First report of Helleborus net necrosis virus isolated from hellebores with black death syndrome in Japan

Helleborus net necrosis virus (HeNNV) in hellebores (Helleborus spp.) has been detected for the first time in Japan. Infected plants had black streaks on fresh leaves and black spots on sepals, which resembled the symptoms of black death disease. The morphology of the virus particles isolated from infected plants was comparable to that of carlaviruses. RT-PCR analysis showed that the entire HeNNV genomic sequence isolated in Japan shared 99% identity with that of HeNNV previously reported in the United States.     

Changes in Microbial Functional Diversity and Activity in Paddy Soils Irrigated with Industrial Wastewaters in Bandung, West Java Province, Indonesia

Characteristics, such as microbial biomass, basal respiration, and functional diversity of the microbial communities, were investigated in paddy soils located in Bandung, West Java Province, Indonesia, that have been heavily polluted by industrial effluents for 31 years. Paddy soil samples (10?C20 cm) were taken from two sites: polluted soils and unpolluted soils (as control sites). The polluted soils contained higher salinity, higher sodicity, higher nutrient contents, and elevated levels of heavy metals (Cr, Mn, Ni, Cu, and Zn) than unpolluted soils. Soil physicochemical properties, such as maximum water holding capacity, exchangeable sodium percentage, sodium adsorption ratio, and swelling factor, in polluted soils were much greater than those in unpolluted soils (P?<?0.05). Changes in the physical and chemical soil properties were reflected by changes in the microbial communities and their activities. BIOLOG analysis indicated that the functional diversity of the microbial community of polluted soils increased and differed from that of unpolluted soils. Likewise, the average rate of color development (average well color development), microbial biomass (measured as DNA concentration), and the soil CO₂ respiration were higher in polluted soils. These results indicate that major changes in the chemical and physical properties of paddy soils following the application of industrial wastewater effluents have had lasting impacts on the microbial communities of these soils. Thus, the increased activity, biomass, and functional diversity of the microbial communities in polluted soils with elevated salinity, sodicity, and heavy metal contents may be a key factor in enhancing the bioremediation process of these heavily polluted paddy soils.     

An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell nuclear transfer enhances full-term development of cloned mouse embryos

In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in vitro development of cloned embryos to the blastocyst stage compared with that of a conventional nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine 12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear reprogramming.     

Mild drying of sandy soil can physically limit the uptake of phosphorus by rainfed lowland rice in northeast Thailand

ABSTRACT

Poor response of rice to phosphorus (P) fertilization and low phytoavailability of soil P have been reported in sandy rainfed fields in northeast Thailand. In order to evaluate the effects of mild soil drying on the uptake of P by rainfed lowland rice, we carried out nutrient omission trials for nitrogen (N) and P at Ubon Ratchathani Rice Research Center under rainfed and flooded conditions. The surface soil was classified as sandy loam. To avoid severe soil drying and drought stress in the rainfed field, soil water potential at a depth of 20 cm was maintained at the field capacity (> ?20 kPa) by flush irrigation. The effects of flooding and drying on the soil properties were also evaluated in the laboratory using soils with diverse textures in and around the center. In the field experiments, the above-ground biomass of rice plants (RD6) did not respond significantly to P fertilization in the rainfed field, although it responded positively to N fertilization. Root length in the surface 10 cm under the rainfed condition was significantly smaller than that under the flooded condition due partly to the increased soil hardness upon drying, but this could not quantitatively explain the large discrepancy of P uptake observed between the rainfed and flooded conditions. Under the rainfed condition, the P uptake did not increase significantly, even when the concentration of soil Bray P was tripled by transferring the surface soil from the flooded to the rainfed field. From the laboratory experiments, it was further suggested that soil P was supplied mainly by diffusion and that the effective diffusion coefficient for P can become less than one-tenth of the value in the flooded field when the sandy soil with clay at around 10% dried to ?100 kPa. Our results suggest that the uptake of P by the rainfed lowland rice grown in sandy soil can be limited physically by mild soil drying that reduces the supply of P to roots by diffusion rather than the chemical extractability of soil P.     

Spatiotemporal distribution of Flavobacterium psychrophilum and ayu Plecoglossus altivelis in rivers revealed by environmental DNA analysis

Outbreaks of bacterial cold-water disease (BCWD), caused by Flavobacterium psychrophilum, are widespread in Japan, especially among ayu Plecoglossus altivelis. There are few investigations of F. psychrophilum in river water, and its seasonal distribution has not been clarified. We aimed to identify the spatiotemporal dynamics of F. psychrophilum and ayu to provide information that is useful for establishing a countermeasure for BCWD. Quantitative analysis of environmental DNA (eDNA) was used to clarify the year-round dynamics of ayu and F. psychrophilum. We sampled river water from the Nagara and Ibi rivers in Japan, and conducted monthly water sampling and eDNA quantification. Changes in the eDNA concentration of ayu were consistent with the known life histories of the fish. There was a strong negative correlation between the eDNA concentration of F. psychrophilum and water temperature, suggesting a strong dependence of F. psychrophilum dynamics in the river on water temperature. Furthermore, relatively high eDNA concentrations were recorded for both organisms in early summer and fall, suggesting that ayu is infected with F. psychrophilum during these seasons when experiencing up- and downmigration, respectively.

     

Effects of feeding high volumes of milk replacer on reproductive performance and on concentrations of metabolites and hormones in blood of Japanese black heifer calves

We evaluated the effects of feeding high volumes of milk replacer on growth and reproductive performances in Japanese black heifers. Fifty-one heifers were fed milk replacer at 9 L/day for 60 days (9 L × 60 days; n = 18) or 41 days (9 L × 41 days; n = 15), or at 7 L/day for 40 days (7 L × 40 days; n = 18). Artificial insemination (AI) was performed on heifers with ≥270 kg body weight and ≥116 cm body height at 300 days of age. The age at the first AI was 0.35 month later for 7 L × 40 days than the other groups (p < .01). However, age at calving did not differ among treatments (22.1 months). The interval from the first AI to pregnancy tended to be ~2 months longer for the 9 L × 60 days than the other groups (p = .07). Our results showed that feeding high volumes of milk replacer may reduce the age at calving via an improved rate of growth. In addition, we propose that feeding a maximum of 7 L milk replacer for 40 days may be the most appropriate rearing regime because the success of pregnancy per AI may be reduced in calves fed a maximum of 9 L for 41 and 60 days.     

Methane and N2O emissions, nitrate concentrations of drainage water, and zinc and copper uptake by rice fertilized with anaerobically digested cattle or pig slurry

We have examined the effects of different types of slurry on CH₄ and N₂O emissions, Zn and Cu contents of rice, and nitrate content of the drainage water. The experiment included four treatments: (1) anaerobically digested cattle slurry (ADCS), (2) ADCS filtered to remove the coarse organic matter fraction, (3) anaerobically digested pig slurry (ADPS), and (4) chemical fertilizer (CF). The application rate was 30?g?NH₄?CN?m^?2. Different amounts of C were incorporated with fertilization: 725?g?C?m^?2 in ADCS, 352?g?m^?2 in filtered ADCS, and 75?g?m^?2 in ADPS. The average CH₄ emissions during a growing period were 304, 359, 452, and 579?mg?m^?2?day^?1 in the CF, ADPS, filtered ADCS, and ADCS treatments, respectively. The CH₄ emission was significantly higher in ADCS than in CF and ADPS. Negligible N₂O emissions were observed during the growing period. Comparable concentrations of Zn and Cu were observed in the rice grain among the treatments. In contrast, their concentrations in the stems and leaves were significantly higher in ADPS than in CF treated rice, although the values were lower than the upper limit of feed additives. Nitrate concentrations in the drainage water were consistently low (0.5?mg?N?L^?1). The present study suggested that ADPS, containing a lower amount of C than ADCS, might be an organic fertilizer in paddy field with comparable environmental impacts to chemical fertilizers (CF), but long-term field studies are needed to better understand the effects of these organic fertilizers.     

Effects of soil temperature and anaerobiosis on degradation of biodegradable plastics in soil and their degrading microorganisms

The bioplastic PHB/HV (copolymer of 3-hydroxybutyrate and 3-hydroxyvalerate) underwent a faster degradation at 30°C than at 52°C in soil under aerobic conditions, while there was no remarkable difference between 30°C and 52°C in the degradation rate of PCL [poly(ε-caprolactone)], PBSA (polybutylene succinate and agipate), and PBS (polybutylene succinate). PHB showed the fastest degree of degradation among the four plastics at 30°C and PBSA the fastest at 52°C. Degradation of all the four plastics was nor observed both at 30°C and 52°C under anaerobic conditions for 50 d. Microorganisms on the degrading plastics appeared to be diverse at 30"C, including bacteria and fungi. However, among the several to ca. 10 kinds of bacterial and fungal strains isolated from the degradation sites of each plastic at 30°C, only one or two fungal strains were able to degrade the respective plastics in vitro. The degraders were identified as Mucor sp. (PHB), Paecilomyces sp. (PCL), Aspergillus sp. (PBSA), and Cunninghamella sp. (PBSA). In contrast, only a single type of fungus was observed at the degradation sites of PCL and PBSA at 52°C. The fungus isolated from PCL and PBSA was identified as Thermomyces sp. This study demonstrated that soil temperature and anaerobiosis exerted significant effects on the degradation of the plastics, and that fungi were mainly responsible for the degradation of the plastics in soil.     

Observation of microbial colonization on the surface of rice roots along with their development and degradation

Microbial colonization on the surface of rice roots along with their development and degradation was examined throughout the growth period of rice plants with the naked eye, by light microscopy, and scanning electron microscopy. Four stages were recognized in the development and degradation processes of rice roots and microbial colonization pattern. At stage I, both crown and lateral roots were undergoing development, white, and covered with a mucigel layer. Microbial colonization was rarely observed. At stage II, lateral roots developed further, and they stained brownish due to the deposition of iron oxides/hydroxides on the root surface. Microbial colonies commonly developed along the grooves between epidermal cells. At stage III, lateral roots ceased to develop, and the root surfaces were widely covered with deposits of iron oxides/hydroxides. Microbial colonies were found inside epidermal and exodermal cells as well as on epidermal cells. At stage IV, the roots became dark brown or transparent. Most of the epidermal and exodermal cells disappeared from the root surface. Roots went through stage I to stage IV as they aged, and respective degradation stages were observed from the nodal roots from higher nodes to the nodal roots from lower nodes. There were fewer microorganisms on the surface of lateral roots than on crown roots. About 50% of the roots were at stages I and II on June 20 and July 20, which corresponded to the early to middle growth stages, while most of the roots were at stages III or IV from August 10.