Real-time PCR genotyping assay for feline erythrocyte pyruvate kinase deficiency and mutant allele frequency in purebred cats in Japan

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.     

Sodium bicarbonate protects uranium-induced acute nephrotoxicity through uranium-decorporation by urinary alkalinization in rats

To evaluate the effectiveness of sodium bicarbonate (SB) in removing uranium and protecting animals from uranium toxicity, we intramuscularly administered 1 mg/kg of uranyl nitrate to 8-wk-old male SD rats, and 20 min after administration of uranyl nitrate, the animals were given a single oral administration of SB at 0.1, 0.3 or 1 g/kg. The SB treatment at a dose of 0.3 g/kg or more raised the pH of the rats’ urine until 4 h after treatment, and it significantly reduced the uranium amounts in the kidneys at 1 day after treatment. In another experiment, rats were intramuscularly administered 1 mg/kg of uranyl nitrate, and 20 min later, the animals were treated with sodium bicarbonate (0.1 or 1 g/kg). The rats were autopsied at 1, 3 and 7 days after uranium treatment. High-dose SB resulted in a significant increase in urinary uranium excretion in the first 24 h and a reduction of uranium deposition in the kidneys and femurs, and it also significantly suppressed uranium-induced renal toxicity, as shown by both histopathology and clinical chemistry at 3 days after uranium treatment. Low-dose SB did not show such marked effects. Our findings demonstrated that the uranium decorporation effect of sodium bicarbonate was observed at the dosage showing urine alkalinization in rats and that decorporation effect of sodium bicarbonate might be beneficial if it is administered immediately after incorporation of soluble uranium.     

Calcium ions are involved in egress of Babesia bovis merozoites from bovine erythrocytes

Bovine babesiosis is a livestock disease known to cause economic losses in endemic areas. The apicomplexan parasite Babesia bovis is able to invade and destroy the host’s erythrocytes leading to the serious pathologies of the disease, such as anemia and hemoglobinuria. Understanding the egress mechanisms of this parasite is therefore a key step to develop new therapeutic strategies. In this study, the possible involvement of Ca²⁺ in the egress of B. bovis merozoites from infected erythrocytes was investigated. Egress was artificially induced in vitro using calcium ionophore A23187 and thapsigargin to increase Ca²⁺ concentration in the cytosol of the parasite cells. The increased intracellular Ca²⁺ concentration following these treatments was confirmed using live cell Ca²⁺ imaging with confocal laser scanning microscopy. Based on our findings, we suggest a Ca²⁺ signalling pathway in the egress of B. bovis merozoites.     

Genetic analysis of fat‐to‐protein ratio,milk yield and somatic cell score of Holstein cows in Japan in the first three lactations by using a random regression model

We estimated the genetic parameters of fat‐to‐protein ratio (FPR) and the genetic correlations between FPR and milk yield or somatic cell score in the first three lactations in dairy cows. Data included 3 079 517 test‐day records of 201 138 Holstein cows in Japan from 2006 to 2011. Genetic parameters were estimated with a multiple‐trait random regression model in which the records within and between parities were treated as separate traits. The phenotypic values of FPR increased soon after parturition and peaked at 10 to 20 days in milk, then decreased slowly in mid‐ and late lactation. Heritability estimates for FPR yielded moderate values. Genetic correlations of FPR among parities were low in early lactation. Genetic correlations between FPR and milk yield were positive and low in early lactation, but only in the first lactation. Genetic correlations between FPR and somatic cell score were positive in early lactation and decreased to become negative in mid‐ to late lactation. By using these results for genetic evaluation it should be possible to improve energy balance in dairy cows.     

Sorption of Cr(VI) on the wood of Japanese larch treated with concentrated sulfuric acid

A carbonaceous sorbent was prepared from the wood of Japanese larch (Larix leptolepis) by dehydration with concentrated sulfuric acid in a 69% yield. The abilities of the sorbent to remove Cr(VI) from aqueous solutions were investigated. Research parameters included the initial solution pH, temperature, and initial concentration of Cr(VI) in solution. The removal of Cr(VI) was highly solution pH dependent and was mainly governed by physicochemical sorption under weak acidic conditions. The equilibrium data fit well in the Langmuir isotherm model. The Langmuir constants were calculated at different temperatures, and the sorption capacity increased with rising temperature, indicating the endothermic nature of the Cr(VI) sorption onto the sorbent. The desorption experiments suggest that the Cr(VI) sorption is generally irreversible, owing to strong interaction of HCrO ₄ ⁻ with the active sites of the sorbent.     

Variations in transpiration rate and leaf cell turgor maintenance in saplings of deciduous broad-leaved tree species common in cool temperate forests in Japan

To clarify mechanisms underlying variation in transpiration rate among deciduous broad-leaved tree species, we measured diurnal changes in stomatal conductance (gs) and leaf water potential, and calculated the maximum transpiration rate (Emax), leaf-specific hydraulic conductance (K(s-l)) and difference between the soil water potential and the daily minimum leaf water potential (Psis - Psi(l,min)). Pressure-volume (P-V) measurements were made on leaves. Saplings of eight broad-leaved tree species that are common in Japanese cool temperate forests were studied. Maximum transpiration rate varied significantly among species. There was a statistically significant difference in Psis - Psi(l,min), but not in K(s-l). Species with large Emax also had large Psis - Psi(l,min) and gs. The results of the P-V analyses showed that species with a large Psis - Psi(l,min) maintained turgor even at low leaf water potentials. The similar daily minimum leaf pressure potentials (Psip) across all eight species indicate that Psip values below this minimum are critical. Based on these results, we suggest that the leaf cell capacity for turgor maintenance strongly affects Psis - Psi(l,min) and consequently Emax via stomatal regulation.     

The process of decline of an endangered tree species,Pinus armandii Franch. var.amamiana (Koidz.) Hatusima, on the southern slope of Mt. Hasa-dake in Yaku-shima Island

The process of decline of an endangered tree species,Pinus armandii var.amamiana, was monitored on the southern slope of Mt. Hasa-dake in Yaku-shima Island from 1994 to 1998. There are 163 trees ofP. armandii var.amamiana over 1.3 m in height. They are distributed on steep ridges and rocks with a thin soil layer mostly consisting of friable granite. During the monitoring period, 21 trees ofP. armandii var.amamiana died and the mortality rate was 12.9%. Dead trees were categorized into three types: standing, uprooted and landslide. The uprooted- and landslide-typed dead trees were found only after a severe typhoon struck Yaku-shima Island. This suggests that the combination of fragile site conditions and severe typhoons play an important role in the process of decline ofP. armandii var.amamiana. The standing-typed dead trees were presumed to have been killed by pine wilt disease, accounting for 71.4% of the dead trees. However,Bursaphelenchus xylophilus, the cause of pine wilt disease, was not detected from any of the wood chips or branch samples from the standing-typed dead trees ofP. armandii var.amamiana. This might indicate that some factor(s) other than pine wilt disease could be responsible for the standing-typed death ofP. armandii var.amamiana in natural habitats. A part of this paper was presented at the 109th Meeting of the Japanese Forestry Society.     

Plus tree of Cryptomeria japonica D. Don with a heterozygous male-sterility gene

To find plus tree clones of Cryptomeria japonica that are heterozygous for a male-sterility gene (Aa), we crossed a homozygous male-sterile tree (aa) with 63 clones. Male sterility in this case is controlled by a recessive allele at a single gene locus and is expressed only in homozygotes. All F₁ seedlings obtained by crossing the male-sterile mother tree and 62 out of the 63 clones produced pollen. In contrast, F₁ seedlings obtained from the crossing between the male-sterile mother tree and a plus tree clone, Ohara 13, produced 64 male-sterile individuals and 52 fertile individuals. The segregation ratio fitted the expected 1 : 1 ratio according to a chi-square test. These results clearly demonstrate that the Ohara 13 clone is heterozygous for a male-sterility gene.     

Effects of a severe typhoon on forest dynamics in a warm-temperate evergreen broad-leaved forest in southwestern Japan

The effects of a typhoon on forest dynamics and the response of major tree species were studied in a warm-temperate evergreen broad-leaved forest in southwestern Japan. The strongest typhoon on record (T9313) passed through this region in 1993. Return periods of typhoons over 30 ms⁻¹ in instantaneous wind velocity and T9313 were estimated to be 2.2 and 104.5 years, respectively. Approximately 10% of all stems suffered some damage from T9313 and annual stem mortality rose from 1.3 to 2.7%. The estimated period that the number of stems would fall below 10% of the initial was four years shorter with T9313-class typhoons than without them. Thus, the disturbance by T9313 was not catastrophic at the site although T9313 was an episodic typhoon. The short-term responses of major tree species to T9313 were classified into four types: 1) blunt-response type with little decrease and recruitment of stems in the DBH ≥ 5 cm class (Distylium racemosum), 2) retreat type with larger decrease than recruitment (e.g. Quercus acuta), 3) sharp-response type with a large decrease and much recruitment (e.g. Cinnamomum japonicum), and 4) advance type with less decrease than recruitment (Eurya japonica). Among the four regeneration types classified by previous studies (climax, light-demanding, subcanopy, and few-sapling), the climax and few-sapling types each showed a specific short-term response, the blunt-response and retreat types, respectively, that explains one aspect of the regeneration strategies of each type. On the other hand, the light-demanding or sub-canopy type showed multiple short-term responses, indicating that each regeneration type contains species with various regeneration strategies. In this paper, I analyzed data from the database that was compiled by the Aya Research Team.