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.

Effect of supplemental vitamin E on antibody titer in Japanese black calves vaccinated against bovine herpesvirus-1.

We investigated the effect of supplemental vitamin E on antibody titer against bovine herpesvirus-1 (BHV-1) in Japanese Black calves after vaccination with modified live virus. Thirty calves kept at the same farm were studied. They were divided into two groups; fifteen calves received 300 IU/day of vitamin E orally from 1 to 3 months of age (VE Group), and the other fifteen calves did not receive vitamin E supplement (Control Group). BHV-1 modified live vaccine was injected twice to all calves when they were 2 and 3 months of age. Following the vaccination, serum vitamin E concentration and neutralizing antibody titer to BHV-1 were measured over time. VE Group showed higher serum vitamin E at 2, 3 and 4 months of age compared to Control Group (P<0.05). The antibody titer in Control Group was the highest at 1 month of age, and it gradually decreased until 4 months of age. VE Group showed increase in antibody titer at 4 months of age resulting in significant difference (P<0.01) from Control Group. This study demonstrated that vitamin E supplementation to Japanese Black calves could increase antibody production after the second modified live BHV-1 vaccination.

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.

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.

Effect of parity on lymphocytes in peripheral blood and colostrum of healthy Holstein dairy cows.

Investigation of the bovine systemic and mammary gland immune cells at calving might provide crucial information about the susceptibility of the mammary gland to infection. This study investigated the leukocyte population and cytokine mRNA levels in peripheral blood mononuclear cells (PBMCs) and colostrum mononuclear cells (CCs) obtained from healthy cows soon after calving. Fifty dairy cows that did not show clinical diseases were divided into 4 groups on the basis of parity: heifer (group 1, n = 10), 2nd calving (group 2, n = 11), 3rd calving (group 3, n = 14), and more than 3rd calving (group 4, n = 15). In the peripheral blood the numbers of CD3(+)TcR1-N12(+), CD3(+), CD4(+), and major histocompatibility complex class II(+)CD14(-) lymphocytes were significantly higher in group 1 than in group 4, whereas in the colostrum the percentages of CD4(+) and CD4(+)CD26(+) lymphocytes and the CD4(+)/CD8(+) ratio were significantly lower in group 1 than in group 4. There were no significant differences in the cytokine mRNA levels of PBMCs among the 4 groups; however, in the CCs the ratio of interferon gamma to interleukin 4 was significantly lower in group 1 than in group 3. These results suggest that the cellular immune function of PBMCs is lower, whereas mammary gland immune cells are more active, in cows with higher parity compared with heifers at calving.

Age-related alterations in peripheral leukocyte population of healthy Holstein dairy cows during the pre-calving period.

Older cows show a high incidence of infectious diseases during the periparturient period. The periparturient infectious diseases are closely associated with the immune function of dairy cows during the pre-calving period. In order to evaluate the relationship between the immune cell population and age in the cows during the pre-calving period, we obtained blood samples from 170 dairy cows during the pre-calving period. We chose only healthy cows, which did not develop clinical diseases within 2 weeks after the calving in this study. The animals were divided into 4 groups based on their parity: in their 1st pregnancy (Group 1), in their 2nd pregnancy (Group 2), in their 3rd calving (Group 3) and in more than 3rd pregnancy (Group 4). The numbers of the peripheral blood CD3(+)TcR1-N12(+) and MHC class-II(+)CD14(-) lymphocytes were significantly higher in Group 1 compared to Group 4. This result indicated that the lower gammadeltaT cells and B cells in older cows compared with heifer during pre-calving period.

Effect of body condition score of the dairy cow on the in vitro immune response of peripheral blood mononuclear cells to progesterone stimulation.

Immune function in cows is closely associated with their physical and hormonal conditions. In order to clarify the relationship between the body condition score (BCS) of lactating dairy cows and the immune response to progesterone (P(4)) in vitro, we examined whether lower BCS in dairy cows affects the responsiveness of peripheral blood mononuclear cells (PBMCs) to P(4) added in to culture medium. Forty-two non-pregnant healthy Holstein dairy cows were examined at 61 to 120 days after calving. The cows were divided into the following two groups; Low BCS group (N=20), which had a BCS of less than 2.25, and a Control group (N=22), which had a BCS over 2.75. PBMCs were stimulated with P(4) (1 microg/mL) and/or phytohemagglutinin (PHA), and the levels of cytokine mRNA were analyzed. In the Low BCS group, a significantly lower IFN-gamma level was stimulated by PHA only compared with the Control group. The combination of P(4) and PHA significant decreased the IFN-gamma/IL-4 ratio in the Control group, but this reaction was not found in the Low BCS group. Our data indicated that expression of IFN-gamma mRNA was basically lower in the low BCS dairy cows and that addition of P(4) did not suppress the cellular immune function in these cows. In this study, we observed that P(4) reduced the cellular immune response in the adequate BCS cows, whereas immunosuppression by P(4) was not found in the PBMCs of the low BCS cows, which already had a lower level of immune function.

Effect of lidocaine hydrochloride on the function of bovine peripheral leukocytes.

To clarify the effect of lidocaine hydrochloride (Lid) on bovine peripheral granulocyte phagocytosis, adhesion molecule expression of leukocytes and peripheral blood mononuclear cell mRNA expression of cytokines were investigated. Lid was added to blood samples at a final concentration of 0 (only PBS; Cont), 0.2 mg/ml or 2.0 mg/ml. Phagocytosis of granulocytes was significantly decreased by addition of 2.0 mg/ml of Lid. CD18 expression of granulocytes and mononuclear cells were significantly reduced by addition of 2.0 mg/ml of Lid. IL-1beta and IL-8 mRNA expressions of mononuclear cells were also significantly reduced by addition of 2.0 mg/ml of Lid other hand. These results suggest that Lid might reduce the protective immunity of cows. On the other hand, reduction of CD18, IL-1beta and IL-8 mRNA expression also indicates that Lid has an anti-inflammatory effect in cows.

Peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis and effect of hoof trimming with antibiotic treatment.

In the present study, 30 cows were used to evaluate the changes in the peripheral blood leukocyte subpopulation of dairy cows with digital dermatitis (DD) following hoof trimming and antibiotic treatment. The cows were divided into two groups; 18 cows (DD group) had DD on both hind feet, and 12 cows (control group) had four feet with no clinical abnormalities. The DD group was further divided into two groups based on the treatment; the antibiotic group (8 cows) was treated with only 2% lincomycin liquid spray once daily for 3 days, and the trimmed group (10 cows) received trimming of hooves as well as treatment with 2% lincomycin liquid spray. The plasma cortisol concentration was significantly higher in both DD groups before treatment than in the control group, and it decreased significantly after hoof trimming in the trimmed group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in both DD groups before treatment was significantly lower than that of the control group. The number of CD3(+), CD4(+), WC1(+) and CD21(+) cells in the trimmed group increased after treatment. These results indicated that cows with DD suffer from stress and reduced number of T and B cells. Treatment of DD with both hoof trimming and 2% lincomycin liquid spray was effective for reducing the stress and bringing the immune cell number back to the normal range.

Effects of active egg white product/ Clostridium butyricum Miyairi 588 additive on peripheral leukocyte populations in periparturient dairy cows.

The leukocyte populations of periparturient dairy cows were analyzed after administration of active egg white/Clostridium butyricum Miyairi additive. Sixty-eight Holstein milking cows were divided into 3 groups. Group A was administered active egg white product (AEWP)/Clostridium butyricum Miyairi 588 (Miyairi 588) additive (n=23). Group B was administered Miyairi 588 only (n=23), and Group C was the control group (n=22). The challenged groups were administered 100 g of AEWP + Miyairi 588, or Miyairi 588 alone, daily for 60 days from 1 month before until 1 month after paturition. Blood samples were collected from all groups three times (1 month before, 1 week after and 1 month after parturition) for analysis of the peripheral leukocyte population. The results showed significantly higher numbers of CD4+ cells in Group A compared with Group C 1 week after paturition. AEWP/Miyairi 588 additive may enhance the number of CD4+ T cells in periparturient dairy cows.

Relationships between the conception rate of estrus synchronization using estradiol benzoate and CIDR (progesterone) and other parameters in holstein lactating dairy cows.

The purpose of this study was to determine the relationships between conception rate and other parameters before estrus synchronization with a Controlled Internal Drug Release Device (CIDR) and estradiol benzoate (EB). In the estrus synchronization program, animals were injected with 2 mg EB and then received a CIDR. Seven days later, the CIDR was removed and the animals were given an injection of Prostaglandin F(2alpha). Twenty-four hours later, they received an injection of 1 mg EB, and they were artificially inseminated 24 h after that. This program was applied to 258 Holstein cows in Tohoku-machi (Aomori, Japan). Blood was collected at the beginning of the program, and the conception rate was determined about 40 days after insemination. The relationships among conception rate, blood biochemical values, age, body condition score and days in milk were statistically analyzed to determine better conditions for cow conception. The conception rate of the cows in the high progesterone group (more than 1 ng/ml, P(4)+) was significantly higher than that of the low progesterone group (less than 1 ng/ml, P(4)-; 47.9% vs. 28.6% P<0.01). In the P(4)- groups, the serum phospholipid level was significantly higher in the conception group than in the non-conception group, and the same tendency was seen in the P(4)+ groups. Blood urea nitrogen (BUN), albumin (Alb), and total cholesterol (TChol) were significantly higher in the conception group compared with the non-conception group, but no with P(4) was observed. We concluded that 1) the conception rate of the P(4)- group was remarkably low, that 2) the low conception rate and low P(4) level was related to a low PL level and that 3) BUN, Alb and TChol were higher in the conception group, although no relation with P(4) was found.

Changes in peripheral blood mononuclear cells after calving in lactating cows with serious decrease of body weight before and after calving.

The aim of this study was to clarify the hormonal and immunological changes in periparturient dairy cows with excessive weight loss following calving. We analyzed leukocyte populations and the peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, IL-4, growth hormone (GH)-receptor (R) and insulin -R and measured the GH and insulin concentrations. Ten dairy cows fed in one herd were used in this study and were divided into two groups. Five cows that experienced loss of body weight following calving of over 10% comprised the Weight Loss Group, and the remaining five cows, which experienced loss of body weight of 5% or less following calving, served as the Control Group. The Weight Loss Group had high serum cholesterol and low blood urea nitrogen levels compared with the Control Group throughout the period of observation. In regard to the leukocyte populations, there were significantly fewer CD4(+)CD45R-T cells in the Weight Loss Group 1 to 2 months after calving compared with the Control Group. The percentage of IFN-gamma/IL-4 among PBMCs in the Weight Loss Group tended to be lower than in the Control Group throughout the observation period. In the Weight Loss Group, the levels of GH-R during month 2 were lower than in the Control Group. We concluded that excessive loss of body weight in dairy cows following calving is associated with depressed cellular immune function.

Effects of mycotoxins on mitogen-stimulated proliferation of bovine peripheral blood mononuclear cells.

The effects of mycotoxins on mitogen-stimulated proliferation of bovine peripheral blood mononuclear cells (PBMCs) were investigated. Aflatoxin B(1) (AFB(1)), deoxynivalenol (DON) and zearalenone (ZEN) were added to cultures of PBMCs, and the proliferation responses were measured using MTT bioassays. Suppression of the proliferation of calf PBMCs by AFB(1) and DON was significantly stronger than that of cow PBMCs, whereas there were no differences in suppressive effects on PBMCs from Holstein and Japanese Black calves and cows. The suppressive effect was greatest in the order of DON, AFB(1) and ZEN, and the effects of DON and AFB(1) seemed to be dose-dependent. The results suggest that some mycotoxins directly suppress proliferation of bovine PBMCs.

Effect of hoof trimming before the dry period on productive performance in perinatal dairy cows.

To investigate the effect of hoof trimming before the dry period, the hooves of 10 cows (trimmed group) were trimmed at 79.6 +/- 8.6 days before parturition and the hooves of 52 cows were left untrimmed (control group). Blood biochemistry and hormone concentrations were investigated for 6 cows from each group. The daily milk yield after parturition in the trimmed group tended to be higher than that of the control group. Between 0 and 30 days after parturition, the levels of non-esterified fatty acids were significantly lower stet, and the plasma glucose and glucose disposal rates were significantly higher in the trimmed group. The plasma cortisol concentration was stable before and after parturition in the trimmed group. Hoof trimming before the dry period appears to reduce stress and maintain the nutritional conditions of perinatal dairy cows.

Comparison of two different nutritive conditions against the changes in peripheral blood mononuclear cells of periparturient dairy cows.

To clarify the relationship between cellular immune status and nutritive condition in periparturient dairy cows, feeding content, blood profiles, and immune condition were observed in cows from two dairy herds with different types of feed content. Immunological analyses such as leukocyte population and peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, TNF-alpha, IL-4, and IL-10, quantified by real-time RT-PCR were performed. With regard to feed content during dry periods, there were six cows in the herd with insufficient non-structural carbohydrate (NFC) intake (group I) and six cows in the herd with sufficient NFC intake (group II). Significantly lower levels of blood glucose were observed in group I between weeks -12 and 16 compared with group II. Serum cholesterol level was significantly lower in group I between weeks 2 and 10 than in group II. The numbers of CD3+ and CD4+ T cells in group I were significantly lower than those in group II in weeks 6 and 14. The numbers of CD21+ B cells were significantly lower in group I than in group II in weeks -16, -12, 2, and 10. On the other hand, the CD4+/CD8+ ratio in group II was significantly higher than group I between weeks 2 and 14. The IFNgamma/IL-4 mRNA rate in group I was significantly lower than group II in week 6. We concluded that cellular immune depression occurrs after calving in dairy cows with low nutritional status in the periparturient period.

Comparison of the insulin reaction of peripheral blood T cells between healthy Holstein dairy cows and JB during the periparturient period.

To compare the changes in the insulin reaction of Holstein dairy cows and Japanese Black cows (JB) during the periparturient period, the insulin resistance test in vivo and lymphocytes proliferation with insulin in vitro were performed. Ten healthy Holstein dairy cows (Holstein group) and 10 healthy JB cows (JB group) used in this study were observed on days 60, 40, and 20 before calving and days 7 and 20 after calving. In insulin resistance reaction in vivo and in vitro, a low insulin-stimulated glucose disposal rate and lymphocyte proliferation with insulin were observed in the Holstein group compared with the JB group during the experimental period. An analysis of the lymphocytes cultured with insulin showed that the percentage of CD4+CD45R- T cells in the Holstein group was significantly lower than that of the JB group before day 20. These findings indicate that T cells reaction to insulin in healthy periparturient Holstein cows is lower than that in Japanese Black.

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.

Evaluation by weight change rate of dairy herd condition.

The purpose of this study was to clarify whether weight change rate (WCR) is useful in evaluating herd health. Five herds (A, B, C, D, and E) were examined monthly for approximately 1 year to measure both WCR and BCS. WCR was calculated using the following equation: WCR (%) = (body weight of current month-body weight of the preceding month)/body weight of current month x 100. In addition, blood profiles were performed in each herd before and after calving. The incidence of periparturient diseases was higher in Herds A, B, and C compared with Herds D and E. Reproductive performance in Herds A and B was rather poor compared with Herds C, D, and E. There were significantly lower WCRs in Herd A compared with Herd C during the period of day -30 to day 30 after calving. According to the blood profiles, high levels of non-essential fatty acids (NEFA) were observed during the period of day -30 to -1 and high beta-hidroxybutyricacid (BHB) and low levels of glucose were observed at day 30 after calving in Herd A. A marked poor condition in Herd A during the peripaturient period was clearly expressed by low WCR in this experiment. WCR can express body condition of herds and is suggested to be a useful parameter in the determination of herd performance in field experiments.

Relationship between leukocyte population and nutritive conditions in dairy herds with frequently appearing mastitis.

To clarify the relationship between cellular immune status and nutritive condition, feeding program, blood profiles, and leukocyte populations were analyzed in two dairy herds experiencing frequent mastitis. Fourteen of the 35 lactating cows in herd A, and 18 of the 50 lactating cows in herd B scored positive on the California Mastitis Test (CMT), and 3 of the 73 lactating cows were CMT positive in herd C, which was the control. All herds were evaluated during five different milking stages, and blood was collected from five cows at each stage. With regard to feed content, the percentages of total digestible nutrients (TDN) and crude protein (CP) were found to be lower in herds A and B than in herd C. Levels of serum total cholesterol and blood urea nitrogen were lower in herds A and B than those in herd C. Neutrophil counts in herds A and B were increased compared to the neutrophil counts in herd C. On the other hand, the numbers of CD3(+) T cells and CD14-MHC class(+) cells were lower in herd A and B than in herd C. A decrease in peripheral lymphocytes and undernourishment were observed in the herds with frequent occurring mastitis.