Role of sulphide reduction by magnesium hydroxide on the sediment of the eutrophic closed bay

This study was focused on the effect of magnesium hydroxide (Mg(OH)₂) for improving the sediment of aquaculture field and eutrophic closed estuaries. Sediment samples were collected from Omura Bay, Nagasaki, Japan. The change in pH, acid volatile sulphides (AVS) and dissolved organic carbon (DOC) in interstitial water of the sediment samples were measured before and after 20‐day incubation under the condition of overlapping seawater at 25°C with 0.3–0.4 mg/L of dissolved oxygen (DO). Hydrogen sulphide (H₂S) and the dsrA gene copy number of Grp3 sulphate‐reducing bacteria (SRB) in the sediment samples were also measured at 7‐day incubation under the same condition. The pH of Mg(OH)₂ treatment group was significantly increased from 7.43 to 8.25; it was higher compared to the control group which pH was maintained around 7.6. Meanwhile, the AVS content of Mg(OH)₂ treatment group was significantly reduced and the value was lower than the control group after the 20‐day incubation. Each concentration of interstitial water DOC in the treatment group and the control group increased after the incubation from initial value of 15.4–40.1 and 29.0  mg/L respectively. Furthermore, simultaneous decrease in H₂S and the dsrA gene copy number of Grp3 SRB was observed at the end of the 7‐day incubation by adding Mg(OH)₂. It is concluded that the loss of activity of SRB was strongly related to the decrease in sulphide (H₂S and AVS) and the increase in DOC was due to the alkalization of the sediment by adding Mg(OH)₂.     

Antibody reactivity in mice and cats to feline enteroepithelial stages of Toxoplasma gondii.

The reactivity of antibodies in mice and cats to feline enteroepithelial stages of Toxoplasma gondii was examined by means of an indirect immunofluorescent antibody test. Mice immunized with feline enteroepithelial stage (FES) parasites produced antibodies not only against FES, but also against tachyzoites, sporozoites/oocysts, tissue cysts and one part of the infected feline enterocytes. After absorption with tachyzoites, the titer of antibodies reactive to enterocytes was significantly reduced. In contrast, the titer of antibodies reactive to FES remained unchanged. The antibodies from cats immunized with FES, reacted specifically to FES, but not to tachyzoites, tissue cysts or enterocytes. These results suggest that FES parasites may have stage-specific antigen(s).     

Implication of C-terminal mutation of PopA of Ralstonia solanacearum strain OE1-1 in suppression of bacterial wilt

Ralstonia solanacearum strain OE1-1 causes bacterial wilt on tobacco plants. The popA -mutant 31b, derived from OE1-1 by insertion of transposon Tn 4431 , did not cause wilt on tobacco plants inoculated through the roots. However, when 31b was directly inoculated into xylem vessels, the tobacco plants wilted, similarly to those inoculated with OE1-1. 31b retained its exopolysaccharide productivity and its type-III secretion function. Furthermore, 31b grew in intercellular spaces and systemically infected tobacco plants, similarly to OE1-1. popA consists of an operon with popB and popC , and suppression of popB and popC expression resulting from polar mutation by transposon insertion did not affect the virulence of 31b. The mutated popA ( popA31b ) was composed of 960 nucleotides, including 39 derived from Tn 4431. A recombinant mutant from OE1-1, where popA31b was introduced by marker exchange, showed the same phenotype as 31b. PopA31b protein was extracellularly secreted by 31b co-cultured with Arabidopsis thaliana . These results suggest that PopA31b extracellularly secreted by 31b in intercellular spaces may be implicated in suppression of disease development, leading to inability of the bacteria to induce wilt on plants. Taken together, interactions between host plants and R. solanacearum existing in intercellular spaces immediately after invasion may be involved in disease development.     

The coat protein of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis> L<Subscript>11</Subscript>Y is associated with virus-induced chlorosis on infected tobacco plants

The L₁₁Y strain of Tomato mosaic virus (ToMV) causes severe chlorosis on infected tobacco leaves. Sequencing analysis for the genome showed that L₁₁Y contained multiple nucleotide changes and that some led to amino acid substitutions, when compared with that of the common L strain of ToMV. The chimeric virus, which has the CP of L₁₁Y in the context of the L strain RNA genome, caused severe chlorosis on infected tobacco plants, suggesting that the CP of L₁₁Y containing three amino acid changes (E33S, A86T and E97K) was the determinant of the chlorosis. Two of these amino acid changes (A86T and E97K) were associated with the induction of chlorosis when present together in the CP. Severe destruction and deformation of chloroplasts and the formation of discrete dark-staining materials adjacent to chloroplasts were observed with electron microscopy in L₁₁Y-infected plants. Fewer virus particles accumulated in the cytoplasm of L₁₁Y-infected plant cells. The level of accumulation of CP subgenomic RNA and CP in the infected protoplasts was similar between L and L₁₁Y. Fewer virus particles accumulated in L₁₁Y-infected protoplasts, and many of them were shorter-than-full-length. The nucleotide sequence data reported is available in DDBJ/EMBL/GenBank databases as accession AB355139.     

Amino acid substitution in the coat protein of Melon necrotic spot virus causes loss of binding to the surface of Olpidium bornovanus zoospores

Melon necrotic spot virus (MNSV) is transmitted by the fungus Olpidium bornovanus. In this study, we used immunofluorescence microscopy to detect MNSV particles over the entire surface of the O. bornovanus zoospore; MNSV particles were not detected on the related fungus O. virulentus, which cannot transmit MNSV. The amino acid substitution Ile → Phe at position 300 in the MNSV coat protein resulted in loss of both specific binding and fungal transmission, while virion assembly and biological aspects were unaffected. Taken together, these results suggest that the MNSV coat protein acts as a ligand to the O. bornovanus zoospore as part of a fungal-vector transmission system.     

A new strain of Melon necrotic spot virus that is unable to systemically infect Cucumis melo

We report a new strain of Melon necrotic spot virus (MNSV) that is unable to systemically infect Cucumis melo. A spherical virus (W-isolate), about 30 nm in diameter like a carmovirus, was isolated from watermelons with necrotic symptoms. The W-isolate had little serological similarity to MNSV, and it did not cause any symptoms in six melon cultivars susceptible to MNSV; however, the host range of the W-isolate was limited exclusively to cucurbitaceous plants, and transmission by O. bornovanus was confirmed. Its genomic structure was identical to that of MNSV, and its p89 protein and coat protein (CP) showed 81.6 to 83.2% and 74.1 to 75.1% identity to those of MNSV, respectively. Analysis of protoplast showed that the W-isolate replicated in melons at the single-cell level. Furthermore, chimeric clones carrying the CP of MNSV induced necrotic spots in melons. These results suggested that the absence of symptoms in melons was due to a lack of ability of the W-isolate to move from cell to cell. In view of these findings, we propose that the new isolate should be classified as a novel MNSV watermelon strain.     

Antimicrobial activity of licorice flavonoids against methicillin-resistant Staphylococcus aureus

Nineteen flavonoids isolated from licorice (Glycyrrhiza glabra, G. inflata and G. uralensis) were tested for their antimicrobial activities against methicillin sensitive Staphylococcus aureus, methicillin resistant S. aureus, Micrococcus luteus, Bacillus subtilis, Escherichia coli, Klebsiella pneumoniae and Pseudomonas aeruginosa.     

Anthocyanins from new red leaf tea 'Sunrouge'

New red leaf tea cultivar 'Sunrouge' (Camellia taliensis × Camellia sinensis), for which an application for registration was made in 2009, is an anthocyanin-rich tea. The anthocyanin content of 'Sunrouge' was the highest among 4 tea cultivars, and was 8.4 times higher than that of 'Yabukita'. We purified and isolated 6 anthocyanins from 'Sunrouge' by chromatography, and identified them by LC/MS/MS and NMR analysis. As a result, the four anthocyanins were identified as delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (2), delphinidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (3), cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)galactopyranoside (4), and cyanidin-3-O-β-D-(6-(E)-p-coumaroyl)glucopyranoside (5), and the other two were estimated as delphinidin-(Z)-p-coumaroylgalactopyranoside (1), petunidin-(E)-p-coumaroylgalactopyranoside (6). Compound 3 was found in tea for the first time. In general, anthocyanins have various bioactivities, including relieving eyestrain and antioxidative effects, so it is expected that drinking 'Sunrouge' tea brings in similar bioactivities.     

Digestibility and metabolizable energy values of processed cassava chips for growing and finishing pigs

Determinations of digestibility of dry matter (DM), digestible energy (DE), and metabolizable energy (ME) in cassava chips with different levels of crude fiber (CF) were measured in growing pigs (20 kg) and finishing pigs (60 kg). The treatments were (1) cassava starch (0% CF), (2) peeled cassava chips (2.5% CF), (3) non-peeled washed cassava chips (3.9% CF), and (4) non-peeled and non-washed cassava chips (5.2% CF). In the growing pigs, peeled cassava chips, non-peeled washed cassava chips, and non-peeled and non-washed cassava chips had DM digestibility of 87.51%, 78.63%, and 73.89%, respectively. Their DE was 3.69, 3.49, and 3.32 Mcal/kg DM, respectively (DE of cassava starch is 3.90 Mcal/kg DM). ME was 3.54, 3.35, and 3.19 Mcal/kg DM, respectively (ME of cassava starch is 3.74 Mcal/kg DM). On the other hand, in the finishing pigs, the digestibility of DM was 89.13%, 80.63%, and 76.13%, respectively. Their DE was 3.72, 3.53, and 3.43 Mcal/kg DM, respectively (DE of cassava starch is 3.91 Mcal/kg DM). ME was 3.57, 3.38, and 3.29 Mcal/kg DM, respectively (ME of cassava starch is 3.75 Mcal/kg DM). These values increased with decreasing CF content, and the peeled cassava chips had the highest values (P < 0.01). These suggest that the digestibility values of DM, DE, and ME of cassava chips is inversely related to the CF content in cassava chips. It is recommended that cassava chips be peeled for better nutrition for growing and finishing pigs.     

Polymorphism of rRNA gene loci in the dog

We examined the number of ribosomal gene (rDNA) loci in the metaphase spreads of 54 dogs by FISH method. We found that in 16 dogs (30%) one or two loci were missing. The total number of rDNA loci was varied from 5 to 7 in males and from 4 to 6 in females. As the male dog consistently bears the rDNA on the Y chromosome, the polymorphism of the rDNA locus was ascribed to the absence of autosomal rDNA loci. Indeed, the frequency of polymorphism is almost equivalent in both sexes in the beagle. In one female beagle dog, remarkable intense fluorescence signals were observed at the four autosomal loci, indicating the in situ amplification of rDNA.