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Interferon and 2',5'-oligo(A) synthetase activities in serum and blood mononuclear leukocytes of cattle after injection of bovine interferon-alpha 1
1990
Perino, L.J. | Short, E.C. Jr | Burge, L.J. | Winter, D.A. | Fulton, R.W.
Cell extracts that were prepared from blood mononuclear leukocytes from 66 samples obtained from 6 clinically normal calves contained mean 2',5'-oligoadenylate (2',5'-oligo[A]) synthetase activity sufficient to synthesize 186 +/- 82 pmol of 2',5'-oligo(A)/h/10(6) cells. Calves had no measurable serum interferon (IFN) activity. Five calves were given IM injections of 10(4), 10(5), 5 x 10(5), 10(6), and 10(7) U of bovine IFN-alpha 1/kg of body weight at 2-week intervals. Five dosing sequences were used with a 5 x 5 Latin square design so that each calf received each dose once. Activity of 2',5'-oligo(A) synthetase increased at 24 hours in response to all dosages of IFN and then declined following first-order kinetics, with an apparent half-life (t1/2) of 2.1 +/- 0.5 days. The area under the concentration-time curve for 2',5'-oligo(A) synthetase increased with dose of IFN more rapidly than did peak response. Serum IFN that was measured at 1-day intervals following administration of IFN was consistently measurable only at dosages above 10(6) U of IFN/kg. The t1/2 for circulating IFN was 12.4 +/- 1.0 hours. Over all dosages, increases in 2',5'-oligo(A) synthetase activity were measurable for 3.5 days longer than were increases in IFN following IM injection of IFN. None of the calves developed detectable anti-IFN antibodies.
Show more [+] Less [-]Pharmacokinetics and metabolic inertness of doxycycline in young pigs
1990
Riond, J.L. | Riviere, J.E.
The disposition of doxycycline hyclate after IV administration of 20 mg/kg of body weight was studied in 6 pigs. Median elimination half-life, estimated in 4 pigs, was 3.92 hours. Mean (+/- SEM) total body clearance was 1.67 +/- 0.18 ml/min/kg, and mean apparent volume of distribution at steady state was 0.53 +/- 0.04 L/kg. In 2 pigs, secondary peaks in the logarithmic serum concentration-time profile suggested discontinuous enterohepatic cycling, and precluded using these pigs in the pharmacokinetic analysis. The extent of doxycycline binding to serum protein was 93.1 +/- 0.2%. Serum or urine from 3 of the pigs was analyzed by use of photodiode array detection and mass spectrometry of a high-performance liquid chromatographic column effluent. These procedures documented lack of doxycycline biotransformation in pigs. It is concluded that, despite an elimination half-life shorter than that reported in other species, doxycycline may be a valuable antimicrobial drug for use in swine practice, pending the development of appropriate formulations.
Show more [+] Less [-]Identification of Bordetella avium antigens recognized after experimental inoculation in turkeys
1990
Hellwig, D.H. | Arp, L.H.
Sera and tracheal washings (TW) were used to identify antigens of Bordetella avium recognized during experimentally induced bordetellosis in young turkeys. Pooled sera and TW were examined for antibody by a microtitration agglutination test and by western immunoblotting. In addition, comparable samples collected from 1-day-old turkeys and uninoculated control turkeys also were examined. At least 8 outer membrane proteins of B avium were recognized in immunoblots of sera and TW from infected turkeys. Reactivity of TW in immunoblots was qualitatively similar but less intense, compared with reactivity of corresponding sera collected on postinoculation (PI) weeks 2, 3, and 4. Molecular weights of the major outer membrane proteins of B avium recognized by sera and TW at PI week 4 were 100,000, 97,000, 36,000, 31,000, 21,000, 18,000, 14,000, and < 14,000. A protein with a molecular weight of 55,000 reacted nonspecifically in all samples tested. Antibody, detectable by microtitration agglutination, was in sera of 1-day-old turkeys and in sera and TW of B avium-infected turkeys during PI weeks 2 to 4.
Show more [+] Less [-]Range of viral neutralizing activity and molecular specificity of antibodies induced in cattle by inactivated bovine viral diarrhea virus vaccines
1990
Bolin, S.R. | Ridpath, J.F.
The range of neutralizing activity to bovine viral diarrhea (BVD) virus and viral protein specificity of antibodies induced by 3 inactivated vaccines were evaluated by use of samples of sera obtained from 13 cattle 14 days after vaccination. Viral neutralizing antibodies were detected in all cattle to each of 10 noncytopathic and 10 cytopathic isolates of BVD virus. A viral-induced polypeptide (53,000 to 56,000 daltons) was detected by radioimmunoprecipitation with serum from all vaccinates. Other viral-induced polypeptides of 115,000, 80,000, 48,000, and 25,000 daltons were precipitated with sera from some vaccinates. Precipitation of those polypeptides was related to the vaccine used. When multiple viral polypeptides were precipitated, the 53,000- to 56 000-dalton polypeptide appeared immunodominant.
Show more [+] Less [-]Ocular and serum disposition kinetics of cloxacillin after topical administration of benzathine cloxacillin and intravenous administration of sodium cloxacillin to calves
1990
Daigneault, J. | George, L.W. | Baggot, J.D.
Disposition kinetics of cloxacillin were examined in calves after topical administration of benzathine cloxacillin and single IV administration of sodium cloxacillin, and the susceptibility of 17 field isolates of Moraxella bovis was measured. For the IV pharmacokinetic phase, sodium cloxacillin was administered at dosage of 10 mg/kg of body weight to male Holstein calves (n = 6, weighing 146 to 170 kg), and serum concentration of cloxacillin was measured thereafter for 10 hours. For the ocular pharmacokinetic phase, 6 calves were given either of 4 benzathine cloxacillin topical formulations consisting of 50-, 125-, 250-, or 375-mg doses. Treatment was repeated every 10 days until all 4 benzathine cloxacillin dosages were tested in the same 6 calves. Blood and tears were collected for 72 hours after each benzathine cloxacillin formulation was administered, and the concentration of cloxacillin in each specimen was measured, using a bioassay. The minimal inhibitory concentration of cloxacillin for 17 field isolates of M bovis was determined by use of an agar pour-plate dilution assay. After single IV administration of sodium cloxacillin, its half-life, body clearance, and volume of distribution were 19.5 +/- 12.8 minutes, 18.3 +/- 2.2 ml/min.kg, and 496 +/- 290 ml/kg, respectively. After topical administration of benzathine cloxacillin, cloxacillin concentration in lacrimal fluid peaked between 30 and 45 minutes and ranged between 963 microgram/ml and 3,256 microgram/ml for the 125- and 375- mg doses, respectively. There was no detectable cloxacillin activity in the lacrimal fluid of any calf by 36 hours after topical administration of benzathine cloxacillin, and cloxacillin was not detected in the serum at any time. The mean lacrimal fluid cloxacillin concentration for the 4 groups during the first 8 hours was not significantly different; however, by 12 hours, the cloxacillin concentration in tears from calves of the 250- and 375-mg groups was significantly (P < 0.05) greater than that in calves of the 50- and 125-mg groups. Cloxacillin concentration greater than or equal to 3.13 microgram/ml was maintained for a significantly (P < 0.05) longer time after treatment, using the 375-mg dose, compared with the 50-mg dose of benzathine cloxacillin. The minimal inhibitory concentration of cloxacillin for 1 isolate was 6.25 microgram/ml, but was less than or equal to 3.13 microgram/ml for 16 other M bovis isolates.
Show more [+] Less [-]Effects of age, sex, and body size on serum concentrations of thyroid and adrenocortical hormones in dogs
1990
Reimers, T.J. | Lawler, D.F. | Sutaria, P.M. | Correa, M.T. | Erb, H.N.
Thyroxine (T4), 3,5,3'-triiodothyronine (T3), and cortisol frequently are quantified in canine serum or plasma samples to aid in the diagnosis of hypothyroidism, hypoadrenocorticism, and hyperadrenocorticism. Many laboratories have established reliable references values for concentrations of these hormones in blood of clinically normal animals. However, nonpathologic factors that affect thyroidal and adrenocortical secretion may lead to misinterpretation of test results when values for individual animals are compared with reference values. The objective of the study reported here was to identify effects of age, sex, and body size (ie, breed) on serum concentrations of T3, T4, and cortisol in dogs. Blood samples were collected from 1,074 healthy dogs, and serum concentrations of the iodothyronines and cortisol were evaluated for effects of breed/size, sex, and age. Mean (+/- SEM) serum concentration of T4 was greater in small (2.45 +/- 0.06 microgram/dl)- than in medium (1.94 +/- 0.04 microgram/dl)- or large (2.03 +/- 0.03 microgram/dl)-breed dogs, the same in females (2.11 +/- 0.04 microgram/dl) and males (2.08 +/- 0.04 microgram/dl), and greater in nursing pups (3.04 +/- 0.05 microgram/dl) than in weanling pups (1.94 +/- 0.05 microgram/dl), rapidly growing dogs (1.95 +/- 0.04 microgram/dl), and young adult (1.90 +/- 0.06 microgram/dl), middle-aged adult (1.72 +/- 0.05 microgram/dl), or old adult (1.50 +/- 0.05 microgram/dl) dogs. Dogs > 6 years old had lower mean serum T4 concentration than did dogs of all other ages, except middle-aged adults. Mean serum T3 concentration in medium-sized dogs (1.00 +/- 0.01 ng/ml) was greater than that in small (0.90 +/- 0.01 ng/ml)- and large (0.88 +/- 0.01 ng/ml)-breed dogs. Serum T3 concentration was lowest in nursing (0.85 +/- 0.01 ng/ml) and weanling (0.77 +/- 0.02 ng/ml) pups, increased in rapidly growing dogs (0.99 +/- 0.01 ng/ml) and young adult dogs (1.10 +/- 0.04 ng/ml), and decreased slightly in middle-aged (0.98 +/- 0.02 ng/ml) and old (1.01 +/- 0.03 ng/ml) adult dogs. Serum T3 concentration was unaffected by sex. Mean serum cortisol concentration was greater in small (1.06 +/- 0.07 microgram/dl)- than in large (0.79 +/- 0.03 microgram/dl)-breed dogs. Serum from nursing pups 0.57 +/- 0.04 microgram/dl) contained less cortisol than did serum from older dogs (mean values greater than or equal to 0.92 microgram/dl). Serum cortisol concentration was not different between males and females. These effects of breed/size and age on serum T3, T4, and cortisol concentrations should be considered when evaluating thyroid and adrenocortical functions in dogs.
Show more [+] Less [-]Methods for detection of immune-mediated neutropenia in horses, using antineutrophil serum of rabbit origin
1990
Jain, N.C. | Vegad, J.L. | Kono, C.S.
Equine neutrophil antibody was raised in rabbits inoculated with equine neutrophils isolated to purity > 99.0%, using Percoll density-gradient sedimentation. Neutrophil antibody was detected by use of agar gel diffusion, leukoagglutination, indirect immunofluorescence, staphylococcal protein A and streptococcal protein G binding, and phagocytic inhibition techniques. Precipitin lines and leukoagglutination were seen in antiserum dilutions of 1:4 and 1:64, respectively. The specific nature of leukoagglutination was characterized by the formation of rosette-like clumps of neutrophils. Specific bright membranous fluorescence was seen in neutrophils treated with the antiserum and exposed to fluorescein-conjugated goat anti-rabbit immunoglobulin, and staphylococcal protein A and streptococcal protein G. Whereas the indirect immunofluorescence and protein G-binding tests were equally sensitive and resulted in titer of 1:256, the protein A-binding test was less sensitive and resulted in titer of only 1:32. Nonspecific binding of protein A and protein G was noticed as uniform or patchy cellular fluorescence in a small number of neutrophils. Treatment of neutrophils with antiserum up to dilution of 1:8 resulted in a significant (P < 0.05) suppression of phagocytosis of opsonized zymosan particles. Thus, protein G-binding and indirect immunofluorescence tests are highly sensitive to detect neutrophil antibody and may be used to diagnose immune-mediated neutropenias in horses and, possibly, in other animal species.
Show more [+] Less [-]Immunohistochemical localization of alpha 2-beta 1-glycoprotein in horses
1990
Winder, N.C. | Pellegrini, A. | Fellenberg, R. von
Alpha 2-beta 1-glycoprotein may be found free in horse serum or complexed with alpha-1-proteinase inhibitor to form pre-alpha 2-elastase inhibitor. There has been little information published concerning alpha 2-beta 1-glycoprotein and its possible tissue sources in horses. A peroxidase-antiperoxidase technique was used to identify alpha 2-beta 1-glycoprotein in buffy coat and bone marrow neutrophils of healthy horses. Macrophages and neutrophils in bronchoalveolar lavage samples from clinically normal horses and from horses with chronic pulmonary disease also were positive for alpha 2-beta 1-glycoprotein. Alpha 2-beta 1-glycoprotein was identified in some instances in normal equine hepatocytes of formalin-fixed liver sections. In formalin-fixed liver sections from horses with chronic, small-airway disease and chronic bronchointerstitial pneumonia, alpha 2-beta 1-glycoprotein was observed in some airway secretions and in macrophages.
Show more [+] Less [-]Effects of ammoniation of endophytic fungus-infested fescue hay on serum prolactin concentration and rectal temperature in beef cattle
1990
A group of 48 crossbred steers (approx 250 kg each) were used to determine the effects of various diets and treatments on serum prolactin concentrations and rectal temperatures. The steers were randomly assigned to groups fed the following: (1) endophyte fungus-infested fescue hay; (2) ammoniated endophyte fungus-infested cattle industry hay; (3) endophyte fungus-infested fescue hay plus 1 g of thiabendazole (TBZ)/9.1 kg of body weight at 7-day intervals; (4) ammoniated endophyte fungus-infested fescue hay plus 1 g of TBZ/9.1 kg at 7-day intervals; (5) ground Bermuda grass hay; and (6) endophyte-fungus-free fescue hay. Blood samples for prolactin determinations and rectal temperatures were obtained biweekly beginning on week 3 and continuing through week 9. A significant correlation (P < 0.05) between low prolactin concentrations and high rectal temperatures in cattle eating endophyte-infested fescue hay was determined; however, as the prolactin concentrations approached normal (control animal) concentrations, this relationship between serum prolactin and rectal temperatures was not observed. Two antifungal agents used in this experiment (thiabendazole and ammonia) appeared to have different effects on the variables measured. Thiabendazole had no significant effect on prolactin concentrations or rectal temperatures of cattle ingesting endophyte-infested fescue hay, whereas ammoniation of this hay induced significantly higher (P<0.05) prolactin concentrations and lower rectal temperatures than in steers receiving only endophyte-infested fescue hay. Therefore, ammonia may be valuable not only as an aid in determining the cause of the fescue problem but also as a practical solution to some of the fescue-related economic problems in the cattleindustry. Also, prolactin concentrations may be valuable in diagnosing fescue-related problems.
Show more [+] Less [-]Failure of pH to predict ionized calcium percentage in healthy horses
1990
Kohn, C.W. | Brooks, C.L.
Blood, serum, and plasma total calcium concentrations and plasma and serum ionized calcium concentrations were anaerobically determined by use of a calcium-specific electrode for samples obtained from 39 healthy horses. Mean (+/- SD) serum ionized calcium concentration was 6.6 +/- 0.3 mg/dl (1.6 +/- 0.1 mmol/L) and the mean serum ionized calcium percentage was 58.2 +/- 3.4%. Serum ionized calcium percentage was not significantly correlated with serum pH. Plasma ionized calcium percentage was weakly correlated with plasma pH (r = - 0.480; P less than or equal to 0.05). Ionized calcium concentration was determined in serum samples manipulated in vitro by additions of 1 to 80 microliter of 0.1N hydrochloric acid or sodium hydroxide to yield 6 to 10 pH values between 6.8 and 8.2. In all horses, the relationship between serum ionized calcium percentage and serum pH at these pH values was then examined by use of a repeated-measures multiple regression analysis. Correlations between serum ionized calcium percentage and adjusted serum pH value for each horse were highly significant (P < 0.05); however, analysis of pooled data from all horses indicated that a statistically significant relationship between serum pH and ionized calcium percentage did not exist. Lack of a significant relationship between these variables was most likely attributable to heterogeneity of variance of ionized calcium percentage among horses, reflecting variation in undefined biochemical constituents of serum that affect the equilibrium of calcium binding. When it is essential to evaluate the calcium status of a horse, direct measurement of serum ionized calcium concentration is recommended.
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