Biomedical subjects

C Burvenich

Publications and source records attributed to C Burvenich.

At least 73 records · Page 4Linked to original sources

PubMed1998

Antibiotics commonly used to treat mastitis and respiratory burst of bovine polymorphonuclear leukocytes.

The in vitro effects of six doses (2 x 10(-3) to 2 x 10(-8) M) of antimicrobial drugs that are frequently used in udder infusions on the capacity of bovine blood polymorphonuclear neutrophilic leukocytes to generate reactive oxygen species were studied by the measurement of luminol-dependent chemiluminescence after stimulation with phorbol 12-myristate 13-acetate. All drugs, except cloxacillin, significantly decreased chemiluminescence at the highest dose. Doxycyline induced the most severe inhibition, followed by neomycin and dihydrostreptomycin. The effect of ampicillin was due to the scavenging of reactive oxygen species and interactions with luminol. The inhibition observed with oleandomycin, neomycin, lincomycin, and dihydrostreptomycin was not due to direct effects on the production of oxidative metabolites but rather to interference with other components involved in the production of light, such as interference with the interaction between luminol and the myeloper-oxidase-H2O2-halide system. The deleterious effects of doxycycline can be explained by several factors: decreased production of superoxide, yellow color, the scavenging of reactive oxygen species, and Ca2+ chelating effect. In conclusion, the results of this study show that antibiotics may affect neutrophil function at concentrations that are reached in the mammary gland after local and repeated administration.

Ampicillin↗

PubMed1998

Reduction of acyloxyacyl hydrolase activity in circulating neutrophils from cows after parturition.

Bovine neutrophils contain the enzyme acyloxyacyl hydrolase, which hydrolyzes the acyloxyacyl linkage of the two nonhydroxylated fatty acyl chains to two 3-hydroxy fatty acids in the highly conserved lipid A part of endotoxins with high specificity. This hydrolysis decreases the toxicity of lipid A, but the immunostimulatory capacity of endotoxins is largely maintained. In two trials, we studied the activity of acyloxyacyl hydrolase in neutrophils that had been isolated from the blood of 18 dairy cows around parturition. Between 10 and 26 d after parturition, the activity of acyloxyacyl hydrolase in neutrophils decreased approximately 20% below prepartum activity. At about 2 mo after parturition, acyloxyacyl hydrolase activity returned to prepartum values. Changes in acyloxyacyl hydrolase activity could not be attributed to changes in binding of lipopolysaccharides by the CD14 molecules on neutrophils or monocytes. We hypothesize that decreased acyloxyacyl hydrolase activity in neutrophils shortly after parturition is a factor that increases the susceptibility of dairy cows to coliform mastitis during early lactation.

Animals↗

PubMed1997

Increased surface expression of CD11b receptors on polymorphonuclear leukocytes is not sufficient to sustain phagocytosis during Escherichia coli mastitis in early postpartum dairy cows.

Phagocytosis, CD11a and CD11b adhesion receptor expression, O2-production and maturity of circulating polymorphonuclear leukocytes (PMN) were studied during acute coliform mastitis in early postpartum dairy cows to obtain a better insight in the role of neutrophils in the pathology of this disease. The mammary gland of twelve newly calved high-yielding dairy cows was experimentally infected with Escherichia coli. Variability in clinical signs of mastitis and inhibition of milk production among cows was very high. There was a significant negative correlation between the number of circulating neutrophils immediately before infection and severity of mastitis represented by the decrease in milk production of non-infected quarters two days after infection. Pre-infection phagocytosis of E. coli, CD11a and CD11b receptor expression, phorbol myristate acetate (PMA)-induced O2-production and maturity of neutrophils on a per cell basis were not related to severity of mastitis. However, significant correlations between severity of mastitis and the total number of phagocytic PMN and mature PMN in blood immediately before infection were found. PMN characteristics responded differently to mastitis depending on the severity of the disease. Neutrophil functions from cows classified as severe (S) and moderate (M) responders to infection of the mammary gland were compared. Surface expression of CD11a receptors on PMN was decreased in all cows 24 h after infection, and this decrease was long-continued in S responders. A biphasic upregulation of the number of CD11b receptors on PMN was observed with a more pronounced response in S cows. PMN phagocytosis was decreased 12 h after infection in S cows and 18 h after infection in S and M cows and was normalized 24 h post-infection. The decrease of phagocytosis coincided with the first peak of CD11b receptor expression. Phorbol myristate acetate (PMA)-induced production of O2-by PMN was decreased for three days after infection in S responders compared to only one day in M responders and was followed by an upregulation. These data demonstrate a complexity in alterations of PMN functions during mastitis and suggest the involvement of differences in systemic factors dependent on severity of mastitis.

Animals↗

PubMed1997

Effect of antibiotics on the phagocytotic and respiratory burst activity of bovine granulocytes.

The influence of antibiotics on respiratory burst (phorbol-12-myristate-13-acetate (PMA)-stimulated luminol-enhanced chemiluminescence) and phagocytosis (flow cytometry) by bovine granulocytes was studied in vitro. Phagocytosis was impaired by 1000 micrograms/ml of oxytetracycline, chloramphenicol, erythromycin and spiramycin. All antibiotics, except sulphadiazine, decreased chemiluminescence at 1000 micrograms/ml or lower concentrations. Enrofloxacin increased chemiluminescence. The inhibition by oxytetracycline and danofloxacin was due to absorption of the light emitted by luminol at 425 nm. Oxytetracycline, ceftiofur, spiramycin and erythromycin affected the myeloperoxidase-H2O2-halide system. Ceftiofur, penicillin and danofloxacin showed scavenging effects on H2O2 and OCI. Penicillin and ceftiofur might interfere with luminol. Chloramphenicol, penicillin and ceftiofur affected the production of superoxide radicals. In summary, the observed effects of antibiotics might be of importance during treatment of infectious diseases in normal and immunocompromised animals. However, before classifying a drug as immunosuppressive, attention has to be paid to possible interference with the chemiluminescence assay.

Animals↗

PubMed1997

Simultaneous flow cytometric measurement of phagocytotic and oxidative burst activity of polymorphonuclear leukocytes in whole bovine blood.

Polymorphonuclear neutrophils (PMN) are important in host defence against bacterial infection in the bovine mammary gland and techniques are needed to evaluate their functional activities. A rapid and sensitive two-color flow cytometric method for simultaneous measurement of phagocytosis rate and oxidative burst activity of bovine PMN in small blood samples is described. The method utilizes the oxidation of intracellular dihydrorhodamine 123 to green fluorescent rhodamine 123 by oxidative burst products that were generated by incubating the PMNs with red fluorescent propidium iodide labeled Staphylococcus aureus. Phagocytosis and oxidative burst both increased with time of incubation and with increasing bacteria concentration. A 20 min phagocytosis time and a ratio of 25 bacteria to one cell were considered optimal for assaying bovine blood PMN activity. To further evaluate the proposed two-color flow cytometric method, blood samples were treated with factors known to interfere with phagocytosis and oxidative burst metabolism of human neutrophils. Incubation of whole bovine blood with cytochalasin B at 10 micrograms ml-1 resulted in an approximate 70% decrease in the phagocytosis rate with a concommitant decrease in oxidative burst activity. Staurosporine (2 micrograms ml-1) inhibited bovine neutrophil oxidative burst formation for 95% while the phagocytosis was unaffected. PMNs in whole blood samples of ten cows differed significantly in their ability to phagocytose Staphylococcus aureus and to produce reactive oxygen products. However, only a weak correlation was detected between the burst activity:phagocytosis ratio of ten individual cows as indicated by the ROD/PI index.

Animals↗

PubMed1997

Influence of antimicrobial agents on bactericidal activity of bovine milk polymorphonuclear leukocytes.

The influence of nine commonly used antibiotics on the respiratory burst activity of bovine milk polymorphonuclear leukocytes (PMNL) of high yielding cows was studied in vitro. Cellular oxidative activity was quantitated after preincubation with drugs at different concentrations and assayed by a PMA (12,13-phorbol myristate acetate)-induced luminol-enhanced chemiluminescence (CL) technique. All antibiotics except sulphadiazine and enrofloxacin decreased CL at the highest concentration. Enrofloxacin significantly increased CL. Oxytetracycline inhibited CL even at low doses. The decreased CL with danofloxacin and oxytetracycline was mainly induced by their color, which caused absorption of the blue light emitted by luminol. Production of superoxide radicals measured by the cytochrome c reduction assay was lowered by danofloxacin, penicillin and chloramphenicol. The decreased CL with ceftiofur was due to inhibition of myeloperoxidase (MPO) activity and to scavenging of reactive oxygen species. Interference with the MPO-H2O2-halide system was also observed with spiramycin, erythromycin and oxytetracycline, while the latter was also observed with penicillin. The stimulatory effect of enrofloxacin might be due to an improvement of the penetration of luminol into the PMNL or to a stimulation of the production of H2O2. Potentiation of the action of PMA by changing the ratio between bound and free intracellular Ca2+ might also be involved. Our results suggest that many antibiotics may affect neutrophil function at concentrations that may be found in milk immediately after intramammary treatment or at concentrations higher than those found in milk after intramammary treatment.

Animals↗

PubMed1997

Elevated levels of beta-hydroxybutyric acid in periparturient cows and in vitro effect on respiratory burst activity of bovine neutrophils.

The in vitro effect of normal (0.01 to 1 mM) and subketotic (1 to 2.5 mM) doses of butyric acid on the respiratory burst activity of bovine polymorphonuclear leucocytes (PMNL) isolated from blood was studied by luminol-enhanced, PMA (phorbol-12-myristate-13-acetate)-induced chemiluminescence (CL). The subketotic concentrations of butyric acid induced a significant inhibitory effect on CL. In a cell free assay, consisting of sonicated cells and H2O2, no changes in activity could be observed. The activity of the myeloperoxidase was not significantly altered as shown by the ortho-dianisidine-oxidation assay. Also, the production of O(2)- measured by the cytochrome c reduction assay was not affected by different doses of butyric acid. Butyric acid had no scavenging effect on hypochlorite. The reason for the inhibitory effect on CL may be a decreased production of H2O2. Indeed, luminol-enhanced CL evaluates the production of H2O2. This could not be confirmed in the other assays mentioned above, because H2O2 was added externally in these assays. In conclusion, because of this inhibitory effect on the respiratory burst activity of PMNL, the elevated blood level of butyric acid after parturition in high yielding cows may be, in part, responsible for the higher susceptibility to local and systemic infections during the postpartum period and during subclinical and clinical ketosis.

3-Hydroxybutyric Acid↗

PubMed1997

Preinfection in vitro chemotaxis, phagocytosis, oxidative burst, and expression of CD11/CD18 receptors and their predictive capacity on the outcome of mastitis induced in dairy cows with Escherichia coli.

Four to 6 wk after parturition, 12 cows in second, fourth, or fifth lactation were experimentally infected in one gland with Escherichia coli. The capacity of chemotaxis, phagocytosis, oxidative burst, and expression of CD11/CD18 receptors to predict the severity of IMI was measured. Bacterial counts in the infected quarter, expressed as area under the curve, and residual milk production in the uninfected quarters were compared to determine severity of the infection. Although these two outcome parameters were highly negatively correlated, regression models with preinfection tests for leukocyte function fitted best with bacterial counts as an outcome parameter. Of the preinfection tests for leukocyte function, chemotaxis best predicted the outcome of the IMI that had been experimentally induced by E. coli. The number of circulating peripheral leukocytes just prior to inoculation was used to predict 52 and 45% of the severity of IMI for bacterial counts and residual milk production, respectively. As a categorical variable, parity predicted 75 and 56% of the severity of IMI expressed as bacterial counts and residual milk production, respectively. Because of the strong effect of parity on the outcome of the experimentally induced mastitis, analysis was performed to discriminate between second parity cows and older cows. Significant differences were found for the number of circulating peripheral leukocytes and for the expression of CD11b/CD18 and CD11c/CD18 receptors between younger and older cows.

Animals↗

PubMed1996

Cell culture system for studying bovine neutrophil diapedesis.

Neutrophils are the major defense against bacterial infection in the bovine mammary gland. Neutrophils migrate from blood into the lumen of the gland in response to inflammatory stimuli. This study describes the development of a system of cell culture that can be used to study neutrophil diapedesis through secretory and ductal mammary epithelial barriers. The culture system consists of successive layers of collagen, fibroblasts, collagen, and a confluent monolayer of secretory or ductal epithelial cells layered on a porous membrane. Confluence was determined by electrical resistance and trypan blue diffusion. Neutrophil diapedesis occurred from the basal to the apical surface of the monolayers. Purified complement C5a, fetal bovine serum that had been activated by zymosan, and fetal bovine serum that had been activated by Escherichia coli induced neutrophil diapedesis. Neutrophil diapedesis was greater across ductal cell monolayers. Blood neutrophils from five cows differed in their ability to migrate through the multilayered culture system in response to C5a. Monoclonal antibodies to C5a blocked diapedesis induced by purified C5a but had no effect on diapedesis induced by fetal bovine serum that had been activated by zymosan or by fetal bovine serum that had been activated by E. coli endotoxin, indicating that factors other than C5a were chemotactic for neutrophils. Monomeric IgG2, immune complexes, and E. coli endotoxin did not induce neutrophil diapedesis.

Animals↗

PubMed1995

The influence of ageing on muscarinic receptors, beta-adrenoceptors and adenylate cyclase activity in the bovine lung.

Muscarinic and beta-adrenoceptors were identified in airway epithelium, smooth muscle and lung parenchyma from Holstein-Friesian calves and cows and were characterized with [3H]quinuclidinyl benzilate and [3H]dihydroalprenolol, respectively. The muscarinic receptor density in the smooth muscle of cows (Bmax = 4803 +/- 245 fmol/mg protein) was 33% greater (p < 0.01) than in calves. Low receptor numbers were detected in the epithelium and parenchyma. In both calves and cows, the density of epithelial beta-adrenoceptors was twice as high as in smooth muscle and parenchyma. The quantity of beta-adrenoceptors in the tracheal epithelium (Bmax = 994 +/- 83 fmol/mg protein) and smooth muscle (Bmax = 492 +/- 41 fmol/mg protein) in cows was respectively 37% (p < 0.001) and 35% (p < 0.01) lower than in calves. Adenylate cyclase (AC) assays indicated that the basal and the (-)-isopropylnoradrenaline- (ISO-) stimulated cAMP production were not significantly different between the calves and cows. After stimulation with NaF, significantly higher cAMP production was found in all tissues from cows. Significant correlations were found between absolute AC responses to NaF and beta-adrenoceptor density in epithelium (r = -0.75, p < 0.001) and smooth muscle (r = -0.63, p < 0.01). It seems that, in older animals, the production of cAMP is independent of the number of receptors, indicating the presence of fully active compensatory mechanisms.

Adenylyl Cyclases↗

PubMed1995

Relationship between numbers of alpha 2- and beta 2-adrenoceptors on blood cells of bulls and milkability of their daughters.

Milk yield and milking time were measured on one occasion for several daughters (n = 6-44) from 16 bulls at morning milkings. Blood from the bulls was collected, and platelets and mononuclear leucocytes were isolated. The alpha 2-adrenoceptors on platelet membranes were identified by binding of [3H]rauwolscine, whereas for the determination of beta 2-adrenoceptors on intact mononuclear leucocytes, [3H]CGP-12177 was used. It was found that mean milk flow rate was highly correlated (P < 0.001) with the alpha 2-adrenoceptor densities on blood platelets. No correlation was found with the beta 2-adrenoceptors on mononuclear leucocytes. It is concluded that estimation of the alpha 2-adrenoceptors on blood platelets from bulls could eventually be used to investigate milking characteristics of cows, and might be useful in the future as a marker in genetic studies.

Animals↗

PubMed1995

Effects of Staphylococcus aureus mastitis after endotoxin application on milk yield and composition during subsequent lactation of guinea-pigs.

The effects of Staphylococcus aureus mastitis on milk yield and composition throughout subsequent lactation in lactating guinea-pigs and the role of endotoxin pretreatment on these phenomena were investigated. Primiparous lactating guinea-pigs were intramammarily inoculated with sterile saline (group 1), S. aureus strain UC 6097 (group 2), or with S. aureus UC 6097 after endotoxin pretreatment (group 3). Clinical signs and survival rate were monitored. During the second lactation, daily milk yield was measured and milk composition was determined. In primiparous inoculated guinea-pigs, moderate (group 3)-to-severe acute mastitis (group 2) was produced. During subsequent lactation, milk yield in the control group peaked on day 5 and then decreased. Concentrations of Na+ and Cl- in milk, and concentrations of fat, gradually increased, but lactose and K+ decreased. After an early decrease, NAGase in milk increased towards the end of lactation. Except for higher NAGase concentrations in group 3, milk yield and composition during the second lactation did not differ significantly between the mastitis and the control groups. Endotoxin pretreatment only plays a role in the determination of the severity of the infection.

Animals↗

PubMed1994

Triacylglycerol fatty acid composition of milk from periparturient cows during acute Escherichia coli mastitis.

Changes in fat concentration and triacylglycerol fatty acid (TGFA) composition were studied in milk from six periparturient cows 1 d before and 20 d after an experimentally induced Escherichia coli mastitis in the fore and rear homolateral quarters. Opposite fore and rear heterolateral quarters remained uninfected and were used as controls. Milk was collected from all individual quarters during the experiment. The fat concentration in milk from infected quarters did not change, but total fat production decreased owing to reduced milk production after the Esch. coli challenge. In milk from the heterolateral uninfected quarters fat concentration rose significantly 48 and 72 h after induction of mastitis, the rise being concomitant with a decrease in milk production. Throughout the experiment similar changes in TGFA composition were observed for both infected and uninfected quarters. There was an increase in all the even, odd-numbered, iso and anteiso short-chain TGFA from day +6 on after induction of mastitis. There was little change in the composition of 16:0 and 18:0 fatty acids, while the long-chain unsaturated fatty acids decreased. Using multivariate analysis, the results are presented visually. The observed changes in the TGFA can be ascribed to changes normally observed in cows' milk soon after parturition.

Animals↗

PubMed1994

Role of the neutrophil leucocyte in the local and systemic reactions during experimentally induced E. coli mastitis in cows immediately after calving.

Mammary leucocytes are the major contributors to natural defence against mastitis after a microorganism has entered the gland. This paper reviews the role of the neutrophil granulocyte during acute coliform mastitis in cows in the periparturient period. Qualitative and quantitative aspects of several neutrophil cell functions before and during experimentally induced infections are briefly discussed.

Animals↗

PubMed1994

Development and maturation of neutrophils.

Inflammatory processes require the activation of immunocompetent cells. In mammals, neutrophil polymorphonuclear leukocytes (PMN) constitute one of the essential body defences against diseases. In this article knowledge of the development and maturation of neutrophils, the control of haematopoiesis and of the factors that regulate neutrophil production is reviewed. As it has recently become apparent that neutrophils can be primed by cytokines to have enhanced functional activity, the future utilization of these growth factors in bovine immunotherapy is briefly discussed.

Animals↗

PubMed1993

Classification of newly calved cows into moderate and severe responders to experimentally induced Escherichia coli mastitis.

In the present study newly calved cows were tentatively classified as moderate and severe responders to experimentally induced Escherichia coli mastitis based upon the reactive oxygen species (ROS)-generating capacity of their blood neutrophils before infection. The groups differed in blood and milk composition prior to infection. This initial classification was supported by the corresponding variation in clinical symptoms and in the changes in milk production and composition measured during mastitis. Responses of newly calved cows to Esch. coli challenge varied from mild to severe symptoms of inflammation in infected glands and differed in the intensity of systemic disturbances and general illness. Losses in milk yield and compositional changes were most pronounced in inflamed glands and in severe responders. In inflamed glands milk yield and composition did not return to preinfection level in either moderate or severe responders. The yields of lactose, alpha-lactalbumin, casein and fat followed the same pattern as milk yield. It is concluded that the severe and long lasting systemic disturbances observed in severe responders can be ascribed to absorption of endotoxin from infected glands into circulation, indicating the important role of endotoxin in the pathology of coliform mastitis in periparturient cows. Evaluation of the ROS-generating capacity of blood neutrophils and blood and milk composition before infection might help to predict the cow's sensitivity to Esch. coli mastitis.

Animals↗

PubMed1993

Effect of clenbuterol on beta-adrenoceptors and adenylate cyclase activity in smooth muscle and epithelium of the trachea of calves.

A potent beta-agonist (clenbuterol) was administered perorally to young calves for 50 days. After this period the animals were slaughtered and beta-adrenoceptor density, ligand affinity, and basal and stimulated adenylate cyclase activities were studied in smooth muscle and epithelium of the trachea. Although the density of lung beta-adrenoceptors was down regulated by clenbuterol, cAMP production remained constant (epithelium) or even increased (smooth muscle). Therefore desensitization of beta-adrenoceptors in the trachea was not observed. This might be a reason for the effectiveness of long-term treatment with beta-agonists.

Adenylyl Cyclases↗

PubMed1993

Preinfection chemotactic response of blood polymorphonuclear leukocytes to predict severity of Escherichia coli mastitis.

Experimental mastitis was induced by inoculating rear right quarters of 10 healthy cows with 10(3) cfu of Escherichia coli. The chemotactic responses of peripheral blood polymorphonuclear leukocytes at d-6, -5, -2, -1, and immediately prior to inoculation were measured. Chemiluminescence of polymorphonuclear leukocytes was measured immediately prior to inoculation. Severity of the experimental mastitis was assessed by bacterial growth in the inoculated quarters. Results of this study indicated that severity of the experimental mastitis may be predicted by the chemotactic response in vitro of polymorphonuclear leukocytes isolated from the peripheral blood at d 2, d 1, and immediately prior to inoculation. The number of circulating polymorphonuclear leukocytes immediately prior to inoculation also showed a negative relationship with the severity of mastitis. No relationship existed between preinfection chemiluminescence of polymorphonuclear leukocytes and the severity of the experimental mastitis.

Animals↗