Furosemide loading test in a case of homozygous solute carrier family 12, member 1 (SLC12A1) mutation (g.62382825G>A,p.Pro372Leu) in Japanese Black cattle

Hydrallantois is the excessive accumulation of fluid in the allantoic cavity in a pregnant animal and is associated with fetal death. We recently identified a recessive missense mutation in the solute carrier family 12, member 1 (SLC 12A1 ) gene (g.62382825G>A, p.Pro372Leu) that is associated with hydrallantois in Japanese Black cattle. Unexpectedly, we found a case of the homozygous risk‐allele for SLC 12A1 in a calf, using a PCR ‐based direct DNA sequencing test. The homozygote was outwardly healthy up to 3 months of age and the mother did not exhibit any clinical symptoms of hydrallantois. In order to validate these observations, we performed confirmation tests for the genotype and a diuretic loading test using furosemide, which inhibits the transporter activity of the SLC 12A1 protein. The results showed that the calf was really homozygous for the risk‐allele. In the homozygous calf, administration of furosemide did not alter urinary Na⁺ or Cl^? levels, in contrast to the heterozygote and wild‐type calves in which these were significantly increased. These results demonstrate that the SLC 12A1 (g.62382825G>A, p.Pro372Leu) is a hypomorphic or loss‐of‐function mutation and the hydrallantois with this mutation shows incomplete penetrance in Japanese Black cattle.     

Preliminary application and evaluation of loop-mediated isothermal amplification (LAMP) for detection of bovine theileriosis and trypanosomosis in Tanzania

The sensitivity of LAMP, PCR and microscopy to detect Theileria spp. and Trypanosoma congolense in field-derived bovine blood samples from Tanzania was evaluated and compared. No parasites were detected by microscopy. Furthermore, no bovine Theileria spp. were detected by LAMP and PCR from all the 24 samples collected from Arusha. Four and one out of 24 samples were positive for Theileria congolense infection by LAMP and PCR respectively while, 18 and nine out of 40 samples from Dar es Salaam were positive by LAMP and PCR for Theileria spp. Infection, respectively. Although all samples from Dar es Salaam were negative for Trypanosoma congolense infections by PCR, 12 out of 40 samples were LAMP positive. Whilst PCR is an established gene amplification method for the detection of Theileria and trypanosome parasites, this study introduces LAMP as an alternative molecular diagnostic tool that could be used in large-scale epidemiological surveys.     

Relation between tacticity and fiber diameter in melt-electrospinning of polypropylene

Electrospun atactic polypropylene (PP) fibers are thicker than those obtained from isotactic PP, although the viscosity of molten PPs is almost same. Thus we focused on the effect of tacticity of PP on fiber diameters. The PP samples with various tacticity were prepared by changing the blend ratio of isotactic PP and atactic PP. Melt-electrospinning is performed by using blended samples, and then electrospun fibers were observed by scanning electron microscope to evaluate fiber diameter of obtained fibers. It is clear that the diameter of electrospun PP fibers decreases as high tacticity content of PP increases. This result suggests that tacticity of samples is an important factor to control the electrospun fiber diameter.     

Identification of SSR markers linked to the Phytophthora resistance gene Rps1-d in soybean

To identify markers for the Phytophthora resistance gene, Rps1‐d, 123 F_{2 : 3} families were produced from a cross between Glycine max (L.) Merr. ‘Tanbakuro’ (a Japanese traditional black soybean) and PI103091 (Rps1‐d) as an experimental population. The results of virulence tests produced 33 homozygous resistant, 61 segregating and 29 homozygous susceptible F_{2 : 3} families. The chi‐squared test gave a goodness‐of‐fit for the expected ratio of 1 : 2 : 1 for resistant, segregating and susceptible traits, suggesting that the inheritance of Rps1‐d is controlled by a monogenic dominant gene. Simple sequence repeat (SSR) analyses of this trait were carried out using the cultivars ‘Tanbakuro’ and PI103091. Sixteen SSR primers, which produced 19 polymorphic fragments between the two parents, were identified from 41 SSR primers in MLG N. Eight SSR markers were related to Rps1‐d, based on 32 of the 123 F_{2 : 3} families, consisting of 16 homozygous resistant and 16 homozygous susceptible lines. The remaining 91 families were analysed for these eight markers, and a linkage map was constructed using all 123 F_{2 : 3} families. The length of this linkage group is 44.0 cM. The closest markers, Sat_186 and Satt152, are mapped at 5.7 cM and 11.5 cM, respectively, on either side of the Rps1‐d gene. Three‐way contingency table analysis indicates that dual‐marker‐assisted selection using these two flanking markers would be efficient.     

Age,growth and age at sexual maturity of fan ray <Emphasis Type="Italic">Platyrhina sinensis</Emphasis> (Batoidea: Platyrhinidae) in Ariake Bay,Japan

Abstract:   Age, growth and sexual maturity of the fan ray Platyrhina sinensis in Ariake Bay, Japan were determined from specimens collected from May 2002 to September 2006. Age determination was conducted by vertebral centrum analysis using soft X-radiography. Annual band pair deposition was determined by marginal increment and edge analyses. The von Bertalanffy growth model best described the overall pattern of growth for both males and females (males L _∝  = 455.2, k  = 0.56, t ₀ = −1.09; females L _∝  = 555.8, k  = 0.28, t ₀ = −1.77; L _∝ is the theoretical asymptotic total length in mm, k is the growth rate coefficient and t ₀ is the theoretical time at zero length). Parameter estimates suggest that females attain a larger asymptotic total length and grow more slowly than males. The observed maximum ages were 5 years for males and 12 years for females. Age at 50% sexual maturity was 2.1 years for males and 2.9 years for females. The results indicate that this species is relatively fast-growing, short-lived and early maturing compared with many batoid species.     

Seasonal changes in oocyte size and maturity of the giant jellyfish, Nemopilema nomurai

To determine the reproductive season of the giant jellyfish Nemopilema nomurai, we investigated gonadal maturity in specimens collected from the East China Sea, Korea Strait, Wakasa Bay, and the Shonai Coast of Yamagata Prefecture. After the sex of the samples was determined, the long axis of at least 256 oocytes from each female was measured. In specimens collected from the coast of Japan in 2005 and in 2006, all gonads were sufficiently developed to determine sex. However, 18 of the 20 specimens from the East China Sea collected in July 2005 were immature, and sex could not be determined. The maximum and third quartile of oocyte length had a significant correlation with days elapsed from 30 June, but they were not related to bell diameter. Observations of gonad tissue sections of specimens collected in Wakasa Bay in 2006 confirmed that oocyte length was a good proxy for female maturity. Male maturity could also be determined. In conclusion, the sex of all of the small-sized medusae collected along the coast of Japan was determinable, and their gonads were at various stages of development up to fully mature. Therefore, the occurrence of small-sized jellyfish during the autumn in Wakasa Bay is not caused by recruitment of young population from the nearby coast.     

Significance of <Emphasis Type="Italic">CmCCD4a</Emphasis> orthologs in apetalous wild chrysanthemum species,responsible for white coloration of ray petals

Most chrysanthemum (Chrysanthemum morifolium Ramat.) flowers have a central capitulum, composed of many disc florets that is surrounded by ray petals. CmCCD4a, a gene that encodes a carotenoid cleavage dioxygenase (CCD), is expressed specifically in the ray petals of chrysanthemum cultivars, and its expression leads to white ray petals as a result of carotenoid degradation. Here, we show that wild chrysanthemums with white ray petals have CmCCD4a orthologs, whereas those with yellow ray petals lack these orthologs, as is the case in chrysanthemum cultivars. CmCCD4a orthologs also exist in some lines of Chrysanthemum pacificum and Chrysanthemum shiwogiku, even though these species lack ray petals. Interspecific hybridization between C. shiwogiku and a yellow-flowered chrysanthemum cultivar showed that the CmCCD4a orthologs from C. shiwogiku lead to the development of white ray petals. This indicates that the translation products of the CmCCD4a orthologs maintain enzymatic activity that can degrade carotenoids in chrysanthemums, irrespective of whether or not the ray petals that CmCCD4a expression actually occurred.     

Mutation of the GABA receptor associated with fipronil resistance in the whitebacked planthopper, Sogatella furcifera

We found the A2′N mutation (index number for M2 membrane spanning region) in the GABA receptor subunit of fipronil-resistant Sogatella furcifera, by analyzing DNA sequences amplified from fipronil-resistant and -susceptible S. furcifera. In order to confirm the role of A2′N mutation in the fipronil resistance, we expressed the wild-type and A2′N mutant Drosophila GABA receptors in Drosophila Mel-2 cells stably. Amino acid sequences of three membrane spanning regions (M1-M3), which are important for binding of fipronil, are conserved between Drosophila and S. furcifera. So the results of A2′N mutant Drosophila GABA receptor suggest the role of A2′N mutation in fipronil-resistant S. furcifera. The membrane potential assay showed that the A2′N mutant Drosophila GABA receptor was not inhibited by fipronil at all, while the IC₅₀ value of fipronil for wild-type Drosophila GABA receptor was 172 nM. These results suggest that A2′N mutation confers the resistance of fipronil in S. furcifera.     

A simple specimen preparation method for histopathological evaluation of vestibular organs

Vestibular organs consist of the maculae staticae, which are located in both the utricle and saccule, as well as the semicircular ducts and their ampullas. There have been no reports on specimen preparation methods for vestibular organs, including maculae staticae or semicircular ducts. In this study, we investigated highly reproducible methods of preparing vestibular organ specimens for histopathological examinations. We established a method that allows researchers to observe the utricle and saccule, including otoliths, the ampulla of a semicircular duct, and parts of semicircular ducts. This highly reproducible method is useful for histopathological analysis of mice with symptoms of abnormal equilibrium caused by medical toxicity and genetic modification.