Method for rapid detection of the PvCesA3 gene allele conferring resistance to mandipropamid, a carboxylic acid amide fungicide, in Plasmopara viticola populations

BACKGROUND: The occurrence of carboxylic acid amide (CAA)‐fungicide‐resistant Plasmopara viticola populations is becoming a serious problem in the control of grapevine downy mildew worldwide. RESULTS: The authors have developed a method, which utilises PCR‐RFLP, for the rapid detection of resistance to the CAA fungicide mandipropamid in P. viticola populations. With this method, a glycine‐to‐serine substitution at codon 1105 of the cellulose synthase gene PvCesA3 of CAA‐fungicide‐resistant P. viticola was easily detected, although no resistant P. viticola was detected from 398 isolates in Japan. CONCLUSION: It is proposed that the PCR‐RFLP method is a reliable tool for the rapid detection of CAA‐fungicide‐resistant P. viticola isolates. Only 4 h was required from the sampling of symptoms to the phenotyping of fungicide resistance. Copyright © 2011 Society of Chemical Industry     

Phenolics composition and antioxidant activity of sweet basil (Ocimum basilicum L.)

The antioxidant activity of a methanolic extract of Ocimum basilicum L. (sweet basil) was examined using different in vitro assay model systems. The crude extract was fractionated on a Sephadex LH-20 column, and six fractions were identified. The DPPH scavenging assay system and the oxidation of the soy phosphotidylcholin liposome model system were used to evaluate the antioxidant activity of each fraction. Fraction IV showed the strongest activity followed by fractions V and VI. Phenolic compounds responsible for the antioxidative activity of the fractions were characterized by atmospheric pressure chemical ionization liquid chromatography-mass spectrometry. The major antioxidant compound in fraction IV was confirmed as rosmarinic acid by (1)H NMR and characteristic fragmentations in the mass spectrum. Moreover, the native of antioxidant activity of rosmarinic acid in the liposome system was examined. The results showed that one rosmarinic acid can capture 1.52 radicals, and furthermore, the existence of a synergistic effect between alpha-tocopherol and rosmarinic acid was revealed.     

Electromyographic activity of cubital joint muscles in horses during locomotion

Electromyographic (EMG) activity of 4 muscles of the cubital joint and the strain of forelimb hooves were recorded telemetrically in 4 Thoroughbreds (with and without a rider) standing, walking, trotting, and cantering. Bipolar fine wire electrodes were inserted into the muscles, and strain gauges were attached to the hoof wall. Motion pictures (16 mm), synchronized with EMG tracings, were taken to obtain kinematic data. When horses were standing, the biceps brachii had tonic activity, but the brachialis and the caput longum and the caput laterale of the triceps brachii had no EMG activity. The biceps brachii had EMG activity during the stance phase. The brachialis had EMG activity from the end of the stance phase to the middle of the swing phase. Unlike the biceps brachii, the brachialis acted as a flexor muscle of the cubital joint during locomotion. The EMG activity of the caput longum of the triceps brachii was detected from midswing phase to early stance phase. The EMG activity of the caput laterale of the triceps brachii began in midswing or late-swing phase and ceased in early stance or midstance phase. During locomotion, caput longum EMG activity always preceded caput laterale activity. When horses were cantering, the brachialis and the caput longum (acting mainly in the swing phase) had an EMG activity phase different from those in leading and trailing forelimbs. These 4 muscles had similar EMG activity patterns during locomotion in horses with and without a rider.     

Prevalence of Giardia intestinalis infection in dogs of breeding kennels in Japan

Giardia intestinalis antigen in fecal samples was examined in 361 dogs of 14 breeding kennels located at various areas in Japan, using a commercial enzyme-linked immunosorbent assay (ELISA) kit. G. intestinalis antigen was detected in 37.4% of the fecal specimens. All of the 14 breeding kennels were positive for G. intestinalis antigen with the range from 6.7 to 59.3%. The prevalence in puppies (54.5%) was significantly (p < 0.01) higher than that in adults (30.9%). There was no difference in prevalence between males and females, and between the puppies from the mother dogs positive and negative for Giardia antigen. In conclusion, G. intestinalis widely invaded the breeding kennels in Japan.     

Renal effects of medetomidine in isoflurane-anesthetized dogs with special reference to its diuretic action

Renal effects of the selective alpha(2)-adrenoceptor agonist, medetomidine, were investigated in anesthetized dogs. Animals were administered medetomidine 20 and 40 microg/kg intravenously (IV) and 80 mug/kg intramuscularly (IM) or 1 ml of saline IV. Urine and blood samples were collected before and at 30, 60, 90 and 120 min following medetomidine injection. Mean arterial blood pressure (MABP), renal blood flow (RBF), glomerular filtration rate (GFR), urine volume (U(v)), urine osmolality (U(osm)), free water clearance (C(H2O)), fractional clearance of sodium (F(Na)), plasma osmolality (P(osm)), plasma glucose levels and plasma antidiuretic hormone (ADH) concentrations were measured. The results showed that IV administration of medetomidine initially increased MABP 5-15 min followed by long-lasting decrease. The initial hypertension was not observed after IM administration, which was accompanied by a more profound hypotensive effects. RBF, GFR, U(v), C(H2O) increased after IV injection and decreased after IM. Medetomidine increased FNa and Posm and decreased U(osm). Plasma glucose levels initially increased and subsequently decreased. Plasma ADH concentration was decreased by IV injection but increased by IM administration. Our data imply that: 1) IV administration of medetomidine at dose rates of 20 and 40 microg/kg results in profound diuresis up to 2 hr; 2) Suppression of ADH release from the CNS is one of the mechanisms of medetomidine-induced diuresis although it may not be the principal one.     

Enzyme-linked immunosorbent assay for detection of canine chromogranin A by use of immunological cross-reactivity of rabbit anti-bovine chromogranin A antibody

Bovine and canine chromogranin A were extracted and purified from each specie's adrenal glands. Isolated bovine 70 kDa protein showed 100% identity to bovine CgA reported previously, whereas isolated canine 68 kDa protein showed 83.3% identity to bovine CgA by the NH(2)-terminal amino acid sequence analysis. Rabbit antibody to purified bovine protein (CgA) was found to immunologically cross-reacted with purified canine protein (CgA). In sandwich ELISA with anti-bovine CgA, concentration-dependent curves were obtained ranging from 0.3 to 20 mug/ml for canine CgA. From these findings, sandwich ELISA with anti-bovine CgA is found to be useful to determine the concentration of canine CgA.     

Comparative studies on lipid analysis and ultrastructure in porcine and southern minke whale (Balaenoptera bonaerensis) oocytes

The present study was conducted to clarify the difference in the color of the cytoplasm in immature follicular oocytes from prepubertal and adult minke whales. The four lipid contents (triglyceride, total cholesterol, phospholipids and non-esterified fatty acids) in vitrified immature oocytes from prepubertal and adult minke whales, and also in fresh and vitrified immature porcine oocytes, were measured. The lipid contents in vitrified-warmed minke whale oocytes were similarly high compared with those in vitrified-warmed porcine oocytes. In particular, the total cholesterol and phospholipid contents in the vitrified immature oocytes from prepubertal and adult minke whales were significantly (P<0.05) higher than those from prepubertal pigs. Furthermore, the distribution of lipid droplets in fresh and vitrified immature oocytes was observed in transmission electron microscopy. Lipid droplets in the prepubertal minke whale oocytes were distributed throughout the cytoplasm. In contrast, adult minke whales had larger lipid droplets which were distributed mainly in the central portion of the cytoplasm. The lipid droplets of immature oocytes from prepubertal pigs were larger than those in minke whale oocytes. These results indicated that the difference in the distribution of the cytoplasmic lipid droplets may result in the difference in the color tone of both prepubertal and adult whale oocyte cytoplasm.