Sister chromatid exchanges in calves with hereditary zinc deficiency (lethal trait A 46)

Frequencies of sister chromatid exchanges (SCEs) in cultured blood lymphocytes are rather lower than higher in calves with hereditary zinc deficiency (lethal trait A 46) than in healthy, normal cows.     

Sister chromatid exchanges in calves with hereditary zinc deficiency (lethal trait A 46)

Summary

Frequencies of sister chromatid exchanges (SCEs) in cultured blood lymphocytes are rather lower than higher in calves with hereditary zinc deficiency (lethal trait A 46) than in healthy, normal cows.     

Postnatal changes in lymphocyte function of dairy calves

Lymphocyte blastogenic response, B-lymphocyte population and antibody-producing activity of lymphocytes were determined to evaluate the lymphocyte function in neonatal calves during the first 4 weeks of life. The mean percentage of B-lymphocytes ranged from 10.2 to 12.5% during the first 14 days of life and from 15.3 to 17.5% in calves from the day 21 to day 28 after birth. The absolute number of B-lymphocytes increased significantly (P less than 0.05) from 370/microliters at birth to 736/microliters on day 28 after birth. The mean stimulation index of blastogenic response, measured by fluorometric assay, ranged from 5.75 to 6.61 with Con A, from 5.29 to 5.98 with PHA and from 1.89 to 2.50 with PWM. The mean (+/- S.D.) number of plaque forming cells ranged from 22.0 (+/- 12.0) to 24.7 (+/- 9.2) in cultured lymphocytes from 5 calves at birth to 14 days after birth and their levels increased markedly from 137.8 (+/- 88.3) to 162.0 (+/- 57.8) in lymphocytes from 20 days to 28 days after birth. The present study showed that antibody-producing activity of lymphocytes is lower in calves within 3 weeks after birth compared to that of calves 3 weeks after birth, indicating that neonatal calf lymphocytes have a low antibody-producing activity at least up to 1 month after birth.     

Vaccination of calves with a modified bacterin or oil-in-water emulsion containing alkali-detoxified Salmonella typhimurium lipopolysaccharide

Twenty-six clinically normal colostrum-fed dairy calves were allotted to 5 groups. Calves of groups 1 and 2 served as nonvaccinated controls and were challenge-exposed with variable numbers of organisms. Group-3 calves were vaccinated SC with a modified Salmonella typhimurium bacterin. The bacterin was composed of killed acid-hydrolyzed S typhimurium G30/C21 (Re-mutant) whole cells coated with alkali-hydrolyzed S typhimurium LT-2 lipopolysaccharide, as antigen, and monophosphoryl lipid A, as adjuvant. Calves of groups 4 and 5 were vaccinated with a 2% mineral oil-in-water emulsion containing lipopolysaccharide as antigen and monophosphoryl lipid A and trehalose 6-6'-dimycolate as adjuvants. Calves of groups 3-5 were vaccinated at 2 weeks of age and again at 4 or 6 weeks of age. Adverse reactions were not observed after vaccination. Calves were challenge-exposed orally at 6 or 8 weeks of age with 1.5 X 10(11) (groups 1 and 4), or 3.0 X 10(11) (groups 2, 3, and 5) colony-forming units of S typhimurium UCD 108-11. Mortality after challenge exposure was 2 of 5 group-1 calves; 4 of 5 group-2 calves; 5 of 6 group-3 calves; 1 of 5 group-4 calves; and 4 of 5 group-5 calves. Statistical difference between calves of similarly challenge-exposed groups was not evident, indicating failure of either vaccine to protect calves of this age from oral challenge exposure with virulent S typhimurium.     

锌中毒对雏鸡外周血T-淋巴细胞的影响

200只1日龄艾维菌肉鸡随机分为4组，即对照组（每千克日粮含锌100mg）、锌中毒Ⅰ组（每千克日粮含锌1500mg）、锌中毒Ⅱ组（每千克日粮含锌2000mg）、锌中毒Ⅲ组（每千克日粮含锌2500mg），每组均饲喂日粮7周。以流式细胞术和酸性α-醋酸萘酯醇（ANAE）染色法观测外周血T-淋巴细胞的动态变化。结果，3个锌中毒组7周龄时，外周血T-淋巴细胞ANAE阳性率显著低于对照组（P〈0．05或P〈0．01），同时3个锌中毒组间比较也呈显著差异（P〈0．05或P〈0．01）。与对照组比较，锌中毒Ⅲ组2至7周龄和锌中毒Ⅱ组6、7周龄CD4^＋T淋巴细胞数量减少，锌中毒Ⅱ组、Ⅲ组2至4周龄和锌中毒Ⅰ组、Ⅲ组7周龄CD8^＋T淋巴细胞数量降低，CD4^＋／CD8^＋比值3个锌中毒组2、4周龄升高，6、7周龄降低。由此表明，锌中毒可抑制T-淋巴细胞的生成，降低其在外周血中的数量。     

Influence of isoprinosine on bovine herpesvirus type-1 infection in cattle

A study was conducted to determine the in vivo efficacy of isoprinosine (ISO) in calves infected with bovine herpesvirus type-1 (BHV-1). Calves were infected with BHV-1 on day 0 and received ISO daily for 14 days. Clinical signs of disease, shedding of BHV-1, lymphocyte proliferative responses to mitogens, interleukin-2 production, and alveolar macrophage bactericidal activity were monitored during the study. Rectal temperatures were increased (P less than 0.05) in BHV-1 and ISO-BHV-1 calves at days 3 to 7 postinfection (PI). Isoprinosine did not influence BHV-1 shedding in calves. Lymphocyte proliferative responses to phytohemagglutinin (PHA) were lower (P less than 0.01) in BHV-1 calves when compared to control or ISO calves at day 4 PI, but ISO did not ameliorate this effect. Interleukin-2 activity was greater (P less than 0.05) in ISO-BHV-1 calves on days 4 and 8 PI in PHA-stimulated lymphocytes and on day 8 PI in concanavalin A-stimulated lymphocytes when compared to control, ISO or BHV-1 calves. Isoprinosine treatment of BHV-1-infected calves tended to decrease alveolar macrophage bactericidal activity. These data suggest that ISO does not reverse BHV-1 suppression of lymphocyte proliferation, but may enhance IL-2 production in BHV-1 infected calves.     

Immunosuppression of lymphocyte blastogenesis in cattle infected with Ostertagia ostertagi and/or Trichostrongylus axei

Twenty 4-month-old calves were infected with O ostertagi and/or T axei and the responses to phytolectins were evaluated. Whole blood cultures were incubated with phytohemagglutinin (PHA), concanavalin A (Con A) and pokeweed mitogen (PWM). The blastogenic response was determined by tritiated thymidine uptake with results presented as counts per minute (cpm), stimulation indices (SI) and a mononuclear cell responsive index determined by dividing the phytomitogen induced cpm by the absolute mononuclear cell number per ul. The control group results were adjusted to 100 percent and changes in the percentage difference by the parasitized calves was determined. There was a decline in lymphocyte responsiveness to PHA beginning at the time of infection. Significant depression of responses to PHA was observed in all parasitized calves 8 weeks after infection although clinical signs of parasitism did not occur. Lymphocyte responses to PW, were not different in infected calves from the control, although the O ostertagi group had significantly higher PWM mean upon than the calves infected with T. axei. A slight depression in response to Con A was also observed at 8 weeks after infection followed by a significant increase after 10 weeks. The immunosuppression appeared to be a feature of gastrointestinal parasitism and related to infections with O. ostertagi and/or T. axei.     

Correlation of macrophage migration-inhibition factor and protection from challenge exposure in calves vaccinated with Salmonella typhimurium

Migration of bovine macrophages under agarose was used to assess cellular immunity in 7 nonvaccinated calves and 9 calves vaccinated with Salmonella typhimurium. The 9 vaccinated calves were allotted to 4 groups. Group I calves were vaccinated twice orally with small doses of virulent S typhimurium; group II calves were vaccinated twice orally with genetically altered aromatic-dependent (aro-) S typhimurium SL3261; group III calves were vaccinated twice IM with small doses of virulent S typhimurium; and group IV calves were vaccinated twice IM with aro- S typhimurium SL1479. Samples of blood were obtained from these calves at 2 weeks after the 2nd vaccinal dose was given, and lymphocytes were harvested, using lymphocyte separation medium. Lymphocytes in serum-free medium were then incubated with S typhimurim antigen for 48 hours. Lymphocytes were then transferred to antigen-free medium and incubated for 48 hours, and the supernatant was assayed for the migration-inhibition factor (MIF). Lymphocyte supernatant was assayed for MIF by incubating it for 48 hours with 2.0 X 10(4) alveolar macrophages in agar wells. The macrophage migration distance was measured and compared with control values. Macrophage migration was inhibited in the presence of supernatant of lymphocytes from vaccinated calves that had been incubated with antigen, indicating the presence of the MIF in the supernatant. Migration distances, as a percentage of control, were 33% for group I calves (oral vaccination, virulent vaccinal organism), 60% for group II calves (oral vaccination, aro- vaccinal organism), 41% for group III (IM vaccination, virulent organism), and 25% for group IV (IM vaccination, aro- vaccinal organism).(ABSTRACT TRUNCATED AT 250 WORDS)     

Hepatic immune response in calves during acute subclinical infection with bovine viral diarrhoea virus type 1

Eight colostrum-deprived calves aged 8-12 weeks were inoculated intranasally with a non-cytopathic strain of bovine viral diarrhoea virus (BVDV) genotype-1 and the effects on the hepatic immune response were studied. Two calves were sacrificed at each of 3, 6, 9 and 14 days post-inoculation (dpi) and two uninoculated animals were used as negative controls. BVDV was detected in hepatic macrophages and monocytes from 3 to 14dpi and in Küpffer cells (KCs) from 6 to 14dpi. Increases in the numbers of MAC387(+) KCs and monocytes, but not interstitial macrophages, differentiated by morphological features, were evident in the liver following inoculation with BVDV. There was a substantial increase in the number of monocytes positive for tumour necrosis factor (TNF)-α, but only small increases in the numbers of TNF-α(+) KCs and interstitial macrophages and interleukin (IL)-6(+) monocytes, KCs and interstitial macrophages. There was an increase in the number of interstitial CD3(+) T lymphocytes in the liver, but no substantial changes in the numbers of circulating CD3(+) T lymphocytes, interstitial or circulating CD4(+) or CD8(+) T lymphocytes, or CD79αcy(+) B lymphocytes. Serum haptoglobin and serum amyloid A increased transiently at 12dpi. Upregulation of some pro-inflammatory cytokines by hepatic macrophages is evident in subclinical acute BVDV type 1 infection in calves.     

Lymphocyte blastogenic responses of calves experimentally infected with Ostertagia ostertagi

Twelve 9-week-old calves were divided into four groups; two groups were maintained helminth-free as controls and the other groups were given Ostertagia ostertagi infective-stage larvae (L3) orally. One group received 100,000 L3 as a single inoculum and the other group received L3 in increasing dosages at weekly intervals for 8 consecutive weeks. The blastogenic responses to concanavalin A (Con A), phytohemagglutinin (PHA), and a soluble larval antigen from O. ostertagi (SLA) of peripheral blood lymphocytes were evaluated using tritiated thymidine incorporation into DNA as a measure of blastogenesis. The responses to Con A of all infected calves were significantly depressed while the responses to PHA were not. SLA, at concentrations of 4 micrograms ml-1 and above, caused blastogenic activity in lymphocytes from uninfected calves. Using SLA at 1 microgram ml-1 in lymphocyte cultures supplemented with autologous serum, an antigen-induced blastogenic response was detected in calves receiving serial inoculations of L3.     

Skin testing, fecal culture, and lymphocyte immunostimulation in cattle inoculated with Mycobacterium paratuberculosis.

Fourteen calves at 21 days of age were experimentally inoculated with 100 mg (wet weight) of Mycobacterium paratuberculosis. Three calves were inoculated orally, 4 intravenously, and 7 subcutaneously. Lymphocyte immunostimulation, fecal culture, and intradermal tuberculin skin testing were done between 112 to 150 days following exposure. Lymphocyte immunostimulation test results, conducted at 112 days after inoculation, showed all animals positive to Mycobacterium avium purified protein derivative. Fecal culture results, taken at 120 days after inoculation, showed that 2 of 3 animals inoculated intravenously were positive, whereas only 2 of 7 inoculated subcutaneously were positive (8 of 14 total were positive). Intradermal skin testing results at 150 days with M avium purified protein derivative showed 13 of the 14 calves were positive. Calves were examined at necropsy 153 days after inoculation, and M paratuberculosis was isolated from tissues of each of the 14 calves.     

Phenotypic comparison of ileal intraepithelial lymphocyte populations of suckling and weaned calves

Ileal intraepithelial lymphocyte (IEL) suspensions from suckling calves (1-3 weeks old) and weaned calves (3-6 months old) were phenotyped to determine whether there were differences in the lymphocyte populations consistent with postnatal maturation of the mucosal immune system. Flow cytometric comparisons of IEL from the two age groups revealed the presence of significantly larger proportions of CD4+ T lymphocytes and CD8+ T cells in the weaned animals. In contrast, there was a significantly larger proportion of B-B2+ IEL in the suckling calves. Freshly isolated IEL from both groups of calves expressed mRNA for TNF-alpha and IFN-gamma, but not IL-4 or IL-10. The B-B2+ IEL population was more closely examined by flow cytometry. These cells co-expressed IgM and CD21. However, they did not express IgA, IgG1, nor any of several additional leukocyte differentiation molecules. Immunohistochemical data confirmed the presence of IgM+ lymphocytes, and the paucity of IgA+ and IgG1+ lymphocytes in suckling calf ileum. However, substantial numbers of IgA+ and IgG1+ cells were observed in weaned calf ileum. Together, the data are consistent with ongoing postnatal maturation of the gut mucosal immune system.     

Ultrasonographic evaluation of thymus in Holstein calves and Heifers and its diagnostic application

This study was performed to evaluate the cross section of the thymus in 46 clinically healthy Holstein calves and heifers from birth to 100 weeks of age and 8 age-matched affected animals with chronic bronchopneumonia by ultrasonography. The cross sectional area of the thymus increased in healthy calves and heifers from birth to 31 weeks of age, peaked around 35 weeks of age, and then decreased until 100 weeks after birth. A positive correlation (r = 0.74, P < 0.05) was found between the area in cross section and the weight of the whole thymus in calves and heifers with chronic bronchopneumonia. The cross- sectional areas of the thymus were significantly (P < 0.05) lowered in animals with chronic bronchopneumonia. The cross sections of thymus in clinically healthy calves at 2 weeks of age and heifers at 34 weeks and 87 weeks of age were hypoechoic, moderately hyperechoic and hyperechoic, respectively. The observation of thymus by ultrasonography seems to be an effective tool for estimating the size of the thymus objectively in Holstein calves and heifers from birth to 100 weeks of age. The results of this study prove the approach to be useful and applicable to monitor the growth of thymus in Holstein calves and heifers and their association with chronic pneumonia.     

Lymphocyte subpopulations and neutrophil function in calves during the first 6 months of life

Fifteen clinically healthy calves were sampled every week during the first 5 weeks of life and thereafter every month until the age of 6 months. The percentages and absolute values of CD4+, CD8+ gammadelta TCR+ and WC1+ T cells, CD21+ B cells and NKp46+ NK cells were determined by flow cytometry, and the expression of the interleukin-2 receptor alpha chain (CD25) was measured to assess the level of activation of the lymphocyte subpopulations. Neutrophil phagocytosis, respiratory burst and bactericidal activity were measured in five different neutrophil function assays. Most of the parameters examined reached a stable level during the first 6 months of life. The proportions of CD4+ and CD8+ lymphocytes remained relatively stable during the study period, while there was a moderate decrease in the relative percentage of gammadelta T cells from birth to approximately 5 months of age. However, the absolute numbers of gammadelta T cells per millilitre of blood remained stable throughout the study period and did not display significant variation with age. The percentage of cells expressing the B-cell maturation marker CD21 increased significantly over the first 5 months of life. The proportion of NK cells showed substantial variation during the study. Marked differences in the relative proportions of the lymphocyte subpopulations were noted between the individual calves, and the individual ranking of the animals was largely maintained over time. CD25 expression was detected on a mean of 6.6% of the CD4+ cells, while a lower percentage of the other lymphocyte subpopulations expressed this receptor. Phagocytic activity was demonstrated in approximately 90% of the neutrophils, and this proportion remained stable during the entire study period, while respiratory burst activity showed a moderate decrease during the first 2 months of life. The present study shows that the T-cell subpopulations are present in peripheral blood of calves at levels comparable with adult values, while the B-cell population increases significantly with age. The decrease in the relative percentage of gammadelta T cells appears to be attributable to an increase in the absolute numbers of CD4+ and CD21+ cells, rather than a change in absolute gammadelta T-cell numbers. Furthermore, the results indicate that the neutrophilic granulocytes are functional and able to mount an effective response in young calves from the first week of life.     

An epidemiologic study of mortality in veal calves subsequent to an episode of zinc toxicosis on a California veal calf operation using zinc sulfate-supplemented milk replacer

Ninety-five 3- to 6-month old male Holstein veal calves were evaluated after an episode of zinc toxicosis, to describe clinical signs and to identify management and/or host-related factors that may have contributed to death. Clinical signs appeared 23 days after feeding of milk replacer commenced. Of 85 calves examined, 64 had pneumonia (75.5%), 62 had ocular signs (72.9%), 46 had diarrhea (54.1%), 34 were anorectic (40.0%), 15 were bloated (17.6%), 8 had cardiac arrhythmias (9.4%), 3 had convulsions (3.5%), and 3 were polydipsic/polyphagic (3.5%). Clinical signs began to appear when calves each were being fed approximately 1.5 to 2.0 g of zinc/day and exposed to a cumulative zinc intake of 42 to 70 g, from a milk replacer containing 706 micrograms of elemental zinc/g of milk replacer. Of 95 calves studied, 1 died before zinc was supplemented, 16 died during the episode, 12 were euthanatized, 1 was lost to follow-up evaluation, 1 was culled, and 64 were slaughtered. Deaths attributable to zinc toxicosis were observed between 25 and 53 days after the milk replacer was supplemented with zinc. Calves died while being exposed cumulatively to 30 to 66 g of zinc. The factors of previous pneumonia severity, age, cumulative daily exposure to zinc, and calf location within a bay were examined for possible associations with mortality, using stepwise logistic regression. Though younger calves tended to have a higher mortality than older calves, neither age category nor severity of pneumonia, before zinc supplementation, accounted for a significant mortality.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lethal acrodermatitis in bull terriers

A lethal syndrome characterized clinically by growth retardation, progressive acrodermatitis, chronic pyoderma and paronychia, diarrhea, pneumonia, and abnormal behavior was observed in 17 related Bull Terrier pups. Median survival time was 7 months. Laboratory evaluation revealed non-degenerative neutrophilia, consistently low activities of serum alkaline phosphatase and alanine transaminase, and frequently, hypercholesterolemia. Lymphocyte blastogenic responses were decreased and there was dysgammaglobulinemia in pups in which quantitative studies of immunoglobulins were made. The mean of plasma zinc concentrations in 5 affected pups was significantly lower than the mean of age- and breed-matched controls. Pathologic findings included parakeratosis, hyperkeratosis, and superficial bacterial infections of the skin. There was severe reduction of lymphocytes in T-lymphocyte areas of lymphoid tissue. Bronchopneumonia and dilatation of the cerebral ventricles were found in most affected pups. Family studies indicated that the syndrome is inherited as an autosomal recessive trait. In spite of its similarities to lethal trait A46 in Black Pied Danish cattle and acrodermatitis enteropathica in man, oral or parenteral treatment with zinc failed to ameliorate the clinical signs of the syndrome.     

Phenotypical characterization of peripheral blood leucocytes in the newborn calf

The present study was undertaken to establish reference values for the composition of blood leucocyte populations in neonatal calves by differential leucocyte counts and immunophenotyping. Neonatal calves 1 h post partum (p.p.) were found to have a very high absolute number of granulocytes while the number of peripheral blood mononuclear cells was lower than in calves aged 3-9 weeks. The relative numbers of T cell subpopulations were similar in newborn and older calves, but newborn calves had lower percentages of B cells and MHC class II positive cells. Within the first 4 h of life the relative numbers of CD2+, CD6+, and CD8+ T cells declined in colostrum-fed as well as in colostrum-deprived calves. In contrast, the percentage of MHC class II positive cells and monocytes increased from 1 h to 4 h p.p. particularly in colostrum-fed calves. Although there is some evidence for immaturity of lymphocytes in neonatal calves, the immune system of these animals seems to be fully present at birth.     

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.     

Inhibition of alpha-glucosidase in cattle by Castanospermum australe: an attempted phenocopy of Pompe''s disease

Pompe's disease is characterised by an absence of lysosomal alpha-glucosidase, but this enzyme is also inhibited by Castanospermum australe seeds. Four calves were fed C. australe seeds at the rate of 0.15 g/kg body weight for periods from 1 to 4 days. Lymphocyte alpha-glucosidase activity was reduced by at least 90%, with the majority of inhibition occurring within 8 h of dosing. Several weeks elapsed before activity returned to normal. Significant inhibition of muscle alpha-glucosidase occurred and the ratio of plasma alpha-glucosidase activity measured at pH 5.6 relative to that at pH 3.7 was depressed. In an attempt to induce Pompe's disease, 2 calves were dosed with 1.2 g C. australe seed/kg body weight/day for 13 months. Lymphocyte and muscle alpha-glucosidase activities were markedly reduced over the entire period of feeding, but the animals showed no clinical signs of disease. Tissue cells were not vacuolated nor did they show any apparent accumulation of glycogen. Despite significant inhibition of alpha-glucosidase in skeletal and cardiac muscle, liver, kidney and brain, it is suggested that there was sufficient residual enzyme to prevent induction of a phenocopy of Pompe's disease.     

Dermal cellular responses of helminth-free and Ostertagia ostertagi-infected calves to intradermal injections of soluble extracts from O. ostertagi L3 larvae

Twelve calves were raised helminth-free until 9 weeks of age when six were orally inoculated with 100,000 Ostertagia ostertagi infective stage larvae (L3). Three uninfected and three experimentally infected calves received intradermal injections of sterile saline and soluble larval extract (SLE) from O. ostertagi L3 with a protein concentration ranging from 1 to 200 micrograms ml-1. Biopsies were performed 48 h post-injection. A kinetic study was performed on the remaining six calves, three infected and three uninfected, using a 100 micrograms ml-1 concentration of SLE and taking biopsies 1, 4, 8, 12, 24, and 72 h post-injection at both the saline and SLE-injected sites. All calves had an immediate wheal and increase in skin thickness at the SLE-injected sites. The numbers of eosinophils infiltrating SLE-injected sites as compared to saline-injected sites were significant in both uninfected and infected calves, but the infected calves had significant numbers to a wider range of SLE concentrations and had significantly higher numbers than uninfected calves in the kinetic study. Infected calves also had significant numbers of basophils in the dose response study at concentrations of 5 and 100 micrograms ml-1 SLE. Neutrophil infiltration was similar in both groups and was significant at SLE-injected sites early in the kinetic study. Detectable mast cells were decreased in SLE-injected sites of infected animals and perivascular accumulation of mononuclear and some polymorphonuclear cells was observed in the deep dermis of infected animals.