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.     

Characteristics and issues related to regional-scale modeling of nitrogen flows

This paper summarizes the characteristics of regional-scale nitrogen (N) flow models. The regional scale is generally considered to be an area that ranges from more than 10 km² to the size of a continent. Parameterization is the key process in creating a regional-scale model. During parameterization, transfer functions that reflect the controlling factors must be created at the target scale because the influence of different factors will change with the size of the scale. Watersheds are the most useful unit for evaluating overall N discharge; however, regional activity data is most often available for municipal units. Thus, municipal units must be reaggregated into watershed units. A longer time period is desirable to normalize seasonal and annual variations at regional scales. Parameters that influence N flow must match the investigated spatial and temporal scales. Given the need to use a range of parameters that vary in terms of the quality of the data, models exhibit inevitable uncertainties. Quantification of the uncertainties and verification of the estimated results are required. Error propagation, the Monte Carlo simulation method and maximum and minimum values have been used to obtain different threshold values of uncertainty. To verify regional-scale N flow models, the following five approaches have been used or proposed: (1) calibration of the model by detailed monitoring at multiple sites, (2) verification of the most important process of the extrapolation mechanisms, (3) verification of the N budget, paying particular attention to water quality, (4) comparison with the results quantified by different models, (5) comparison with aerial or satellite image analysis. As regional-scale modeling of N flow will become more important in the future, it is important to develop models than can accurately estimate N dynamics at this scale.     

Nitrogen uptake by sago palm (Metroxylon sagu Rottb.) in the early growth stages

Previous trials have revealed variable responses of sago palm ( Metroxylon sagu Rottb.) to fertilizer application, particularly nitrogen (N). In the present study, we quantified the fertilizer use efficiency (FUE) of sago palm for the first time using ¹⁵N-labeled fertilizer in pot and field experiments. The pot experiment was conducted in Japan using a 2:1 mixture of sand to Philippine soil. The field experiment was conducted in Leyte in the Philippines. Both experiments consisted of three replicates in each of three treatments: control, ¹⁵N urea at 50 kg N ha⁻¹ and ¹⁵N urea at 100 kg N ha⁻¹. The N uptake of sago palm increased significantly, but inconsistently with increasing N application. The few instances of a significant increase in N uptake did not translate into significant improvements in growth parameters, except for the number of leaflets in the pot experiment. The FUE values for sago seedlings (< 6 months) in the pot experiment treated with 50 and 100 kg N ha⁻¹ were 10.5 and 13.2%, respectively, whereas for the 2-year-old sago palms in the field, the corresponding FUE values were 14.8 and 12.0%. The FUE values were similar at the two levels of N application in both experiments. Sago growth parameters appeared to be insensitive to N application, suggesting that the form of N and the timing of N fertilization are important factors for sago palms. Therefore, the use of N fertilizer in sago production can only be justified after determining and fully understanding the response of sago palm to N application.     

Element interconnections in Lotus japonicus: A systematic study of the effects of element additions on different natural variants

Lotus japonicus was used to study the distribution and interconnections of 15 elements in plant tissues, including essential and non-essential elements: boron (B), sodium (Na), magnesium (Mg), potassium (K), calcium (Ca), manganese (Mn), iron (Fe), nickel (Ni), copper (Cu), zinc (Zn), arsenic (As), strontium (Sr), molybdenum (Mo), cadmium (Cd) and cesium (Cs). Large amounts of B and Ca accumulated in plant leaves, while Fe, Na, Ni, As and Cd tended to mainly occur in the roots, and Mo was the only element to accumulate in the stems. The elemental compositions within plants were severely disturbed by treatment with toxic elements. Competition between element pairs in the same group (e.g. K and Cs; Ca and Sr) was not found. Iron, Cu and Zn accumulation were induced by Cd and Ni addition. When natural variants grew in a nutrition solution with subtoxic levels of As, Cd, Cs, Ni, Mo and Sr, intriguing relationships between the elements (such as Fe, As and K; Mg and Ni; Mn and Ca) were revealed using principal-component analysis. This study on the plant ionome offers detailed information of element interactions and indicates that chemically different elements might be closely linked in uptake or translocation systems.     

T4-type bacteriophage communities estimated from the major capsid genes (g23) in manganese nodules in Japanese paddy fields

The present study compared the capsid gene ( g23 ) of T4-type bacteriophages (phages) in Mn nodules with those in the plow layer soil and subsoils of two Japanese paddy fields by applying the primers MZIA1bis and MZIA6 to DNA extracts from the nodules and soils. The deduced amino acid sequences of the g23 genes in the Mn nodules were similar to those in the plow layer soil and in the subsoils. This result indicated that similar T4-type phage communities developed at these sites and that the diversity of T4-type phage communities was wide enough to cover those in the plow layer soil and in the subsoils. The majority of g23 clones formed several clusters with the clones and phages obtained from far-apart paddy fields, and the sequences of two clones were completely identical to a phage and a clone from other paddy fields at the nucleotide or amino acid level, indicating horizontal transfer of g23 genes between those paddy fields. A clone with a long nucleotide residue (686 bp) and a distribution remote from the other clones in the phylogenetic tree indicated that there were many uncharacterized, novel g23 genes in the paddy fields.     

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.     

Allometric equations for accurate estimation of above-ground biomass in logged-over tropical rainforests in Sarawak, Malaysia

Although allometric equations can be used to accurately estimate biomass and/or carbon stock in forest ecosystems, few have been developed for logged-over tropical rainforests in Southeast Asia. We developed allometric relationships between tree size variables (stem diameter at breast height (dbh) and tree height) and leaf, branch, stem and total above-ground biomass in two logged-over tropical rainforests with different soil conditions in Sarawak, Malaysia. The study sites were originally classified as mainly lowland dipterocarp forest and have been selectively logged in the past 20 years. In total, 30 individuals from 27 species were harvested to measure above-ground parts. The correlation coefficients for the allometric relationships obtained for total above-ground biomass as a function of dbh had high values (0.99), although the relationships for leaf biomass had a relatively low coefficient (0.83). We also found relatively high coefficients for allometric relationships between tree height and plant-part biomass, ranging from 0.82 to 0.97. Moreover, there were no differences for allometric equations of total above-ground biomass between study sites. A comparison of equations of above-ground biomass in various previously reported tropical rainforests and pan-tropic general equations imply that our allometric equations differ largely from the equations for tropical primary forests, early successional secondary forest, and even for the general models. Therefore, choosing the biomass estimation models for above-ground biomass in the logged-over forests of Southeast Asia requires careful consideration of their suitability.     

中国和日本农业活动对流域氮素收支的影响

To analyze the effect of agricultural activity on nitrogen(N) budget at the watershed scale,a comparative study was conducted at two Japanese watersheds,the Shibetsu River watershed(SRW) and Upper-Naka River watershed(UNRW),and one Chinese watershed,the Jurong Reservoir watershed(JRW).The total area and the proportion of agricultural area(in parentheses) of the watersheds were 685(51%),1 299(21%),and 46 km 2(55%) for SRW,UNRW,and JRW,respectively.The main agricultural land use in SRW was forage grassland,while paddy fields occupied the highest proportion of cropland in UNRW(11% of total area) and JRW(31% of total area).The farmland surplus N was 61,48,and 205 kg N ha 1 year 1 for SRW,UNRW,and JRW,respectively.The total input and output for the whole watershed were 89 and 76,83 and 61,and 353 and 176 kg N ha 1 year 1 for SRW,UNRW,and JRW,respectively.The proportion of discharged N to net anthropogenic N input was 31%,37%,and 1.7% for SRW,UNRW,and JRW,respectively.The two watersheds in Japan showed similar proportions of discharged N to those of previous reports,while the watershed in China(JRW) showed a totally different characteristic compared to previous studies.The high N input in JRW did not increase the amount of discharged N at the outlet of the watershed due to high proportions of paddy fields and water bodies,which was an underestimated N sink at the landscape scale.     

Bacillary hemoglobinuria in Japanese black cattle in Hiroshima, Japan: a case study

Three Japanese Black cows housed with 6 other cows exhibited main clinical symptoms of severe hemoglobinuria. Hematological analyses conducted after antibiotic therapy demonstrated severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. One of the three cows died. Based on the above analyses and observation of typical clinical symptoms, a speculative diagnosis of bacillary hemoglobinuria was made, and immediate high-dose antibiotic treatment improved the general conditions of the surviving animals. Blood samples from the other 2 cows were collected sequentially after antibacterial therapy. Clostridium haemolyticum was detected by a nested polymerase chain reaction analysis of the blood samples. The cows were diagnosed with the second recorded occurrence of bacillary hemoglobinuria in Japan.     

Economic valuation of cultivated lands as nitrogen removal/effusion sites by newly proposed replacement cost method

To valuate the multifunctionality economically is effective to make it possible to realize the value for the nation and to compare functionalities among countries of the world. In this paper, the external economies of paddy fields and fallow paddy fields including wetlands as N removal function sites, and of upland fields and orchards as pollution sites are valuated by the newly proposed replacement cost method, by replacing them with construction costs of water quality improvement facilities. In addition, we discuss an agricultural land-use scenario in which cultivated land has no net negative economic effect on the water environment. The results showed that (1) paddy fields and fallow paddy fields including wetlands were respectively valued at 1.2×10³ and 2.81×10³ JPY m⁻² on average as the N removal sites, (2) upland fields had 0.32×10³ JPY m⁻² on average of economic value, and suggested that paddy fields have an external economic value that compensates for the negative external economic value of upland fields 3.65 times their size.