Phylogenetic groups and cephalosporin resistance genes of Escherichia coli from diseased food-producing animals in Japan

A total of 318 Escherichia coli isolates obtained from different food-producing animals affected with colibacillosis between 2001 and 2006 were subjected to phylogenetic analysis: 72 bovine isolates, 89 poultry isolates and 157 porcine isolates. Overall, the phylogenetic group A was predominant in isolates from cattle (36/72, 50%) and pigs (101/157, 64.3%) whereas groups A (44/89, 49.4%) and D (40/89, 44.9%) were predominant in isolates from poultry. In addition, group B2 was not found among diseased food-producing animals except for a poultry isolate. Thus, the phylogenetic group distribution of E. coli from diseased animals was different by animal species. Among the 318 isolates, cefazolin resistance (minimum inhibitory concentrations: ≥32 μg/ml) was found in six bovine isolates, 29 poultry isolates and three porcine isolates. Of them, 11 isolates (nine from poultry and two from cattle) produced extended spectrum β-lactamase (ESBL). The two bovine isolates produced bla_CTX-M-2, while the nine poultry isolates produced bla_CTX-M-25 (4), bla_SHV-2 (3), bla_CTX-M-15 (1) and bla_CTX-M-2 (1). Thus, our results showed that several types of ESBL were identified and three types of β-lactamase (SHV-2, CTX-M-25 and CTX-M-15) were observed for the first time in E. coli from diseased animals in Japan.     

Molecular cloning of canine interleukin-31 and its expression in various tissues

The newly discovered cytokine, interleukin-31 (IL-31), belongs to the short-chain cytokine group. It was reported that transgenic expression of IL-31-induced pruritus, similar to atopic dermatitis, in mice, further, excessive amounts of IL-31 was also expressed in the skin from human patients with atopic dermatitis as compared to that from normal people. In this study, canine IL-31 was molecularly cloned from concanavalin A-stimulated canine peripheral blood mononuclear cells (PBMCs), and its nucleotide sequence was determined. Canine IL-31 contains 4 alpha-helix structures characteristic of the IL-31 family, and the amino acid identity of canine IL-31 with those of human or mouse is 54% and 28%, respectively. Furthermore, we detected low levels of canine IL-31 in the thymus, testis, spleen, and kidneys, but not in the skin of atopic dogs.     

Isolation of some glycosides as aroma precursors in young leaves of Japanese pepper (Xanthoxylum piperitum DC.).

To clarify the formation mechanism for the major alcoholic aroma compounds in young leaves of Japanese pepper, the glycosides were isolated as aroma precursors. The presence of glycosides of the main alcoholic aroma constituents was indirectly determined by enzymatic hydrolysis and trifluoroacetylation (TFA) of the glycoside-containing fraction. After Amberlite XAD-2 column chromatography, ODS flash chromatography, and high-performance liquid chromatography (HPLC), two new compounds, namely, (3S,6S)-cis-linalool-3,7-oxide beta-D-glucopyranoside and 2-methylpropanyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, were isolated. In addition, (3S,6R)-cis-linalool-3,6-oxide beta-D-glucopyranoside, which absolute configuration was the first determined, and six known glycosides, citronellyl beta-D-glucopyranoside, linalyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, (Z)-3-hexenyl beta-D-glucopyranoside, benzyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside, dendranthemoside A, and 3,6-dihydroxy-5,6-dihydro-beta-ionol 9-beta-D-glucopyranoside, were isolated. All of these glycosides were isolated for the first time from the leaves of Japanese pepper. Their structures were established on the basis of spectral data and chemical evidence. The ratios of stereoisomers of the aglycon moieties of citronellyl beta-D-glucopyranoside and linalyl 6-O-beta-D-apiofuranosyl-beta-D-glucopyranoside were investigated by a chiral GC analysis and compared with those of free citronellol and linalool in the aroma concentrate.     

Nutrient and bulk density characteristics of soil profiles in six land use systems along topo-sequences in inland valley watersheds of Ashanti region,Ghana

The bulk density and nutrient distribution in soil profiles as affected by various land use systems were studied. Mean bulk densities of the various soil profiles, 0-20, 20-40, and 4060 cm in the selected five land use systems along the upland / lowland topo-sequences were, 1.26, 1.73, and 1.44 Mg m^-3 for the primary forest (PF) plot in the upland area, 1.13, 1.51, and 1.50 Mg m^-3 for the cacao farm (CP) plot, 1.16, 1.63, and 1.26 Mg m^-3 for the mixed cropping (MC) plot, 1.39, 1.61, and 1.50 Mg m-a for the fallow (Fallow) plot, all three land use systems being in the upland / fringe areas, and 1.46, 1.72, and 1.60 Mg m^-3 for the lowland traditional rice farming (TR) plot, respectively. The very high bulk density of the second layer, especially in the PF plot, may be related to the hardening of the iron / quartz stone layer due to prolonged dry conditions, which may affect plant growth and hydrological cycles in the benchmark inland valley watersheds. The TC contents of the topsoil samples were 43, 32, 24, 26, 18, and 15 g kgm^-1 for the PF, CP, Fallow, MC, TR, and sawah (Sawah) plots, respectively. The C / N ratios of the topsoil samples were 10, 10, 9.8, 9.1, 8.8, and 8.8 for the CP, Fallow, MC, Sawah, TR, and PF plots, respectively, displaying a higher natural fertility, especially for the N supply in the PF plot in the upland area. The available phosphorus levels were low even for the topsoil samples, 1.8 to 3.4 mg kg^-1 for the upland soils while the lowland TR and Sawah plots recorded values of 4.4 and 4.9 mg kg^-1, respectively. The contents of exchangeable K, 0.1-0.4 cmole kg^-1, of the topsoil samples were in the range of those of normal inland valley soils in West Africa. In contrast to the content of available P, although the MC plot showed a depletion, the PF, Fallow, and CP plots showed a replenishment of available K through biological nutrient cycling. The levels of exchangeable Ca and Mg as well as eCEC, were also relatively high compared to those of mean inland valley soils in West Africa. The major component of eCEC was Ex. Ca. The levels of Ex. Ca in the topsoil samples were 11.1, 9.1, 7.8, 4.7, 5.1, and 3.9 cmol_c kg^-1 for the PF, CP, Fallow, MC, TR, and Sawah plots, respectively. The distinctive feature of Ex. Mg was its large distribution in the upper soil profiles in the CP plot, indicating that in the cacao farms, Ex. Mg was enriched, probably due to the high level of Mg in the leaf litter. The lowland soils, normally, should have been enriched in exchangeable bases such as Ca, K, and Mg, leading to a higher eCEC value and higher general fertility through the process of geological fertilization, Le., nutrient flows from upland to lowland areas, as in the case of lowland sawah in monsoon Asia. At the benchmark sites, however, the effects of the geological fertilization process were not evident. This is one of the most important characteristics of West African inland valley watersheds.     

Epidemiological survey of Babesia gibsoni infection in dogs in eastern Japan

To determine the distribution of Babesia gibsoni infection in dogs in the eastern part of Japan, an epidemiological survey of dogs suspected of having B. gibsoni infection was attempted using the polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Thirty-five of 115 such dogs (30.4%) were positive by PCR and/or ELISA. The 35 positive dogs consisted of 28 Tosa dogs, 4 American Pit Bull Terriers, and 3 mongrel dogs in Aomori, Fukushima, Ibaraki, Gunma, Chiba, Tokyo, Kanagawa, and Nagano Prefectures. The positive dogs had a significantly lower rate of tick exposure and a higher rate of bites by other dogs. Twenty-two of 35 B. gibsoni-positive dogs were infected with hemoplasma, and the rate of infection was significantly higher than that of B. gibsoni-negative dogs.     

Effect of relationships among clinical mastitis incidence,reproductive performance,and culling rate on the lifetime of dairy cows at Hiroshima University Farm

Farm managers' decision to cull dairy cows is based on the cows' milk production, history of disorder(s), and reproductive performance, each of which affects dairy cows' lifetime (herd life and productive lifespan). We investigated the relationships among the incidence of clinical mastitis (CM), the reproductive performance, and the culling rate. We also assessed the effects of these relationships on the lifetimes of dairy cows, using the records made before and after the introduction of an automatic milking system (AMS) at Hiroshima University Farm. Milk yield, CM incidence density, and culling rate of dairy cows increased after the AMS introduction. The CM incidence was associated with an elongation of the calving interval in cows with the same parity. CM in the 1st parity might have caused the reductions of the cows' lifetime and their parity at culling. A higher age at first calving (AFC) was associated with an increase in culling rate but did not lead to a significant decrease in lifetime. Investigations of the factors mediating CM in the 1st parity or AFC with CM incidence or culling rate in the later stages might contribute to the control of lifetime of dairy cows.     

<Emphasis Type="Italic">In silico</Emphasis> genomic subtraction guides development of highly accurate,DNA-based diagnostics for <Emphasis Type="Italic">Ralstonia solanacearum</Emphasis> race 3 biovar 2 and blood disease bacterium

New rapid diagnostic methods are urgently needed to discriminate the quarantine pathogen Ralstonia solanacearum (Rs) race 3 biovar 2 (R3B2) from other populations of Rs that lack the adaptation to cause bacterial wilt disease in temperate regions. We used an in silico bioinformatic approach to identify several genome sequences potentially specific to R3B2 strains. Primer sets were designed to PCR-amplify sequences in these regions, and four sets were ultimately shown to be >99% accurate for detection of R3B2 strains. On the basis of these results, several primers were designed to enable development of a loop-mediated isothermal amplification assay that was rapid, technologically simple, and essentially 100% accurate for identification of R3B2 when applied to a comprehensive collection of geographically diverse Rs strains. We fortuitously found that a sequence in one of the “R3B2-specific” regions has ~90% identity to a sequence present in strains of the blood disease bacterium (BDB), a member of the Rs species complex that infects banana. Alignments of these sequences allowed design of a second PCR primer set that proved 100% accurate for identification of BDB strains when tested on the 22 BDB strains available to us. These results demonstrate the power of in silico genomic subtraction for rapid identification of population-specific DNA sequences and for the development of simple, reliable detection methods for Rs subpopulations.     

Isolation and characterization of lectins from the AG-D group of binucleate <Emphasis Type="Italic">Rhizoctonia</Emphasis> species

Isolates from 18 anastomosis groups (AGs) of binucleate Rhizoctonia were screened for lectin activity. Eight AGs (AG-B, AG-D, AG-F, AG-G, AG-H, AG-I, AG-R and AG-U) had low to moderate lectin activities. Among these, members of AG-D and AG-I had the highest activity. Partially purified lectins from AG-D preferentially agglutinated human blood type A to type B and O. Mucin and galactose were the most potent inhibitors among the tested carbohydrates. The molecular masses of these lectins ranged from 12.7 kDa for the monomer to 62 kDa for the pentamer type. Proline, alanine, glutamic acid, aspartic acid, leucine, threonine, serine and tyrosine were the major amino acid components of these lectins. Lectins from AG-D were stable at 4–50°C and from pH 6.0 to 10.0. When assayed with isoelectric focusing, these lectins gave bands at pI 9.30. Specificity of lectins from AG-D to galactose and its derivatives suggest a possible recognition role in this fungal species.     

Appearance of race Pfs:5 of spinach downy mildew fungus, Peronospora farinosa f. sp. spinaciae, in Japan

The races for the causal agent of spinach downy mildew Peronospora farinosa f. sp. spinaciae were identified by inoculation of race-differential cultivars. One isolate was identified as Pfs:5s and the others belonged to a new race. This is the first report of race Pfs:5 and another new race in Japan.