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Immunomodulatory effects of tulathromycin on apoptosis, efferocytosis, and proinflammatory leukotriene B4 production in leukocytes from Actinobacillus pleuropneumoniae–or zymosan-challenged pigs
2015
Duquette, Stephanie C. | Fischer, Carrie D. | Williams, Alison C. | Sajedy, Saman | Feener, Troy D. | Bhargava, Amol | Reti, Kristen L. | Muench, Gregory P. | Morck, Douglas W. | Allison, Jim | Lucas, Merlyn J. | Buret, Andre G.
OBJECTIVE To investigate the anti-inflammatory and immunomodulatory properties of tulathromycin in vitro and in experimental models of Actinobacillus pleuropneumoniae–induced pleuropneumonia and zymosan-induced pulmonary inflammation in pigs. ANIMALS Blood samples from six 8- to 30-week-old healthy male pigs for the in vitro experiment and sixty-five 3-week-old specific pathogen–free pigs. PROCEDURES Neutrophils and monocyte-derived macrophages were isolated from blood samples. Isolated cells were exposed to tulathromycin (0.02 to 2.0 mg/mL) for various durations and assessed for markers of apoptosis and efferocytosis. For in vivo experiments, pigs were inoculated intratracheally with A pleuropneumoniae, zymosan, or PBS solution (control group) with or without tulathromycin pretreatment (2.5 mg/kg, IM). Bronchoalveolar lavage fluid was collected 3 and 24 hours after inoculation and analyzed for proinflammatory mediators, leukocyte apoptosis, and efferocytosis. RESULTS In vitro, tulathromycin induced time- and concentration-dependent apoptosis in neutrophils, which enhanced their subsequent clearance by macrophages. In the lungs of both A leuropneumoniae– and zymosan-challenged pigs, tulathromycin promoted leukocyte apoptosis and efferocytosis and inhibited proinflammatory leukotriene B4 production, with a concurrent reduction in leukocyte necrosis relative to that of control pigs. Tulathromycin also attenuated the degree of lung damage and lesion progression in A pleuropneumoniae–inoculated pigs. CONCLUSIONS AND CLINICAL RELEVANCE Tulathromycin had immunomodulatory effects in leukocytes in vitro and anti-inflammatory effects in pigs in experimental models of A pleuropneumoniae infection and nonmicrobial-induced pulmonary inflammation. These data suggested that in addition to its antimicrobial properties, tulathromycin may dampen severe proinflammatory responses and drive resolution of inflammation in pigs with microbial pulmonary infections.
Show more [+] Less [-]Effect of exercise on serum markers of muscle inflammation in Spanish Greyhounds
2015
Luca, Vanesa | Barrera, Rafael | Duque, Francisco J. | Ruiz, Patricia | Zaragoza, Concepcion
OBJECTIVE To investigate effects of exercise on hematologic and biochemical values (especially markers of inflammation and muscle damage) in Spanish Greyhounds used for hunting without previous training. ANIMALS 32 Spanish Greyhounds and 31 dogs of other breeds. PROCEDURES Hematologic variables and concentrations of C-reactive protein (CRP) and other biochemical markers were compared in samples obtained from Spanish Greyhounds 24 hours after exercise (eg, a hunting race) and 2 months after exercise (ie, at rest) and from non–Spanish Greyhounds at rest. All dogs were healthy. Hematologic and biochemical analyses were performed within 24 hours after samples were obtained, and results were compared by means of a Student t test. RESULTS CRP concentration and muscle enzyme (creatine kinase, lactate dehydrogenase, and aspartate aminotransferase) activities were significantly higher and serum iron concentration was significantly lower for Spanish Greyhounds after exercise than at rest. The WBC and neutrophil counts were significantly higher after exercise then at rest. Plasma alanine transaminase activity and total protein, calcium, and phosphorus concentrations were significantly higher after exercise than at rest. Spanish Greyhounds at rest had higher RBC counts, PCVs, and hemoglobin concentrations and lower WBC, neutrophil, and lymphocyte counts, compared with values for non–Spanish Greyhounds at rest. CONCLUSIONS AND CLINICAL RELEVANCE Exercise of Spanish Greyhounds without prior training activated an acute-phase response represented by an increase in serum CRP concentration and decrease in serum albumin and iron concentrations. These changes, along with leukocytosis and neutrophilia, were indicative of a subclinical inflammatory state in Spanish Greyhounds.
Show more [+] Less [-]Evaluation of serum amyloid A and haptoglobin concentrations as prognostic indicators for horses with inflammatory disease examined at a tertiary care hospital
2015
Westerman, Trinia L. | Tornquist, Susan J. | Foster, Crystal M. | Poulsen, Keith P.
OBJECTIVE To evaluate use of serum amyloid A (SAA) and haptoglobin concentrations as prognostic indicators for horses with inflammatory disease in regard to euthanasia, complications, and hospitalization duration and cost. ANIMALS 20 clinically normal horses and 53 horses with inflammatory disease. PROCEDURES Total WBC count, neutrophil count, and fibrinogen, SAA, and haptoglobin concentrations were determined for clinically normal horses and horses with suspected inflammatory disease. Clinicopathologic values at admission were compared to test the use of SAA and haptoglobin concentrations in predicting euthanasia, complications, and hospitalization duration and cost. Haptoglobin and SAA concentrations of 22 horses were monitored during hospitalization to test the use of serial measurements in predicting survival and complications. RESULTS Neutrophil count and SAA and haptoglobin concentrations were significantly different at admission for horses with inflammatory disease, compared with those for clinically normal horses. Horses with colitis and peritonitis had significantly higher SAA and haptoglobin concentrations than clinically normal horses. A moderate positive correlation (r = 0.355) between hospitalization duration and haptoglobin concentration was identified. Horses with an increase in SAA concentration between 24 and 72 hours after admission, compared with admission SAA concentration, were significantly more likely (OR, 7.0; 95% confidence interval, 1.1 to 45.9) to be euthanized or develop complications. CONCLUSIONS AND CLINICAL RELEVANCE Concentrations of SAA and haptoglobin at admission were not significantly correlated with outcome in horses with inflammatory conditions. Acute-phase proteins likely have more utility in serial analysis rather than testing at a single time point for horses with inflammatory conditions.
Show more [+] Less [-]Evaluation of the ability of a gravitational filtration system to enhance recovery of equine bone marrow elements
2015
Mundy, Lauren N. | Ishihara, Akikazu | Wellman, Maxey L. | Bertone, Alicia L.
OBJECTIVE To assess efficiency of gravity filtration to enhance recovery of equine bone marrow elements including stem and progenitor cells. ANIMALS 12 healthy adult horses. PROCEDURES Bone marrow aspirates were collected from the fifth sternebral body and filtered by gravitational flow to obtain bone marrow elements. Raw and harvested bone marrow and marrow effluent were evaluated for WBC and platelet counts, automated and cytomorphologic cell differential counts, mesenchymal stem cell CFUs, cell viability, and differentiation capacity. Isolated cells were analyzed for CD90 and major histocompatibility complex (MHC) class I and II antigens. RESULTS Mean cell viability of harvested bone marrow was 95.9%. Total WBCs and platelets were efficiently captured on the filter (> 95%), and mean recovery in harvested bone marrow was 30%. Cytologic cell differential counts indicated that the percentage of neutrophils was significantly less and the progenitor cell population was significantly higher and concentrated 1.56-fold in harvested bone marrow, compared with results for raw bone marrow. Flow cytometry and cell culture were used to characterize harvested bone marrow cells as positive for expression of CD90 and negative for MHCI and MHCII, which indicated stem cells with a multipotent phenotype that differentiated into chondrocytes, osteocytes, adipocytes, and tenocytes. CONCLUSIONS AND CLINICAL RELEVANCE Gravitational filtration of bone marrow efficiently yielded platelets and cells and produced a progenitor-enriched, leukocyte-reduced product, compared with raw bone marrow.
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