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inflammation. These findings should pave the way toward in vivo experiments to test the effects of fully oxidized β-carotene in feedlot cattle. ABBREVIATIONS BAL Bronchoalveolar lavage EBTr Bovine embryonic tracheal FITC Fluorescein

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in American Journal of Veterinary Research

Abstract

Objective

To determine the role of tissue factor (TF) in the coagulation events leading to intra-alveolar fibrin deposition and intravascular thrombosis associated with pneumonic pasteurellosis in cattle.

Animals

Healthy 2- to 4-week-old male Holstein calves.

Procedures

Blood and bronchoalveolar lavage samples were collected before and at 1, 2, 4, and 6 hours after inoculation of saline solution or Pasteurella haemolytica. Total leukocyte count, platelet count, plasma total protein concentration, prothrombin time, and partial thromboplastin time were measured in blood samples. Total nucleated cell count, total protein concentration, and procoagulant activity were measured in bronchoalveolar lavage samples. Additionally, platelet survival in blood, platelet accumulation in affected lung tissue, and gross and microscopic lung lesions were determined.

Results

Administration of TF monoclonal antibodies (MAB) TF1-1F7 prevented the decrease in platelet survival and the increase in bronchoalveolar lavage fluid TF-dependent procoagulant activity observed in calves not treated with MAB TF1-1F7 antibody, but did not attenuate the increase in lavage fluid neutrophil numbers and total protein concentration. MAB TF1-1F7 administration reduced the percentage of lung affected by pneumonic lesions from 51.81 % to 10.40% and attenuated intra-alveolar deposition of fibrin, neutrophils, and erythrocytes.

Conclusion

Intra-alveolar fibrin deposition and activation of coagulation in cattle with pneumonic pasteurellosis is, at least in part, mediated by TF.

Clinical Relevance

Treatments that neutralize TF activity may attenuate lung injury in cattle with pneumonic pasteurellosis (Am J Vet Res 1997;58:28–33)

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in American Journal of Veterinary Research

SUMMARY

Alpha 2-(β1-glycoprotein may be found free in horse serum or complexed with α-1-proteinase inhibitor to form pre-α2-elastase inhibitor. There has been little information published concerning α2-β1-glycoprotein and its possible tissue sources in horses. A peroxidase-antiperoxidase technique was used to identify α2-(β1 -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 α2-(β1-glycoprotein. Alpha 2-β1-glycoprotein was identified in some instances in normal equine hepatocytes of formalin- fixed liver sections. In formalin-fixed lung sections from horses with chronic, small-airway disease and chronic bronchointerstitial pneumonia, α2-β1-glycoprotein was observed in some airway secretions and in macrophages.

Free access
in American Journal of Veterinary Research

Abstract

Objective

To investigate the effects of anti-inflammatory drugs on lipopolysaccharide-induced procoagulant activity of bovine alveolar macrophages.

Design

Procoagulant activity was induced in bovine alveolar macrophages from 4 healthy Holstein calves aged 6 to 16 weeks by incubation with lipopolysaccharide. 3 anti-inflammatory drugs were used at 4 concentrations and 3 times to pretreat the alveolar macrophages. Results were analyzed to determine whether drug, concentration, or exposure period had a significant (P> 0.05) effect.

Procedure

Bovine alveolar macrophages, harvested by volume-controlled bronchoalveolar lavage, were pretreated for 30, 60, or 120 minutes with an anti-inflammatory compound (dexamethasone, flunixin meglumin, or phenylbutazone) at several concentrations (0, 1, 10, and 100 μM). Bovine alveolar macrophages were exposed to lipopolysaccharide (Escherichia coli O55:B5) in the presence and absence of fetal bovine serum for 4 hours. Procoagulant activity was measured, using a chromogenic assay.

Results

None of the drugs was associated with a modification of procoagulant activity expression.

Conclusion

Use of these 3 anti-inflammatory drugs is unlikely to modify the extent of the fibrinous reaction commonly observed in cases of acute bovine respiratory tract disease complex.

Clinical Relevance

The alveolar macrophage has a key role in fibrin production. Assuming in vivo events mimic the in vitro model, is appears unlikely that administration of anti-inflammatory drugs will reduce the procoagulant activity of the bovine alveolar macrophages and the directly associated pulmonary fibrosis. (Am J Vet Res 1996; 57:659–663)

Free access
in American Journal of Veterinary Research

Abstract

Objective

To characterize the relation between bronchoalveolar and blood eosinophil numbers, serum total IgE concentration, and nonspecific airway reactivity in healthy dogs.

Animals

26 healthy Beagles.

Procedure

Prior to measurement of nonspecific airway responsiveness, dogs were anesthetized and bronchoscopy was performed to recover bronchoalveolar lavage (BAL) fluid. Repeated measurements were made in 6 dogs.

Results

The percentage of blood eosinophils varied between 0 and 13 (mean ± SD, 5.6 ± 3.6) %, the percentage of eosinophils in BAL fluid ranged between 0 and 63.5 (8.8 ± 12.9) %, and total serum IgE concentration was 0.1 to 107.5 (23.4 ± 29.1) U/ml. A strong association was evident between numbers of blood eosinophils and total serum IgE concentration (R 2 = 0.413, P < 0.001), and a trend toward an association between numbers of blood eosinophils and numbers of eosinophils in BAL fluid was apparent (R 2 = 0.110, P = 0.053). Significant associations were not found between any other aspects of the blood and BAL fluid cell profiles and total serum IgE concentration or airway reactivity. Serum total IgE concentration was not associated with airway reactivity. Further, in dogs examined on repeated occasions, variation in BAL fluid eosinophil numbers was not associated with any change in serum total IgE concentration or airway reactivity.

Conclusions

Neither numbers of bronchoalveolar or blood eosinophils nor serum total IgE concentration have a significant role in determining airway reactivity in healthy dogs. (Am J Vet Res 1997;58:34–39)

Free access
in American Journal of Veterinary Research

SUMMARY

Total protein concentration was determined in serum, bronchoalveolar lavage (bal) fluid, and nasal flush fluid obtained from specific-pathogen-free cats from birth to maturity and from adult conventionally raised cats. Protein components were analyzed by Immunoelectrophoresis and isoelectric focusing. Albumin, and α-, β-, and γ-globulins were among the proteins identified in bal fluid, and their isoelectric point ranged from 3.1 to 5.1. γ-Globulin was not detected in serum or bal fluid of newborn kittens before they had ingested colostrum. By day 3 after ingestion of colostrum, IgG was detected in high concentration in serum and was the predominant immunoglobulin in serum and bal fluid of older cats. Nasal flush fluid from cats > 6 months old contained albumin, and α-, β-, and γ-globulins, with IgA being the predominant immunoglobulin. Total protein concentration in nasal flush fluid increased progressively with increasing age, and albumin was the predominant protein. Protein concentration was significantly (P < 0.01) higher in bal fluid from conventionally raised adult cats than in that from specific-pathogen-free cats.

Free access
in American Journal of Veterinary Research

Abstract

Objective

To characterize cytokine profiles and lymphocyte subpopulations in lung parenchyma and bronchoalveolar lavage (BAL) fluid from normal bovine lungs.

Animals

Eight 12- to 18-month-old cattle.

Procedure

Cell populations in BAL fluid and collagenase-digested lung parenchyma were analyzed by flow cytometry and monoclonal antibodies. Proportions of total cell populations were determined, using Giemsa-stained cytospots. Distribution of lymphocytes within the lung parenchyma was analyzed by immunohistochemistry, and cytokine mRNA species in the parenchyma were characterized by use of reverse transcriptase-polymerase chain reaction analysis.

Results

Cytokine profiles indicated high amounts of mRNA for interleukins 6 and 10 and transforming growth factor β. In the BAL fluid and lung parenchyma, macrophages were the predominant cell type, although the proportion was lower in the parenchyma. Lymphocytes made up approximately 3% of both cell populations. Common to both lung compartments was the predominance of CD2+ and γδ T cells over B lymphocytes. There were more CD8+ T cells than CD4+ T cells in both compartments. The γδ cells made up approximately 9% of the lymphocyte populations. Two-color flow cytometry revealed CD8+ γδ T cell and CD8+CD5 populations that were unique to BAL fluid. In the BAL fluid and parenchyma, most CD4+ and CD8+T cells expressed high amounts of CD44, a characteristic of memory T cells. The γδ T cells were CD44lo, as were B cells in the lung parenchyma. The B cells from BAL fluid expressed high amounts of CD44. Immunohistologic analysis of lung tissue revealed bronchus-associated lymphoid tissue structures with distinctive germinal center organization of B cells encompassed by CD4+ T cells.

Conclusions

Results provided normal values for comparison with those of other species and with the bovine respiratory tract response to disease. (Am J Vet Res 1997;58:969–975)

Free access
in American Journal of Veterinary Research

SUMMARY

Five newborn isolation-reared colostrum-deprived calves were inoculated orally and intranasally when they were 20 to 30 hours old and challenge exposed when they were 21 days old with a suspension of virulent bovine coronavirus (bcv). Blood, feces, nasal swabs, tears, saliva, and bronchoalveolar lavage (bal) fluids were collected from each calf prior to inoculation and then weekly for 5 postinoculation weeks. An elisa was used to quantitate the immunoglobulin isotype titers of bcv antibodies in all samples, An immunoblot assay was used to determine the antibody isotype responses to bcv structural proteins in all the samples, except saliva.

At postinoculation days 2 to 3, all calves had severe watery diarrhea, shed bcv in their feces, and had evidence of bcv replication in their upper respiratory tract. After challenge exposure, no calves became ill and no evidence of bcv replication in the respiratory or intestinal tracts was detected. At postinoculation week 1, IgM responses to the N protein were seen in mucosal secretions (except nasal fluid) and feces. At postinoculation weeks 2 and 3, IgA was predominant in mucosal secretions and feces directed toward all the bcv proteins (except the E2 protein in bal fluid). After challenge exposure, an increase (or failure to decrease) in most IgA and some IgG1 titers to bcv proteins was seen. The increases in IgA titers were to all viral proteins in all mucosal secretions and feces, except to the N and E1 viral proteins in feces. The IgGl titer increases were to the E2 proteins in tears and bal fluid and to the E3 protein in bal fluid. In serum, IgM to the N and E3 proteins appeared first, followed by IgGl to mainly the N and E2 proteins, and then more moderate and slower IgG2 and IgA responses. Challenge exposure resulted in an increase (or failure to decrease) in IgGl reactions to all bcv proteins; in IgG2 reactions to E2 and E3 proteins; in IgA reactions to E1, E2, and E3 proteins; and IgM reactions to the N protein only. The N protein elicited the greatest antibody responses, followed by the E2 and E3 proteins, and least by the E1 protein in serum, feces, or mucosal secretions. The E1 and E2 proteins elicited no detectable IgM responses in serum or mucosal secretions.

Our findings indicate that there may be local antibody production at mucosal sites of viral replication, as well as antibody production at associated systemic sites resulting in serum antibodies. Immunoglobulin A antibodies on mucosal surfaces to some or all of the bcv proteins may be important in protection from bcv reinfection.

Free access
in American Journal of Veterinary Research

SUMMARY

Ten colostrum-deprived calves were assigned to 1 of 2 treatment groups (5 calves/group), and fed colostrum that had either low (naturally infected cows) or high (immunized cows) antibody titers to bovine coronavirus (bcv). All calves were inoculated orally and intranasally with virulent bcv when they were 24 to 48 hours old and challenge exposed 21 days later. Blood, feces, nasal secretions, tears, saliva, and bronchoalveolar lavage (bal) fluids were collected weekly from each calf for 5 weeks after inoculation. The titers to whole bcv or the relative amounts of isotype-specific antibodies to bcv structural proteins were evaluated in these samples by ELISA or immunoblotting, respectively.

Both pools of colostrum contained primarily IgG1, IgG2, and IgA antibodies to the E2 and E3 bcv proteins. Calves fed the high-titer colostrum had correspondingly higher amounts of passive IgG1 and IgA antibodies to whole bcv and to the E2 and E3 bcv proteins in serum, feces, and bal fluid at postinoculation week 1 than those calves fed low-titer colostrum. Active IgG1, IgA, and IgM antibody responses in serum and active IgA and IgM antibody responses in most mucosal secretions to whole bcv and to the E2 and E3 proteins were lower or delayed in calves fed high-titer colostrum, compared with responses in calves fed low-titer colostrum. In contrast, increased responses to the bcv N protein were observed in all samples (except in serum and bal fluid) in the calves fed high-titer colostrum, compared with calves fed low-titer colostrum. Upon challenge exposure, responses to E2 and E3 bcv proteins in serum and bal fluid were lower in the group fed high-titer colostrum, compared with those in the group fed low-titer colostrum.

Our findings indicate that the level of passive immunity in calves at the time of bcv inoculation can influence the development of active antibody responses in serum, feces, and mucosal secretions to whole bcv and to some bcv proteins individually.

Free access
in American Journal of Veterinary Research

J 2014 ; 46 : 276 – 288 . 10.1111/evj.12204 3. Bullone M , Lavoie JP . Science-in-brief: equine asthma diagnosis: beyond bronchoalveolar lavage cytology . Equine Vet J 2017 ; 49 : 263 – 265 . 10.1111/evj.12679 4. Barton AK

Full access
in American Journal of Veterinary Research