Serum IgG and IgM antibody response in cattle to antigens of Pasteurella haemolytica

The serum IgG and IgM antibody responses of 48 cattle vaccinated with live Pasteurella haemolytica (LIVE), formalin-killed P. haemolytica in Freund's incomplete adjuvant (FIA), or formalin-killed P. haemolytica in aluminum hydroxide adjuvant (ALH) to a variety of P. haemolytica antigens were evaluated. Enzyme-linked immunosorbent assays (ELISAs) were used to determine the sequential and day 21 IgG and IgM antibody responses to whole P. haemolytica (WB), a capsular carbohydrate-protein subunit (CPS) extracted from the organism, P. haemolytica capsular carbohydrate (CC), and P. haemolytica leukotoxin (LT). LIVE and FIA vaccinates developed generally higher IgG and IgM responses to all antigens compared to ALH vaccinates. LIVE vaccinates developed IgG responses to LT which were significantly higher (P less than 0.05) than all other vaccinates. In contrast, FIA vaccinates developed significantly higher IgG responses to CPS than all other vaccinates. On the basis of the ELISA results, similar or cross reacting antigenic sites were present in preparations containing surface antigens (WB, CPS and CC), but not LT. Disease resistance, as determined by experimental lesions induced in the 48 calves by transthoracic challenge with P. haemolytica, was significantly greater in the LIVE and FIA vaccinates compared with ALH vaccinates. No significant difference in resistance was detected between LIVE and FIA vaccinates. Lesions in ALH vaccinates were not significantly different than those in phosphate-buffered saline (PBS) controls. Increased IgG responses to all antigens were significantly associated with resistance to experimental disease; however, IgG responses to CPS were most highly correlated with resistance. The only IgM response which was significantly correlated with resistance was the response to CPS. These studies indicate that serum IgG antibody responses to various surface antigens of P. haemolytica, as well as LT, can enhance resistance to experimental pneumonic pasteurellosis. Serum IgM responses, however, do not appear to play a major role in resistance to experimental disease.     

Serum neutralizing antibody titers in dairy cattle administered an inactivated vesicular stomatitis virus vaccine

Two doses of a formalin-killed, cell culture-derived vesicular stomatitis virus (vsv)-New Jersey serotype vaccine were administered intramuscularly, 30 days apart, to all lactating and nonlactating cows in a 350-cow dairy herd. Serum specimens were obtained serially from 96 cows before vaccination and at 30, 52 and 80 days after vaccination and from 24 of these cows 175 days after vaccination. Serum neutralizing antibody titers to vsv-New Jersey serotype were determined from serum-dilution, plaque-reduction tests. Serum neutralizing antibody titers also were determined during the same period for 67 nonvaccinated heifers in the herd. Peak group geometric mean serum neutralizing antibody titers of 1:530.46 +/- 1.14 (group geometric mean titer log10, 2.725 +/- 0.055) developed 21 days after the second vaccination, but decreased to a low value of 1:65.36 +/- 1.38 (group geometric mean titer log10, 1.815 +/- 0.142) by 175 days after vaccination. The nonvaccinated group had no detectable antibody titer to vsv-New Jersey serotype throughout the study. All serum specimens from the vaccinates and controls were negative for heterologous reactivity to vsv-Indiana serotype.     

Serum antibody isotype responses of Fasciola-infected sheep and cattle to excretory and secretory products of Fasciola species

This study investigated the immunoglobulin isotype responses of sheep and cattle chronically infected with Fasciola hepatica and Fasciola gigantica to adult F. hepatica excretory/secretory products (Fh-ES) or F. gigantica excretory/secretory products (Fg-ES), respectively. An antibody enzyme-linked immunosorbent assay (Ab-ELISA) was used to determine serum antibody (total Ig, IgG(1), IgM, IgG(2) and IgA) responses. At necropsy, the mean number of flukes recovered was lower in cattle than in sheep. All F. hepatica and F. gigantica infected sheep and cattle showed an increased total Ig levels from 3 to 4 weeks post-infection (wpi). Among isotypes IgG(1) was most dominant while IgM was the earliest (2 wpi) to be detected in both sheep and cattle infected with both F. hepatica and F. gigantica animals. IgG(2) response was early (2 wpi) in sheep infected by F. hepatica but there was no response in sheep infected with F. gigantica. There was a late and strong IgG(2) response in cattle infected with both flukes. The IgA isotype showed an early and a clear biphasic response in sheep with F. hepatica but was less pronounced in F. gigantica infected sheep. While IgA response to Fh-ES was noticed 5 wpi in F. hepatica infected cattle, it appeared much later (21 wpi) in those infected with F. gigantica. The dominance of IgG(1) isotype in infected sheep and cattle suggest an associated Th2 response. This early response to adult Fasciola spp. ES antigen suggests an early exposure to the antigen presumably through the cross-reacting ES products of juvenile flukes. There is clearly difference in IgG(2) isotype response in cattle (resistant) compared to sheep (susceptible). The late IgG(2) response in cattle may suggest late Th1 involvement in bovine cellular responses to adult Fh-ES/Fg-ES.     

Serum IgG and total protein concentrations in dairy calves fed two colostrum replacement products

OBJECTIVE: To evaluate effects of 2 commercially available colostrum replacement products on serum IgG and total protein concentrations in dairy calves. DESIGN: Prospective clinical trial. ANIMALS: 84 Holstein bull calves from a single dairy. PROCEDURES: Calves were randomly assigned to be given 4 quarts of colostrum (group 1; n = 21), 2 packages of a colostrum replacement product (product A; group 2; 21), 1 package of a different colostrum replacement product (product B; group 3; 21), or 2 packages of product B (group 4; 21). Treatments were given within 3 hours after birth, and blood samples were collected 24 hours later and submitted for determination of serum total protein and IgG concentrations. RESULTS: Group 1 calves had significantly higher serum total protein and IgG concentrations than did calves in the other 3 groups. However, the percentage of calves with adequate passive transfer (ie, serum IgG concentration > 1,000 mg/dL) was not significantly different among groups 1 (90%), 3 (81%), and 4 (95%). In contrast, only 10% of calves in group 2 had adequate passive transfer. It was predicted that calves fed product B that had serum total protein concentrations > 5.2 g/dL would have serum IgG concentrations > 1,000 mg/dL at least 90% of the time. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that product B could be considered as an alternative to colostrum in dairy calves, but product A failed to routinely provide adequate serum IgG concentrations when fed according to label directions.     

Serum isocitrate dehydrogenase during polybrominated biphenyl toxicosis in dairy cattle

Bovine serum isocitrate dehydrogenase (sICDH) was investigated in dairy cattle as a clinical measurement indicative of hepatic injury. Conditions for optimization of isocitrate dehydrogenase assays for bovine serum are described. Assays of sICDH in normal cattle show average activities of .814 (SD = .202) units/ml serum with a range of .316 to 1.268 for 83 samples taken from 32 animals. Investigation of sICDH in pregnant dairy cattle experimentally dosed with polybrominated biphenyl (PBB) showed no discernible elevations until doses were sufficient to cause toxicosis (25,000 mg PBB/d). Cows lethally dosed with 25,000 mg PBB/d had moderate elevations of sICDH (approximately a twofold increase) concomitant with severe toxicosis in some but not all animals. This PBB dose also caused abortion or fetal death in pregnant animals; elevation of sICDH in these animals was coincident with fetal trauma. This suggests that sICDH may be influenced by fetoplacental contributions in pregnant animals. Non-pregnant cows, intoxicated with PBB, had minimal sICDH elevation as compared with 10-fold in a calf with experimentally induced hepatotoxicity (thioacetamide). This observation was consistent with histopathological findings of minimal, if any, hepatic involvement in dairy cattle lethally intoxicated with PBB. Serum isocitrate dehydrogenase appears to be a useful adjunct to the ordinary complement of serum chemistries used for clinical diagnosis; however, it does not appear to reflect exclusively hepatic injury.     

Serum antibody levels to Taenia saginata in cattle grazed on Scottish pastures

The serum antibody levels to Taenia saginata of three groups of cattle were assessed by an enzyme-linked immunosorbent assay (ELISA). The first group of cattle were from four farms which had a confirmed T saginata cysticercosis outbreak, all of which had cattle classed as infected by ELISA. The second group were from four farms where sewage sludge had been applied to pasture subsequently grazed by the cattle. One of these farms had cattle classed as infected by ELISA. The control cattle, which were all classed as uninfected by ELISA, came from five farms whose pasture had not been treated with sewage sludge. In a wider survey, involving sera from 47 additional farms, the majority could not be distinguished from the control farms in the earlier survey. However, samples from three of the farms had a similar number of positives to two of the known infected farms in the initial survey. Since the ELISA assay may indicate infected herds, farms such as these warrant further investigation.     

Serum biochemical parameters and embryo production during superovulatory treatment in dairy cattle

The objective of this study was to determine the relationship between the number of transferable embryos (TE) and various blood chemistry parameters as a reflection of the metabolic state of cows after superovulatory treatment. Forty-nine Holstein cows were subjected to superovulatory treatment for commercial embryo production. At the time of embryo harvest, individual blood samples were taken from cows for biochemical analysis. All embryos including dead ones as well as non-fertilized oocytes were counted in uterine lavage. Feed samples collected daily for a period of two weeks before embryo harvest, were analyzed for mycotoxins: vomitoxin, zearalenone and T-2 toxin. On average, cows produced 9.45 ± 5.60 embryos and oocytes of which 5.27 ± 4.20 were TE, 0.37 ± 0.80 were dead embryos and 3.82 ± 3.78 were non-fertilized oocytes. Higher concentrations of Mg and K were associated with a higher production of TE (p = 0.005 and p = 0.043, respectively) and higher activity of creatinine kinase was associated with a lower production of TE (p = 0.011).     

Serum biochemical parameters and embryo production during superovulatory treatment in dairy cattle

The objective of this study was to determine the relationship between the number of transferable embryos (TE) and various blood chemistry parameters as a reflection of the metabolic state of cows after superovulatory treatment. Forty-nine Holstein cows were subjected to superovulatory treatment for commercial embryo production. At the time of embryo harvest, individual blood samples were taken from cows for biochemical analysis. All embryos including dead ones as well as non-fertilized oocytes were counted in uterine lavage. Feed samples collected daily for a period of two weeks before embryo harvest, were analyzed for mycotoxins: vomitoxin, zearalenone and T-2 toxin. On average, cows produced 9.45 ± 5.60 embryos and oocytes of which 5.27 ± 4.20 were TE, 0.37 ± 0.80 were dead embryos and 3.82 ± 3.78 were non-fertilized oocytes. Higher concentrations of Mg and K were associated with a higher production of TE (p = 0.005 and p = 0.043, respectively) and higher activity of creatinine kinase was associated with a lower production of TE (p = 0.011).     

Distribution of T and B lymphocytes in mammary dry secretions, colostrum and blood of adult dairy cattle

The distribution of lymphocyte subpopulations from dry secretions, colostrum and blood from 10 healthy adult Hostein-Fresian cows was studied using the TH21A and B26A mouse monoclonal antibodies (MAb) to adult bovine B and T lymphocytes, respectively. The mammary gland lymphocytes (MGL) were isolated from composite sample of all four quarters by density centrifugation over discontinuous gradient of ficoll-diatrizoate. The peripheral blood lymphocytes (PBL) were purified using the ficoll-thrombin method. Isolated PBL and MGL were analyzed using the two fluorochromes method (TFM) and laser flow cytometry (LFC). The mean viability of isolated PBL and MGL from dry secretions and colostrum after the TFM and LFC were 92.4% +/- 3.2%, 91.4% +/- 6.0% and 87.1% +/- 6.1%, respectively. There was a good correlation between the two MAbs and the percentage of surface immunoglobulin (SIg) positive cells in the peripheral blood using the TFM. The PBL yielded a mean percentage of 21.2% B cells, 66.4% T cells and 9.4% "Null cells" (TH21A+; SIg-). The TFM on MGL from dry secretions and colostrum indicated two distinct patterns (group I and II) of SIg and reactivity to MAb markers (p less than 0.001). The MGL data included in group I and group II were gathered from both colostral and dry secretions. In comparison to the distribution of lymphocyte subsets within peripheral blood the mean percentages of B cells, T cells and "Null cells" in the mammary gland were respectively, 2.8%, 88.1% and 5.4% for group I and 3.5%, 89.0% and 15.1% for group II. In the mammary secretions, the use of SIg alone was not considered to be a good marker for B cells; in four animals a mean percentage of 15.6% (13.9/89.0 X 100) of the mammary gland T lymphocytes were also SIg+. Of the TH21A+ MGL, only 18.8% were SIg+ in group II compared with 34.1% for MGL from group I and 69.3% for the PBL. Marked differences in cell size distribution and cell surface antigen density were found when PBL and MGL from dry secretions were compared by LFC using the B26A MAb. The results of this study demonstrate a difference in the percentages of peripheral blood and mammary gland B and T lymphocytes and confirm previous findings in which the T lymphocytes were found to represent the major subpopulation of lymphocytes in bovine mammary secretions. This may represent an essential event in the adoptive transfer of cellular immunity through the colostrum in cattle.     

The association between antibody titres againstCampylobacter fetus and reproductive efficiency in dairy cattle

The relationship between the antibody titres againstCampylobacter fetus and various indices of reproductive efficiency was studied in a cross-sectional study of 178 dairy cows from three California Dairy Herd Improvement Association herds. Blood samples were collected from the lactating cows during December 1986. Enzyme-linked immunosorbent assays were used to determine the antibody titres of the cow againstCampylobacter fetus, Haemophilus somnus andLeptospira hardjo and were classified as either negative or positive. The status of a cow as either negative or positive againstCampylobacter fetus andHaemophilus somnus represents serological evidence of natural exposure to the corresponding bacterial agents. However, the status againstLeptospira hardjo was assumed to reflect a vaccinal titre since all the cows studied had been routinely vaccinated against this organism in September 1986. The data on demographic and reproductive parameters pertained only to the current lactation of the cows and were obtained from Dairy Herd Improvement Association individual cow records of December 1986. Five indices of reproductive efficiency were used, namely the recent calving interval, the calving-to-conception interval, the calving-to-last-service interval, the number of services per conception, and the number of services since last calving. The serological status againstHaemophilus somnus, Leptospira hardjo and other covariates suggested by the results of previous studies were included in modelling the relationships of interest. Stepwise multiple linear regression analyses were carried out to study the adjusted relationship ofCampylobacter fetus with each measure of reproductive efficiency.Multivariate analyses revealed that the adjusted relationship forCampylobacter fetus with all five measures of reproductive efficiency was non-significant (p > 0.05). Among the covariates,Leptospira hardjo had a strong and independent relationship with recent calving interval, the unstandardized partial regression coefficient being -0.77. The possible biological mechanisms of these associations are discussed.Abbreviations BMDP biomedical computer programs - DHIA dairy herd improvement association - ELISA enzyme-linked immunosorbent assay - ICR individual cow record - PSS physiological saline solution - SD standard deviation; Other abbreviations are shown in Table I     

Serum complement-fixing antibody to Pasteurella haemolytica A1 and its effect on experimentally induced pneumonic pasteurellosis in calves

Ninety-three calves comprising 16 experimental groups were exposed to viral (bovine herpesvirus-1 or parainfluenza-3 virus) and Pasteurella haemolytica aerosols. Serum samples from these calves were tested before and after exposure for antibodies to P haemolytica by a modified direct complement-fixation test. At slaughter of the calves, the extent of pneumonia produced was estimated for each calf and compared with the results of the modified direct complement-fixation tests. The extent of pneumonia was not related (P greater than 0.05) to the amount of anti-P haemolytica antibody produced by either naturally occurring or experimentally induced infection.     

The association between antibody titres againstCampylobacter fetus and milk production efficiency in dairy cattle

The association between serological evidence of exposure toCampylobacter fectus and milk production performance was studied in 178 lactating cows from three California Dairy Herd Improvement Association herds using a cross-sectional study design in December 1986. ELISAs were used to determine the antibody titres againstCampylobacter fetus, Haemophilus somnus andLeptospira hardjo, which were classified as either negative or positive. The status of a cow as negative or positive againstC. fetus andH. somnus represents the serological evidence of natural exposure to the corresponding bacteria. However, the status againstL. hardjo was assumed to be the level of vaccinal titre against this organism since all the cows studied had been vaccinated against this agent. The data on demographic and productivity variables relating to the current lactation of the cows were obtained from Dairy Herd Improvement Association individual cow records for December 1986. Four measures of milk production efficiency for the current lactation were used. The status againstL. hardjo and other covariates suggested by previous studies were included in modelling the relationships of interest. Stepwise multiple linear regression was used to study the adjusted relationship ofC. fetus with each measure of milk production efficiency.Multivariate analyses revealed that the adjusted relationships ofC. fetus with the test-day's milk production, the extended 305-day milk production and the relative value of milk production were not significant (p>0.1). However, after adjusting for possible covariates,C. fetus-positive cows had an average of 7.43% lower mature equivalent milk production thanC. fetus-negative cows (p=0.02). Among the covariates, the serological status againstL. hardjo had a strong and independent relationship with the percentage mature equivalent milk production, the unstandardized partial regression coefficient being 4.53.We conclude from this cross-sectional study that the association ofC. fetus seropositivity with one index of milk production is the first indication that latentC. fetus infection may be associated with subclinical mastitis, perhaps through a hypersensitivity reaction. However, this needs further investigation.Abbreviations BMDP Biomedical Computer Programs - DHIA California Dairy Herd Improvement Association - ELISA enzyme-linked immunosorbent assay; other abbreviations are shown in Table I     

Serotype-specific resistance to nasal colonization by Pasteurella haemolytica in cattle

Vaccination of calves with a Pasteurella haemolytica serotype 1 antigen preparation elicited a serotype-specific inhibition of nasal colonization by P haemolytica under field conditions. Inhibition was evidenced by a low frequency of nasal colonizations and by relatively few P haemolytica serotype 1 organisms isolated from vaccinated calves. The study comprised 3 field trials, each on a separate year, and included 480 calves.     

奶牛源微小隐孢子虫的分子鉴定及动物感染试验

采用饱和蔗糖溶液漂浮法检查商丘市某奶牛场牛新鲜粪便样本的隐孢子虫卵囊,用18SrRNA基因对隐孢子虫进行PCR扩增和限制性片段长度多态性分析;基于GP60基因位点对微小隐孢子虫进行基因亚型鉴定。结果显示:103份样本中有50份为隐孢子虫阳性,42份经形态学鉴定为安氏隐孢子虫,8份形态学未能鉴定到种。经限制性片段长度多态性分析,7个分离株为微小隐孢子虫,1个分离株为牛隐孢子虫;序列比对分析,7个微小隐孢子虫均为人兽共患基因亚型IIdA19G1。接种1头3日龄犊牛1×106个卵囊,潜隐期为3d,显露期为14d,于感染后第7天和第10天出现2个排卵囊高峰期,收集到大量纯卵囊。     

Serum antibody response to soluble extract of the third-larval stage of Ostertagia ostertagi in cattle

Serum IgG, IgM, and IgA antibody responses against L₃ antigens of Ostertagia ostertagi were monitored by enzyme-linked immunosorbent assay (ELISA) after one, two or multiple sequential inoculations of this nematode in calves. Following the first infection, antibody levels did not change. After a second inoculation, IgG increased significantly (P < 0.05) after 2 months. IgG was not significantly increased 1 month after challenge inoculation. IgM and IgA antibody levels did not change following the first or second inoculations of L₃. IgG antibody levels rose only slightly following multiple sequential inoculations with infectious L₃.

Results indicate that calves with ostertagiasis have very weak serum antibody responses to L₃, and these appear to be of little value in detection of the infection in these animals.     

Comparison of three methods of feeding colostrum to dairy calves

Absorption of colostral immunoglobulins by Holstein calves was studied in 3 herds in which 3 methods of colostrum feeding were used. Failure of passive transfer, as determined by calf serum immunoglobulin G1 (IgG1) concentration less than 10 mg/ml at 48 hours of age, was diagnosed in 61.4% of calves from a dairy in which calves were nursed by their dams, 19.3% of calves from a dairy using nipple-bottle feeding, and 10.8% of calves from a dairy using tube feeding. The management factor determined to have the greatest influence on the probability of failure of passive transfer in the herds using artificial methods of colostrum feeding (bottle feeding or tube feeding) was the volume of colostrum fed as it affected the amount of IgG1 received by the calf. In dairies that used artificial feeding methods, failure of passive transfer was infrequent in calves fed greater than or equal to 100 g IgG1 in the first colostrum feeding. In the dairy that allowed calves to suckle, prevalence of failure of passive transfer was greater than 50% even among calves nursed by cows with above-average colostral IgG1 concentration. Analysis of the effect of other management factors on calf immunoglobulin absorption revealed small negative effects associated with the use of previously frozen colostrum and the use of colostrum from cows with long nonlactating intervals.