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Serum sensitivity of field isolates and laboratory strains of Brucella abortus
1995
Eisenschenk, F.C. | Houle, J.J. | Hoffmann, E.M.
Objective-To evaluate the ability of bovine complement to kill a variety of field isolates and laboratory strains of Brucella abortus. Design-The experimental approach was to determine the sensitivity of B abortus isolates to killing by bovine serum, and to document the role of complement in brucellacidal activity. Sample population-Six laboratory isolates and 12 field isolates of B abortus were tested. Procedure-The ability of B abortus to survive exposure to undiluted bovine serum for 2 hours at 37 C was assessed. The role of complement in killing was determined by examining the ability of heat (56 C for 60 minutes) and cobra venom factor to obliterate the activity in serum, and by detecting binding of the ninth component of bovine complement to serum-sensitive target cells. Results-Isolates of B abortus that were resistant to the bactericidal activity of normal bovine serum were revealed. These included field isolates and laboratory strains. Furthermore, the study confirmed earlier reports that bovine serum-mediated killing of B abortus is caused by the complement cascade. Conclusions-Some isolates of B abortus, like other gram-negative bacteria, were resistant to complement-mediated killing. Resistance was associated with smooth colony morphology. Isolates lacking detectable O antigen were serum sensitive.
Показать больше [+] Меньше [-]Detection of bovine immunodeficiency virus in blood and milk-derived leukocytes by use of polymerase chain reaction
1995
Nash, J.W. | Hanson, L.A. | St Cyr Coats, K.
Bovine immunodeficiency virus (BIV) is prevalent in beef and dairy cattle, yet the mode(s) of BIV transmission are undefined. Using polymerase chain reaction, which specifically targeted a 235-bp, highly conserved region of the BIV pol gene, BIV-infected leukocytes were detected in the blood and milk of BIV-seropositive cows. These data confirm the presence of BIV in milk and identify the potential for lactogenic transmission of this virus.
Показать больше [+] Меньше [-]Passive immune status at postpartum hour 24 and long-term health and performance of calves
1995
Wittum, T.E. | Perino, L.J.
We quantified the effect of passive immune status on pre- and postweaning health and growth performance of calves raised in a beef production environment. Blood samples were collected at postpartum hour 24 from 263 crossbred calves for determination of plasma protein (PP) and serum IgG concentrations. Serum IgG concentration was classified as adequate (> 1,600 mg/dl), marginal (800 to 1,600 mg/dl), or inadequate (< 800 mg/dl). Plasma protein concentration was classified as adequate (greater than or equal to 4.8 g/dl) or inadequate (< 4.8 g/dl). Morbidity and mortality events in the study population were monitored from birth to weaning, and after weaning throughout the feeding period. The lowest concentrations of serum IgG and PP were observed among calves that experienced morbidity or mortality prior to weaning. Calves that experienced morbidity in the feedlot had lower 24-hour PP values, but had IgG concentration similar to that in calves that were not observed to be ill during the feeding period. Calves classified as having inadequate IgG concentration were at greater risk of preweaning mortality (odds ratio [OR] = 5.4), neonatal morbidity (OR = 6.4), and preweaning morbidity (OR = 3.2), compared with calves classified as having adequate IgG concentration at 24 hours. Calves classified as having inadequate PP concentration at 24 hours had a greater risk of morbidity (OR = 3.0) and respiratory tract morbidity (OR = 3.1) while in the feedlot, compared with calves classified as having adequate PP concentration. The effects of 24-hour passive immune status on calf growth were indirect through effects on morbidity outcomes. Morbidity during the first 28 days of life was associated with a 16-kg lower expected weaning weight. Respiratory morbidity in the feedlot resulted in a 0.04-kg lower expected mean daily gain. Thus, passive immune status at postpartum hour 24 was an important determinant of health before and after weaning, and was indirectly associated with calf growth during the same periods.
Показать больше [+] Меньше [-]Improved early and long-term detection of bovine lentivirus by a nested polymerase chain reaction test in experimentally infected calves
1995
Suarez, D.L. | Maaten, M.J. van der | Whetstone, C.A.
A nested polymerase chain reaction (PCR) test was developed to examine infection with the bovine lentivirus, bovine immunodeficiency-like virus (BIV), in cattle. Primers were designed to amplify 2 separate regions of the pol and env segments of the BIV genome. Two calves were experimentally infected with an isolate derived from the original strain of BIV, R29, or with a recent field isolate, FL491. Serial blood samples were collected and examined by virus isolation, protein immunoblot, and nested PCR. The nested PCR test detected BIV infection by 3 days after inoculation, earlier than the other 2 methods, and continued to identify infected cattle 9 to 15.5 months after inoculation, even when results from virus isolation and serology became negative. Nested PCR also detected multiple-size env products in samples obtained later in the infection from the calf that received FL491, giving evidence that viral quasispecies were selected during in vivo replication of the virus. Results indicated that the nested PCR test is more sensitive than virus isolation or serology for the detection of BIV infection in cattle.
Показать больше [+] Меньше [-]Administration of recombinant human interleukin 1 receptor antagonist during endotoxin-induced mastitis in cows
1995
Shuster, D.E. | Kehrli, M.E. Jr
The role of interleukin 1 (IL-1) as an inflammatory mediator during mastitis and the therapeutic effect of recombinant human IL-1 receptor antagonist (IL-1ra) for bovine mastitis was studied. Cows were intramammarily infused with lipopolysaccharide (25 g) in 1 mammary gland. Half the cows also received infusions of 5 mg of IL-1ra into the same mammary gland just prior to endotoxin infusion and 4, 8, and 12 hours later. After endotoxin infusion, tumor necrosis factor and high IL-1 bioactivity were detected in whey from infused glands. Vascular permeability changes and neutrophil accumulation in milk paralleled the appearance of cytokines. A systemic reaction, characterized by pyrexia and an increase in blood cortisol concentration, also were observed. Milk yield was inhibited and milk composition was altered in infused and noninfused glands. The increase in IL-1 bioactivity in milk after endotoxin infusion was almost completely prevented in glands receiving IL-1ra. However, IL-1ra had no effect on local inflammation, systemic reaction, or impairment in productive performance. These results indicate that IL-1 does not mediate its effect within the milk compartment, and suggest either that IL-1 is not critical to the mastitic response or that intramammary infusion of IL-1ra does not place the antagonist where IL-1 interacts with its receptor.
Показать больше [+] Меньше [-]Comparison of the sensitivity of the caudal fold skin test and a commercial gamma-interferon assay for diagnosis of bovine tuberculosis
1995
Whipple, D.L. | Bolin, C.A. | Davis, A.J. | Jarnagin, J.L. | Johnson, D.C. | Nabors, R.S. | Payeur, J.B. | Saari, D.A. | Wilson, A.J. | Wolf, M.M.
A study to determine and compare the sensitivity of the caudal fold tuberculin test (CFT) and a commercial gamma-interferon (gamma-IFN) assay for diagnosis of bovine tuberculosis was conducted. A dairy herd with approximately a third of the cattle infected with Mycobacterium bovis was chosen for this study. All cattle from this herd were slaughtered, and tissue specimens for bacteriologic culturing and histologic examination were collected. Results of the CFT and gamma-IFN assay were compared with results of bacteriologic culturing and histologic examination to determine test sensitivity. Results were analyzed, using each of the following 4 standards to classify cattle as infected: positive test result by bacteriologic culturing only; histologic examination only; bacteriologic culturing and histologic examination; and bacteriologic culturing or histologic examination. Sensitivity of the CFT ranged from 80.4 to 84.4%, depending on the standard of comparison. Sensitivity of the gamma-IFN assay ranged from 55.4 to 97.1%, depending on the standard of comparison and on the method of interpretation. The CFT was significantly (P < 0.001) more sensitive than the gamma-IFN assay for diagnosis of bovine tuberculosis when the gamma-IFN assay was conducted and interpreted as instructed by the manufacturer. Maximum overall sensitivity was achieved when results of the CFT and gamma-IFN assay were interpreted in parallel.
Показать больше [+] Меньше [-]Bovine immunodeficiency virus in stud bull semen
1995
Bovine immunodeficiency virus (BIV), a lentivirus, is prevalent in dairy and beef cattle in southeastern United States and may be associated with a lymphoproliferative disease. The mode(s) of BIV transmission are undefined. Because artificial insemination is a common practice in dairy production, contaminated stud semen could serve as an important source of infection if the virus is harbored in seminal fluids. To evaluate this possibility, we procured 11 cryopreserved semen specimens from a stud semen repository. Leukocytes were purified from the specimens, and the leukocyte DNA was used as template in a polymerase chain reaction procedure that targeted a 235-base pair, highly conserved domain of the BIV pol gene. The target sequence was amplified from the seminal leukocyte DNA of 9 of the specimens (82%), and nucleotide sequencing confirmed the BIV specificity of the fragment. This finding provides evidence that stud bull semen may serve as an important reservoir of BIV, suggesting the possibility that artificial insemination of dairy cows may have a major role in transmission and wide-spread dissemination of this bovine lentivirus.
Показать больше [+] Меньше [-]Enzyme-linked immunosorbent assay for serum apolipoprotein B-100, a major triglyceride-transport protein in dairy cows
1995
Yamamoto, O. | Oikawa, S. | Katoh, N.
An ELISA was developed to determine serum concentration of apolipoprotein B-100, a major triglyceride-binding protein in very low-density lipoproteins and a putative maker for hepatic lipidosis of dairy cows. Serum apolipoprotein B-100 was prepared electrophoretically, and antibodies to this protein were raised in rabbits. The antiserum prepared was further purified by affinity chromatography, using bovine serum albumin-Sepharose 4B, to remove antibodies to albumin. For the ELISA, addition of 2-mercaptoethanol to the coating buffer (50 mM sodium carbonate, pH 9.6) was required to evaluate apolipoprotein B-100 concentration in serum. The ELISA developed was sensitive (detection limit was 300 to 400 ng/ml of serum) and reliable (coefficients of variance were in the range of 3.3 to 7.6%). By use of the established ELISA, the serum apolipoprotein B-100 concentration was found to be significantly (P < 0.01) lower during the early lactating stage than during other stages of lactation. Reduced hepatic synthesis or secretion of apolipoprotein B-100 during the early lactating stage, together with the excess uptake by the liver of serum nonesterified fatty acids, is suggested to be relevant in the accelerated accumulation of triglycerides in the liver of dairy cows during the periparturient period.
Показать больше [+] Меньше [-]Ceftiofur distribution in serum and milk from clinically normal cows and cows with experimental Escherichia coli-induced mastitis
1995
Erskine, R.J. | Wilson, R.C. | Tyler, J.W. | McClure, K.A. | Nelson, R.S. | Spears, H.J.
Eight Holstein cows, 4 inoculated intracisternally in 1 quarter of the mammary gland with Escherichia coli and 4 noninfected controls, were administered ceftiofur sodium (3 mg/kg of body weight, IV, q 12 hours) for 24 hours, beginning at 14 hours after inoculation of infected cows. All challenge-exposed cows became infected, with mean +/- SEM peak log10 bacterial concentration in milk of 5.03 +/- 0.69 colony-forming units/ml. The infection resulted in systemic signs (mean peak rectal temperature, 41.5 +/- 0.3 C; anorexia; signs of depression) and local inflammation (mean peak albumin concentration in milk, 7.89 +/- 1.71 mg/ml). Ceftiofur was detectable in milk from all challenge-exposed cows, compared with only 1 of 4 noninfected cows, and the mean period after inoculation that ceftiofur was detectable in milk was longer (P < 0.05) in infected (147.7 +/- 27.5 hours) than noninfected cows (1.3 +/- 1.3 hours). However, maximal ceftiofur concentration attained in milk for all cows was 0.28 micrograms/ml, and was 0.20 micrograms/ml or less for all but 2 milk samples collected for 10 days after challenge exposure. Mean serum concentration of ceftiofur peaked at 1.0 +/- 0.3 micrograms/ml and 0.7 +/- 0.1 micrograms/ml for infected and noninfected COWS, respectively. After each ceftiofur dose, mean peak and trough concentrations of ceftiofur in serum did not differ between groups; however, concentration of ceftiofur in serum was higher at 7 hours after each dose in noninfected cows, suggesting more rapid clearance of the drug in infected cows. Ceftiofur was not detected in serum (< 0.05 micrograms/ml) of any cow at or after 120 hours following inoculation of infected cows. Storage of serum samples at -20 C for 3 weeks resulted in a 98.8% decrease in ceftiofur activity, compared with that in fresh serum samples. Eighty-seven percent of this loss occurred 30 minutes after mixing serum and ceftiofur; thus, about 13% of the original activity was lost in storage. Storage of milk samples under similar conditions did not result in loss of ceftiofur activity. Despite acute inflammation, the dosage of ceftiofur used in this trial would not result in drug concentrations in milk above FDA safe concentrations, or above the reported minimum inhibitory concentration for coliform bacteria.
Показать больше [+] Меньше [-]Regulation of neutrophil adhesion molecules and shedding of Staphylococcus aureus in milk of cortisol- and dexamethasone-treated cows
1995
Burton, J.L. | Kehrli, M.E. Jr
The effects of 3 days of glucocorticoid administration on bovine blood neutrophil expression of L-selectin and CD18, and on the health status of mammary glands subclinically infected with Staphylococcus aureus were measured in 9 lactating Holsteins. The experiment was a 3 x 3 Latin square cross-over design, with 3 glucocorticoid treatments switched among groups of 3 cows/treatment during 3 periods. Treatments consisted of a vehicle (control, 10 ml of excipient/cow/d), cortisol (7.5, 15, and 7.5 mg/cow on days 1, 2, and 3, respectively), and dexamethasone (0.04 mg/kg of body weight/cow/d for total daily dosages that ranged from 21.6 to 33.2 mg). Blood samples for immunostaining and flow cytometric analysis of L-selectin and CD18 and leukograms, as well as foremilk samples for determination of S aureus shedding somatic cell counts, protein and fat percentages, and daily milk yields were collected repeatedly before, during and after treatment days. Dexamethasone caused a profound, acute, short-lived down-regulation of L-selectin on neutrophils, which correlated in time to leukocytosis, mature and immature neutrophilias, increased shedding of S aureus in infected glands, and onset of high percentages of fat and protein and decreased milk yields. Dexamethasone also caused profound but delayed down-regulation of neutrophil CD18, which reached nadir simultaneously with reappearance of L-selectin-bearing neutrophils, normalized blood neutrophil counts, markedly high foremilk somatic cell counts and protein percentage, decreased S aureus shedding in milk, and finally, expression of clinical mastitis in some infected quarters. Each of these variables had returned to control (vehicle) values by the ninth (and last) sample collection day. Although cortisol treatment also decreased expression of L-selectin and CD18 on neutrophils, dosages used in this study were not sufficient to alter the number of circulating cells or to convert subclinical mammary gland infections to clinical mastitis. These results suggest that mammary gland health status can be altered by sudden exposure of blood neutrophils to glucocorticoids, because these steroid hormones caused profound down-regulation of the adhesion molecules that direct neutrophil margination and migration through the vascular endothelium. The results also reinforce the potential disease risk of treating infected animals with potent synthetic glucocorticoids, such as dexamethasone.
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