Effect of calcium concentration in nutrient solution before and after inoculation with Ralstonia solanacearum on resistance of tomato seedlings to bacterial wilt

We previously reported that calcium (Ca) nutrition in tomato (Lycopersicon esculentum Mill.) significantly affected the resistance to bacterial wilt caused by Ralstonia solanacearum Smith. To elucidate the mechanisms underlying the Ca-dependent resistance, the effect of the Ca concentration in the nutrient solution applied before and after inoculation with the pathogen on the resistance of tomato seedlings to bacterial wilt was studied. One week before inoculation, seedlings were transferred to nutrient solutions containing Ca at concentrations of 0.4, 4.4, or 20.4 mM. Soon after inoculation, the seedlings that were treated with each concentration of Ca before inoculation were transferred to solutions containing the same three concentrations of Ca. Although the disease development was not affected by the concentration of Ca in the solution before inoculation, a higher concentration of Ca after inoculation reduced the disease severity. This result suggests that the concentration of Ca in the host, especially in the cell walls, before infection may not be directly involved in the Ca-dependent resistance of tomato seedlings to bacterial wilt.     

Effect of fungicide thiram on microbial communities of rice seed surface estimated by culture-dependent and culture-independent (PCR-RFLP) methods

Coating of rice seeds with fungicide Thiram improved the seed germination capability over a long period of time (11 weeks) under low temperature conditions (4 and 8°C), which simulated the sowing of rice seeds in the winter season (the farmer's slack season). To analyze the effect of Thiram on the community structure of microorganisms on the rice seed surface, culture-dependent and culture-independent (PCR-RFLP) methods were applied. PCR-RFLP patterns of 16S rDNA showed that the bacterial communities on the rice seed surface were different between coated and uncoated treatments under 8°C conditions, but that they were very similar under 4°C conditions. PCR-RFLP patterns of 18S rDNA revealed the remarkable effect of Thiram on the fungal community structure under both 4 and 8°C conditions. Although the fungal communities were quite different between coated and uncoated seeds at the beginning of incubation, the fungal communities on the coated seed surface became similar to those of uncoated seeds along with the duration of the incubation period. As the dominance percentage of Fusarium spp. among the isolates increased with the duration of the incubation period for both coated and uncoated seeds, Fusarium was considered to be a responsible for the poor germination of rice seeds that were sown in the winter season.     

Nutrient leaching from the plow layer by water percolation and accumulation in the subsoil in an irrigated paddy field

To estimate the impact of water percolation on the nutrient status in paddy fields, the seasonal variations of the concentrations of cations, anions, inorganic carbon (IC), and of dissolved organic carbon (DOC) in percolating water that was collected from just below the plow layer (PW-13) and from drainage pipes at the 40 em depth (PW-40), as well as in irrigation water were measured in an irrigated paddy field. Total amounts of Ca, Mg, K, Fe, and Mn leached from PW-13 during the period of rice cultivation were estimated to range from about 390 to 770, 65 to 130, 33 to 66, 340 to 680, and 44 to 87 kg ha^-1, respectively. Amounts of losses that were estimated from the differences between the input by irrigation water and the output by percolation water from the plow layer corresponded to 11 to 26, 22 to 47,5.9 to 12, and 13 to 26% of exchangeable Ca and Mg, amorphous Fe, and easily reducible Mn in the plow layer, respectively. The concentrations of Ca, Mg, K, Fe, and Mn in PW13 were higher than those in PW-40. The amounts of these nutrients that were retained in the subsoil between the 13 em and 40 em soil depth corresponded to 83, 86, 61, 99, and 89% of the amounts that percolated from the plow layer, respectively. Total amounts of IC and DOC that percolated from the plow layer ranged from 750 to 1,500 and 85 to 170 kg-C ha^-1, which corresponded to 5.0 to 10.0% and 0.6 to 1.1% of the total carbon content in the plow layer, respectively. Eighty eight % of IC in the percolating water from the plow layer was also retained in the subsoil.     

Chemical characteristics and potential source of fulvic acids leached from the plow layer of paddy soil

To understand better the chemical characteristics and source of dissolved organic matter (DOM) leached from the plow layer of rice paddies, a lysimeter study was conducted, which simulated submerged paddy topsoil during rice growth. The fulvic acid (FA) fraction in the percolation water from the lysimeter was collected by adsorption onto insoluble polyvinylpyrrolidone (PVP), and the temporal variations in its elemental composition, molecular size distribution, IR spectra, ¹³C CPMAS NMR spectra, and δ¹³C values were investigated. The proportion of the FA fraction to bulk DOM varied greatly, but the chemical characteristics of the FA fraction did not change appreciably during the experimental period. Thus, it is considered that the changes in the DOM composition in percolation water were mainly due to the differing contributions of the FA fraction. Further, to investigate the source of the FA fraction in the leachate, the chemical characteristics of the FA fraction in the leachate were compared with those extracted from the plow layer soil. A sequential extraction of the FA fraction was conducted using a sequence of water, 0.25 M Na₂SO₄, 0.1 M Na₄P₂O₇ (pH 7.0), 0.1 M Na₄P₂O₇ (pH 10.5), and NaBH₄+0.1 M Na₄P₂O₇ (pH 10.5). It was found that the water- and 0.25 M Na₂SO₄-extractable fractions, which were most mobile, were not the only source of the FA fraction in the leachate. The small molecular size sub-fraction of the NaBH₄+0.1 M Na₄P₂O₇ (pH 10.5)-extractable FAs, most of which are probably bound mainly to iron oxides, are considered to be another source of the FA fraction leached from the plow layer of paddy fields.     

Therapeutic effect of anti-TNF-alpha antibody and levofloxacin (LVFX) in a mouse model of enterohemorrhagic Escherichia coli O157 infection

The ability of an anti-TNF-alpha antibody to confer protection against enterohaemorrhagic Escherichia coli (EHEC) O157 was investigated in germfree IQI mice. The use of an antibiotic levofloxacin (LVFX) alone or with the antibody was also studied. Protection included an increase in survival rate. Treatment with the anti-TNF-alpha antibody inhibited the histological signs associated with EHEC infection but did not prevent the colonization of EHEC or production of Shiga toxin (Stx). No clinical signs were observed and EHEC was completely eliminated in the mouse model receiving both anti-TNF-alpha antibody and LVFX. Anti-TNF-alpha antibody suppressed inflammatory cytokine response in the mouse kidney and brain by EHEC infection.     

Physiopathological Effects of the Administration of Chronic Cyanide to Growing Goats – A Model for Ingestion of Cyanogenic Plants

Ingestion of cyanogenic plants, such as cassava and sorghum, has been associated with goitre and tropical pancreatic diabetes in both humans and animals. Thus, the objective of the present study was to determine the toxic effects on the thyroid and pancreas in growing goats of prolonged exposure to potassium cyanide (KCN). Thirty-four male goats were divided into five groups dosed with KCN at 0 (control), 0.3, 0.6, 1.2 or 3.0 mg/kg daily for 5 months. Blood samples were obtained in order to determine the glucose, cholesterol, thyroxine (T₄), triiodothyronine (T₃) and thiocyanate concentrations and for haematological studies; pancreas and thyroid gland were collected for histopathological study. The group receiving the highest dose of cyanide showed lower body weight gains and carcase weights and a decrease in plasma T₃ concentrations compared to the control group. Reabsorption vacuoles in follicular colloid and normocytic normochromic anaemia were observed in the experimental animals. Inhibition of peripheral conversion of T₄ to T₃ is suggested. However, no diabetogenic effects were observed.     

Changes in ascorbic acid content in primary cultured rat hepatocytes exposed to 2,2'-azobis (2-amidinopropane) dihydrochloride, a radical initiator

Changes in ascorbate content in primary cultured rat hepatocytes exposed to oxidative stress derived from water soluble radical initiator 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) were examined. Cells were exposed to 0.05 and 5 mg/ml of AAPH as 'mild' and severe' oxidative stresses, respectively. Lipid peroxidation in hepatocytes was induced by 'severe' oxidative stress, but not by 'mild' oxidative stress. Ascorbate decreased at 6 hr after administration of both mild' and severe' oxidative stresses, and recovered to the control level after a further 6 hr. In cells treated with 'severe oxidative stress, however, total ascorbate (reduced form plus oxidized form) had increased 24 hr after administration. These results indicated that consumption alone did not account for the increase of ascorbate in hepatocytes under oxidative stress.     

Oocyte transport: Developmental competence of bovine oocytes arrested at germinal vesicle stage by cycloheximide under air

The effects of the medium (TCM 199 or SOFaa) and temperature (20 or 39 C) during meiotic arrest by cycloheximide (CHX) under air on the developmental competence of bovine oocytes after in vitro maturation (IVM) and fertilization (IVF) were investigated. Oocytes were maintained in meiotic arrest by 10 microg/ml CHX in a 50-microl droplet of 25-mM HEPES-buffered TCM 199 (H199) at 39 C or synthetic oviduct fluid (HSOFaa) at 20 or 39 C in air for 24 h. After release from the arrest, the oocytes was matured and fertilized in vitro and their developmental competence was examined. The developmental rate of oocytes arrested in HSOFaa at 20 C to the blastocyst stage was similar to that of non-arrested oocytes but was significantly higher (P<0.05) than that of oocytes arrested at 39 C in H199 or in HSOFaa. In consideration of oocyte transport conditions, we also investigated the meiotic arrest of oocytes maintained in a 0.25-ml straw by CHX individually with 10 microl HSOFaa or as a group (40-50 oocytes) with 170-200 microl HSOFaa at 20 C in air for 24 h. After release from meiotic arrest, the developmental competence of these oocytes was assessed similarly. The developmental rate of oocytes treated with CHX individually was similar to that of those treated with CHX in 50-microl droplet of HSOFaa at 20 C. However, the developmental rate of oocytes treated with CHX as a group was lower than that of oocytes treated with CHX in a 50-microl droplet. Five blastocysts developed from oocytes maintained in meiotic arrest in a plastic straw were transferred to five recipient heifers. Consequently, three recipients became pregnant and 2 calves were delivered. The results of the present study indicate that bovine oocytes treated with CHX in HSOFaa at 20 C under air retain the same developmental competence as non-arrested oocytes.