen from each region (i.e. 18 cows per sampling) was sampled in March, June, September and December. Number of calvings (1, 2, ≥3) and lactation duration (early: <100, middle: 101–200, and late: >201 days) were balanced between regions. Cortisol was extracted from hair by methanol, and its level was determined with a cortisol immunoassay kit. A multi‐way analysis of variance revealed that the effects of month of hair sampling (P < 0.001) and its combination with region (P < 0.05) were significant. In a multiple comparison test, significant differences (P < 0.01) in hair cortisol level (pg/mg of hair) were found between June (13.0 ± 1.0) and the other 3 months, and between September (1.6 ± 0.2) and December (4.5 ± 0.3). The rise in cortisol level from March to June was more intense in the cold‐temperate region. These results demonstrate the necessity of considering seasonal variations in each climatic region when we use hair cortisol level as an indicator of stress.     

Proton Budgets of Forest Ecosystems on Volcanogenous Regosols in Hokkaido,Northern Japan

The proton budgets of deciduous and coniferous forest ecosystems on volcanogenous regosols in Hokkaido, northern Japan, were studied by measuring the biogeochemical fluxes (atmospheric deposition, canopy leaching, vegetation uptake and leaching from soil) at each site during a three year period. The proton budgets were developed for individual compartments of the ecosystem: vegetation canopy, organic and mineral soil layers. At both sites, atmospheric S deposition was the dominant proton source in the vegetation canopy. In organic horizons, dissociation of weak acids (bicarbonate and/or organic acids) and vegetation uptake of base cations were the dominant proton sources, and the net mineralization of base cations was the dominant proton sink. Atmospheric acid deposition was almost neutralized in the forest canopy and organic horizon. At both sites, weathering and/or ion exchange of base cations and protonation of weak acids (mainly bicarbonate) were the dominant proton sinks in the mineral soil. In both organic and mineral soil, internal proton sources (mainly vegetation uptake of base cations and dissociation of weak acids) exceeded external proton sources, indicating that acid deposition was not the main driving force of soil acidification in the studied forest ecosystems.     

Reduced tillage and increasing cropping intensity in the Great Plains conserves soil C

Concern about soil organic matter losses as a result of cultivation has been voiced consistently since the early part of the 20th century. Scientists working in the US. Great Plains recognized that organic matter losses from an already small pool could have major negative consequences on soil physical properties and N supplying capacity. The advent of reduced- and no-till systems has greatly improved our ability to capture and retain precipitation in the soil during the non-crop periods of the cropping cycle, and has made it possible to reduce fallow frequency and intensify cropping systems. The purpose of this paper is to summarize the effects of reduced tillage and cropping system intensification on C storage in soils using data from experiments in North Dakota, Nebraska, Kansas, Colorado, and Texas. Decades of farming with the wheat (Triticum aestivum L.)–fallow system, the dominant farming system in the Great Plains, have accentuated soil C losses. More intensive cropping systems, made possible by the greater water conservation associated with no-till practices, have produced more grain, produced more crop residue and allowed more of it to remain on the soil surface. Combined with less soil disturbance in reduced- and no-till systems, intensive cropping has increased C storage in the soil. We also conclude that the effects of cropping system intensification on soil C should not be investigated independent of residue C still on the surface. There are many unknowns regarding how rapidly changes in soil C will occur when tillage and cropping systems are changed, but the data summarized in this paper indicate that in the surface 2.5 cm of soil, changes can be detected within 10 years. It is imperative that we continue long-term experiments to evaluate rates of change over an extended period. It is also apparent that we should include residue C, both on the surface of the soil and within the surface 2.5 cm, in our system C budgets if we are to accurately depict residue–soil C system status. The accounting of soil C must be done on a mass basis rather than on a concentration basis.     

Comparison of the activities of enzymes related to glycolysis and gluconeogenesis in the liver of dogs and cats.

Activities of enzymes related to glucose metabolism were measured in canine and feline liver. There were no significant differences in plasma glucose and immunoreactive insulin concentrations between dogs and cats. Glucokinase activities were absent in feline liver, however, activities of other glycolytic enzymes such as hexokinase, phosphofructokinase and pyruvate kinase, were significantly higher than those in canine livers. Activities of rate limiting enzymes of gluconeogenesis such as pyruvate carboxylase, fructose-1, 6-bisphosphatase and glucose-6-phosphatase in feline livers were significantly higher than those in canine livers.     

Comparison of morphological and DNA-based techniques for stomach content analyses in juvenile chum salmon <Emphasis Type="Italic">Oncorhynchus keta</Emphasis>: a case study on diet richness of juvenile fishes

To identify the stomach contents of marine organisms, morphological observations are commonly conducted. However, the results obtained by this traditional method are frequently biased, as it is difficult to detect partially digested, soft-bodied organisms. To resolve this, recent studies have used DNA-based (DNA barcoding) analyses to examine the diet breadth of marine organisms. Here, we compared the prey richness of juvenile fishes using morphological observations and DNA-based analyses, with a focus on juvenile chum salmon Oncorhynchus keta. A higher number of prey taxa were successfully identified using DNA-based analyses than morphological observations. However, we also noticed several shortcomings of the DNA-based analyses, as reported in other diet-analysis studies that used molecular techniques. For example, the degree of digestion among prey taxa might have resulted in differential sensitivity to DNA detection. Additionally, several prey taxa could not be precisely identified, as the sequence data for some of the targeted organismal groups are unavailable in public gene databases. Remarkably, it is also possible that DNA-based analyses detected secondary prey, and therefore, the richness of prey taxa was likely overestimated. Thus, dietary analyses of juvenile fishes need to be carefully conducted, considering the respective advantages and disadvantages of DNA-based and morphological techniques.     

Vitamin contents of eggs that produce larvae showing a high survival rate in the Japanese eel Anguilla japonica

This paper describes the relationship between the egg vitamin concentrations and the egg quality in the Japanese eel Anguilla japonica . No notable relation was found between any vitamin and the fertilization rate. Hatching and survival rates of larvae, however, significantly increased with an elevated level of egg vitamin C (VC). In contrast to VC, the relation between vitamins E (VE) and A (VA) concentrations and survival rate showed a clear peak, with a reduced survival rate at both higher and lower vitamin concentrations. The ratio of VE to lipid or highly unsaturated fatty acid (HUFA) in eggs positively correlated with hatching and survival rates of larvae. High-quality (HQ) eggs were determined as eggs that produced larvae having a survival rate higher than 80% at 8 days post hatch, and low-quality (LQ) eggs were determined as eggs that did not hatch. The level of VC of HQ was significantly higher than LQ. The results of this study suggest that HQ eggs, which produce larvae having a high survival rate, must have high levels of VC and VE/HUFA ratio and contain optimum levels of VA and VE in Japanese eel.     

Release of Bound Lipids in Cereal Starches Upon Hydrolysis by Glucoamylase

The raw starch granules from corn, rice, and wheat were hydrolyzed by practically pure glucoamylase (Rhizopus niveus). The bound lipids remaining in the residual starches were investigated, of which the major components of the lipids, free fatty acids (FFA) in corn starch, FFA and phospholipids (PL) in rice starch, and PL in wheat starch were determined. In each case, the bound FFA and PL were decreased to some extent during the initial stage of hydrolysis. During the later stages, the FFA continued to gradually decrease, while the level of PL stabilized. It was interesting that some of the bound lipids were released from the granules upon glucoamylase hydrolysis, differing from the model amylose-lipid complexes. Furthermore, the structures of the residual starches were investigated. The blue value and λ_max of the starches were increased by partial hydrolysis of the starch granules using practically pure glucoamylase. Two gel-permeation chromatography analyses revealed that the relative amount of amylose fraction was increased by glucoamylase hydrolysis, and also that the increments were reduced by the defatting of bound lipids. The results suggest that the increase in amylose fraction is attributable to the existence of bound lipids in the granules.     

Quality Characteristics of Waxy Hexaploid Wheat (Triticum aestivum L.): Properties of Starch Gelatinization and Retrogradation

The viscoelastic properties and molecular structure of the starch isolated from waxy (amylose-free) hexaploid wheat (WHW) (Triticum aestivum L.) were examined. WHW starch generally had lower gelatinization onset temperature, peak viscosity, and setback than the starch isolated from normal hexaploid wheat (NHW). Differential scanning calorimetry (DSC) showed that WHW starch had higher transition temperatures (T_o, T_p, and T_c) and enthalpy (ΔH) than NHW starch. However, when compared on the basis of amylopectin (AP) content, ΔH of WHW starch was almost statistically identical to that of its parental varieties. Typical A-type X-ray diffraction patterns were observed for the starches of WHW and its parental varieties. Somewhat higher crystallinity was indicated for WHW starch. WHW starch was also characterized by having greater retrogradation resistance. The high-performance size-exclusion chromatography (HPSEC) of amylopectin showed that each amylopectin yielded two fractions after debranching. Although WHW amylopectin had somewhat long B chains, little difference was observed in the ratio of Fr.III/ Fr.II between WHW and its parental varieties.     

Availability of potato protein concentrate as an alternative protein source to fish meal in greater amberjack (Seriola dumerili) diets

Potato protein concentrate (PPC) is a promising candidate as a fish meal (FM) substitute because it has high protein and essential amino acid content. In the present study, we replaced FM in greater amberjack diets with PPC to investigate the effect on growth and feed utilization. Four isonitrogenous, isolipidic and isocaloric experimental diets were prepared by substituting 0, 20, 40 and 60% of FM protein with PPC (Control, P20, P40 and P60 respectively). The in vitro protein digestibility of protein in PPC was 88.8%, relative to 100% protein in the FM. The in vitro protein digestibility of protein in the experimental diets also decreased with increasing PPC and was lowest at 84.2% in P60. After the 7‐week feeding trial, final body weight, weight gain and thermal growth coefficient tended to decrease with increasing PPC and were significantly lower in P60 than in the control (p < .05). Further, fish fed with diets P40 and P60 showed significantly lower feed conversion and protein efficiency ratios than the control group (p < .05). In conclusion, the results suggest that PPC can replace up to 20% of FM in the diet of greater amberjack without compromising the growth performance or feed efficiency.