Cover Image,Volume 16, Issue 3

The cover image, by K. Sakai et al., is based on the Original Article Association of tumour‐infiltrating regulatory T cells with adverse outcomes in dogs with malignant tumours, DOI: 10.1111/vco.12383 .

     

Development of two immunochromatographic tests for the serodiagnosis of bovine babesiosis

In this study, we developed two immunochromatographic tests (ICTs), which are nitrocellulose membrane-based immunoassays for the convenient and rapid serodiagnosis of bovine babesiosis caused by Babesia bovis (BoICT) and Babesia bigemina (BiICT). The efficacy of two ICTs was evaluated using 13 positive sera from experimentally infected cattle with B. bovis or B. bigemina. Clear results showed that the BoICT and ELISA detected antibodies in sera collected from 14 to 93 days post-infection, while BiICT and ELISA detected from 13 to 274 days post-infection. In additon, non-infected cattle, Neospora caninum, and Cryptosporidium parvum were negative in two ICTs. To evaluate the field utility of the ICTs, we tested 186 field bovine sera collected from cattle living in Yanbian (China) and Mato Grosso do Sul (Brazil). The results of ICTs were compared to those of classical serodiagnostic methods, enzyme-linked immunosorbent assay (ELISA) and the indirect immunofluorescence assay (IFAT). The overall concordances of BoICT were determined as 92.5 and 90.3% when the results of ELISA and IFAT were set as the reference standards, respectively. In contrast, those of BiICT showed 96.8 and 92.5% relative to the results of standard ELISA and IFAT, respectively. Conventional and rapid diagnosic devices for bovine babesiosis may provide a valuable tool in clinical and field applications.     

Effect of Amylose Content on Gelatinization,Retrogradation, and Pasting Properties of Starches from Waxy and Nonwaxy Wheat and Their F1 Seeds

We studied the effect of amylose content on the gelatinization, retrogradation, and pasting properties of starch using wheat starches differing in amylose content. Starches were isolated from waxy and nonwaxy wheat and reciprocal F1 seeds by crossing waxy and nonwaxy wheat. Mixing waxy and nonwaxy wheat starch produced a mixed starch with the same amylose content as F1 seeds for comparison. The amylose content of F1 seeds ranged between waxy and nonwaxy wheat. Nonwaxy‐waxy wheat had a higher amylose content than waxy‐nonwaxy wheat. Endothermic enthalpy and final gelatinization temperature measured by differential scanning calorimetry correlated negatively with amylose content. Gelatinization onset and peak temperature clearly differed between F1 and mixed starches with the same amylose content as F1 starches. Enthalpy for melting recrystallized starches correlated negatively with amylose content. Rapid Visco Analyser measurement showed that F1 starches had a higher peak viscosity than waxy and nonwaxy wheat starches. Mixed starches showed characteristic profiles with two low peaks. Setback and final viscosity correlated highly with amylose content. Some of gelatinization and pasting properties differed between F1 starches and mixed starches.     

Work-related increases in titer of Campylobacter jejuni antibody among workers at a chicken processing plant in Miyazaki prefecture,Japan, independent of individual ingestion of edible raw chicken meat

Workers in poultry abattoirs may be frequently exposed to Campylobacter jejuni, which is a leading cause of bacterial food poisoning in Japan. The present study was conducted to measure the titers of IgG and IgA antibodies against C. jejuni among 104 female workers in a chicken processing plant in Miyazaki prefecture, Japan. Information regarding habitual ingestion of raw chicken meat and potential occupational risk factors was collected using a questionnaire. Acid extracts of four C. jejuni strains representing the genotypes most dominant in Miyazaki were used as antigens. The levels of both immunoglobulins measured by ELISA were not correlated with ingestion of edible raw chicken meat, the amount consumed in one sitting, or its frequency. Although age was correlated with antibody levels, the length of employment was not. Furthermore, the IgG and IgA levels in workers at the evisceration step were significantly higher than those at other locations in the plant. To identify the bacterial proteins recognized by the workers’ IgG and IgA antibodies, Western blotting followed by LC/MS was conducted. Flagellin was identified as the common protein recognized in the sera of workers for whom ELISA demonstrated both the highest and lowest antibody levels. We concluded that the titers of IgG and IgA against C. jejuni in workers at the processing plant had been increased by occupational exposure to Campylobacter, regardless of raw chicken meat ingestion.     

Isolation and pathogenic characterization of an OB1 variant of Babesia rodhaini which has a glycophorin A-independent pathway to murine red blood cells

Recent studies using several Babesia spp. have demonstrated that these species commonly recognize host sialic acids of red blood cells (RBCs) for their invasion. Glycophorin A (GPA), which is a major carrier of the sialic acids on RBCs, is a possible invasive receptor for Babesia parasites. In the present study, a variant of Babesia rodhaini was successfully isolated from a GPA homozygous knockout (GPA^−/−) mouse infected with an Australian strain of B. rodhaini which had originally been unable to replicate in GPA^−/− mice. The isolated parasite (designated as an OB1 variant) caused lethal infection to wild-type mice, as in the case of the parent Australian strain. However, although the growth of the OB1 variant in GPA^−/− mice was comparable with that in wild-type mice at 1–4 days after infection, the growth was significantly inhibited from day 5 onward, leading to the eventual survival of the GPA^−/− mice. Resistance of GPA^−/− mice against OB1 infection was lost by splenectomy, although the cytokine responses to the infection in the sera of GPA^−/− mice were similar to those of wild-type mice. The autoantibody levels to GPA-defective RBCs in the sera of GPA^−/− mice were depressed at a lower level at 0–2 days after infection than those of wild-type mice, while the levels of GPA^−/− mice progressively increased and reached comparable levels to those of wild-type mice at day 3 or later. These results indicate that the isolated OB1 variant has a GPA-independent invasion pathway into murine RBCs and suggest that the resistance of GPA^−/− mice against infection with the OB1 variant may be attributed to the effective clearance of the parasitized RBCs lacking GPA in the spleen, possibly mediated by preferential autoantibody binding to the RBC membrane.     

Effects of sika deer (Cervus nippon) and dwarf bamboo (Sasamorpha borealis) on seedling emergence and survival in cool-temperate mixed forests in the Kyushu Mountains

Effects of sika deer (Cervus nippon) and dwarf bamboo (“sasa;” Sasamorpha borealis) on seedling emergence and survival were investigated in cool-temperate mixed forests in the Kyushu mountain range, Japan. We compared the effects of sika deer between two sites with different sika deer densities. One site (no-sasa forest) has sparse cover of S. borealis and a high density of sika deer, and the other site (sasa forest) has dense cover of S. borealis with a low density of sika deer. In the no-sasa forest, more seedlings emerged and the survival rate of the seedlings was higher compared with the sasa forest. Compared with the sasa forest, the canopy in the no-sasa forest was more open, the organic layer was shallower and drier, the mean daily soil temperature was higher, and soil temperature fluctuated more widely. Those environmental parameters did not differ between unfenced and fenced plots. The comparison between the sasa and no-sasa forests suggested that removal of S. borealis by sika deer had an indirect positive effect on seedling emergence and survival. However, the fenced-plot experiments indicated that sika deer inhibited seedling emergence and survival. These contradictory conclusions could result from the much larger negative effects of S. borealis on seedling emergence and survival compared with the effects of sika deer browsing.     

Daily growth rate model of Japanese anchovy larvae Engraulis japonicus in Hiuchi-nada Sea, central Seto Inland Sea

In the present study, we developed a larval anchovy growth model in relation to sea temperature and food availability via food consumption and metabolic process terms, based on biological data from previous laboratory experiments and field surveys from 2003 to 2006 in Hiuchi-nada Sea, central part of Seto Inland Sea, Japan. To investigate when food shortage for larval anchovy and then recruitment failure occur in Hiuchi-nada Sea, anchovy food requirements were estimated by using the growth model, and we compared the food requirement with anchovy food availability. We applied an estimation method for growth model parameters, Hewett?CJohnson p and Q ₁₀, by minimizing the sum of squares of difference between mass-specific growth rates estimated by the models and those by otolith growth analysis. Parameter p was 0.86, slightly higher than typical values, and Q ₁₀ was 2.11, close to the value used for the biological model of larval northern anchovy. Food shortage for anchovy larvae did not occur in Hiuchi-nada Sea, although it was indicated that low food availability led to a low reproductive success rate. The newly developed growth model is considered optimal at present and useful to link environmental conditions and larval growth.     

Seasonality of factors controlling N mineralization rates among slope positions and aspects in cool-temperate deciduous natural forests and larch plantations

This study aimed to evaluate the spatial patterns of soil nitrogen (N) transformations in relation to slope aspect and position, and to investigate the main factors controlling N transformation patterns during both the growing and dormant seasons in cool-temperate deciduous natural forests and larch plantations in eastern Hokkaido, northern Japan. Net rates of N mineralization (NR_minN) and of nitrification (NR_nit) in surface soils on north-facing and lower slopes were higher than those on south-facing and upper slopes, whereas the net rate of ammonium-N production (NR_amm) on south-facing and upper slopes was higher than that on north-facing slopes in both the natural forests and larch plantations. Both NR_minN and NR_nit were higher in the growing than in the dormant season, whereas NR_amm was higher in the dormant season. The soil C/N ratio, water content, soil pH and frequency of freeze–thaw cycles were important variables affecting N transformation patterns in any season. In relation to seasonality, the solar radiation index, daily temperature range and earthworm biomass were important controlling factors only during the growing season, and watershed area and soil N concentration only during the dormant season, suggesting that biological control accompanied with wet–dry events were important factors affecting N transformations during the growing season, but that run-off water and chemical controls were important determinants of spatial variation in N transformations during the dormant season.     

Effects of topography and planted trees on the distribution of naturally regenerated broad-leaved trees in a 140-year-old <Emphasis Type="Italic">Cryptomeria japonica</Emphasis> plantation in northern Kyushu,Japan

We investigated factors affecting the distribution of naturally regenerated broad-leaved trees in a 140-year-old Cryptomeria japonica plantation. We used path analysis to examine the relationship among microtopography, the biomass of planted trees, and the biomasses of canopy and subcanopy trees of broad-leaved species. The study plot was divided into three topographic types (ridge, slope, and valley), and we discuss how the different topographic types are affected. For all topographic types, the biomass of canopy trees of broad-leaved species decreased with convexity. For slope and valley topographies, the biomass of subcanopy trees of broad-leaved species also decreased with convexity. For ridge topography, the biomass of subcanopy trees of broad-leaved species increased with the biomass of planted trees, and decreased with the biomass of canopy trees of broad-leaved species. These results suggest the effects of microtopography on the biomass of subcanopy trees were much larger than the effects of canopy trees for slope and valley topographies, while the effects of microtopography were smaller for ridge topography.