Rapid and reliable detection of phytoplasma by loop-mediated isothermal amplification targeting a housekeeping gene     

Kyoko Sugawara  Misako Himeno  Takuya Keima  Yugo Kitazawa  Kensaku Maejima  Kenro Oshima  Shigetou Namba 《Journal of General Plant Pathology》2012,78(6):389-397

Phytoplasmas are plant pathogenic bacteria that infect more than 700 plant species. Because phytoplasma-resistant cultivars are not available for the vast majority of crops, the most common practice to prevent phytoplasma diseases is to remove infected plants. Therefore, developing a rapid, accurate diagnostic method to detect a phytoplasma infection is important. Here, we developed a phytoplasma detection assay based on loop-mediated isothermal amplification (LAMP) by targeting the groEL gene and 16S rDNA. We designed 19 primer sets for the LAMP assay and evaluated their amplification efficiency, sensitivity, and spectra to select the most suitable primer sets to detect Candidatus Phytoplasma asteris. As a result, DNA was efficiently amplified by one of the primer sets targeting the groEL gene, and LAMP assay sensitivity with this primer set was 10-fold higher than that of the polymerase chain reaction. Moreover, the groEL gene was successfully amplified from several strains of Ca. Phytoplasma asteris by this primer set, indicating that the groEL gene can be used as a LAMP assay target gene for a broad range of phytoplasma strains. Additionally, a simple DNA extraction method that omits the homogenizing and phenol extraction steps was combined with the LAMP assay to develop a simple, rapid, and convenient diagnostic method for detecting phytoplasma.  相似文献   

A newly established bovine intestinal epithelial cell line is effective for in vitro screening of potential antiviral immunobiotic microorganisms for cattle     

Chiba E  Villena J  Hosoya S  Takanashi N  Shimazu T  Aso H  Tohno M  Suda Y  Kawai Y  Saito T  Miyazawa K  He F  Kitazawa H 《Research in veterinary science》2012,93(2):688-694

We evaluated whether a bovine intestinal epithelial (BIE) cell line could serve as a useful in vitro model system for studying antiviral immune responses in bovine intestinal epithelial cells (IECs) and for the primary screening of immunobiotic microorganisms with antiviral protective capabilities. Immunofluorescent analyses revealed that toll-like receptor 3 (TLR3) was expressed in BIE cells, and the results of real-time quantitative PCR showed that these cells respond to stimulation with poly(I:C) by up-regulating pro-inflammatory cytokines and type I interferons. In addition, we demonstrated that BIE cells are useful for the primary screening of immunobiotic lactic acid bacteria strains which are able to beneficially modulate antiviral immune responses triggered by TLR3 activation in bovine IECs. The characterization of BIE cells performed in the present study represents an important step towards the establishment of a valuable bovine in vitro system that could be used for the development of immunomodulatory feed for bovine hosts.  相似文献   

A cytolethal distending toxin gene-based multiplex PCR assay for detection of Campylobacter spp. in stool specimens and comparison with culture method     

Shiramaru S  Asakura M  Inoue H  Nagita A  Matsuhisa A  Yamasaki S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(7):857-862

In this study, we evaluated the applicability of cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR for the direct detection and identification of Campylobacter jejuni, C. coli and C. fetus from stool specimens of patients with gastroenteritis in comparison to culture methods. A total of 711 stool specimens were examined for the isolation or detection of campylobacters by using Skirrow's selective agar culture plates, a filtration method and the multiplex PCR assay. Forty-one and 36 C. jejuni strains were isolated by culture and filtration methods, respectively. In addition, 2 and 3 C. coli strains were isolated by Skirrow and the filtration methods, respectively. However, when the multiplex PCR was employed, the cdtB genes of C. jejuni and C. coli were detected in 45 and 4 stool samples, respectively, and 9 C. jejuni PCR-positive samples by multiplex PCR were negative by culture method. Sequence analysis of the PCR products obtained from 8 stool specimens from which campylobacters were not isolated by culture method but the sequences exactly matched with that of the cdtB gene of C. jejuni strain 81-176. None of the remaining stool samples which were culture negative for campylobacters produced any amplicon. Stool samples were defined as Campylobacter-positive if detected by any method. The sensitivity of the multiplex PCR was 83%, which was higher than Skirrow (74%) and filtration method (66%). These data indicate that cdtB gene-based multiplex PCR is a rapid and more sensitive method to identify the most important species of Campylobacter for human diseases. (248).  相似文献   

Trapped neutrophil syndrome in a Border Collie dog: clinical, clinico-pathologic, and molecular findings     

Mizukami K  Shoubudani T  Nishimoto S  Kawamura R  Yabuki A  Yamato O 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(6):797-800

Trapped neutrophil syndrome (TNS) is an autosomal recessive inherited neutropenia known in Border Collies since the 1990's. Recently, the causative mutation has been identified in the canine VPS13B gene and a DNA-based diagnosis has now become available. The present paper describes clinical and clinico-pathologic findings in a Border Collie with TNS that was molecularly diagnosed for the first time in Japan. In a 10-week-old male Border Collie with microgenesis and symptoms related to recurrent infections, a hematological examination revealed severe leukopenia due to neutropenia, suggesting the dog to be affected by inherited neutropenic immunodeficiency. Direct DNA sequencing demonstrated that the dog was homozygous for the causative mutation of TNS and both its parents were heterozygous carriers. In addition, a simple and rapid polymerase chain reaction-based length polymorphism analysis coupled with microchip electrophoresis was developed for the genotyping of TNS. This assay could discriminate clearly all genotypes, suggesting that it was suitable for both individual diagnosis and large-scale surveys for prevention.  相似文献   

Plasma Profiles of Glucose, Insulin and Lipids in the Male WBN/Kob-Lepr(fa) Rat, a New Model of Type 2 Diabetes with Obesity     

Kaji N  Okuno A  Ohno-Ichiki K  Oki H  Ishizawa H  Shirai M  Asai F 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2012,74(9):1185-1189

Plasma profiles of glucose, insulin and lipids were examined in the male WBN/Kob-Lepr(fa) (fa/fa) rat, a new model of type 2 diabetes (T2D), in comparison with age-matched original male WBN/Kob (lean) rats. The fa/fa rats developed hypertriglycemia, obesity and hyperglycemia from 5, 7, and 9 weeks of age, respectively. Plasma insulin levels in fa/fa rats were significantly higher than those in lean rats at 5 weeks of age, but after 11 weeks of age gradually declined to the levels in lean rats. HOMA-IR, a measure of insulin resistance status, showed that fa/fa rats had insulin resistance. The fa/fa rat has the potential to become an important animal model of T2D with obesity.  相似文献   

Effects of soil scraping on the reclamation of tsunami-damaged paddy soil     

Dong-Jin Kang  Akira Endo  Young-Jin Seo 《Journal of Crop Science and Biotechnology》2013,16(3):219-223

We investigated the possibility of tsunami-damaged paddy soil remediation in Japan using a scraping method. We collected undisturbed soil blocks to maintain the soil structure from paddy damaged by salt from the tsunami. Scraping treatments to remove the top 2 cm of the soil surface were conducted once, twice, and thrice after drying naturally in a greenhouse environment, as well as a noscraping control. Electrical conductivity (EC) of the soil dramatically decreased to < 1.36 dS m^?1 in the residual soils after scraping. The EC values of the residual soils were significantly lower after being scraped twice or thrice compared with a single scraping. Similarly, the mean NaCl removal rates were significantly higher with two (91%) or three scrapings (97.2%) compared with a single scraping (73.3%). The growth of rice plants was almost normal in the residual soils based on a visual score (approximately 3 on average) using a standard evaluation system at 4 weeks after transplantation. There were no significant differences in the grain yields from the residual soils with different scraping frequencies, and the EC values of all the residual soils were < 1.36 dS m^?1. This study clearly indicates that there were reductions in the EC and good plant growth performance in the residual soils after scraping, and there was also a high rate of NaCl removal. Therefore, scraping may be a useful method for the remediation of salt-damaged paddy soils when irrigation or culvert systems are affected severely by tsunamis and earthquakes.  相似文献   

Fine mapping of quantitative trait loci affecting organ weights by mouse intersubspecific subcongenic strain analysis     

Md. Bazlur Rahman Mollah  Akira Ishikawa 《Animal Science Journal》2013,84(4):296-302

The genetic architecture of organ weights is not well understood. In this study, we fine‐mapped quantitative trait loci (QTLs) affecting organ weights by characterizing six intersubspecific subcongenic mouse strains with overlapping and non‐overlapping genomic regions on chromosome 2 derived from wild Mus musculus castaneus. QTLs for heart, lung, spleen and kidney weights were revealed on a 6.38‐Mb genomic region between two microsatellite markers, D2Mit323 and D2Mit472. Effects of the castaneus alleles at the organ weight QTLs were all opposite in direction to a body weight QTL previously mapped to the same genomic region. In addition, new QTLs for lung and kidney weights were revealed on a different 3.57‐Mb region between D2Mit205 and D2Mit182. Their effects were dependent on that of another body weight QTL previously mapped to that genomic region. The organ weight QTLs revealed were all duplicated in independent analyses with F₂ intercross populations between subcongenic strains carrying these QTLs and their background strain. The findings suggested that organ weights are not exclusively regulated by genetic loci that commonly influence overall body weight and rather that there are loci contributing to the growth of specific organs only.  相似文献   

Empirical Percentile Growth Curves with Z-scores Considering Seasonal Compensatory Growths for Japanese Thoroughbred Horses     

Tomoaki ONODA  Ryuta YAMAMOTO  Kyohei SAWAMURA  Harutaka MURASE  Yasuo NAMBO  Yoshinobu INOUE  Akira MATSUI  Takeshi MIYAKE  Nobuhiro HIRAI 《Journal of Equine Science》2013,24(4):63-69

Percentile growth curves are often used as a clinical indicator to evaluate variations of children’s growth status. In this study, we propose empirical percentile growth curves using Z-scores adapted for Japanese Thoroughbred horses, with considerations of the seasonal compensatory growth that is a typical characteristic of seasonal breeding animals. We previously developed new growth curve equations for Japanese Thoroughbreds adjusting for compensatory growth. Individual horses and residual effects were included as random effects in the growth curve equation model and their variance components were estimated. Based on the Z-scores of the estimated variance components, empirical percentile growth curves were constructed. A total of 5,594 and 5,680 body weight and age measurements of male and female Thoroughbreds, respectively, and 3,770 withers height and age measurements were used in the analyses. The developed empirical percentile growth curves using Z-scores are computationally feasible and useful for monitoring individual growth parameters of body weight and withers height of young Thoroughbred horses, especially during compensatory growth periods.  相似文献   

Physiological responses of young thoroughbred horses to intermittent high-intensity treadmill training     

Hajime Ohmura  Akira Matsui  Tetsuro Hada  James H Jones 《Acta veterinaria Scandinavica》2013,55(1):59

Background

Training of young Thoroughbred horses must balance development of cardiopulmonary function and aerobic capacity with loading of the musculoskeletal system that can potentially cause structural damage and/or lameness. High-speed equine treadmills are sometimes used to supplement exercise on a track in the training of young Thoroughbreds because the horse can run at high speeds but without the added weight of a rider. We tested the hypothesis that intermittent high-intensity exercise on a treadmill of young Thoroughbred horses entering training can enhance development of aerobic capacity (Vo₂max) and running performance more than conventional training under saddle, and do so without causing lameness.

Results

Twelve yearling Thoroughbreds trained for 8 months with conventional riding (C) only, conventional riding plus a short (2 month, S) interval of once-per-week high-intensity treadmill exercise, or a long (8 month, L) interval of once-per-week high-intensity treadmill exercise. Three treadmill exercise tests evaluated Vo₂max, oxygen transport and running performance variables in June of the yearling year (only for L), October of the yearling year and April of the 2-year-old year. No horses experienced lameness during the study. Aerobic capacity increased in all groups after training. In both October and April, Vo₂max in L was higher than in C, but did not differ between L and S or S and C. Running speeds eliciting Vo₂max also increased in all groups after training, with S (809 ± 3 m/s) and L (804 ± 9 m/s) higher than C (764 ± 27 m/s). Maximum heart rate decreased for all groups after training. Hematocrit and hemoglobin concentration increased for L throughout training.

Conclusions

Young Thoroughbred horses can increase aerobic capacity and running performance more than by strictly using track training under saddle with the addition of intermittent high-intensity treadmill exercise, and they can do so without experiencing lameness. This finding suggests that young racehorses might be able to achieve higher aerobic fitness during training without subjecting their musculoskeletal systems to increased loading and risk of developing lameness. The findings of this preliminary study do not indicate a specific protocol to best achieve this goal.  相似文献   

Effects of Exposure to Nanoparticle-rich Diesel Exhaust on Pregnancy in Rats     

ChunMei LI  Xuezheng LI  Akira K. SUZUKI  Yonghui ZHANG  Yuji FUJITANI  Kentaro NAGAOKA  Gen WATANABE  Kazuyoshi TAYA 《The Journal of reproduction and development》2013,59(2):145-150

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