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.     

Effect of modification of ovary preservation solution by adding glucose on the maturation and development of pig oocytes after prolonged storage

Oocytes lose their developmental competence during prolonged storage of the ovary. In the present study, we supplemented the preservation solution for pig ovaries (phosphate buffered saline, PBS) with glucose and preserved the ovaries for 6 h at 25 C. Subsequently, we examined the glucose concentration of the follicular fluid (FF), pH of the FF, survival rate of the granulosa cells, and maturation and developmental competence of oocytes after storage. During storage, the glucose concentration of the FF (2.1 mM), pH of the FF (7.4), and survival rate of the granulosa cells (69.5%) rapidly decreased (glucose concentration: under 1.1 mM; pH: 6.8; and survival rate: 43%). On the other hand, when the preservation solution was supplemented with glucose (15 mM), the glucose concentration of the FF increased and the survival rate of the granulosa cells improved, although the pH of the FF decreased further (from 6.8 to 6.6). In addition, supplementation with glucose significantly improved the rates of oocytes at metaphase II (0 h: 65.0%; 6 h without glucose: 23.8%; and 6 h with glucose: 43.8%) and attenuated the decline in the rates of fertilization and development that resulted from prolonged storage, although there were no significant differences. In conclusion, modification of the preservation solution by the addition of glucose increased the glucose concentration of the FF and improved the rate of maturation of pig oocytes.     

Induction of estrus by prostaglandin F2alpha administration in the vaginal vestibules of miniature pigs

Estrus induction is an important step in embryo production. It has been difficult to induce estrus in miniature pigs by intramascular (i.m.) injection of prostaglandin F2alpha (PGF2(2alpha)) in the early luteal stage of the estrous cycle. In the present study, we injected two different doses of PGF2(2alpha) i.m. and into the submucosa of the vaginal vestibule (i.ves.) of miniature pigs, and examined the effect of these treatments on estrus induction. Fifteen miniature pigs were divided into five experimental groups (control, saline injected i.m.; PGF2alpha treated, 1.0 or 1.5 mg of PGF2alpha injected twice i.m. or i.ves.), and the estrus length and concentrations of 17beta-estradiol (E2) and progesterone (P) in the blood were examined. Estrus length was significantly shortened by a large amount of PGF2alpha injected i.ves. In addition, the concentration of P in the blood significantly decreased after two injections of PGF2alpha (i.m. or i.ves.). These results suggest that in miniature pigs, administration of at least 3.0 mg of PGF2alpha is required for the induction of luteolysis and injection of PGF2alpha into the vaginal vestibule is a useful method of estrus induction.     

Induction of feline immunodeficiency virus from a chronically infected feline T-lymphocyte cell line

The infection of the feline T-lymphocyte cell line FeT-J with the feline immunodeficiency virus (FIV) Petaluma strain led to the establishment of nonvirus-producing cells. One clone (C15) obtained by limiting dilution was found to express FIV in response to chemical inducers of retroviruses. The chemical treatment of C15 cells led to not only FIV protein synthesis but also an augmentation of viral production. Examination of the C15 cell derivatives obtained by recloning revealed that 10-40% of treated cells constitutively expressed FIV antigens, whereas 100% with expressed FIV antigen in response to the inducer. Chemical induction resulted in more than a 100-fold increase in infectious viral production. The results suggest that a majority of FeT-J cells that are infected with FIV exist in a non-productive state. Establishing a cell line that can be non-productively infected by FIV may help determine the mechanisms of FIV latency.     

Effect of Maternal Age on the Ratio of Cleavage and Mitochondrial DNA Copy Number in Early Developmental Stage Bovine Embryos

Age-associated deterioration in both the quality and quantity of mitochondria occurs in older women. The main aim of this study was to examine the effect of age on mitochondrial DNA copy number (mtDNA number) in early developmental stage bovine embryos as well as the dynamics of mtDNA number during early embryo development. Real-time PCR was used to determine mtDNA number. In vitro-produced embryos 48 h after insemination derived from Japanese black cows, ranging in age from 25 to 209 months were categorized based on their cleavage status. There was an overall negative relationship between the age of the cow and cleavage status, to the extent that the ratio of embryos cleaved over the 4-cell stage was greater in younger cows. The mtDNA number did not differ among the cleaved status of embryos. In the next experiment, oocytes collected from each donor cow were divided into 2 groups containing 10 oocytes each, in order to compare the mtDNA number of mature oocytes and early developmental stage embryos within individuals. Upon comparing the mtDNA number between oocytes at the M2 stage and early developmental stage 48 h post insemination, mtDNA number was found to decrease in most cows, but was found to increase in some cows. In conclusion, age affects the cleaving ability of oocytes, and very old cows (> 180 months) tend to have lower mtDNA numbers in their oocytes. The change in mtDNA number during early development varied among individual cows, although overall, it showed a tendency to decrease.     

Extensive study on forest runoff water chemistry over east Asia

The chemistry of runoff waters from 13 forested watersheds in six regions (four regions from Japan, one from southern China and one from northern Thailand) was evaluated. The Cl⁻ concentrations in runoff waters were higher in those watersheds which had closed canopies and were nearer to the ocean. The NO₃ ⁻ concentrations were higher in those watersheds having the developed soils and high moisture conditions, but were lower in tropical and subtropical regions for those watersheds which had high rates of nitrogen uptake and for watersheds with large areas of saturated soils. The SO₄ ²⁻ concentrations were affected by SO₄ ²⁻ adsorption properties of the soils: at Shibecha, Jiulianshan, and Chiang Mai with high adsorption capacities SO₄ ²⁻ concentrations in streams were low. High SO₄ ²⁻ concentrations were found at Mt. Hiei and Kagawa due to the weathering of sulfur minerals and high levels of atmospheric sulfur deposition. Within regions SO₄ ²⁻ concentrations were inversely related to NO₃ ⁻ concentrations. A comparison among the watersheds suggested H⁺ consumption in deeper soil that increased pH and HCO₃ ⁻ concentrations of the runoff waters of some watersheds. Anion concentrations increased with low H.I. (Humidity Index) values suggesting that dry conditions increased concentrations due to high rates of evapotranspiration.     

Reduced sensitivity to fenarimol in Japanese field strains of Venturia nashicola

Monoconidial strains of Venturia nashicola Tanaka et Yamamoto were isolated from Japanese or Chinese white pear trees which had never been treated with sterol demethylation inhibitors (DMIs) and their baseline sensitivities to fenarimol were determined by mycelial growth tests on fungicide-amended culture media. Strains were also obtained from Japanese pear orchards, which had been intensively treated with DMIs for several years and monitored for the shifts of fenarimol sensitivity in comparison with the baseline sensitivity. Results suggested slight shifts to lower fenarimol sensitivity in strains isolated from DMI-treated Japanese pear orchards. However, in inoculation tests on pear seedlings, fenarimol still provided adequate control of V. nashicola strains with reduced sensitivity to fenarimol in vitro, suggesting that the performance of this fungicide will still be maintained in the field. © 1998 Society of Chemical Industry     

Milking performance evaluation and factors affecting milking claw vacuum levels with flow simulator

Milking performance of milking machines that matches the production capability of dairy cows is important in reducing the risk of mastitis, particularly in high‐producing cows. This study used a simulated milking device to examine the milking performance of the milking system of 73 dairy farms and to analyze the factors affecting claw vacuum. Mean claw vacuum and range of fluctuation of claw vacuum (claw vacuum range) were measured at three different flow rates: 5.7, 7.6 and 8.7 kg/min. At the highest flow rate, only 16 farms (21.9%) met both standards of mean claw vacuum ≥35 kPa and claw vacuum range ≤ 7 kPa, showing that milking systems currently have poor milking performance. The factors affecting mean claw vacuum were claw type, milk‐meter and vacuum shut‐off device; the factor affecting claw vacuum range was claw type. Examination of the milking performance of the milking system using a simulated milking device allows an examination of the performance that can cope with high producing cows, indicating the possibility of reducing the risk of mastitis caused by inappropriate claw vacuum.     

Bacillary hemoglobinuria in Japanese black cattle in Hiroshima, Japan: a case study

Three Japanese Black cows housed with 6 other cows exhibited main clinical symptoms of severe hemoglobinuria. Hematological analyses conducted after antibiotic therapy demonstrated severe anemia, and biochemical analyses indicated both severe hemolysis and disruption of hepatic function. One of the three cows died. Based on the above analyses and observation of typical clinical symptoms, a speculative diagnosis of bacillary hemoglobinuria was made, and immediate high-dose antibiotic treatment improved the general conditions of the surviving animals. Blood samples from the other 2 cows were collected sequentially after antibacterial therapy. Clostridium haemolyticum was detected by a nested polymerase chain reaction analysis of the blood samples. The cows were diagnosed with the second recorded occurrence of bacillary hemoglobinuria in Japan.