Detecting endotoxin activity in bovine serum using an automated testing system

The aim of the present study was to compare the ability of the commercially available portable test system (PTS^TM) to detect endotoxin activity in bovine serum, with that of the traditional LAL-kinetic turbidimetric (KT) and chromogenic (KC) assays. Prior to testing, serum samples, which were obtained from endotoxin-challenged cattle, were diluted 1:20 in endotoxin-free water and heated to 80°C for 10 min. The performance of the PTS^TM was not significantly different from that of the traditional LAL-based assays. The results using PTS^TM correlated with those using KT (r²=0.963, P<0.001) or KC assays (r²=0.982, P<0.001). Based on these findings, the PTS^TM could be applied as a simplified system to assess endotoxin activity in bovine serum.     

Prolonging hypothermic storage (4 C) of bovine embryos with fish antifreeze protein

Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming.     

Development of a quantification assay for the cysts of the toxic dinoflagellate <Emphasis Type="Italic">Alexandrium tamarense</Emphasis> using real-time polymerase chain reaction

ABSTRACT:   The cysts of toxic dinoflagellate Alexandrium tamarense are the seed population for the bloom responsible for paralytic shellfish poisoning (PSP). However, it is impossible to identify the Alexandrium spp. cyst on the basis of morphological features. In this study, we prepared A. tamarense cysts by sexual conjugation in laboratory conditions and developed an efficient DNA extraction method for polymerase chain reaction (PCR) assay. Using the A. tamarense cysts, we established the identification and quantification method showing the species specificity and the high sensistivity for A. tamarense cysts using real-time PCR. This assay was also able to detect and quantify the A. tamarense cysts accurately when mixed with excess cysts of A. catenella (Whedon and Kofoid) Balech prepared by conjugation experiment.     

Characterisation of starch properties and physical characteristics in buckwheat (Fagopyrum esculentum Moench.) mutant lacking accumulation of ‘granule-bound starch synthase a’

The concentration of amylose, which is synthesised using granule-bound starch synthase, affects the physical properties of food. However, no studies have focused on starch properties and physical characteristics of low-amylose buckwheat. Here, we hypothesised that low-amylose buckwheat would be useful to produce new buckwheat products because low-amylose characteristics change the texture of buckwheat food. In this study, we bred relatively low-amylose buckwheat compared to wild type and investigated the causative genes of the traits, starch properties and physical properties of noodles. In the GBSSa mutant, the amylose concentration was lower than that in the wild type. Compared with the wild type, the mutant exhibited the following traits: Amylose concentration decreased by approximately 2%, setback in the Rapid Visco Analyzer decreased by 30 points and the physical characteristics of noodles in the sensory analysis were soft and sticky. These results suggest that this trait may be useful for changing the texture of foods. In addition, the mutant is promising for producing new foods with physical characteristics that are different from those of the wild type.     

Modeling of Salt Sorption in Volcanic Ash Soil

Groundwater pollution by nitrate from agricultural fields is a worldwide problem. To improve the understanding of nitrate transport processes through volcanic ash soils overlying groundwater aquifers, salt sorption experiments were performed. Salt sorption is the process that involves the simultaneous use of cation and anion adsorption mechanisms without changing the pH. Prepared six different concentrations of potassium chloride (KCl) solutions were mixed with soil samples. Anion and cation contents of the liquid and solid phases were determined. Experiments were performed in triplicate of six kinds of KCl solutions for three types of soils. Thus, total 54 of samples were analyzed. Relationships between ionic strength and sorption capacity increments of cations and anions were determined. The results show that if ionic strength of the infiltrating solution to the volcanic ash soil is known, the ECEC (effective cation exchange capacity) and EAEC (effective anion exchange capacity) can be reliably estimated. The resulting prediction equations can be included to improve reactive transport simulation models. Consequently, it will be possible to better understand solute transport with changing sorption capacity for the solid soil phase.     

Land cover change detection based on satellite data for an arid area to the south of Aksu in Taklimakan desert

An experiment is made to detect the land-cover change in the area located to the south of Aksu in the northern Taklimakan desert through analyses of satellite data pixel by pixel basis. The analyzed data are those observed in the late summer and early autumn of 1973, 1977, 1993 and 1995. As a parameter of land-cover, SAVI (Soil Adjusted Vegetation Index) derived from the data of Landsat MSS and JERS-1 OPS (Optical Sensor) is used. The result indicates the increase of vegetation in the oasis areas, confluent area of the Yarkant and Kashgar Rivers and around reservoirs while little change occurs in the desert area. The 1973 satellite image shows the abundant flow in the Yarkant River while the river is almost dried up in the satellite images of later years. The trend of the decrease in the Hotan River flow is recognized although not so dramatic as that of the Yarkant River.     

Effect of supplementation with the reduced form of coenzyme Q10 on semen quality and antioxidant status in dogs with poor semen quality: Three case studies

Oxidative stress owing to an imbalance between reactive oxygen species and antioxidants, such as coenzyme Q10 (CoQ10), is a major contributor to male infertility. We investigated the effects of the reduced form of CoQ10 (ubiquinol) supplementation on semen quality in dogs with poor semen quality. Three dogs received 100 mg of ubiquinol orally once daily for 12 weeks. Semen quality, serum testosterone, and seminal plasma superoxide dismutase (SOD) activity were examined at 2-week intervals from 2 weeks before ubiquinol supplementation to 4 weeks after the treatment. Ubiquinol improved sperm motility, reduced morphologically abnormal sperm, and increased seminal plasma SOD activity; however, it had no effect on testosterone level, semen volume, and sperm number. Ubiquinol supplementation could be used as a non-endocrine therapy for infertile dogs.     

Comparative Analysis of Japanese and Foreign Strains of <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> Based on 16S Ribosomal RNA Gene Sequences

We determined nearly the complete sequences of the 16S ribosomal RNA gene (rDNA) for Japanese strains of R. solanacearum. The comparison of 1471 nucleotide positions separated the Japanese strains into two groups, group 1 with biovars 1, 2, 3 and 4 strains which belonged to race 1, and group 2 with biovar 2 strains corresponding to race 3. Group 1 strains all had identical sequences, and strains representing the four biovars within the group did not differ from each other. Group 2 strains had characteristic nucleotides which differed at seven positions from group 1 strains. Comparative analysis of Japanese and foreign strains based on 16S rDNA sequences showed that Japanese group 1 was closely related to Asian and Australian biovars 3, 4 and 5, and belonged to the known division 1. Japanese group 2 was homogeneous to Indonesian biovars 2 and N2 in subdivision 2b. Since the differences in the nucleotides corresponded to restriction sites for the AluI, RFLP analysis of PCR-amplified 16S rDNA efficiently differentiated not only Japanese group 1 from group 2, but also differentiated three types of foreign strains which differed in biovar and geographic origin. Received 26 July 1999/ Accepted in revised form 19 November 1999