Relationship between transportation stress and polymorphonuclear cell functions of bottlenose dolphins, Tursiops truncatus

Dolphins in a captive environment are exposed to various kinds of stresses. Handling and transportation are stressful events for terrestrial mammals, and such stress may affect immune system function and increase susceptibility to infectious diseases. The same phenomenon could occur in dolphins, however, few studies have reported this in dolphins. The objective of this study was to evaluate the relationship between stress and polymorphonuclear (PMN) cell function of dolphins during transportation. Four bottlenose dolphins (Tursiops truncatus) were transported for 6 hr by truck. Serum cortisol levels, leukograms, phagocytosis, and superoxide production of PMN cells were evaluated during handling and transportation compared to resting values. The mean serum cortisol level was significantly increased during handling and transportation (p<0.05) when compared with the resting values. White blood cell (WBC) counts, eosinophil counts, phagocytosis, and superoxide production of PMN cells during handling and transportation stages decreased significantly in comparison with the resting stage (p<0.05). The concentration of serum cortisol was significantly correlated with the results of the WBC counts, eosinophil counts, superoxide production, and phagocytosis (p<0.01, p<0.05, p<0.05, and p<0.001, respectively). The present results indicate that handling and transportation are stressful events for dolphins and could affect their PMN cell functions, thereby leading to the impairment of the immune system.     

First application of PCR-Luminex system for molecular diagnosis of fungicide resistance and species identification of fungal pathogens

Fungicide resistance in plant pathogens is often caused by a single point mutation in a gene encoding fungicide target proteins. Such is the case for resistance to MBI-D (inhibitors of scytalone dehydratase in melanin biosynthesis) fungicides in rice blast fungus (Magnaporthe oryzae), which is caused by a mutation in the scytalone dehydratase gene that results in a replacement of valine with methionine at codon 75 of the fungicide target protein. PCR-Luminex, a novel system developed for high-throughput analysis of single nucleotide polymorphisms (SNPs) was successfully introduced to diagnose MBI-D resistance using specific oligonucleotide probes coupled with fluorescent beads. The PCR-Luminex system was further tested for its potential in identifying species causing Fusarium head blight on wheat. Four major pathogens, Fusarium graminearum (=F. asiaticum), F. culmorum, F. avenaceum, and Microdochium nivale, known to cause the disease, were tested, and the species were identified using the PCR-Luminex method. So far, this report is the first on the application of the DNA-based PCR-Luminex system in the area of crop protection and/or agricultural sciences.     

Spinal epidural empyema concurrent with sequestrum in a cat: a case report

A 5-month-old intact female mixed cat presented with repetitive paraplegia and drainage of pus from the back despite continuous antibiotic medication. Neurologic examination was consistent with below T3-L3 myelopathy. Computed tomography and magnetic resonance imaging revealed a contrast-enhanced mass in the L1-3 spinal canal, and bone fragments in the T13 and L1 spinal canal. Spinal epidural empyema was suspected, and hemilaminectomy was performed for T12-L2 on the right side and T11-12 on the left side. Bone fragments were diagnosed as sequestrum infected with Bacteroides sp. The cat recovered enough to ambulate next day. One month after surgery, there was no deficit in neurological function. This is the first report of spinal epidural empyema concurrent with sequestrum in a cat.     

Genetic studies for breeding of rice cultivars with superior grain appearance and lodging resistance from the rice cultivar ‘Emi-no-kizuna’

ABSTRACT

We conducted a quantitative trait locus (QTL) analysis of grain appearance (GA) and agronomic traits of rice, using 128 recombinant inbred lines derived from ‘Emi-no-kizuna’ and ‘Tomohonami’. We detected two promising QTLs associated with GA: qGA4 on chromosome 4 and qGA8 on chromosome 8. qGA4 contributed highly to the greater percentage of perfect grains of the Emi-no-kizuna genotype. In the same region, we detected other QTLs associated with panicle number and spikelet number per panicle. In near-isogenic lines (NILs) in which Emi-no-kizuna alleles were introgressed in the genomic region of only the semi-dwarf 1 (sd1) locus (NIL_1) and both the sd1 locus and qGA8 (NIL_2), respectively, the percentage of perfect grains was significantly higher and the percentages of milky white, basal white, and white back grains were significantly lower than in Tomohonami; and the percentages of milky white and white back grains of NIL_2 were significantly lower than those of NIL_1. These results suggest that introgression in the sd1 region could improve GA, and that the addition of qGA8 could further improve GA. The culm lengths of the NILs were significantly shorter than that of Tomohonami, indicating improved lodging resistance. Grain weight of NIL_2 was significantly smaller than that of NIL_1, suggesting that the effect of qGA8 could be pleiotropic, or the gene that underlies qGA8 could be linked with genes associated with grain weight.

Abbreviations: ANOVA: analysis of variance; AT20: mean air temperature in the 20 days after heading; BW: basal white grain; CL: culm length; DAH: days after heading; GA: grain appearance; GW: 1000-grain weight; LOD: logarithm of odds; MW: milky white grain; NIL: near-isogenic line; PG: perfect grain; PL: panicle length; PN: panicle number; PTSN: putative total spikelet number; PVE: percentage of phenotypic variation explained; QTL: quantitative trait locus; RIL: recombinant inbred line; SN: spikelet number per panicle; SNP: single nucleotide polymorphism; WB: white back grain     

Establishment of a potency test by ELISA for a rabies vaccine for animal use in Japan

The ELISA we developed was able to determine the antigen content and was suitable for a potency test, and we described a relative potency assay method which determines the potency of test vaccines by comparing the ELISA value of a test vaccine to that of a reference vaccine. In the present study, we standardized the reference vaccine used for determining the potencies of test vaccines, and established a potency test by ELISA. We evaluated the proposed reference vaccine by the neutralizing antibody responses in dogs after vaccination, by the challenge protection test in guinea pigs (GP potency test), which is the earlier official potency test used in Japan, and by the NIH potency test, which is widely used throughout the world. The results showed that a 4-fold dilution of the proposed reference vaccine induced sufficient immunity in dogs. A 3-fold dilution of the proposed reference vaccine passed the GP potency test. The international units (IU) calibrated by the NIH potency test were 3.7 IU/dose. From the results and the WHO recommendation that veterinary rabies vaccines should have a potency of at least 1.0 IU/dose, we determined to dilute the proposed reference vaccine by 3 fold and regarded it as the reference vaccine. Finally, we confirmed that there is a good agreement between the results of the potency test by ELISA and the results of the GP potency test. The establishment of the potency test by ELISA has made it possible to monitor the potency in the production process and has contributed to the stable production of the vaccine.     

Role of sodium bicarbonate on the initiation of sperm motility in the Japanese eel

ABSTRACT:   In order to find out the role of sodium bicarbonate (NaHCO₃) on the initiation of sperm motility in the Japanese eel Anguilla japonica , interactions were investigated between NaHCO₃ and various reagents (K⁺ channel blocker 4-aminopyridine [4-AP], ammonium chloride [NH₄Cl], sodium acetate and calcium chloride [CaCl₂]) that could regulate internal factors (intracellular K⁺, intracellular pH [[pH]_i] and intracellular Ca²⁺) in sperm motility. Contradictory effects of NaHCO₃ were observed (i.e. an inhibitory effect when 4-AP was absent and a promoting effect when 4-AP was present). Sodium bicarbonate inhibited the initiation of sperm motility in the Japanese eel. However, NaHCO₃ restored the motility of immotile sperm that 4-AP inhibited. The inhibitory effect of NaHCO₃ disappeared with the addition of NH₄Cl, which raised [pH]_i, but the promoting effect was not affected by [pH]_i. Although NaHCO₃ recovered motility in the presence of 4-AP, this recovery was also observed with the addition of CaCl₂ instead of NaHCO₃. In the initiation of sperm motility in the Japanese eel, two roles for NaHCO₃ are suggested: an inhibitory role relating to the regulation of [pH]_i and a promoting role relating to the uptake of another initiation factor, which could be Ca²⁺.     

Identification and use of a wild plant with antimicrobial activity against Ralstonia solanacearum, the cause of bacterial wilt of potato

Fresh aerial tissue and roots of 14 wild plants in Okinawa prefecture were investigated for their antimicrobial activity against Ralstonia solanacearum , which causes bacterial wilt of potato. A 70% aqueous ethanol extract of fresh aerial tissue of Geranium carolinianum L. showed strong antimicrobial activity against R. solanacearum . This extract also showed antimicrobial activity against the pathogens causing common scab of potato and soil rot of sweet potato. The antimicrobial substance could be extracted with hot water, and was effective against R. solanacearum in soil. In the field test, a treatment combining incorporation of dried aerial tissue into the soil and solarization was highly effective for control of bacterial wilt of potato. These findings suggest that G. carolinianum L. could be used as a biological agent for the control of bacterial wilt of potato.     

Relationships of culm surface area to other culm dimensions for bamboo, Phyllostachys pubescens

We determine the relationships of culm surface area to other culm dimensions for one of the largest bamboo species, Phyllostachys pubescens Mazel ex Houzeau de Lehaie. A total number of 150 sample culms were collected from a stand of P. pubescens in Mt. Toshima, Kumamoto Prefecture, western Japan. The culm surface area for each sample was calculated, and then the relationships of the culm surface area to basal area and product of diameter at breast height (dbh) and culm height were analyzed. The relationship between culm surface area and basal area could be described successfully by the power equation, whereas there was a linear relationship of culm surface area to product of dbh and height. Although the regression equations determined here would be useful in estimating culm surface area of P. pubescens, it is necessary to select an appropriate equation depending on the purpose and available time and labor.