Experimental infection of cats with Chlamydophila felis

Cats experimentally infected with a British isolate of Chlamydophila felis (C. felis), B166 strain, by droplet into the eye and nose developed conjunctivitis, mild rhinitis and fever. The chlamydophila were first isolated from conjunctiva, nictitating membrane and then from lung, tonsil, liver, spleen, kidney, nasal and vaginal swabs and blood. These results indicate that C. felis B166 strain first infected and replicated in the conjunctiva and nictitating membrane in cats with symptoms which were mostly limited to conjunctivitis, and then pervaded the whole body by bacteremia.     

Evaluation of recent changes in genetic variability in Japanese thoroughbred population based on a short tandem repeat parentage panel

The integrity of thoroughbreds is maintained under strict regulation involving DNA parentage testing, which is robust in a population with high genetic variability. The genetic variability of the thoroughbred population is possibly fluctuating because of selective breeding that has focused on adaptations for racing performance. To monitor genetic variability within the population and the effectiveness of short tandem repeat (STR) parentage testing, we investigated allele frequencies and the exclusion probability (PE) of 16–17 loci of a parentage panel in the Japanese thoroughbred population over 15 years. Expected heterozygosities (He) of 14 loci indicated a decreasing trend, and the average He of the population decreased significantly. Low genetic variability was possibly induced by a decrease in population size and a selective breeding bias. Four loci showed both a significant increase in allele frequency and a significant decrease in He; it is assumed that those loci were affected by positive selection for racing performance. There was a significant decrease in the PE because of the changes in genetic variability; however, it has remained over 0.99995. The current STR panel is still effective for parentage control, but it will be necessary to continuously monitor genetic variability, which has decreased over 15 years.     

Advanced treatment technique for swine wastewater using two agents: Thermally polymerized amorphous silica and hydrated lime for color and phosphorus removal and sulfur for nitrogen removal

The efficacy of advanced treatment of swine wastewater using thermally polymerized, modified amorphous silica and hydrated lime (M‐CSH‐lime) for color and phosphorus removal and sulfur for nitrogen removal was examined with a demonstration‐scale treatment plant. The color removal rate was approximately 78% at M‐CSH‐lime addition rates of > 0.055 wt/v%. The ‐P removal rate exceeded 99.9% with > 0.023 wt/v%. pH of the effluent from the M‐CSH‐lime reactor increased with the addition rate till a maximum value of 12.7, which was effective in disinfection. The recovered M‐CSH‐lime would be suitable as a phosphorus fertilizer because the total P₂O₅ content was approximately 10%. The nitrogen oxide (NOx‐N) removal rate by sulfur denitrification increased to approximately 80% when the NOx‐N loading rate was around 0.1 kg‐N/ton‐S/day. It was suggested that the combination of the two processes would be effective in the advanced treatment of swine wastewater.     

Design and storage stability of reference materials for microfluidic quantitative PCR-based equine gene doping tests

One method of gene doping in horseracing is administering of exogenous genetic materials, known as transgenes. Several polymerase chain reaction (PCR)-based methods have been developed for detecting transgenes with high sensitivity and specificity. However, novel designs for reference materials (RMs) and/or positive template controls (PTCs) are necessary for simultaneous analysis of multiple transgene targets. In this study, we designed and developed a novel RM for simultaneously detecting multiple targets via microfluidic quantitative PCR (MFQPCR). Twelve equine genes were selected as targets in this study. A sequence region including primers and probes for quantitative PCR was designed, and a 10 bp sequence was inserted to allow the RM to be distinguished from the original transgene sequences. The sequences of individual detection sites were then connected for 12 genes and cloned into a single plasmid vector. We performed fragment size analysis to distinguish between the PCR products of the original transgene sequence and those of the RM, enabling identification of RM contamination. PTCs diluted to 10,000, 1,000, 100, and 10 copies/µl with horse genomic DNA from RM were stably stored at 4°C for 1 year. As digital PCR enabled absolute quantification, the designed substances can serve as an RM. These findings indicate that the RM design and storage conditions were suitable for gene doping tests using MFQPCR.     

Flexible Alumina-Silica Nanofibrous Membrane and Its High Adaptability in Reactive Red-120 Dye Removal from Water

Flexible and self-standing membrane composed of ultrafine alumina-silica nanofibers (NFs) has been successfully fabricated by the electrospinning method, and further used as an adsorbent for the adsorptive decolorization of Reactive Red-120 (RR-120) dye from an aqueous system. Effects of pH, adsorbent dosage, and contact time on adsorption have been studied. The adsorption of RR-120 on the NFs was found to be highly pH dependent and the optimum pH was found to be 3. The adsorption equilibrium data was explained well with the Langmuir isotherm model, and the maximum sorption capacity was found to be 884.95 mg/g, which was several folds higher than the adsorption capacity of a number of recently studied potential adsorbents. After adsorption, the NF mat could be separated from the liquid phase conveniently and reused. The sorption kinetics was found to follow an intraparticle diffusion model. The high adsorption performance, excellent flexibility, easy recovery, and reuse characteristic of the alumina-silica NF membrane all favor its practical application in environmental remediation.     

Breed Differences in Dopamine Receptor D4 Gene (DRD4) in Horses

Genetic polymorphisms in genes related to neurotransmitters or hormones affect personality or behavioral traits in many animal species including humans. In domestic animals, the allele frequency of such genes has been reported to be different among breeds and it may account for breed differences in behavior. In this study, we investigated breed differences in horses in the dopamine receptor D4 gene (DRD4), which has been reported to affect horse personality. We collected samples from seven horse breeds including those native to Japan and Korea, and compared the sequence of the DRD4 exon3 region among these breeds. We found that there were two types of polymorphisms (VNTR and SNPs) in the exon3 region, and some of them seemed to be breed-specific. In addition, we found that the allele frequency of G292A, reported to be associated with horse personality, differed greatly between native Japanese horses and Thoroughbred horses. The frequency of the A allele which is associated with low curiosity and high vigilance, was much lower in native Japanese horses (Hokkaido, 0.03; Taishu, 0.08) than in Thoroughbreds (0.62). This difference may account for breed differences in personality or behavioral traits. Further studies of the function of these polymorphisms and their effect on behavior are indicated.     

Heritability estimates of fractures in Japanese Thoroughbred racehorses using a non‐linear model

Thoroughbred racehorses are produced by mating small numbers of Arabian stallions and native British mares, and have been improved by selection of horseracing performance for about 300 years. While these improvements led to good performance as racehorses, they exposed horses to numerous medical disorders, aggravated by extensive exercise. Fractures are frequent medical disorders in Thoroughbred racehorses. In this study, fracture heritability was estimated using 3,927 Japanese Thoroughbred racehorses to elucidate the risk of racehorse fractures. The heritability estimates of all examined fractures were low (h² = 0.06), while those of fractures in carpal bone and carpus (carpal bone plus distal radius) were moderate (h² = 0.37, 0.24, respectively). Fracture occurrence age for carpal bone and distal radius was both 3.3 years old and was younger than that for other fractures. These results indicated that a larger proportion of the variation in the studied population was due to genetic factors for carpal fractures than for other fractures, while the fractures at other bones were largely affected by environmental factors, correlated with the athlete period (number year in racing). These findings contribute to develop a management plan for suppressing racehorse fractures and improving horseracing safety.     

Polymorphism identification, RH mapping, and association analysis with the anxiety trait of the equine serotonin transporter (SLC6A4) gene

Equine anxiety trait is considered an important temperament in various situations, including riding, training, and daily care. This study examined the polymorphism of the equine serotonin transporter (SLC6A4) gene as a candidate genetic element influencing equine anxiety trait. The sequence of the coding region of this gene was highly homologous with those of other mammals, and four single nucleotide polymorphisms were found by comparing the sequences of ten genetically unrelated thoroughbred horses. Radiation hybrid mapping revealed that this gene was located 26.92 cR from neurofibromin 1 on ECA 11. Using two-year-old thoroughbred horses (n=67), the association of these polymorphisms with the anxiety trait was examined, but no significant association was identified between each haplotype of the serotonin transporter gene and the anxiety score.     

Effects of deer and mice on seedling survival in a temperate mixed forest of Japan

To detect the magnitude of indirect positive effects of deer and mice on seedling survival of some woody species in a Japanese temperate forest, we analyzed the data from an earlier field experiment using a hierarchical Bayesian approach. The forest studied was inhabited by sika deer (Cervus nippon) and mice (Apodemus spp.), and the floor was covered with dwarf bamboo, Sasa nipponica, which negatively affected tree seedlings and was in turn negatively affected by deer and mice. The field experiment was designed as the combination of exclusion or removal of these factors: deer, mice and dwarf bamboo. A hierarchical Bayesian model was constructed and the parameters were estimated by the Markov chain Monte Carlo method. The model successfully showed the indirect positive effects of deer and mice, which improved the survival rate of five cohorts of the seedlings of three tree species by ameliorating the direct negative effect of dwarf bamboo. The cohorts studied were formed by the seedlings of Abies homolepis that emerged in 1997 and 2002, those of Fraxinus lanuginosa f. serrata that emerged in 1998 and 2002, and those of Fagus crenata that emerged in 1999. The positive indirect effect was especially large in F. crenata, which is known to be severely affected by dwarf bamboo. The total effect of deer was shown to be positive for all cohorts except the A. homolepis cohort that emerged in 1997, at which time the dwarf bamboo had not yet fully recovered from the browsing pressure of deer. The total effect of mice was shown to be positive for all of the cohorts. We conclude that these positive effects were due to the large negative effect of dwarf bamboo on the seedlings.