Modification of the hyperosmotic infiltration method of vaccination against vibriosis in cultured ayu, Plecoglossus altivelis Temminck and Schlegel

Abstract. A vaccine solution of a formalin-killed culture of Vibrio anguillarum cells was observed to be toxic to young ayu when administered by the hyperosmotic infiltration method. The toxin was present in the culture broth. After the toxin was removed from the broth by centrifugation, the fish were dipped in 5.32% NaCl solution for 2 min and then in a solution containing precipitated cells for 3 min. The immunized fish were protected against vibriosis when challenged one month after immersion. The bacterin was administered to ayu by a further two methods, both using lyophilized whole cells of formalin-killed V. anguillarum. In one method, the fish were placed in a 5.32% NaCl solution for 2 min and then in a solution containing lyophilized cells at 2 g/l of well water for 3 min (two-step immersion). In the other method, the fish were placed in a 5.32% NaCl solution containing lyophilized cells also at 2 g/l for 3 min (one-step immersion). A high level of protection against artificial challenge was achieved with either method. No agglutinating antibodies to V. anguillarum were detected in either the serum or mucus of fish dipped in a vaccine solution, a supernatant, or a precipitated solution, one month after immersion. On the other hand, serum titres were detected in fish vaccinated by injection, although no titres were detected in mucus. LD₅₀ values are presented for the virulence of the V. anguillarum strain. Compared to the original strain, virulence increased after the third passage in ayu, but decreased after the thirteenth passage in medium.     

Canopy structure and productivity of Festuca avundinacea Schreb. swards during vegetative and reproductive growth

Canopy structure, productivity and their relationships were examined in 2-year-old swards of fourteen tall fescue ( Festuca arundinacea Schreb.) strains during the vegetative and reproductive growth stages. During the vegetative growth stage morphological characters, particularly tiller size, were closely associated with productivity. Swards with large tillers showed an effective distribution of the incoming light energy within the canopy and hence low extinction coefficient ( K ) value and high productivity at complete light interception. On the other hand, although there was no apparent correlation between K and the productivity or the whole crop during the reproductive growth stage, the productivities of the reproductive and vegetative tillers were positively and negatively related to K respectively. Leaf area index of the reproductive tillers and their position in the canopy had marked effects on the distribution of the incoming light energy within the canopy and on the productivity of both types of tillers. The productivity of the vegetative and the reproductive tillers is discussed in terms of the effect of the competition for incoming light energy between both types of tillers.     

Dynamics of CD3+ T-cell Distribution Throughout the Estrous Cycle and Gestation in the Bovine Endometrium

T cells are the dominant lymphocytes in the endometrium and are considered to play a crucial role in implantation and in the maintenance of gestation through cytokine production and immune regulation. The mechanisms underlying immunoregulation at the feto-maternal interface are still obscure for this complex system. Understanding the role of T cells is a key factor in understanding the endometrial immune system. In this study, the distribution of endometrial CD3⁺ T cells in bovines was examined by immunohistochemical analysis. The estrous cycle and gestation was divided into 4 stages, and the number of CD3⁺-positive T cells was counted in each stage. CD3⁺ cells were found in the endometrium in significant numbers throughout the estrous cycle and were mostly located in the subepithelial area. The number of CD3⁺ cells significantly increased in the early and mid-luteal phases but decreased after implantation with the progression of gestation. No T cells were found in the placentome or specifically in the tissues near the fetus, including the trophoblastic area. In addition, very few T cells were found in stromal regions close to the myometrium of the endometrium. These findings suggest that downregulation of bovine endometrial CD3⁺ T-cell functions is closely related to the successful maintenance of gestation in a spatiotemporal manner.     

Effect of estrus synchronization treatment after luteolysis on Holstein heifers as embryo transfer recipients

This study was designed to evaluate the effect of estrus synchronization treatments on recipient heifers for embryo transfer (ET). Holstein heifers were separated into the following three groups: (i) an administration of 50 µg GnRH (gonadotropin‐releasing hormone) analog was given to heifers at a random stage of the estrus cycle, followed 7 days later by two administrations of 7.5 mg prostaglandin F2 alfa analog (PG) as control; (ii) another administration of 100 µg GnRH was given to the control group at 48 h after the administration of PG as the second GnRH group; and (iii) an administration of 0.75 mg estradiol benzoate (E2) was given to the control group at 24 h after the administration of PG as the E2 group. Each method caused estrus synchronization. Fresh embryos were nonsurgically transferred into the suitable recipients that had a functional corpora lutea (CL) 7 days after estrus. The E2 group showed a significantly higher (P < 0.01) rate of estrus synchronization (98.9%) at 1–3 days after PG administration and the final pregnancy rate of the E2 group (50.6%) was also significantly higher than the other groups (37.1%, P < 0.05 and 30.9%, P < 0.01, respectively). These findings demonstrate that E2 administration 24 h after PG protocol is effective for estrus synchronization of Holstein heifers, thus improving the productivity of ET.     

Increase of ruminal fiber digestion by cellobiose and a twin strain of Saccharomyces cerevisiae live cells in vitro

This experiment was designed to investigate the effects of different concentrations of cellobiose (CB) or a twin strain of Saccharomyces cerevisiae live cells (YST) (20, 40 and 60 mg/60 mL), and CB + YST (60 + 20, 60 + 40, 60 + 60 mg/60 mL) on mixed ruminal microorganism fermentation in vitro. Ruminal fluid was collected from a cow, mixed with phosphate buffer (1:2) and incubated (60 mL) anaerobically at 38°C for 24 h with or without supplement plus 400 mg (dry matter [DM] basis) substrate (hay plus concentrate, 1.5:1). The medium pH numerically decreased with CB and CB + YST, but was unchanged with YST. The total volatile fatty acid and proportion of propionate increased (P < 0.05) in all cases. The proportion of acetate decreased (P < 0.05) with CB and CB + YST, but increased (P < 0.05) with YST and that of butyrate increased (P < 0.05) with CB and CB + YST, but decreased (P < 0.05) with YST. Ammonia‐N decreased (P < 0.05) with CB and CB + YST, but was unchanged with YST. The number of protozoa was unchanged, and that of cellulolytic bacteria increased (P < 0.05) in all cases. Total gas production increased (P < 0.05) in all cases. Methane decreased, hydrogen was unchanged by YST and both gases were unchanged by CB and CB + YST. The in vitro disappearance of DM and neutral detergent fiber increased (P < 0.05) by 11.2% and 8.9%, 9% and 8.5%, and 12.1% and 10.2% in the case of CB, YST and CB + YST, respectively. Therefore, the dietary supplementation of CB and/or YST may improve ruminal fermentation and digestibility.