Changes in myoglobin content and proximate compositions of the dorsal ordinary muscles of full-cycle cultured Pacific bluefin tuna Thunnus orientalis (Temminck et Schlegel) with body size

We investigated the changes in myoglobin (Mb) content and proximate compositions of the cephalad parts of the dorsal ordinary muscles (Ce‐DOM) of full‐cycle cultured (FC) Pacific bluefin tuna [body length: 42.6–115.4 cm, body weight (BW): 1.7–38.4 kg, killed from July 2004 to October 2005]. The Mb content of Ce‐DOM increased (P<0.01) until a BW of 7.0 kg was reached. However, the Mb content of Ce‐DOM had a wide variety above a BW of 7.0 kg. On the other hand, the moisture, protein and ash contents of Ce‐DOM of FC tuna decreased (P<0.05 and 0.01) with an increase in BW. However, the lipid content of Ce‐DOM increased gradually above about a BW of 20.0 kg. Furthermore, the Mb content in protein of the Ce‐DOM of FC tuna increased with an increase in BW. These results indicated that the increase in Mb content of Ce‐DOM of FC tuna was not related to changes in proximate composition between a BW of 1.7 and 38.4 kg.     

Addition of oxytocin to semen extender improves both sperm transport to the oviduct and conception rates in pigs following AI

Oxytocin (OXT) contained in boar semen is known to produce uterine contraction; therefore, we hypothesized that the co‐injection of OXT with sperm would improve artificial insemination (AI) using liquid or frozen‐thawed boar sperm. We initially examined whether OXT added to semen extender improved sperm transport to the oviduct. Although the addition of OXT did not affect the fresh or frozen‐thawed sperm motility or acrosomal integrity, it significantly increased the number of sperm in the oviduct at 6 h after AI injection with OXT, as compared with the control (P < 0.05). Moreover, some sperm were observed in the sperm reservoir of the isthmus in the OXT treatment group, whereas few sperm were observed in the control. When OXT was added to the semen extender immediately prior to AI, the conception rates were significantly higher in both fresh semen and frozen‐thawed semen than in the control group (P < 0.05: liquid, 87.5% vs. 70.5%; frozen‐thawed, 89.8% vs. 75.0%). From these results, we concluded that the addition of OXT to the semen extender assisted in sperm transportation from the uterus to the oviduct, which resulted in improved reproductive performance.     

Protein Requirements of the Freshwater Prawn Macrobrachium rosenbergii Evaluated by the Factorial Method

The dietary protein requirement of the freshwater prawn Macrobrachium rosenbergii to obtain the maximum body protein retention was assessed by the estimated protein needed daily for maintenance (M) and body protein increment (G). The body protein increment was estimated by quantifying body nitrogen increments when the prawn was fed high‐protein diets composed primarily of casein. The protein required for maintenance was estimated by regressing weight gains in feeding experiments using graded dietary levels of casein. The true daily retention (R) or increase in body protein of the prawn corresponds to the sum of M and G. The values of M, G, and R determined in the present study were 3.86, 0.24, and 4.10 g/kg body weight/d, respectively. On the basis of the daily body protein increment and the net protein utilization of casein (58%), dietary protein requirement of the juvenile M. rosenbergii for maximum body protein retention was suggested to be about 7.1 g/kg body weight/d.     

Photoperiod influences the growth, food intake, feed efficiency and digestibility of red sea bream (Pagrus major)

Two consecutive trials were conducted to investigate the effects of photoperiod manipulation on growth rate, food intake and feed conversion efficiency (Trial 1), and the digestibility of nutrients and energy (Trial 2) in red sea bream, Pagrus major (body weight 19–120 g). Fish were exposed to four photoperiods (6L:6D, 12L:12D, 16L:8D and 24L:0D) with light intensity 1500 lx on the water surface. The fish were fed with a commercial diet to apparent satiation. In Trial 2, 0.5% chromic oxide (Cr₂O₃) was used as an inert marker. Significantly higher weight gain and specific growth rates were observed in fish exposed to a 24L:0D photoperiod followed by 16L:8D, 6L:6D and 12L:12D photoperiods (P < 0.05). Food intake and feed conversion efficiency (FCE) were also significantly higher in fish exposed to 24L:0D followed by 16L:8D, 6L:6D and 12L:12D photoperiods (P < 0.05). Fish exposed to 24L:0D and 16L:8D photoperiods showed significantly higher lipid and energy digestibility than those exposed to a 12L:12D photoperiod (P < 0.05). The results demonstrated that the enhancement of growth performances under 16L:8D and 24L:0D photoperiods were attributed to improved appetite, greater food intake and higher feed conversion efficiency as well as higher digestibility.     

Viability of maternally heat-stressed mouse zygotes in vivo and in vitro

Mammalian preimplantation embryos are susceptible to heat stress. This present study examined how maternal heat stress affects the development of mouse zygotes in vivo and in vitro. In Experiment 1, zygotes collected from female mice that were heat‐stressed for 12 h on day 1 of pregnancy were cultured in vitro. Maternally heat‐stressed zygotes developed normally to the two‐cell stage, but the majority of embryos failed to develop into morulae or blastocysts. In Experiment 2, pregnant mice were heat‐stressed on day 1 or from day 1 to day 3 of pregnancy. The number of living fetuses on day 14 of pregnancy was lower in heat‐stressed mice than in non‐stressed mice, but the difference was significant only in successively heat‐stressed mice. These results demonstrate that maternally heat‐stressed zygotes have reduced in vitro viability, but this phenomenon does not necessarily lead to embryo loss in the maternal environment.     

Elevational Patterns of Acid Deposition into a Forest and Nitrogen Saturation on Mt. Oyama, Japan

Virgin fir trees have been dying on Mt. Oyama, which is located in the southwestern part of Kanto Plain, although the frequency of death seems to be reducing recently. We report elevational patterns of acid deposition in precipitation and throughfall under fir and cedar canopies and nitrogen saturation in the forest ecosystem on Mt. Oyama. The deposition fluxes of major inorganic ions in precipitation were nearly constant regardless of elevation except for hydrogen and ammonium ions, whereas the deposition fluxes of all major inorganic ions in throughfall among cedar increased. The 5-year average of annual nitrate deposition in precipitation from 1994 to 1998 showed 19.3 – 23.5 kg ha^?1 yr^?1 (annual inorganic total N deposition: 9.6 – 10.7 kgN ha^?1 yr^?1) at four sites ranging in elevation from 500 to 1252 m, whereas the deposition in both cedar and fir throughfall was over 6 times greater than that in precipitation. The average soil surface nitrate concentration in 1998 was 140 µg g^?1 (the range: 21.1 – 429 µg g^?1, n=80) and the 7-year average of nitrate concentration in stream water from 1992 to 1998 was 4.81 mg L^?1 (the range: 2.38 – 20.6 mg L^?1, n=317). Our results indicate that nitrogen saturation is occurring in the forest ecosystem because of high N deposition, probably via acid fog, on Mt. Oyama.     

Nuclear transfer using a bovine mammary epithelial cell line (BMEC)

The developmental potential of nuclei from a bovine mammary epithelial cell line (BMEC) in nuclear transfer was investigated. For nuclear transfer donors, BMEC cells (passage 15) were cultured for 4–5 days after seeding at cell densities of 1.0 × 10⁵ cells/cm² (high‐density group) or 0.8 × 10⁴ cells/cm² (low‐density group). First, the effective electric stimulation for fusion of enucleated oocytes with BMEC cells was examined. Fusion rates reached maximum with two DC pulses of 30 V/150 µm for 20 µs in the high‐density group and with two DC pulses of 25 V/150 µm for 10 µs in the low‐density group. The fusion rate (37.5%) in the high‐density group was significantly (P < 0.005) lower than in the low‐density group (71.4%). Second, the in vitro developmental potential of nuclear transfer embryos derived from BMEC cells was examined. In the high‐density and low‐density groups, 18.8% and 24.1% of fused oocytes, respectively, developed to blastocyst stage. The results of this study indicate that nuclei from BMEC cells support the development of nuclear transfer embryos to the blastocyst stage and that the efficiency of oocyte–cell fusion is affected by the culture conditions of the donor BEMC cells before nuclear transfer.     

Epidemiology, pathology, and immunohistochemistry of layer hens naturally affected with H5N1 highly pathogenic avian influenza in Japan

Epidemiology, pathology, and immunohistochemistry were investigated in layer hens affected with H5N1 highly pathogenic avian influenza, which occurred for the first time in 79 years in Japan. The farm, which had a total of 34,640 chickens, experienced up to 43.3% mortality before the chickens were depopulated. Clinically, the affected chickens exhibited mortality without apparent clinical signs. Histologically, hepatocytic necrosis; necrosis of ellipsoids and follicles with fibrin in the spleen; necrosis with glial nodules in the brain stem, cerebrum, and cerebellum; necrosis of acinar cells in the pancreas; and necrosis of lymphoid tissues in intestinal lamina propria were seen. Occasionally, mild bronchiolitis, degeneration of smooth muscle fibers in the cecum, and mild tubulonephrosis were noted. Immunohistochemically, influenza virus antigens were detected often in the liver and spleen, heart, intestine, gizzard, proventriculus, and oviduct. In addition, antigens were seen also in the brain, kidney, pancreas, and ovary, but seldom in the lung and trachea. Virus antigen was mainly detected in the capillary endothelium and parenchymal cells. This suggests that virus excretion from the respiratory tract was not as prevalent as that from the digestive tract in the present cases.     

Effects of heat stress on the redox status in the oviduct and early embryonic development in mice

This study examined the association between redox status in the oviduct and early embryonic death in heat-stressed mice. In Experiment 1, non-pregnant mice were heat-stressed at 35 C with 60% relative humidity for 12, 24, or 36 h, and the maternal redox status was verified by measuring the levels of reactive oxygen species (ROS) and free radical scavenging activity (FRSA) in the oviduct, and thiobarbituric acid reactive substances (TBARS) and glutathione peroxidase (GSH-Px) activity in the liver. In Experiment 2, zygotes were collected from mice heat-stressed for 12 h on the day of pregnancy, and their developmental abilities were assessed in vitro, along with the intensity of DNA damage at the 2-cell stage. The TBARS value and GSH-Px activity in the liver, and ROS level in the oviduct were significantly higher in heat-stressed mice, and this increase appeared to depend on the duration of the heat stress. Maternal heat stress significantly reduced the percentage of zygotes that developed to the morula and blastocyst and the total cell number in the blastocyst. In addition, DNA damage at the 2-cell stage was significantly higher in maternally heat-stressed embryos. These results suggest that heat stress induces systemic changes in redox status in the maternal body, and the resultant increase in oxidative stress in the oviduct is possibly involved in heat stress-induced early embryonic death .