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Pharmacokinetic behavior of doxycycline after intramuscular injection in sheep
2012
Castro Robles, Luis J. | Sahagún Prieto, Ana M. | Diez Liébana, M Jose | Fernández Martínez, Nelida | Sierra Vega, Matilde | García Vieitez, Juan J.
Objective: To determine the pharmacokinetics of a commercial formulation of doxycycline hyclate after IM administration of a single dose to sheep. Animals: 11 healthy domestic sheep. Procedures: For each sheep, doxycycline was administered as a single dose of 20 mg/kg, IM. Blood samples were obtained prior to and for 84 hours after doxycycline administration. Plasma concentrations of doxycycline were determined via high-performance liquid chromatography with UV detection. Pharmacokinetic data were analyzed with noncompartmental methods. Results: Mean ± SD values for pharmacokinetic parameters included maximum plasma concentration (2.792 ± 0.791 μg/mL), time to reach maximum plasma concentration (0.856 ± 0.472 hours), mean residence time (91.1 ± 40.78 hours), elimination half-life (77.88 ± 28.45 hours), and area under the curve (65.67 ± 9.877 μg•h/mL). Conclusions and Clinical Relevance: Results indicated that doxycycline had prolonged absorption and elimination in sheep after IM administration. A daily dose of 20 mg/kg would be sufficient to reach effective plasma concentrations against Chlamydia spp (minimum inhibitory concentration, 0.008 to 0.031 μg/mL) and Staphylococcus aureus (minimum inhibitory concentration, 0.12 μg/mL). Doxycycline administered IM could be an option for therapeutic use in sheep, although further studies are needed.
Mostrar más [+] Menos [-]Effect of a trivalent vaccine against Staphylococcus aureus mastitis lymphocyte subpopulations, antibody production, and neutrophil phagocytosis
2005
Lee, J.W. | O'Brien, C.N. | Guidry, A.J. | Paape, M.J. | Shafer-Weaver, K.A. | Zhao, X.
The effect of a novel bovine mastitis trivalent vaccine, containing Staphylococcus aureus capsular polysaccharide type 5 (T5), 8 (T8), and 336 (T336), on lymphocyte subpopulations, antibody production, and neutrophil phagocytosis was evaluated. Twenty pregnant heifers were immunized with either the trivalent alone, trivalent emulsified in Freund's incomplete adjuvant (FICA), trivalent in aluminum hydroxide, or adjuvant only (FICA). Immunization was done 30 d before the expected calving date followed by 2 boosts in a 2-week interval. Compared to FICA, serum antigen-specific immunoglobulin (Ig)G1 and IgG2 were significantly increased in all the vaccinated groups before parturition and sustained until 3 wk postpartum. In comparison with the trivalent alone, formulation with either adjuvant enhanced production of IgG2, but not IgG1. Immune sera, which contained the highest amount of antibodies, slightly increased neutrophil phagocytosis to the 3 serotypes of killed S. aureus, but most of the differences were not significant due to large variation between the cows. The percentage of CD4+ lymphocyte was significantly higher in vaccinated groups than that of FICA 4 wk after the primary immunization. In comparison with FICA, cows inoculated with trivalent vaccine and adjuvants had an increased percentage of CD8+ lymphocytes at 2 time points, 2 wk before and after calving. Our results indicated that the whole cell trivalent vaccine, with or without adjuvants, is able to elicit antibody responses specific to the 3 capsular polysaccharide antigens. The increase of T8-specific IgG2 was more noticeable when the vaccine was emulsified with adjuvants.
Mostrar más [+] Menos [-]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.
Mostrar más [+] Menos [-]Functional variation in endogenous and exogenous immunoglobulin binding to bovine neutrophils relative to parturition
1993
Berning, L.M. | Paape, M.J. | Peters, R.R.
Ten healthy first- and second-lactation Holstein cows were observed from 1 week before to 1 week after calving and at postpartum day 30 to determine polymorphonuclear neutrophil (PMN) functional variation and immunoglobulin binding profiles. Blood and mammary PMN were obtained 3 times weekly and within 24 hours of calving. Functional traits measured included phagocytosis of Stapbylococcus aureus and in vitro chemotaxis through micropore filters in a Boyden chamber. Additionally, PMN were evaluated for endogenous binding of IgG1, IgG2, IgA, and IgM before and after in vitro chemotaxis. Exogenous binding of the same isotypes was determined after incubation in pooled colostrum, purified immunoglobulin, and pooled sera. Phagocytosis results indicated a significant and transient increase in percentage of milk PMN with associated, rather than phagocytosed, bacteria for 1 week after calving. Blood PMN phagocytosis was not significantly different during this period. Though total chemotaxis was essentially unchanged, the percentage of PMN that were unable to complete migration increased substantially on the day of calving, an effect that disappeared by postpartum day 4. A significant (P < 0.01) positive correlation (r = 0.29) between percentage of PMN migrating completely through the micropore filter and percentage of blood PMN with associated bacteria was observed. Changes were not observed in endogenous immunoglobulin binding, with the exception of a peak in relative fluorescence intensity for IgG1 on the day of calving; this disappeared within 2 days after calving. Correlations between relative intensities of IgG2, and IgM, and percentage of mammary neutrophils phagocytosing were 0.37 and 0.70. Exogenous binding of antibody to blood neutrophils before chemotaxis was generally accomplished most effectively by pooled colostrum, whereas use of pooled sera markedly reduced binding and percentage and intensity of IgM in all cases. Binding of all isotypes was slightly higher before than after calving. Incubation of blood neutrophils in isotypes G1, G2, A, and M after chemotaxis yielded lower immunoglobulin binding among all isotypes, particularly IgM. Fluctuations in neutrophil function were observed immediately around parturition, and these changes correlated strongly with endogenous immunoglobulin-binding profiles.
Mostrar más [+] Menos [-]Cytologic responses of Staphylococcus aureus-infected mammary glands of heifers to interferon gamma and interleukin-2 treatment
1993
Quiroga, G.H. | Sordillo, L.M. | Adkinson, R.W. | Nickerson, S.C.
Cytologic and bacteriologic responses, and changes in cytokine activity were evaluated in secretions of Staphylococcus aureus-infected mammary glands after treatment of heifers with recombinant bovine interferon gamma (rbIFN gamma) or interleukin 2 (rbIL-2). Two groups of 4 heifers each, experimentally infected with 10(7) colony-forming units (CFU) of S aureus, were injected in 2 quarters via the teat canal, with 10(5) U of rbIFN gamma (trial 1) or 7.5 X 10(5) U of rbIL-2 (trial 2) 2 weeks after experimentally induced infection; control quarters received phosphate-buffered saline solution. Mammary secretion samples were taken on days 0, 1, 2, 3, 4, 7, and 14 after cytokine infusion. Secretions were diluted 1:10 and used to perform somatic cell counts (SCC), differential cell counts, and CFU enumerations, and to determine the number of leukocytes expressing major histocompatibility complex class-II (MHC II) antigens. In addition, mammary secretion samples taken on days 0, 1, and 2 were processed to obtain skimmed milk for evaluation of rbIFN gamma- and rbIL-2-like activities. Treatment with rbIFN gamma did not influence SCC, or differential or bacteria counts, or the number of leukocytes expressing MHC II antigens. However, rbIL-2 stimulated leukocytosis, which may have reduced bacteria counts early in the trial; treatment with this cytokine also increased the neutrophil, macrophage, lymphocyte, and eosinophil counts in secretions. Similarly, numbers of MHC II-positive leukocytes were greater in rbIL-2-treated quarters vs controls. Compared with day 0, IFN gamma-like activity was increased on only day 1 in both trials. Interleukin-2-like activity was not influenced in the rbIFN gamma trial, but was increased on days 1 and 2 in the rbIL-2 trials. Results indicated that neither cytokine may have had a major influence on the course of established S aureus infections. However, the increased SCC in rbIL-2-treated quarters may have accounted for the reduction in CFU throughout the trial after treatment with this cytokine. Greatest cytokine-like activity was observed on day 1; however, the consequences of cytokine activity, such as the sustained eosinophilia after rbIL-2 treatment, were detected over the 14-day trial period, indicating possible prolonged action.
Mostrar más [+] Menos [-]Experimentally induced Staphylococcus aureus mastitis in selenium-deficient and selenium-supplemented dairy cows
1990
Erskine, R.J. | Eberhart, R.J. | Scholz, R.W.
Ten Holstein cows were fed a selenium-deficient (SeD) diet containing 0.04 mg of Se/kg of dry matter for 3 months before and throughout their first lactation. A selenium-supplemented (SeS) group of 10 cows was fed an additional 2 mg of Se/head/d to increase dietary Se concentration of the dry matter to approximately 0.14 mg/kg of body weight. An intracisternal challenge exposure of 40 to 60 colony-forming units (CFU) of Staphylococcus aureus was administered into 1 or 2 quarters of the udder of each trial cow at about the twenty-second week of lactation. Blood Se concentration (microgram/ml +/ - SEM) at the time of challenge exposure was 0.035 +/- 0.002 in SeD and 0.139 +/- 0.006 in SeS cows. Infections were established in 14/16 of the challenge-exposed quarters in SeD and 16/19 of the challenge-exposed quarters in SeS cows. The infection in 1 quarter of each Se group cleared without treatment by the end of the 8-week trial period. Log10 peak bacterial concentrations in milk from infected SeD quarters (5.04 +/- 0.25 CFU/ml) were higher (P < 0.05) than those of infected SeS quarters (4.40 +/- 0.12 CFU/ml). Log10 peak somatic cell count (SCC) in milk from infected SeD quarters (7.18 +/- 0.08 cells/ml) did not differ from that of SeS quarters (7.17 +/- 0.05 cells/ml). Peak bacterial concentrations were attained sooner (P < 0.05) in SeD quarters (9.5 +/- 4.0 days) than in SeS quarters (20.7 +/- 3.1 days). Similarly, peak SCC were reached earlier (P < 0.05) in SeD (4.3 +/- 1.1 days) than in SeS quarters (13.3 +/- 3.8 days). The Se groups did not differ significantly with respect to peak milk concentrations of bovine serum albumin or IgG1. Throughout the 8-week trial, the Se groups did not differ significantly in milk bacterial concentration, SCC, bovine serum albumin, or IgG1. Selenium status did not affect the percentage of challenge exposures resulting in infection, duration, or severity of experimentally induced S aureus mastitis.
Mostrar más [+] Menos [-]Comparison of the effect of polysulfated glycosaminoglycan, corticosteroids, and sodium hyaluronate in the potentiation of a subinfective dose of Staphylococcus aureus in the midcarpal joint of horses
1989
Gustafson, S.B. | McIlwraith, C.W. | Jones, R.L.
Four groups of 8 horses each had 1 midcarpal joint injected with 33 colony-forming units (CFU) of viable Staphylococcus aureus plus: 1 ml of saline solution (group 1, control), 250 mg of polysulfated glycosaminoglycan (PSGAG, group 2), 100 mg of methylprednisolone acetate (group 3), or 20 mg of sodium hyaulronate (group 4). Horses were euthanatized, and samples were obtained on the basis of clinical signs of septic arthritis that were nonresponsive to phenylbutazone administration. One group-1 horse, all 8 group-2 horses, 3 group-3 horses, and 4 group-horses 4 were culture-positive for S aureus and had clinical signs, results of synovial fluid analysis, and histopathologic findings that were consistent with sepsis. The addition of 250 mg of PSGAG increased the development of sepsis significantly (P = 0.001), compared with results in control horses. Differences in the development of sepsis between horses injected with methylprednisolone acetate or sodium hyaluronate and control horses were not significant.
Mostrar más [+] Menos [-]Further investigations into the potentiation of infection by intra-articular injection of polysulfated glycosaminoglycan and the effect of filtration and intra-articular injection of amikacin
1989
Gustafson, S.B. | McIlwraith, C.W. | Jones, R.L. | Dixon-White, H.E.
Polysulfated glycosaminoglycan (PSGAG) recently have been reported to potentiate the infectivity of Staphylococcus aureus in horses with experimentally induced septic arthritis. Four groups of 8 horses each had 1 midcarpal joint injected with approximately 33 viable colony-forming units (CFU) of S aureus plus either 1 ml of saline solution (group 1), 250 mg of PSGAG (group 2), 250 mg of PSGAG passed through a 0.6-micrometer filter (group 3), or 250 mg of PSGAG plus 125 mg of amikacin (group 4). Horses that developed clinical signs consistent with sepsis were euthanatized, and samples were collected at necropsy. Horses that survived had samples obtained by use of arthroscopy at days 13 and 14 after injection. Staphylococcus aureus was isolated from 1 group-1 horse, 8 group-2 horses, and 7 of 7 group-3 horses that met protocol, but was not isolated from any group-4 horses. All 16 aforementioned horses had clinical signs, results of synovial fluid analysis, and gross pathologic and synovial membrane histopathologic findings that were consistent with septic arthritis. Polysulfated glycosaminoglycan (250 mg) increased the infectivity of 33 CFU of S aureus (P = 0.001); filtering the PSGAG had no effect. Intra-articular injection of 125 mg of amikacin immediately after inoculating the joint with 33 CFU of S aureus significantly (P = 0.001) decreased potentiation of infection by the PSGAG.
Mostrar más [+] Menos [-]Sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood
1988
Paape, M.J. | Miller, R.H.
A study was conducted to examine sources of variation introduced into a phagocytosis assay as a result of the isolation of neutrophils from bovine blood, including variation attributable to isolation of neutrophils from blood, variation between duplicate determinations of percentage phagocytosis, and the variation in the ability of neutrophils isolated from blood (over repeated collections from the jugular vein) to phagocytose. For the phagocytosis assay, jugular venous blood from each of 4 cows was divided into 2 equal portions. The neutrophils were isolated by lysis of red blood cells with 0.2% sodium chloride. The neutrophils (2 X 10(7)) were incubated in duplicate with 32P-labeled Staphylococcus aureus ([32P]SA; 2 X 10(8)) inskimmed milk samples (2.5% final concentration) prepared from 4 cows. This process was repeated thrice on neutrophils isolated from 4 cows at 2-week intervals. The proportions of variation in percentage of 32P-labeled S aureus phagocytosed between duplicate neutrophil isolations and between duplicate assay determinations were 0 and 1%. Differences among skimmed milk sources and among runs, using blood neutrophils taken at different times from the same donor cow, accounted for 62 and 36% of the total variation. The results indicated that variation arising from blood neutrophil isolation introduced into a phagocytosis assay within a single-day trial is of no concern. The large variation among skimmed milk sample sources indicated differences among cows in the ability of their milk to support phagocytosis. The variation in neutrophil isolations over time for any cow was considered too large to allow for evaluation of physiologic and environmental effects on phagocytosis of neutrophils isolated from blood.
Mostrar más [+] Menos [-]Prevalence and characterization of Staphylococcus aureus in young goats
1988
Centorbi, O.N.P. de | Cuadrado, A.M.A. de | Carrizo, S.C.C. de | Milan, M.C.C. de | Gimenez, D.F. | Bergdoll, M.S.
Thirty-six Staphylococcus aureus isolates recovered from 35 of 204 young goats at slaughter were characterized. All isolates were susceptible to cephalothin, clindamycin, chloramphenicol, gentamicin, kanamycin, and amikacin. All but 2 were susceptible to erythromycin and tetracycline, and 19 and 20 were susceptible to penicillin and ampicillin, respectively. Thirteen isolates were classified as biotype A, 9 isolates were classified as biotype B, 8 isolates were clssified as biotype C, and 6 isolates were classified as intermediate between B and C or were not biotypable. Six biotype A isolates were enterotoxigenic; 4 produced enterotoxin B, 1 produced enterotoxin C, and 1 produced enterotoxin D. Two biotype B strains produced enterotoxin B, and all 8 biotype C isolates produced enterotoxin C and the toxic shock syndrome toxin-1.
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