Morphological and molecular characterization of <Emphasis Type="Italic">Oidium</Emphasis> subgenus <Emphasis Type="Italic">Reticuloidium</Emphasis> (powdery mildew) newly occurred on cucumber in Japan

In 2002, a powdery mildew with catenate conidia lacking fibrosin bodies was found on cucumber in a greenhouse in Kanagawa Prefecture, Japan. Morphological observation revealed that the fungus belongs to Oidium subgenus Reticuloidium, anamorph of the genus Golovinomyces. Molecular phylogenetic analyses of the nucleotide sequences of the rDNA ITS regions and D1/D2 domains of the 28S rDNA indicated that the fungus belongs to the clade of G. orontii with other Golovinomyces fungi from a wide range of host plants, suggesting that the fungus was newly transported from abroad. Because there has been no prior report of cucumber powdery mildew caused by Reticuloidium, further research on the physiology, epidemiology, control and resistant cucumber varieties is required.     

Purification and characterization of two anionic trypsins from the hepatopancreas of carp

SUMMARY: Two trypsins, designated as trypsin A and trypsin B, have been purified from the hepatopancreas of carp. The purification procedures consisted of ammonium sulfate fractionation, and chromatographies on DEAE-Sephacel, Ultrogel AcA54 and Q-Sepharose. Trypsin A was purified to homogeneity with the molecular mass of approximately 28 kDa, while trypsin B gave two close bands of 28.5 kDa and 28 kDa on sodium dodecylsulfate polyacrylamide gel electrophoresis both under reducing and non-reducing conditions. On native-PAGE, both trypsin A and trypsin B showed a single band. Trypsin A and trypsin B revealed optimum temperature of 40°C and 45°C, respectively, and shared the same optimum pH 9.0 using Boc-Phe-Ser-Arg-MCA as substrate. Both enzymes were effectively inhibited by trypsin inhibitors and their susceptibilities were similar. The NH₂-terminal amino acid sequences of trypsin A and trypsin B were determined to 37th and 40th amino acid residue, respectively. Their sequences were very homologous, but not identical to that of a trypsin-type serine proteinase from carp muscle and these of other trypsins. Immunoblotting test using the antibody raised against trypsin A cross-reacted with trypsin B positively.     

Seasonality of serum levels of vitellogenin in male Japanese whiting, Sillago japonica, reared under natural temperature and photoperiod

ABSTRACT:   Because blood vitellogenin (Vg) has been considered a biomarker for environmental estrogens, the basal levels of Vg and 17β-estradiol (E₂) were determined in male Japanese whiting reared under natural conditions. Serum levels of Vg and E₂ were measured and gonadal development was assessed by gonadosomatic index (GSI) and histological observation in 8–10 male fish at monthly intervals throughout the annual reproductive cycle. Serum E₂ was <60 pg/mL throughout the study period. In contrast, serum Vg exhibited seasonal changes: serum levels of Vg gradually increased from April to May (mean 63 ± 13 ng/mL and 124 ± 48 ng/mL in April and May, respectively), and then reached a peak value (mean 352 ± 68 ng/mL) in June. Thereafter, serum Vg gradually decreased, reaching undetectable levels (<50 ng/mL) in October. Serum levels of Vg tended to increase in the male fish in which the GSI was >1%. Histological observation revealed that testes in such male fish were in active spermatogenesis and then all of the testes of male fish in which serum Vg decreased to ND levels were regressed. These results suggest that Vg productive potency (sensitivity to estrogens) may increase in the spermatogenic stage, resulting in production of Vg in response to very low levels of natural or xenobiotic estrogens.     

Inhibitory effect of isoprothiolane on metabolism of a phosphoramidate by isolates of Pyricularia oryzae cav. in relation to fungicide sensitivity

The inhibitory effect of isoprothiolane(diisopropyl 1,3-dithiolan-2-ylidenemalonate), a fungicide for rice blast control, on the metabolism of dibutyl N-methyl-N-phenylphosphoramidate (BPA) by 20 isolates of Pyricularia oryzae was examined in relation to sensitivity of the isolates to the reference fungicide IBP(S-benzyl diisopropylphosphorothiolate). The isolates were divided into five groups based on the modes of BPA metabolism and the inhibition of BPA metabolism by isoprothiolane. Every isolate in groups I and II, which was either a field isolate or a stock culture, decomposed BPA rapidly and produced both hydroxylated and N-demethylated BPA as metabolites. BPA decomposition by these isolates was strongly inhibited by isoprothiolane, resulting in the decreased production of both metabolites in group I and of the hydroxylated metabolite in group II. These isolates were almost equally sensitive to isoprothiolane. Isolates in groups III, IV, and V were all obtained from selection of the fungus mutants found growing on media containing isoprothiolane. Isolates in group III, derived by plating large numbers of conidia, did not decompose BPA to any extent. Mutants of groups IV and V were obtained from fast-growing sectors on agar containing isoprothiolane. Both these groups decomposed BPA, but isolates belonging to group IV produced copious amount of N-demethylated BPA whereas isolates in group V did not. BPA metabolism by these in vitro mutants in groups III, IV, and V was not inhibited by isoprothiolane. Thus, the inhibitory effect of isoprothiolane on BPA metabolism was correlated with sensitivity of an isolate to isoprothiolane. The inhibitory effect of IBP on BPA metabolism was not always correlated with the sensitivity of an isolate to IBP.     

Prevention of adverse reactions following milbemycin D administration to microfilaremic dogs infected with Dirofilaria immitis

Some adverse reactions such as shock-like reaction and dirofilarial hemoglobinuria (caval syndrome) occasionally occurred in microfilaremic dogs following milbemycin D (Milbe) administration. This study was carried out to seek the prevention of these adverse reactions. In two groups containing 16 and 9 dogs respectively which were administered either chlorpheniramine maleate (1 mg/kg) or indomethacin (2.5 mg/kg) simultaneously with Milbe (1 mg/kg), the incidence of clinical signs such as the pale color of the visible mucous membranes, respiratory disorders, caval syndrome and shock-like reaction as well as changes in clinical parameters such as RBC and WBC counts, WBC profile and serum total protein, were almost equal to that observed in the group administered Milbe alone. In 41 dogs administered prednisolone (1 mg/kg) simultaneously with Milbe (1 mg/kg), no shock-like reaction was observed. Changes in clinical parameters were different from those in the group administered Milbe alone, whereas some clinical signs of adverse reactions, including caval syndrome, were observed. These results indicated that prednisolone was effective for prevention of the shock-like reaction in microfilaremic dogs induced by Milbe.     

Reducing effect of dietary anacardic acid on body fat pads in rats

We have previously shown that anacardic acid has an uncoupling effect on oxidative phosphorylation in rat liver mitochondria using succinate as a substrate (Toyomizu et al. 2000). In the present study, in order to clarify whether or not anacardic acid could be used advantageously as a special feed/food supplement to reduce fat deposition through the uncoupling action, two sets of experiments were conducted to determine quantitatively the effect of dietary anacardic acid (0.1% w/w) supplementation. More specifically, effects on growth, feed efficiency, fattening, and levels of several constituents of blood serum in rats fed normal and low protein–high carbohydrate (CHO) diets were examined. There were no significant differences in bodyweight gain, feed consumption and feed efficiency among the experimental groups. For the total fat pad content, including inguinal and epididymal fat, significant interaction was shown between both treatments: dietary anacardic acid at 0.1% w/w significantly decreased the total fat pad content in rats fed the CHO diet, but not in rats fed the normal diet. Weights of heart, spleen and brown adipose tissue were not affected by either the dietary treatment or anacardic acid, while both liver and kidney weights decreased with feeding of anacardic acid at 0.1% w/w, but were not affected by the CHO diet. Anacardic acid supplementation in the diet had no effect on serum glutamic oxaloacetic transaminase, alkaline phosphatase or lactate dehydrogenase levels, suggesting that the dysfunction of liver or kidney may not be induced by dietary anacardic acid. The results of the present study reveal a unique function of anacardic acid in that, for dietary conditions enhancing body fat deposition, that is consumption of a diet high in carbohydrates, dietary anacardic acid has the potential to decrease body fat deposition. A possible mechanism for differences observed in anacardic acid‐induced regulation of body fat pad content between rats fed the normal and CHO diets, based on uncoupling action of anacardic acid on the mitochondrial oxidative phosphorylation, is discussed.