Studies on microörganisms and their activities in soil Part. 2. Virgin and cultivated soil of the same origin

The nature of soil is modified differently depending upon the artificial condition such as its utilization or management. It is therefore expected that the microbiological characteristic of soil is changed also. Greaves¹⁾ and Williams²⁾ reported that the reclamation of virgin soil brought about a change of bacterial count. Suzuki et al³⁾ observed that the kind of fungi differed between a virgin and a cultivated soil and that the vegetative mycelium was numerous in the former than in the latter. On the other hand, according to W aksman and Starkey⁴⁾, the bacterial count differed depending upon the fertility of soil. Singh⁵⁾ reported also that the number of fungi and actinomycetes was higher in a fertile than in an infertile soil. Lochhead⁶⁾, and Lochhead and Chasen studied the bacterial flora of a fertile (long-continued application of manure) and an infertile (no fertilizer for many years) soil and found that a certain difference could be observed when morphological, physiological and nutritional classification are tried.     

Chemical forms and stability of selenium in soil

The chemical forms of selenium (Se) in soils derived from different parent rocks, and the stability of Se species in soils were investigated. Around 40% of the total Se occurred as organic Se on the average. Total selenite content was higher than that of total selenate. The content of organic Se decreased with heating of the soils at high temperatures, and most of it disappeared at heating temperatures above 500°C. However, the results of incubation experiments indicated that Se species were stable in soil, and that their chemical forms did not change appreciably with the variation of soil conditions. In soluble Se, organic Se accounted for about 50%, and in contrast to the total Se, the selenate content was higher than that of selenite.     

Effect of burning on soil organic matter content and N mineralization under shifting cultivation system of Karen people in Northern Thailand

In the traditional shifting cultivation system practiced by the Karen people in northern Thailand, the effects of burning on the content of extractable organic matter, microbial biomass, and N mineralization process of the soils were studied. Five plots (5×5 m² quadrat) with 0, 10, 20, 50, and 100 Mg ha^-1 of slashed materials were arranged and burned. Ten to 20 Mg ha^-1 of slashed biomass corresponded to the amount commonly burned by the Karen people. During the burning process, the soil temperature at the depth of 2.5 cm in the 100 Mg ha^-1 plot almost evenly increased to 300°C while the temperature in the 10 to 50 Mg ha^-1 plots increased with large variations from 50 to 300°C. Burning caused a conspicuous increase in the contents of organic C and (organic + mineral)-N extracted at room temperature and a simultaneous decrease in the contents of microbial biomass C and N, especially in the soil of the 100 Mg ha^-1 plot. In the rainy season, the values of the changes induced by burning reverted to the values recorded before burning, except for the microbial biomass in the 100 Mg ha^-1 plot, which still remained lower. Based on an incubation experiment, N mineralization rate was higher in the soils taken just after burning, especially in the 100 Mg ha^-1 plot, than in the soils taken during the rainy season. However, the soil in the 100 Mg ha^-1 plot was considered to have the lowest ability to supply mineral N among the soils in the rainy season. Burning of 10 to 20 Mg ha^-1 biomass corresponding to the values recorded in Karen peoples' shifting cultivation system was more compatible with soil ecology in terms of N supply at the initial stage of crop growth and of microbial biomass recovery during the rainy season, compared to the burning of 100 Mg ha^-1 biomass corresponding to the value recorded in a natural forest. Thus, the shifting cultivation system implemented by the Karen people can be considered to be a well-balanced agricultural system.     

Sexing of Dog Sperm by Fluorescence In Situ Hybridization

Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97% hybridization efficiency and a good preservation of sperm morphology. There was no significant difference between the theoretical ratio (50:50) and the observed ratio of X- and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that the FISH protocol worked reliably for both unsorted and sexed sperm samples.     

Good Thermally Conducting Material Supports Follicle Morphologies of Porcine Ovaries Cryopreserved with Ultrarapid Vitrification

Effects of supporting materials during vitrification procedure on the morphologies of preantral follicles of pig ovaries were assessed. Ovarian cortical sections of prepubertal pigs were randomly allocated to 5 groups. The sections were vitrified ultrarapidly with 5 different vitrification devices. The sections were put on 4 fine needles (Cryosupport), on a thin copper plate, or on a carbon graphite sheet or were sandwiched between copper plates or between carbon graphite sheets before cooling. The cooling and warming rates with the graphite sheets were significantly higher than those with the copper plates (P<0.05). A total of 3,064 follicles were analyzed following HE staining after vitrification with 5 different devices. The morphologies follicles vitrified on the Cryosupport or on the graphite sheet were well preserved compared with those vitrified on the copper plate or between copper plates (P<0.01). The morphologies of follicles vitrified between copper plates were mostly damaged (P<0.05). Taken together, good thermally conducting material supports follicle morphologies of ovaries cryopreserved with ultrarapid vitrification.     

Evaluation of the PATHFAST Chemiluminescent Enzyme Immunoassay for Measuring Progesterone in Whole Blood and Serum of Mares

Evaluation of a new chemiluminescent enzyme immunoassay, the PATHFAST assay system (PATHFAST), for measurement of circulating progesterone in mares was performed. Five mares at the mid-luteal stage were administrated a single i.m. injection of prostaglandin F2α analog (PGF2α; cloprostenol 250 μg/ml), and then blood samples were collected from the jugular vein at 0, 15, 30 and 45 min, at one-hour intervals until 24 and at 48 hr via a catheter in the jugular vein. To monitor the physiological changes in circulating progesterone in mares after induced luteolysis, concentrations of progesterone in whole blood and serum samples were measured by PATHFAST. In addition, concentrations of progesterone in serum samples measured by PATHFAST were compared with those measured by radioimmunoassay (RIA) and enzyme immunoassay (EIA). Using PATHFAST, the serum concentrations of progesterone in mares correlated highly with those of whole blood samples (r=0.9672, n=88). The serum concentrations of progesterone as measured by PATHFAST correlated well with RIA (r=0.9654, n=88) and EIA (r=0.9323, n=112). An abrupt decline in circulating progesterone in whole blood samples was observed within 2 hr (50%), followed by a gradual decline until 48 hr later. The results for progesterone in whole blood samples correlated highly with those in serum samples, and the declining pattern paralleled that of the serum samples. These results demonstrated that PATHFAST is useful in the equine clinic as an accurate diagnostic tool for rapid assay of progesterone within 26 min, using unextracted whole blood.     

Primary analysis of DNA polymorphisms in the TRIM region (MHC subregion) of the Japanese quail,Coturnix japonica

Based on sequences of two cosmid clones from Japanese quail (Coturnix japonica, Coja), we confirmed that the syntenic cluster, GNB2L1～BTN1～BTN2, is located in the quail TRIM subregion of the quail major histocompatibility complex (MHC Coja) region. These cosmids also included four CjBG loci and one CjLEC locus; therefore, the quail TRIM subregion was thought to be adjacent to the BG/LEC subregion. We then identified three polymorphic markers – CjHEP21, CjTRIM39.2 and CjBTN2 – in the TRIM subregion that may be useful for the functional analysis of the MHC‐Coja region. We examined MHC‐Coja sequences from 321 individual quails sampled from 11 inbred strains, and we found eight alleles for each of the three genes – CjHEP21, CjTRIM39.2 and CjBTN2. These polymorphisms represent the first avian DNA markers in the TRIM subregion. Additionally, we discovered a quail‐specific VNTR (variable number of long tandem repeats, 133–137 bp) in intron 7 of CjBTN2. We identified 25 haplotypes in the sample of 321 quail; these haplotypes comprised combinations of all 24 alleles of the three polymorphic genes. We suggest that there are two recombination hotspots, one between each pair of adjacent loci. All strains, except AMRP, contained multiple haplotypes; the AMRP strain contained a single, apparently fixed haplotype.