Detection of Nesopora caninum-Specific DNA from Cerebrospinal Fluid by Polymerase Chain Reaction in a Dog with Confirmed Neosporosis

A one-month male Greyhound dog presented with a swinging gait of the hindlimbs, and later developed muscular atrophy of the femoral region and hyperextension of hindlimbs. The dog had positive serum IFAT titers to Neospora caninum, but a negative titer in the cerebrospinal fluid (CSF). N. caninum-specific DNA was amplified from the CSF using a semi-nested polymerase chain reaction assay. Clusters of protozoa in biopsied muscle fibers were subsequently confirmed as N. caninum tachyzoites by immunohistochemical examination. Early recognition and treatment are necessary for effective recovery of clinical canine neosporosis, but antemortem diagnosis is difficult. We suggest that the detection of parasite deoxyribonucleic acid in the CSF is a useful antemortem diagnostic method in facilitating treatment of this disease.     

The occurrence of Listeria monocytogenes in imported ready-to-eat foods in Japan

Quantitative analyses of Listeria monocytogenes in imported ready-to-eat (RTE) foods sold at retail stores in Japan were performed. Of the 77 non-cooked meat products, 6 samples (7.8%) tested positive. The levels of contamination of 4 of the samples were below 100 colony-forming units (CFU)/g, which is the microbiological criterion for L. monocytogenes in RTE foods as determined by Codex. However, Listeria cells at levels of 100 and 400 CFU/g were detected in a salami sample and a raw ham sample, respectively. All of the 70 cheese samples and the 3 samples made from raw ham and cheese showed negative test results. These results suggest that imported RTE foods are potential sources of the causative agent of listeriosis.     

Phylogenetic relationships of three species within the family Heligmonellidae (Nematoda; Heligmosomoidea) from Japanese rodents and a lagomorph based on the sequences of ribosomal DNA internal transcribed spacers, ITS-1 and ITS-2

Nematodes of the family Heligmonellidae (Heligmosomoidea; Trichostrongylina) reside in the digestive tracts of rodents and lagomorphs. Although this family contains large numbers of genera and species, genetic information on the Heligmonellidae is very limited. We collected and isolated adult worms of three species in Japan that belong to the family Heligmonellidae, namely Heligmonoides speciosus (Konno, 1963) Durette-Desset, 1970 (Hs) from Apodemus argenteus, Orientostrongylus ezoensis Tada, 1975 (Oe) from Rattus norvegicus and Lagostrongylus leporis (Schulz, 1931) (Ll) from Pentalagus furnessi, and sequenced the entire internal transcribed spacer regions, ITS-1 and ITS-2 of ribosomal DNA. ITS-1 of Hs, Oe and Ll was 426, 468 and 449 bp in length, and had a G+C content of about 41, 41 and 37 %, respectively. ITS-2 of Hs, Oe and Ll was 297, 319 and 276 bp in length and had a G+C content of about 38, 40 and 28%, respectively. The data of Hs, Oe and Ll were compared with those of two other known species within the family Heligmonellidae, Calorinensis minutus (Dujardin, 1845) (Cm) and Nippostrogylus brasiliensis (Travassos, 1914) (Nb), and with those of two species of Heligmosomidae (Heligmosomoidea), Heligmosomoides polygyrus bakeri and Ohbayashinema erbaevae. Phylogenetic analysis placed Hs, Oe and Ll in the same clade with Cm and Nb, forming a Heligmonellidae branch in both ITS-1 and ITS-2, separate from the Heligmosomoidea branch. These results demonstrated that the ITS-1 and ITS-2 sequences are useful for differentiating the Heligmonellidae nematode species. This study is the first to describe the ITS-1 and ITS-2 sequences of Hs, Oe and Ll.     

Absorption and urinary excretion of quercetin,rutin, and alphaG-rutin,a water soluble flavonoid,in rats

Quercetin, rutin, alphaG-rutin (a water soluble flavonoid), and a mixture of rutin and alphaG-rutin were administered to rats by a single gastric intubation, and their absorption and urinary excretion were examined. The plasma and 24 h urinary levels of aglycons (quercetin and tamarixetin/isorhamnetin) were measured by HPLC after deconjugation with beta-glucuronidase/sulfatase treatment. alphaG-rutin was absorbed more rapidly than quercetin or rutin, and the plasma concentrations of quercetin and tamarixetin/isorhamnetin reached the highest peak level 30 min after dosing. Quercetin, rutin, and the mixture of rutin and alphaG-rutin showed the first peak level 8 h, 8 h, and 30 min after dosing, respectively. The area under the concentration-time curve (AUC) for quercetin in rats administered alphaG-rutin was approximately 4.5- and 2-fold higher than those in rats administered quercetin and rutin, respectively, and was almost the same as that in rats administered a mixture of rutin and alphaG-rutin. The highest 24 h urinary excretion was observed in alphaG-rutin-administered rats. These results suggest that alphaG-rutin is absorbed more efficiently than either quercetin or rutin and that a high plasma concentration can be maintained by supplying rutin and alphaG-rutin in combination.     

Isolation,structural elucidation,and inhibitory effects of terpenoid and lipid constituents from sunflower pollen on Epstein-Barr virus early antigen induced by tumor promoter,TPA

Eight fatty acid esters of triterpene alcohols (1-8), four free triterpene alcohols (9, 12, 17, and 18), four diterpene acids (19-22), two tocopherol-related compounds (23 and 24), four estolides (25-28), three syn-alkane-4,6-diols (29-31), one 1,3-dioxoalkanoic acid (32), and one aliphatic ketone (33), along with the mixture of free fatty acids, were isolated from the diethyl ether extract of the pollen grains of sunflower (Helianthus annuus). Among these compounds, 14 (2-8, 12, 23, 25-28, and 33) were new naturally occurring compounds, and their structures were determined on the basis of spectroscopic methods. Twenty-four terpenoids and lipids (1-4, 6-9, 12, and 19-33) and six free triterpene triols (10, 11, and 13-16), derived from their fatty acid esters (2, 3, and 5-8) by alkaline hydrolysis, were evaluated with respect to their inhibitory effects on the induction of Epstein-Barr virus early antigen (EBV-EA) induced by the tumor promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), in Raji cells, which is known to be a primary screening test for antitumor promoters. Among the 30 compounds tested, 21 compounds possessing a di- or a polycyclic ring system in the molecule (1-4, 6-16, and 19-24) showed potent inhibitory effects on EBV-EA induction (91-100% inhibition at 1 x 10(3) mol ratio/TPA).     

Analysis of Proton Generation and Consumption of Forest Surface Soils in Hokkaido, Northern Japan

We determined proton budgets of surface soils in a deciduous forest (Df) and a coniferous forest (Cf) of Volcanogenous Regosols in Tomakomai, Hokkaido of northern Japan. The total H⁺ source was 12.9 and 11.6 kmol_c ha^?1 y^?1 at Df and Cf respectively, and the external H⁺ was 1% at Df and 2% at Cf. The primary H⁺ sources were vegetation uptake of base cations and nitrification, while the major H⁺ sinks were release of base cations and NO₃ ⁺ uptake by vegetation. Leaching incubation experiments using A horizon soils including Df and Cf with NH₄ ⁺ solutions (5.3, 15.9 mg N L^?1) showed that H⁺ from nitrification was generally higher in the Df soil than Cf soil, and nitrification of Tomakomai Df soil was the highest in both treatments. Results of multiple regression analyses suggested that pH_kCl and exchangeable Ca²⁺ contributed to the H⁺ generation via nitrification. Leaching experiments with dilute HCl (pH 3.3) revealed that cation release (mainly Ca²⁺) occurred, and the proportion of release by decrease of exchangeable cations was higher than that by mineral weathering. Mineral weathering in the Tomakomai soil was higher than the other soils.     

Determination and quantification of γ-glutamyl-valyl-glycine in commercial fish sauces

It was recently reported that kokumi substances such as glutathione are perceived through the calcium-sensing receptor (CaSR). In addition, screening by the CaSR assay and sensory evaluation revealed that γ-glutamyl-valyl-glycine (γ-Glu-Val-Gly) was a potent kokumi peptide. In this study, the quantities of γ-Glu-Val-Gly in various commercial fish sauces originating from Vietnam (Nuoc Mum), Thailand (Nampra), China (Yu-lu), Korea, Japan (Shottsuru and Ikanago-shoyu), and Italy (Garum) were investigated using a LC/MS/MS method followed by derivatization with 6-aminoquinoyl-N-hydroxysuccinimidyl-carbamate (AQC). The analyses revealed γ-Glu-Val-Gly at concentrations ranging from 0.04 to 1.26 mg/dL, indicating that γ-Glu-Val-Gly is widely distributed among various commercial fish sauces.     

Growth and yield of corn grown on two alluvial soils of Louisiana applied with different phosphorus rates

The influence of soil properties on phosphorus (P) availability of Louisiana alluvial soils is not fully understood. A pot experiment was conducted in 2011 to evaluate the effect of different P fertilizer rate (0, 34, 67, 101 and 134 kg P₂O₅ ha^?1) on corn growth and development on Perry clay and Commerce silt loam (sil) soils and relate Mehlich-3 and Bray-2 soil test P values with yield, total biomass, and P uptake of corn. The Bray-2 P values were six times higher than Mehlich-3 P values for Commerce sil while they were similar for Perry clay. Bray-2 and Mehlich-3 extractable-P of both soils increased with increasing P rate but only corn grown on Perry clay responded to P rate (P < 0.05). Implementation of appropriate testing procedure for estimating plant-available P in soil is an important component of effective P fertilization guidelines for corn.     

Gene expression profiles of yeast Saccharomyces cerevisiae sod1 caused by patulin toxicity and evaluation of recovery potential of ascorbic acid

Patulin (PAT) is a fungal secondary metabolite and exhibits various toxicities including DNA damage and oxidative stress. These toxicities are eased by ascorbic acid (AsA). Although a number of studies regarding the mitigating effect of AsA against PAT toxicity have been reported, a comprehensive study about gene expressions is currently underway. Here, we carried out a detailed evaluation of PAT toxicity by co-incubation with AsA using the superoxide dismutase (SOD) mutant. DNA microarray results extracted the alterations in iron transporter and Fe/S cluster assembly genes; some of the genes that constitute the cellular iron transporter systems remained dysfunctional even in the presence of AsA. Meanwhile, AsA treatment reduced the alterations of G1/S phase cell cycle regulation genes. These results suggest that oxidative stress-derived DNA damage still exists, although AsA treatment effectively reduces PAT toxicity. This implies that a combined condition is required for complete blockade of PAT toxicity.