Pathological and phylogenetic features of prevalent canine distemper viruses in wild masked palm civets in Japan

Ten wild masked palm civets infected with canine distemper virus (CDV), captured in Japan from 2005 to 2007, were histopathologically and phylogenetically analyzed. Phylogenetic analysis based on the amino acid sequences of the H protein of two CDV isolates from masked palm civets revealed that the two isolates were classified into the clade of recent isolates in Japan. Histopathologically marked lesions of virus encephalitis were present in the brain, whereas gastrointestinal lesions were absent or at a mild degree. The distribution of the lesions resembles that of recent CDV cases in dogs. Therefore, recent CDV infections in masked palm civets could be caused by recently prevalent CDV in dogs. The possibility of the masked palm civet as a spreader of CDV among wildlife is also discussed.     

CYP2D-related Metabolism in Animals of the Canoidea Superfamily – Species Differences

CYP2D-related drug metabolism in liver microsomes from animals of the Canoidea super family, i.e. mink (Mustela vison), bears (Ursus arctos), foxes (Vulpes vulpes) and dogs, were investigated. Propranolol, bunitrolol and imipramine, which are typically substrates of CYP2D subfamilies, were used in the experiment. All the animals of the Canoidea superfamily that were tested lacked the ability to catalyse 7-hydroxylation of propranolol, which is one of the major metabolic pathways in rats. Stereoselectivity of propranolol metabolism was towards (S)-propranolol in all the reactions of the animals tested with the exception of mink, which showed a selective tendency towards (R)-propranolol in N-dealkylation. As far as metabolic patterns of (R)- and (S)-propranolol are concerned, bears, foxes and dogs are alike, but minks are somewhat different. Liver microsomes from mink showed, among the animals of the Canoidea superfamily, the lowest propranolol hydroxylase activity at 4- and 5-positions and imipramine 2-hydroxylation and {N-}demethylation activities. We could not detect bunitrolol 4-hydroxylation in mink liver microsomes at the low substrate concentration used. We conclude that mink have the lowest activity of CYP2D-related xenobiotic metabolism among the Canoidea superfamily.     

Involvement of NbNOA1 in NO production and defense responses in INF1-treated Nicotiana benthamiana

During defense responses, plant cells produce nitric oxide (NO), which may control many physiological processes. In a previous study, we reported that nitrate reductase (NR) is responsible in part for INF1 elicitor-induced NO production in Nicotiana benthamiana, but the possibility remains that other NO-generating system(s) contribute to NO production. In mammalian cells, NO production is catalyzed by NO synthase (NOS). However, NOS-like enzyme(s) have never been identified in plants, and only the gene for Arabidopsis thaliana nitric oxide-associated 1 (AtNOA1) has been identified as a putative regulator of NOS activity in plants. In this study, we cloned NbNOA1, a homolog of AtNOA1, from N. benthamiana and investigated its involvement in NO production induced by INF1. The NbNOA1 gene was silenced by a virus-induced gene-silencing (VIGS) technique. NbNOA1-silenced plants had yellowish leaves. Silencing NbNOA1 partially decreased INF1-induced NO production, while overexpressing NbNOA1 did not affect NO production. Silencing NbNOA1 suppressed INF1-induced PR1a gene expression and increased susceptibility to Colletotrichum lagenarium. These results suggest that NbNOA1 is involved in INF1-mediated NO production and is required for defense responses. The nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under accession number AB303300.     

An avirulence gene to rice cultivar K60 is located on the 1.6-Mb chromosome in <Emphasis Type="Italic">Magnaporthe oryzae</Emphasis> isolate 84R-62B

A Japanese differential rice cultivar K60 was tested with 114 F₁ cultures of Magnaporthe oryzae from a cross between isolates 84R-62B and Y93-245c-2. Segregation patterns of avirulence and virulence in the progeny suggested that avirulence on cv. K60 was controlled by a single gene derived from 84R-62B and tentatively named AvrK60. In the F₁ population, AvrK60 cosegregated with avirulence gene AvrPik on a small 1.6-Mb chromosome of 84R-62B and with the 1.6-Mb chromosome itself. Therefore, we suggest that, along with AvrPik, AvrK60 is also located on the 1.6-Mb chromosome of 84R-62B.     

Comparisons among MRI signs,apparent diffusion coefficient,and fractional anisotropy in dogs with a solitary intracranial meningioma or histiocytic sarcoma

Although MRI has become widely used in small animal practice, little is known about the validity of advanced MRI techniques such as diffusion‐weighted imaging and diffusion tensor imaging. The aim of this retrospective analytical observational study was to investigate the characteristics of diffusion parameters, that is the apparent diffusion coefficient and fractional anisotropy, in dogs with a solitary intracranial meningioma or histiocytic sarcoma. Dogs were included based on the performance of diffusion MRI and histological confirmation. Statistical analyses were performed to compare apparent diffusion coefficient and fractional anisotropy for the two types of tumor in the intra‐ and peritumoral regions. Eleven cases with meningioma and six with histiocytic sarcoma satisfied the inclusion criteria. Significant differences in apparent diffusion coefficient value (× 10^?3 mm²/s) between meningioma vs. histiocytic sarcoma were recognized in intratumoral small (1.07 vs. 0.76) and large (1.04 vs. 0.77) regions of interest, in the peritumoral margin (0.93 vs. 1.08), and in the T2 high region (1.21 vs. 1.41). Significant differences in fractional anisotropy values were found in the peritumoral margin (0.29 vs. 0.24) and the T2 high region (0.24 vs. 0.17). The current study identified differences in measurements of apparent diffusion coefficient and fractional anisotropy for meningioma and histiocytic sarcoma in a small sample of dogs. In addition, we observed that all cases of intracranial histiocytic sarcoma showed leptomeningeal enhancement and/or mass formation invading into the sulci in the contrast study. Future studies are needed to determine the sensitivity of these imaging characteristics for differentiating between these tumor types.     

Effect of sampling size on the determination of accurate pesticide residue levels in Japanese agricultural commodities

The uncertainty in pesticide residue levels (UPRL) associated with sampling size was estimated using individual acetamiprid and cypermethrin residue data from preharvested apple, broccoli, cabbage, grape, and sweet pepper samples. The relative standard deviation from the mean of each sampling size (n = 2(x), where x = 1-6) of randomly selected samples was defined as the UPRL for each sampling size. The estimated UPRLs, which were calculated on the basis of the regulatory sampling size recommended by the OECD Guidelines on Crop Field Trials (weights from 1 to 5 kg, and commodity unit numbers from 12 to 24), ranged from 2.1% for cypermethrin in sweet peppers to 14.6% for cypermethrin in cabbage samples. The percentages of commodity exceeding the maximum residue limits (MRLs) specified by the Japanese Food Sanitation Law may be predicted from the equation derived from this study, which was based on samples of various size ranges with mean residue levels below the MRL. The estimated UPRLs have confirmed that sufficient sampling weight and numbers are required for analysis and/or re-examination of subsamples to provide accurate values of pesticide residue levels for the enforcement of MRLs. The equation derived from the present study would aid the estimation of more accurate residue levels even from small sampling sizes.     

Identification of a Germicidal Compound against Picornavirus in Bamboo Pyroligneous Acid

The germicidal activity of pyroligneous acid (PA) against a picornavirus, encephalomyocarditis virus (EMCV), was analyzed, and the component responsible for its disinfectant activity was identified. Bamboo PA (BPA) inactivated EMCV, but neutralization of BPA abolished this activity. Using liquid-liquid phase extraction and silica gel column chromatography, the hydrophobic active fraction of BPA was separated and its 12 major components were identified. The active fraction was reconstructed by mixing synthetic chemicals at the determined concentrations, and a subtraction series of one chemical from the complete mixture was prepared. An in vitro virus assay demonstrated that phenol was the sole germicidal component, and acetic acid augmented the phenol's inactivating activity resulting in >5-log decrease in EMCV infectivity. Considering the low environmental risk of PA, these findings suggest that BPA is a potentially useful agent for preventing viral epidemics in agricultural and human environments.     

Pigs as an experimental model for systemic Mycobacterium avium infectious disease

Mycobacterium avium complex (MAC) is an opportunistic pathogen in AIDS patients and pigs, and causes dissemination through primary intestinal lesions. However, its pathogenesis is not well understood. In this article, we hypothesize that pigs can provide a suitable experimental model of disseminated MAC disease. We compared the initial route of infection, the characteristics of the pathogenic strains, the immunological status of the hosts, and the histological characteristics. The route of infection and infective strains are similar in AIDS patients and pigs. Pigs can respond to infection by the formation of systemic epithelioid granuloma with sufficient cell-mediated immunity. However, there are differences in immunological status and histological features between AIDS patients and pigs. Therefore, pigs might be used as an appropriate animal model because of their good cell mediated immunity triggered by systemic mycobacterial infection. In conclusion, MAC infections in AIDS patients and pigs show similarities in terms of the initial route of infection and the genetic characteristics of the pathogenic strains.     

Uptake by dietary exposure and elimination of aflatoxins in muscle and liver of rainbow trout (Oncorhynchus mykiss)

Uptake and elimination of aflatoxins (AFs) by rainbow trout ( Oncorhynchus mykiss ) during a long-term (21 days) dietary exposure were studied to assess contamination by AFs in aquaculture fish fed AF-containing feed. The uptake factor (UF) of aflatoxin B(1) (AFB(1)) in muscle ranged from 0.40 × 10(-3) to 1.30 × 10(-3). AFB(1) concentrations in liver were 165-342 times higher than in muscle. AFs from feed were more highly accumulated in liver than in muscle. Aflatoxicol (AFL) and aflatoxin M(1) (AFM(1)) were detected in muscle and liver and also in the rearing water. AFL concentrations were higher than AFM(1) by 2 orders of magnitude in muscle, and AFL was a major metabolite of AFB(1). The elimination rate constants (α) of AFB(1) and AFL in muscle (1.83 and 2.02 day(-1), respectively) and liver (1.38 and 2.41 day(-1), respectively) were very large. The elimination half-life (t(1/2)) of AFB(1) was 0.38 days (9.12 h) in muscle and 0.50 days (12.00 h) in liver. The elimination half-life of AFL in muscle and liver was 0.34 day (8.16 h) and 0.29 day (6.96 h), respectively. These data show that AFs are eliminated rapidly and are not biomagnified in fish. Thus, AFB(1) concentration in muscle of fish fed AFB(1)-containing feed (ca. 500 μg/kg) decreased to below the detection limit (20 ng/kg) of the most sensitive analytical method at 1.54 days (36.96 h) after the change to uncontaminated feed.