Ontogeny of testicular inhibin and activin in ducks: An immunocytochemical analysis

The ontogeny of testicular inhibin/activin in ducks was investigated. Testicular localization of three inhibin/activin subunits (α, β_A and β_B) was determined in embryonic and newly hatched ducks from 12 days of incubation to 1 day of age, in immature ducks and in adult ducks. In the duck embryonic testis, positive α‐subunit immunostaining was first detected in the Leydig cells and Sertoli cells on day 15 of incubation, whereas β_A‐subunit and β_B‐subunit immunostaining were found in Sertoli cells and primary germ cells on day 18 of incubation. In 1 month old ducks, intense staining of α‐subunit was present in the seminiferous epithelium consistent with localization in Sertoli cells and primary germ cells, and the immunostaining of the β_A‐ and β_B‐subunit was also present in Sertoli cells and primary germ cells. Specific immunostaining with inhibin/activin α‐, β_A‐ and β_B‐subunits antisera occurred in Sertoli cells in the adult duck testes. In conclusion, it was shown that, in the duck testis, the majority of α‐, β_A‐ and β_B‐subunits are colocalized in Sertoli cells with a certain degree of staining in germ cells and the α‐subunit is present in Leydig cells of embryonic testes before day 18 of incubation. These results indicate that Sertoli cells and possibly germ cells in the embryonic testes of late stage of incubation and newly hatched ducks, immature ducks and mature ducks may produce bioactive inhibin dimers, inhibin A and inhibin B, as a possible regulator of follicle‐stimulating hormone secretion. Free inhibin/activin subunits and their dimers may also play an autocrine/paracrine role in the development of the testis and spermatogenesis. Furthermore, early onset of the α‐subunit in duck testes indicates that it may have an autocrine/paracrine effect on steroid hormones, which is important for sex differentiation.     

Induction of Systemic Resistance in Cucumber against Several Diseases by Plant Growth-promoting Fungi: lignification and Superoxide Generation

Five fungal isolates (Trichoderma, Fusarium, Penicillium, Phoma and a sterile fungus) from zoysiagrass rhizosphere that promote plant growth were tested for their ability to induce systemic resistance in cucumber plants against Colletotrichum orbiculare. Roots of cucumber plants were treated with these fungal isolates using barley grain inocula (BGI), mycelial inocula (MI) or culture filtrate (CF). Most isolate/inoculum form combinations significantly reduced the disease except BGI of Trichoderma. These fungal isolates were also evaluated for induction of systemic resistance against bacterial angular leaf spot and Fusarium wilt by treatment with BGI. Penicillium, Phoma and the sterile fungus significantly reduced the disease incidence of bacterial angular leaf spot. Phoma and sterile fungus protected plants significantly against Fusarium wilt. Roots treated with CFs of these fungal isolates induced lignification at Colletotrichum penetration points indicating the presence of an elicitor in the CFs. The elicitor activity of CFs was evaluated by the chemiluminescence assay using tobacco callus and cucumber fruit disks. The CFs of all isolates elicited conspicuous superoxide generation. The chemiluminescence activity of the CF of Penicillium was extremely high, and its intensity was almost 100-fold higher than that of other isolates. The chemiluminescence activity was not lost following treatment with protease or autoclaving or after removal of lipid. The MW 12,000 dialyzed CF fraction was highly effective in eliciting chemiluminescence activity. Chemiluminescence emission from cucumber fruit disks treated with Penicillium was the same as that obtained from tobacco callus, except that the lipid fraction also showed a high activity. Both the MW 12,000 fraction and the lipid fraction induced lignification in the epidermal tissues of cucumber hypocotyls.     

Development and promotion of laborsaving application technology for paddy herbicides in Japan

Total labor time on paddy rice has been decreasing year by year with the development and introduction of appropriate herbicides, especially ‘one‐shot' herbicides. However, in the case of application of granules, it is still difficult for a farmer to apply herbicides while carrying heavy power backpack sprayers. New formulation recipes and different application technology such as a throw‐in type formulation on jumbo granules or flowable has improved heavy workloads in comparison with granule application by using heavy power backpack sprayers. In addition, other advantageous points such as application volume and elimination of the drift problems of this new application technology were introduced and confirmed. As a result of this introduction, these formulations were used in 830 000 ha and reached 30% of the total treatment area in paddy rice in 2000.     

Biocontrol ofRhizoctonia damping-off of cucumber by non-pathogenic binucleateRhizoctonia

Three isolates of binucleateRhizoctonia (BNR) were tested for biological control of damping-off of cucumber seedlings caused byRhizoctonia solani AG 2-2 and AG 4. BNR isolates L2 (AG Ba) and W1 and W7 (AG A) provided protection of 58 to 71% against virulent isolate C4 of AG 4 and 64 to 75% protection against virulent isolate RH 65 of AG 2-2. Varying protection was provided to the seedlings by the BNR isolates against the virulentR. solani from the two AGs depending on their combination. The BNR isolates did not vary in providing protection to the seedling when tested against virulent C4 when both isolates were inoculated using three different methods,viz. in water agar, combination of water agar and soil and using soil alone. Protection of 58 to 71 % was provided by the isolates when inoculation was done on the hypocotyl using water agar, 62.8 to 75% using the combination of water agar and soil, and 75 to 85% when inoculation of both isolates was done in soil. Pre-incubation of BNR W7 or delayed inoculation of C4 (from 0.5 day to longer duration) using the different methods provided an increased protection to the seedlings to give complete inhibition of damping-off disease. Simultaneous inoculation of both BNR W7 and C4 using the three methods failed to provide protection to the seedlings. Among the BNR isolates, BNR W7 showed plant growth promotion in terms of significant increase in plant height (P=0.01) and fresh weight (P=0.05).     

Isolation and purification of a protective protein antigen of Erysipelothrix rhusiopathiae

The rapid growth and high survival rate of Erysipelothrix rhusiopathiae was determined using a culture of the bacterium in tryptic soy broth supplemented with 0.3% Tris-hydroxymethyl aminomethane and 0.1% Tween 80 (TT-TS broth). High concentrations of 64, 66 and 43 kDa proteins, which are associated with protection against E. rhusiopathiae infection in mice, were obtained by alkaline treatment of whole cells using 0.05-1 N NaOH. The supernatant of alkaline treated cells (alkaline extract; AE) was stable at alkaline or neutral pH. However, aggregates appeared at neutral pH in the absence of sodium dodecyl sulphate (SDS). A high yield of 64, 66 and 43 kDa proteins was obtained from strain Agata (serovar 5). The proteins were eluted from gel bands following SDS-polyacrylamide gel electrophoresis (SDS-PAGE) of the AE from strain Agata and designated P64 and P43. The amounts of P64 and P43 isolated were 0.7 and 0.3 mg/16 g of wet bacteria, respectively. In a mouse protection test, 50% protective doses (PD50) of P64 and P43 were 0.58 and 0.63 microgram, respectively. Upon Western blotting of the AE, both anti-P64 and anti-P43 antibodies reacted with the 64 and 43 kDa proteins. From these results, it is suggested that P64 is the most effective protective antigen and that P43 (43 kDa protein) is a degradation product of P64. Therefore, the 64 kDa structural proteins are associated with the induction of a protective activity against E. rhusiopathiae infection in mice.     

T-cell lymphoma in a savanna monkey (Cercopithecus aethiops) probably related to simian T-cell leukemia virus infection

Lymphoma was seen in an 11-year-old female savanna monkey (Ceropithecus aethiops). The superficial inguinal and visceral lymph nodes were markedly enlarged, and their architecture was completely effaced by neoplastic cells. The neoplastic cells, which were highly pleomorphic, resembled those in adult T-cell lymphoma-leukemia in humans. Ultrastructurally the neoplastic cells were characterized by nuclear irregularity and clustered dense bodies, and almost all cells showed positivity for CD3. The animal had been reared with her family, and her mother and 2 brothers had antibodies reactive to human T-cell leukemia virus. This virus serologically cross-reacts with simian T-cell leukemia virus, which may be the causative agent of the present neoplasm.     

Energy utilisation of carbohydrate, fat and protein sources in newly hatched broiler chicks

1. TME, TMEn and metabolisability (TME/gross energy) of energy-yielding foodstuffs were determined in 1, 3 and 10 d old broiler chicks, using the assay method developed by Murakami et al. (1994), in order to characterise energy utilisation during the 10 d after hatching. 2. TME, TMEn and metabolisability of dextrin and starch were low in chicks aged 1 d, and increased with age up to 10 d. Energy values of glucose and maltose at days 1 and 3 could not be determined because of the sudden death of birds soon after the feeding. TMEn and metabolisability of cereal sources were lower in d-old chicks than in those aged 3 and 10 d. At all ages, maize was better utilised than wheat and sorghum. 3. In the fat sources (coconut oil, beef tallow and safflower oil) no age dependency was observed in TME, TMEn and metabolisability. 4. Bioavailability of soyabean meal and fish meal was lower at 1 d than at 3 d and 10 d. Energy utilisation from casein was the highest among the protein sources tested and it was not age dependent. 5. It is concluded that energy utilisation of carbohydrate and protein sources during 10 d post-hatch tended to increase with age. Among the energy yielding foodstuffs fat sources seem to be better utilised, with no age dependency.     

Physicochemical properties of akabane virus: A member of the simbu arbovirus group of the family bunyaviridae

The multiplication of Akabane virus was not inhibited in the presence of 5-iodo-2′-deoxyuridine, indicating the presence of RNA. The virus was considered to have an envelope, as it was sensitive to ether and chloroform. It was readily inactivated by deoxycholate and trypsin, but was not precipitated by protamine sulphate. The virus was very labile at pH 3 and also rather heat-labile. Akabane virus was readily filtered through membrane filters of 200 or 100-nm pore size, but not through 50-nm filters. Equilibrium centrifugation in a CsCl density gradient gave a peak of infectivity and hemagglutinin at a density of 1.22 g/ml. The peak fractions thus obtained contained numerous virus particles, roughly spherical, variable in size, 70 to 130 nm in diameter, and mostly having a ragged, closely adherent envelope with projections, when examined, following phosphotungstic acid negative staining, in an electron microscope.