Changes in peripheral leukocyte populations of weak calf syndrome of Japanese black calves

To clarify the cellular immune condition in Japanese Black (JB) calves with a weak syndrome, peripheral leukocyte populations were analyzed by flow cytometry. Twenty JB calves were divided into two groups based on clinical observations; one group of calves was weak, because they had experienced an onset of diarrhea within 3 days after birth and needed treatment (Group 1 ;n=10), and the other group of calves was healthy (Group 2; n=10). With regard to leukocyte populations, CD8(+) cells and gamma delta T cells in Group 1 were markedly lower than those in Group 2 during the experimental periods. It is possible that immune-insufficiency might be based on T lymphocyte function in weak syndrome JB calves during the growth process.     

Relationship between diarrhea and peripheral leukocyte population in neonatal Japanese black calves

Neonatal Japanese Black (JB) calves show a high incidence of diarrhea. The objective of this study was to analyze the immune cell populations of neonatal JB calves in detail and examine its correlation with the incidence of diarrhea immediately after birth. Understanding the immune cell populations is helpful in clinics in order to determine the condition of the immune system for prevention of diseases. Blood samples were obtained from JB calves on the day of birth. The peripheral leukocyte populations were analyzed separately for calves that had diarrhea within 2 weeks after birth (diarrhea group; n = 26) and for calves without diarrhea (control group; n = 74). The numbers of the peripheral blood CD3(+)TcR1-N12(+) and CD8(+) T cells were significantly lower in the diarrhea group compared with the control group. These findings suggest that the congenital lower peripheral γδ and CD8(+) T cells results in a high risk of diarrhea in neonatal JB calves.     

Effect of supplemental vitamin E on the peripheral blood leukocyte population in Japanese Black calves

We investigated the effect of supplemental vitamin E on the peripheral blood leukocyte population in Japanese Black calves. Twenty-six calves kept at the same farm were studied. They were divided into two groups; thirteen calves received 300 IU/day of vitamin E orally from 1 to 3 months of age (VE group), and the other thirteen calves did not receive the vitamin E supplement (control group). The VE group showed a higher serum vitamin E concentration at 2 and 3 months of age compared with the control group (P<0.01). The numbers of CD3⁺ cells and CD4⁺ cells were higher in the VE group than in the control group, and the difference was statistically significant at 3 months of age (P<0.05). The numbers of CD21⁺ cells were higher in the VE group than in the control group, and the difference was statistically significant at 2 months of age (P<0.05). The numbers of CD335⁺ cells tended to be higher in the VE group than in the control group. The numbers of CD8⁺ cells and CD14⁺ cells tended to be higher in the VE group than in the control group at 3 and 4 months of age. This study demonstrated that the supplementation of suckling Japanese Black calves with vitamin E might affect the numbers of some immune cell types in the peripheral blood.     

Changes in leukocyte populations in pulmonary lavage fluids of calves after inhalation of Pasteurella haemolytica

The distribution of leukocytes in bovine bronchoalveolar lavage fluids was determined in 15 calves at various times after aerosol exposure to Pasteurella haemolytica. For comparison, 10 calves were exposed to aerosols of phosphate-buffered saline solution; 15 calves, to Staphylococcus epidermidis; and 10 calves, to Salmonella typhimurium endotoxin. At 10 minutes after inhalation exposure for each group, the predominant cell type was the macrophage. Macrophages remained the predominant cell type throughout each lavage interval for calves exposed to phosphate-buffered saline solution and Staph epidermidis. For calves exposed to P haemolytica, there was a decrease in the percentage of macrophages detectable by 30 minutes after exposure, with a corresponding increase in the percentage of neutrophils. Sixty minutes after the inhalation exposure to P haemolytica, the percentages of macrophages and neutrophils in the lavage fluid were equal. By 240 minutes after exposure to P haemolytica, greater than 90% of the cells in the lavage fluids was neutrophils. The increase in the percentage of neutrophils in lavage fluids from calves exposed to S typhimurium endotoxin was similar to that seen for the calves exposed to P haemolytica.     

Peripheral mononuclear cell response in Japanese black calves after oral administration of IFN-alpha

To investigate the effects of oral administration of an interferon (IFN)-alpha drug on the immune reaction of healthy Japanese Black (JB) calves, peripheral leukocyte populations and their ability to produce cytokine mRNA were analyzed after oral administration of IFN-alpha. Fourteen calves fed in one herd were divided into two groups; seven calves were orally administered 0.1 g/day of IFN-alpha from the day of birth to day 5 on each day (group 1, N=7), and the other seven calves were used as the control (group 2, N=7). Blood samples were collected from the jugular veins of all calves before administration and in weeks 1, 2, 4, 8, and 16 after birth. The number of MHC class II+CD14+ monocytes in the leukocytes population of group 1 increased gradually after birth, and significantly higher numbers were detected in week 4 compared with group 2. MHC class II-CD14+ monocytes in group 1 peaked in week 1, and a significant increase was detected compared with group 2. The level of IL-12 in the cytokine mRNA of group 1 increased gradually between weeks 1 and 2, and a significantly higher level of IL-12 was found compared with group 2. These results suggest that oral administration of IFN-alpha induces activation of the monocyte functions in JB calves.     

Comparison of the peripheral blood leukocyte population between Japanese Black and Holstein calves

Japanese black (JB) calves have greater susceptibility to infectious diseases compared to Holstein (Hol) calves. In order to clarify the differences in cellular immune status between JB and Hol calves, the leukocyte population and lymphocyte proliferative ability were analyzed. In total 200 healthy calves, 1 day to 14 weeks of age, were examined: 105 JB and 95 Hol calves. Lower numbers in peripheral blood and percentage in peripheral blood mononuclear cells of CD3⁺TcR1‐N12⁺ T cells and major histocompatibility complex class‐II⁺CD14^‐ B cells were observed in the JB compared to the Hol. The percentage of TcR1‐N12⁺CD25⁺ T cell in the JB was significantly lower than that of the Hol at 4–6, and 8–10 weeks. Interleukin (IL)‐2 sensitivity in the JB was lower than that in the Hol, and significant differences were observed in age groups of 6–8 weeks and 10–14 weeks. These findings indicated that the lower numbers of γδ T cells and B cells in the JB compared to the Hol might be associated with the specificity of the immune systems in JB calves.     

Interaction of GH polymorphism with body weight and endocrine functions in Japanese black calves

We assessed the interaction of GH gene polymorphisms (AA, AB and BB genotypes) with body weight and measures of endocrine function in Japanese black calves at 10 months of age. The average body weight for the BB genotype (281+/-5 kg) was significantly lower (P=0.0017, ANOVA) than those for the AA (324+/-9 kg) and AB (317+/-7 kg) genotypes. Plasma concentrations of insulin and IGF-I were greater for the AA genotype than for the AB genotype, and AB and BB genotypes, respectively. There were significant differences in the triglyceride and cholesterol concentrations among the GH genotypes. The area under the basal GH concentration was significantly greater (P=0.0314) for the AA genotype than for the two other genotypes. The incremental area over the basal GH concentrations in response to intravenous GHRH injection (0.4 microg/kg BW) was significantly smaller (P=0.0005) for the BB genotype than for the two other genotypes. In addition, linear regression analysis between GH incremental area induced by GHRH and body weight demonstrated that there was a positive linear correlation (r=0.6496, P<0.002) for incremental areas less than 600 ng min/ml, but a negative correlation (r=0.6473, P<0.05) for incremental areas over 600 ng min/ml. These findings indicate that the GH genotypes of the animals could be associated with difference in the GH response in Japanese black cattle at 10 months of age. We also observed a relationship between genotype and animal performances, but other studies on more animals in different conditions must be realized to make a definite conclusion.     

Pathological changes of renal tubular dysplasia in Japanese black cattle

Pathological studies were conducted on 91 Japanese Black cattle with a hereditary disease which induced growth retardation, long hooves and renal failure. In calves one to two months old, no gross abnormalities were observed in the kidneys, but microscopical examinations revealed immature epithelia which were arranged irregularly and not attached to the basement membranes in some proximal tubules. In animals three to 36 months old, the kidneys had shrunk perceptibly and had grey-white radial streaks; microscopically they showed severe interstitial fibrosis with round-cell infiltration in the outer zone of the medulla and cortex, and reductions in the numbers of glomeruli and tubules. In the fibrotic areas there were immature epithelia with an irregular arrangement, and the basement membrane of the tubules was thickened. It was concluded that renal tubular dysplasia was the primary lesion of the disease, and that interstitial fibrosis and reductions in the numbers of nephrons were secondary lesions.     

Characteristics of developmental changes in the kinetics of glucose and urea in Japanese Black calves: comparison with Holstein calves

The current study was conducted to clarify the characteristics of glucose and urea kinetics in male Japanese Black calves, using a glucose and urea dilution method with stable isotopes, at preweaning (3 wk of age) and postweaning (13 and 26 wk of age) stages, in comparison with the kinetics of glucose and urea in male Holstein calves. Six Japanese Black and 6 Holstein calves were arranged in a 2 (breed) x 3 (stage) factorial block design. These 12 suckling calves were fed only whole milk, offered concentrate and or chardgrass hay after 3 wk of age, and weaned at 6 wk of age. Under steady-state conditions, glucose challenges (1.0 mg/kg of BW; [U-13C]d-glucose) and urea challenges (both 2.2 mg/kg of BW; [13C]urea and [15N2]urea) were performed at the 3 stages examined. There were no significant differences in plasma glucose concentrations between Japanese Black and Holstein calves at any stage, but the glucose concentrations at 3 wk of age were greater (P <0.05) than those at 13 wk of age in both breeds. The glucose pool size in Japanese Black calves was smaller (P <0.05) than that in Holstein calves at all stages. Within each breed, there were no significant differences between the glucose pool sizes at 3 and 13 wk of age, but the glucose pool size was larger (P <0.05) at 26 wk of age. Moreover, Japanese Black calves had greater glucose irreversible loss rates (P <0.01) than Holstein calves at 3 wk of age, and the glucose irreversible loss rates were less (P <0.05) on and after 13 wk of age in both breeds. Japanese Black calves had greater plasma urea N concentrations (P <0.05) than Holstein calves at all stages, and in both breeds, the urea N concentrations increased (P <0.05) with age. The urea pool size was smaller (P <0.01) in Japanese Black calves than in Holstein calves at all stages, and in both breeds, the urea pool size increased (P <0.05) with age. In comparison with Holstein calves, Japanese Black calves had greater urea irreversible loss rates (P <0.05) on and after 13 wk of age and greater urea recycling rates (P <0.05) at 26 wk of age. In addition, in both breeds, urea irreversible loss rates and urea recycling rates increased (P <0.05) with age. We conclude that Japanese Black calves have partially different glucose and urea kinetics from Holstein calves and that the kinetics of these metabolites in both Japanese Black and Holstein calves are strongly influenced by weaning.     

营养水平对肉犊牛营养物质消化率和血清指标的影响

研究不同能量、蛋白质水平的代乳品对南方地区哺乳期肉用犊牛营养物质消化和血清指标的影响,进而确立代乳品能量和蛋白质参数,为南方地区肉犊牛的饲料配制提供数据支持。采用2×2因子设计,犊牛代乳品的能量水平为16、14 MJ/kg,粗蛋白水平为25%、22%,共计4种。选取新生犊牛40头,21日龄断母乳并根据日龄和体重一致原则分为4个处理组,每组10头犊牛,分别饲喂上述4种代乳品,于77~85日龄进行消化试验并静脉采集血液。结果显示相同能量水平,犊牛对代乳品中粗脂肪和钙的表观消化率随粗蛋白水平的升高而提高(P0.05);较低粗蛋白水平代乳品,随其能量水平的升高,粗蛋白消化率显著提升(P0.05);低能低蛋白组(DE:14 MJ/kg,CP:22%)营养物质消化率略小于其它各组。饲喂低能量水平代乳品时,随其粗蛋白水平的升高,犊牛血清中尿素氮含量显著降低(P0.05);而饲喂高能量水平代乳品时,随其粗蛋白水平的升高,犊牛血清谷草转氨酶含量显著提高(P0.05);代乳品能量和粗蛋白水平对犊牛其它血清指标均无显著影响(P0.05)。高能量水平(DE:16 MJ/kg)有助于犊牛粗蛋白质消化吸收,高蛋白水平(CP:25%)有助于犊牛对粗脂肪和钙的利用,低能低蛋白水平(DE:14 MJ/kg,CP:22%)日粮不利于犊牛营养物质消化吸收。     

Congenital myopathy in Japanese Black calves

Two Japanese Black bull calves from a dam showed muscular weakness and became recumbent after birth. At necropsy, skeletal muscles, including face, neck, body and proximal and distal forelimb and hindlimb were extremely pale in color and edematous. Histopathological examination of skeletal muscles revealed degenerative changes as follows: replacement of muscle with fat, variation in muscle fiber diameter, internal nuclei, central core-like structures, and vacuolar and hyaline degeneration of muscle fibers. Ultrastructurally, the lesions were characterized by focal myofibrillar disorganization with streaming or irregularity of Z bands. The present cases seem to be classified as congenital myopathy based on pathological alterations and age of onset.     

Whole blood leukocyte vs. separated mononuclear cell blastogenesis in calves: time-dependent changes after shipping.

The blastogenic response of peripheral blood mononuclear cells to mitogenic stimulation by concanavalin A was lower (P less than 0.01) after transporting 60 dairy calves 480 km than it was either one or two weeks later. The response was similar for phytohemagglutinin. There was a decrease (P less than 0.05) in the number of peripheral blood monocytes and neutrophils two weeks after shipping. The transportation of calves did not affect plasma IgG1 or IgM level. The mitogenic stimulation of peripheral blood leukocytes by both phytohemagglutinin and concanavalin A in whole blood cultures was more variable than with the culture of peripheral blood mononuclear cells. Technique variation, which was defined as the coefficient of variation among quadruplicate cultures, was greater than 20% for while blood assays and less than 10% for cultures of peripheral blood mononuclear cells. The variation among different calves tested at the same time and the variation within single calves tested at different times were also lower in peripheral blood mononuclear cell cultures than in whole blood mononuclear cell cultures than in whole blood assays. It is suggested that the variation among replicate cultures be reported in blastogenesis studies.     

Effect of aspirin and indomethacin on changes caused by Pateurella multocida toxins in calves

Acetylsalicyclic acid acid (50 mg/kg die) and indomethacin (1.5 mg/kg die) were administered to calves three days prior to intravenous application of endotoxin. Such treatment resulted in delayed occurence of clinical symptoms and higher survival rates, as compared to untreated controls. Parameters relating to coagulation physiology, changed by endotoxin, were not controllable with statistical security by administration of acetylsalicyclic acid indomethacin.     

Effect of blood derivatives with immunomodulators on the immunoprotein profile in calves under clinical conditions

The data on the selected humoral and cellular parameters of the immuno-protein profile were evaluated in calves in a critical period of the availability of immunoglobulins (Ig), in dependence on the administration of the blood derivative with levamisole (group AL) and on the administration of levamisole solution (group L). The preparations were administered intramuscularly at a rate of 10 mg levamisole per 1 kg of live weight. No statistical significance was recorded in the comparison of the humoral parameters after repeated administration (interval of five days) of the Algalev (AL) and levamisole (L) preparations. During the same investigation, the administration of AL produced a significant increase in the T lymphocytes of the blood both in comparison with the T lymphocyte concentration before treatment (P less than 0.05 on the 5th, 7th, 11th, 14th and 20th day of experiment) and in comparison with the calves of group L (P less than 0.05 on the 7th, 11th, 14th, and 20th day); in the case of B lymphocytes an increase was recorded only in comparison with the initial value (P less than 0.01 on the 2nd, 5th and 7th day). Administration of levamisole alone led to a positive change in T lymphocyte concentration (P less than 0.05 on the 5th, 7th, 11th, 15th and 20th day) but not in the concentration of the B cells (P greater than 0.05). In another experiment, administration of the AL preparation (repeatedly after six days) led to a significant increase in the percentage of T lymphocytes (P less than 0.01 the 8th day and P less than 0.05 the 2nd, 6th, 15th, 21st and 35th day) and B lymphocytes (P less than 0.01 the 15th and 21st day), in comparison with their levels before administration as well as in comparison with the control (T lymph.: P less than 0.01 the 6th day and P les than 0.05 the 8th, 15th, 21st and 35th day; B lymph.: P less than 0.05 the 15th day). The maximum percent proportion of T lymphocytes after administration of the AL preparation was recorded on the seventh and eighth day after the first administration.     

Effect of vaccination on cell populations in lung washes from calves after infection with respiratory syncytial virus

The inflammatory response in the air-passages of the lungs of calves after intranasal inoculation with respiratory syncytial virus (RSV) was compared in RSV-vaccinated and control animals. Total cells recovered from lung washings remained the same; however, the fold by eight days after infection and the type of cells changed from a predominance (85 per cent) of macrophages to equal proportions of macrophages and neutrophils (45 per cent) during the course of infection. The absolute numbers of neutrophils rose by 15-fold. In contrast, when RSV-vaccinated calves were challenged, the total number of cells recovered from lung washings remained the same; however, the numbers of macrophages decreased and the numbers of neutrophils increased by fivefold. Cytological studies of the lung washings revealed no evidence of an exacerbated inflammatory response in RSV-vaccinated calves. Levels of virus replication were significantly reduced in RSV-vaccinated compared with control animals.     

Idiopathic thrombocytopenia in Japanese black cattle

A female Japanese Black cow showed hemorrhage under the mucous membrane of the eye conjunctiva and the vagina, and thrombocytopenia in peripheral blood. The clinical manifestations repeated spontaneously during long-term observation. The clinical findings were stable except for severe periodic bleeding. Prothrombin time, activated partial thromboplastin time and fibrinogen levels were normal. The anti-platelet antibody in the serum was analyzed by indirect platelet coagulation tests. Because the platelets in the micro-plate wells of the subject series showed coagulation up to 1:128, it was judged that anti-platelet antibody was present in the serum. A number of megakaryocytes were recognized in smear samples obtained from bone marrow of the scapula immediately after euthanasia. Therefore, this case was diagnosed as idiopathic thrombocytopenia.     

Effects of peripheral blood polymorphonuclear leukocyte function and blood components in Japanese black steers administered ACTH in a cold environment.

An examination of the effects of artificial stress induced by adrenocorticotropin (ACTH) on total and differential leukocyte counts, plasma cortisol levels, metabolic profiles and peripheral blood polymorphonuclear leukocyte (PMN) function was performed on Japanese Black steers kept in a cold environment, with the following regimes; 1) -5 degrees C x ACTH (100 IU/day for 3 days), 2) 0 degrees C x ACTH, 3) 15 degrees C x ACTH and 4) 15 degrees C x PBS. Blood samples were collected before and at 1, 2, 24, 48, 72 and 96 hr prior to the application of the stressor. The plasma cortisol level was found to greatly increase at one hr after the first treatment of ACTH, particularly so in animals exposed to -5 degrees C. Total leukocytes (-5 degrees C and 0 degrees C experiments, respectively), the monocytes (-5 degrees C), neutrophils and eosinophils (-5 degrees C, 0 degrees C and 15 degrees C, respectively) obviously increased just after the first administration, although lymphocyte counts at -5 degrees C were inversely related to those described above. All of these tendencies were augmented by the cold environment except for eosinophils. The chemiluminescent (CL) response of PMN decreased in the ACTH-administered steers at an early stage of post-administration, however, it tended to recover from the lower-than-base value in the cold-affected steers. ACTH administration resulted in higher plasma glucose (Glu) compared to a control, although only steers housed at -5 degrees C evidently showed lower plasma inorganic phosphorus (IP). No abnormal serum acute phase protein, or immunosuppressor, was noted. ACTH thus appears not only to promote physiological reactions but also to temporarily suppress PMN cellular immune function in Japanese Black steers. Although, a cold environment rapidly restored the CL activity to over the pre-administrational value, suggesting that a vital response was activated by crymo-stimuli.