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Abstract

Objective—To compare results of intradermal tests (IDT) for environmental allergens at 30 minutes and 4, 6, and 24 hours after injection in horses without atopy and horses with atopic dermatitis (AD) or recurrent urticaria (RU).

Animals—22 horses without atopy, 10 horses with RU, and 7 horses with AD.

Procedure—In all horses, medical history was obtained, and results of physical examination, hematologic examination, serum biochemical analyses, examination of bronchoalveolar lavage fluid, and IDT with 73 allergens were examined.

Results—Horses with AD or RU had a significantly greater mean number of positive reactions for IDT, compared with horses without atopy. Horses with AD had a significantly greater number of positive reactions than horses without atopy in every allergen group at all time periods, except for molds at 4 and 24 hours. Horses with RU had a significantly greater number of positive reactions than horses without atopy in every allergen group, except for molds at 30 minutes and 4 and 6 hours, trees at 4 and 6 hours, and grasses at 4 hours.

Conclusions and Clinical Relevance—A significantly greater number of positive reactions for IDT in horses with AD or RU, compared with horses without atopy, provides evidence of type-I IgE-mediated hypersensitivity for these diseases. Evaluation of results of IDT performed in horses with AD or RU is useful in determining specific allergens for the formulation of immunotherapy along with providing identification of allergens that could be useful when creating avoidance strategies. (Am J Vet Res 2001;62:1051–1059)

Full access
in American Journal of Veterinary Research

Abstract

Objectives—To determine collagenase activity and evaluate matrix metalloproteinase (MMP)-8 and MMP-13 in horses with chronic obstructive pulmonary disease (COPD).

Animals—12 horses with COPD and 12 healthy control horses.

Procedure—Collagenase activity was determined by use of an assay for degradation of type-I collagen. Western immunoblot analysis was used to identify interstitial collagenases MMP-8 and MMP-13 in tracheal epithelial lining fluid (TELF). Immunocytochemistry and in situ hybridization were used to determine cellular expression of these 2 collagenases in cells in bronchoalveolar lavage fluid (BALF).

Results—Collagenase activity was approximately 7 times higher in samples obtained from horses with COPD, compared with control horses. During stabling, horses with COPD had significantly higher collagenase activity than after being maintained on summer pasture, when activity was similar to that of control horses. Immunoreactivity of MMP-8 and MMP-13 was significantly increased in TELF of horses with COPD, compared with healthy horses. In TELF, a positive correlation was detected between immunoreactivity of MMP-8 and MMP-13 and the amount of degradation of type-I collagen. Macrophages and epithelial cells were the major cellular sources of MMP-8 and MMP-13.

Conclusions and Clinical Relevance—Increased collagenase activity in TELF indicates active ongoing disease and, thus, may reflect lung tissue changes in horses with COPD. Measurements of collagenase activity and MMP immunoreactivity may provide additional diagnostic tools to identify the active phase of chronic lung disease. (Am J Vet Res 2001;62:1142–1148)

Full access
in American Journal of Veterinary Research

Abstract

Objective—To determine concentrations of nitric oxide (NO) in plasma and bronchoalveolar lavage fluid (BALF) and localize nitric oxide synthesis in the lungs of horses with summer pasture-associated obstructive pulmonary disease (SPAOPD).

Animals—7 adult horses with SPAOPD and 6 clinically normal adult horses.

Procedure—Severity of SPAOPD was determined by use of clinical scores, change in intrapleural pressure (ΔPpl) during tidal breathing, cytologic analysis of BALF, and histologic evaluation of lung specimens obtained during necropsy. Nitric oxide concentrations in plasma, BALF, and epithelial lining fluid (ELF) were determined by use of a chemiluminescent method. Inducible nitric oxide synthase (iNOS) and nitrotyrosine (NT) were localized in formalin-fixed lung specimens by use of immunohistochemical staining, and nicotinamide adenine dinucleotide phosphate diaphorase (NADPHd) activity was localized in cryopreserved specimens by use of histochemical staining.

Results—Plasma concentration of NO in affected horses was slightly but not significantly greater than concentration in nonaffected horses. Nitric oxide concentrations in BALF or ELF did not differ between groups. Immunoreactivity of iNOS in bronchial epithelial cells of 3 of 5 lung lobes was greater in horses with SPAOPD, compared with nonaffected horses. However, staining for NT and NADPHd activity did not differ between groups.

Conclusions and Clinical Relevance—Expression of iNOS was greater in bronchial epithelial cells of horses with SPAOPD, compared with nonaffected horses, suggesting that NO may play a role in amplifying the inflammatory process in the airways of horses with this disease. (Am J Vet Res 2001;62:1381–1386)

Full access
in American Journal of Veterinary Research

Objective

To examine use of corticosteroids in treating dogs with airway obstruction secondary to hilar lymphadenopathy caused by chronic histoplasmosis.

Design

Retrospective study.

Animals

16 dogs.

Procedure

Records for dogs with airway obstruction examined from January 1979 through December 1997 were reviewed. Dogs were included in the study if they had hilar lymphadenopathy documented radiographically and bronchoscopically, had serum antibodies against Histoplasma capsulatum, and did not have organisms in any cytologic or histologic samples. Dogs were assigned to groups on the basis of treatment given (5 dogs, corticosteroids only; 5 dogs, corticosteroids and antifungal medication; 6 dogs, antifungal medication only).

Results

Clinical signs resolved in < 1 week in dogs treated only with corticosteroids. In dogs treated with corticosteroids and an antifungal medication, improvement was evident in a mean of 2.6 weeks. In 5 of 6 dogs treated with only an antifungal medication, clinical signs resolved in a mean of 8.8 weeks. Dogs receiving corticosteroids did not develop active or disseminated histoplasmosis.

Clinical Implications

Corticosteroids can be used successfully in the treatment of dogs with hilar lymphadenopathy secondary to histoplasmosis. Affected dogs must be carefully evaluated for active infection. Specimens obtained by means of bronchoalveolar lavage, tracheal washing, or other methods should be examined to exclude the possibility of an active infection, which could result in corticosteroid-induced dissemination of disease. (J Am Vet Med Assoc 1999;214–1345-1348)

Free access
in Journal of the American Veterinary Medical Association

Abstract

Objective

To characterize surfactant protein D (SP-D) isolated from bronchoalveolar lavage fluid (BALF) of healthy horses.

Sample Population

BALF from 10 Thoroughbreds (5 males, 5 females; 26 to 40 months old) without history or clinical signs of respiratory tract disease.

Procedure

BALF was obtained and centrifuged at 33,000 × g. The supernatant was applied to a mannose-Sepharose 6B affinity column in the presence of calcium, and the bound protein fraction was analyzed by use of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoblot analysis; amino acid composition was determined and partial sequencing was done. Phospholipid binding and liposome aggregation assay were performed, using purified proteins.

Results

The protein isolated by use of mannose affinity matrices was SP-D. It bound carbohydrates and phosphatidylinositol, which are the characteristic features of SP-D isolated from other animal species. Amino acid analysis and partial primary sequence of the isolated protein indicated high homology with rat and human SP-D. Furthermore, immunoblot analysis indicated that equine SP-D reacted with human and rat SP-D-specific antibodies.

Conclusion and Clinical Relevance

SP-D exists in equine lungs; its measurement may be useful in evaluating equine lung disease. (Am J Vet Res 1999;60:368–372)

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

A study was conducted to investigate whether aspiration of amniotic fluid is associated with a deleterious effect on absorption of colostral immunoglobulins or on blood gas and acid-base values of healthy newborn calves. Fourteen calves purchased from commercial sources were transported to a research facility immediately after birth and fed colostrum with known concentrations of immunoglobulins. Blood samples for gas analyses were collected within 5 hours of birth, 24 hours later, and prior to euthanasia. Between 3 and 5 days of age, calves were euthanatized by an overdose of barbiturates. Eleven calves had evidence of bronchoaspiration of amniotic fluid, as determined by presence of meconium, squamous epithelium, or keratin in histologic sections of fixed lung or by cytologic analysis of bronchoalveolar lavage fluid. Blood gas tensions and pH were within reference ranges in 11 of 14 calves. Aspiration of amniotic fluid could not be linked to any specific changes in blood gas tensions, add-base status, or absorption of colostral immunoglobulins. Presence of keratin and meconium in the lungs often was accompanied by mild exudative alveolitis and focal atelectasis. It was concluded that aspiration of small amounts of amniotic fluid with or without meconium is common in calves and is not associated with hypoxemia, respiratory acidosis, or failure of passive transfer.

Free access
in American Journal of Veterinary Research

SUMMARY

Pneumonic pasteurellosis was experimentally induced in calves by inoculation of 5 × 108 Pasteurella haemolytica organisms into the right diaphragmatic lung lobe. Blood and bronchoalveolar lavage fluid samples were obtained prior to inoculation and at postinoculation hour (pih) 2, 4, and 6. Calves developed acute lung injury, characteristic of pneumonic pasteurellosis. Lesions were found only in the right diaphragmatic lobe. By pih 4, significant (P < 0.01) increases were detected in lavage fluid total cell count, neutrophil count, total protein and albumin concentrations, and alkaline phosphatase (alp) and lactic dehydrogenase (ld) activities. Myeloperoxidase and elastase activities did not increase. Neutrophil depletion ameliorated the lung lesions and prevented the increase in lavage fluid cell count, total protein, and albumin concentrations and alp and ld activities. Treatment with the iron chelator, deferoxamine mesylate-hydroxyethyl starch, attenuated the increase in total protein and albumin concentrations and alp and ld activities at pid 4, but not pih 6. Treatment with a neutrophil function inhibitor, pentoxifylline, prevented the increase in lavage fluid neutrophil numbers, but accentuated the increase in total protein and albumin concentrations, and alp, ld, myeloperoxidase, and elastase activities.

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