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.     

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.     

Comparison of carcass and meat characteristics of Brahman grade cattle (Bos indicus) and crossbred water buffalo (Bubalus bubalis) fed on high roughage diet

The objective of this study was to compare the carcass and meat quality in crossbred cattle and crossbred water buffalo at the same younger age and fed with high roughage in the Philippines. Ten crossbred cattle and 10 crossbred water buffalo, with an average age of 22 months (18–24 months) were used in this experiment. The animals were fed a similar diet, which consisted of 85% Napier or Para grass and 15% concentrate mixture on a dry matter basis, for 180 days before slaughter. Slaughter weight of the crossbred water buffalo was higher (P < 0.05) than that of the cattle, although the dressing percentage was significantly (P < 0.01) lower for the former than for the latter. The estimated lean yield was higher (P < 0.05) in crossbred cattle than crossbred water buffalo. The muscle fiber diameter of loin in buffalo meat was smaller (P < 0.05) than that in cattle. In regard to carcass and meat quality, the crossbred water buffalo was comparable or slightly superior to the crossbred cattle in tenderness, sensory score of color (P < 0.01) and flavor (P < 0.05). These results clearly indicate that at a younger age and even under high roughage‐based fattening rations, crossbred water buffalo are able to produce good quality meat, which will be by no means inferior to that of crossbred cattle in the Philippines.     

Analysis of 46-kDa protein in the perivitelline membrane of Japanese quail (Coturnix japonica)

The extracellular matrix surrounding the oocyte before ovulation is called the perivitelline membrane (PL) in avian species. The PL is constructed with two major glycoproteins, ZPC and ZP1, which are synthesized in the ovarian granulosa cells and the liver, respectively. Although the properties of the major components in the PL have been examined, knowledge about the nature of its minor constituents is lacking. In this study we focused on PL protein, which migrates at 46‐kDa in the gel of SDS‐PAGE. N‐terminal sequence analysis demonstrated that the 46‐kDa protein is the C‐terminal fragment of ZP1. Analysis of lysylendopeptidase digests or cyanogens bromide‐degraded fragments of ZP1 confirmed this postulate. Western blot analysis using antiserum against 46‐kDa protein indicated the absence of 46‐kDa protein in the serum. Moreover, small immunoreactive bands, thought to be cleaved fragments of ZP1, were detected in the PL lysate by western blot analysis using antiserum against the N‐terminal peptide of ZP1. These results indicated that the N‐terminal proteolytic processing of ZP1 might take place after the arrival of ZP1 at the ovary, and the resulting product, 46‐kDa protein, is incorporated into the PL.     

Involvement of gacA Gene in the Suppression of Tomato Bacterial Wilt by Pseudomonas fluorescens FPT9601

Pseudomonas fluorescens FPT9601, a plant growth-promoting rhizobacterium (PGPR) isolated from tomato rhizosphere, can protect tomato (Lycopersicon esculentum Mill) from bacterial wilt disease caused by Ralstonia solanacearum. This strain produces antibiotics 2,4-diacetylphloroglucinol (2,4-DAPG) and hydrogen cyanide (HCN). It also produces proteases and uncharacterized siderophores (Sid). A mutant strain SM2214, obtained by Tn5 insertion, did not produce 2,4-DAPG, HCN or proteases, but overproduced Sid. Marker-exchange mutagenesis confirmed that a single transposon insertion caused the multiple phenotypic changes of this mutant. Complementation of the mutant with a 1.3-kb DNA fragment that was amplified from genomic DNA of the wild-type P. fluorescens strain by PCR could restore the lost functions of the mutant strain. Nucleotide sequencing revealed that the fragment contained a 642-bp open reading frame (ORF) highly homologous to the regulator responser gene gacA. The in vitro anti-bacterium test and plant protection experiment under greenhouse conditions indicated that the gacA gene played an important role in the suppression of tomato bacterial wilt disease. Received 20 November 2000/ Accepted in revised form 19 January 2001     

Identification of herds with cattle persistently infected with bovine viral diarrhea virus by virological evaluation of three calves

For the identification of herds with cattle persistently infected (PI) with bovine viral diarrhea virus, 1,272 animals from 20 herds were subjected to serum neutralizing (SN) test using the Nose strain and virus isolation. Eighteen PI cattle were detected from 5 herds. On the phylogenetic tree based on the nucleotide sequences of the 5' untranslated region, the isolates from the PI cattle were classified into genotypes-1a or -1b. Of 3 unvaccinated calves aged 6 to 12 months selected from each herd, the probabilities of obtaining 2 or more non-PI cattle with SN antibody titers of 64 or more (P(SN)), one or more PI cattle (P(VI)), and either of the conditions (P(Total)) were calculated using the hypergeometric probability model. P(Total) for the 5 herds with PI cattle was 1.000. P(SN) for 3 herds with many PI cattle within the selected age group was as low as 0.500 or less, and P(VI) was as high as 0.886 or more. P(SN) in the 2 other herds with few PI cattle was 1.000, and P(VI) was as low as 0.375 or less. P(Total) in 13 of 15 herds without PI cattle was 0.000, and was 0.714 or 0.774 for the 2 other herds. These results suggest that herds with PI cattle can be predicted with high accuracy when both SN test and virus isolation are performed on only 3 unvaccinated calves aged 6 to 12 months selected from a herd.     

Wind tunnel study on aerodynamic characteristics of shrubby specimens of three tree species

With the popularization of green roofing systems, their wind resistant performance is becoming more and more important especially in typhoon areas. A prerequisite study was carried out to determine the aerodynamic characteristics of suitable trees to provide basic scientific data. Shrubby specimens of three tree species appropriate for intensive green roofs with different aerodynamic forms were tested through a boundary layer wind tunnel experiment, including one deciduous tree, one coniferous tree and one broadleaf evergreen tree. Drags and overturning moments under different wind speeds were measured; wind-speed-specific frontal areas and tip displacements were recorded. After initial tests, trees were pruned to investigate the effect of crown porosity, and the effects of view angle and turbulence intensity were also considered. The results showed that even considering frontal area reduction due to increasing wind speeds, drag coefficients decreased with increase in wind speed for deciduous and coniferous trees. Drag coefficient characteristics for different trees agreed well with those obtained from previous researches. The effect of crown porosity varied among tree species, but view angle and turbulence intensity did not significantly affect it.     

Benzodiazepines inhibit the acetylcholine receptor-operated potassium current (IK.ACh) by different mechanisms in guinea-pig atrial myocytes

The anticholinergic effects of 7 benzodiazepines, bromazepam, camazepam, chlordiazepoxide, diazepam, lorazepam, medazepam and triazolam, were compared by examining their inhibitory effects on the acetylcholine receptor-operated potassium current (I(K).(ACh)) in guinea-pig atrial myocytes. All of these benzodiazepines (0.3-300 μM) inhibited carbachol (1 μM)-induced I(K).(ACh) in a concentration-dependent manner. The ascending order of IC(50) values for carbachol-induced I(K).(ACh) was as follows; medazepam, diazepam, camazepam, triazolam, bromazepam, lorazepam and chlordiazepoxide (>300 μM). The compounds, except for bromazepam, also inhibited I(K).(ACh) activated by an intracellular loading of 100 μM guanosine 5'-[γ-thio]triphosphate (GTPγS) in a concentration-dependent manner. The ascending order of IC(50) values for GTPγS-activated I(K).(ACh) was as follows; medazepam, diazepam, camazepam, lorazepam, triazolam chlordiazepoxide (>300 μM) and bromazepam (>300 μM). To clarify the molecular mechanism of the inhibition, IC(50) ratio, the ratio of IC(50) for GTPγS-activated I(K).(ACh) to carbachol-induced I(K).(ACh), was calculated. The IC(50) ratio for camazepam, diazepam, lorazepam, medazepam and triazolam was close to unity, while it for chlordiazepoxide could not be calculated. These compounds would act on the GTP binding protein and/or potassium channel to achieve the anticholinergic effects in atrial myocytes. In contrast, since the IC(50) ratio for bromazepam is presumably much higher than unity judging from the IC(50) values (104.0 ± 30.0 μM for carbachol-induced I(K).(ACh) and >300 μM for GTPγS-activated I(K).(ACh), it would act on the muscarinic receptor. In summary, benzodiazepines had the anticholinergic effects on atrial myocytes through inhibiting I(K).(ACh) by different molecular mechanisms.