Phenotypic analysis of peripheral blood and bronchoalveolar lavage fluid lymphocytes in control and chronic obstructive pulmonary disease affected horses, before and after 'natural (hay and straw) challenges'.

Phenotypic analysis of lymphocytes in peripheral blood (PB) and bronchoalveolar lavage fluid (BALF) of control and chronic obstructive pulmonary disease (COPD) affected horses, both before and after 'natural (hay and straw) challenge', were performed using immunofluorescent labelling with monoclonal antibodies and flow cytometry. BALF lymphocytes were shown to be predominantly EqCD5+ cells, approximately half of which were also EqCD8+, with a smaller proportion of B cells. In comparison with PB, BALF contained higher proportions of EqCD5+ cells and EqCD8+ cells and a lower proportion of B cells. Horses with asymptomatic COPD had a higher proportion of BALF B cells and a lower proportion of BALF EqCD5+CD8- cells (considered to be predominantly EqCD4+ cells) than controls. Hay and straw challenge increased the ratio of BALF EqCD5+CD8- cells and reduced the ratio of BALF EqCD8+ cells in COPD affected horses, but not in controls. This finding is similar to the pulmonary recruitment of CD4+ cells reported for human asthmatics following allergen challenges.

Evaluation of urea and albumen as endogenous markers of dilution of equine bronchoalveolar lavage fluid.

The urea and albumen dilution techniques for standardising the variable concentrations of pulmonary epithelial lining fluid (PELF) in bronchoalveolar lavage fluid (BALF) samples were evaluated in horses. Both techniques proved satisfactory and were of equal accuracy. Albumen adjusted BALF cell counts were significantly higher than PELF cell counts. BALF from normal horses contained, on average, 0.4 per cent PELF (range 0.1 to 1.0 per cent), as determined by the urea dilution technique.

Comparison of cellular and molecular components of bronchoalveolar lavage fluid harvested from different segments of the equine lung.

A comparison was made of the cellular and molecular components of bronchoalveolar lavage fluid (BALF) harvested from the left and right diaphragmatic lobes, the accessory lobe of the right lung and the apical lobe of the left lung, of seven control horses and six horses with symptomatic chronic obstructive pulmonary disease (COPD). Neither control nor symptomatic COPD affected horses showed significant regional differences in BALF recovery volumes, total and differential BALF cell counts, albumen adjusted total and absolute BALF cell counts, total and absolute pulmonary epithelial lining fluid (PELF) cell counts, and BALF albumen and urea concentrations. This suggests that the composition of PELF is uniform throughout the lungs of normal horses and horses with symptomatic COPD and that a single BALF sample is representative of the entire lung in these horses.

Effect of stabling on bronchoalveolar cells obtained from normal and COPD horses.

Bronchoalveolar lavages (BAL) were performed before and after 3 weeks of housing in 5 horses suffering from COPD and 5 normal horses. In the two groups, the total number of cells recovered remained unchanged after stabling. The most common cell populations in BAL fluid of control animals were alveolar macrophages (46.4%) and lymphocytes (44.9%). The percentage of neutrophils increased after stabling from 8.7% to 27.6%. In COPD horses, lymphocytes predominated (40.7%) in animals at pasture with neutrophils increasing from 29.4% to 71.6% after stabling. After fractionation by Percoll density gradient, alveolar macrophages and neutrophils from normal and COPD horses had a similar density distribution. After stabling, these cells from normal horses were increased in the low density layers, while those from COPD horses were predominantly in the hyperdense layers. Therefore, BAL cells obtained from COPD animals at pasture and after stabling differ from those of control horses in the same environment, not only in their populations but also in their buoyant densities. These differences could be related to different states of cellular activation and perhaps be responsible for disease activity in the COPD horses.

Bronchoalveolar lavage in the evaluation of pulmonary disease in the dog and cat. State of the art.

Bronchoalveolar lavage is a diagnostic procedure used to obtain specimens representative of disease processes involving the deep lung. Saline is instilled into an airway in sufficient volumes to bathe the alveoli dependent on that airway. The saline is retrieved by suction along with cellular and acellular material lining the epithelial surfaces of the lung. Cytologic and microbiologic evaluation of the fluid can be used to characterize pulmonary diseases in the dog and cat.

Changes in blood and bronchoalveolar lavage fluid components in calves with experimentally induced pneumonic pasteurellosis.

Pneumonic pasteurellosis was experimentally induced in calves by inoculation of 5 x 10(8) 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 less than 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 mesylatehydroxyethyl 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.

Immunohistochemical localization of alpha 2-beta 1-glycoprotein in horses.

Alpha 2-beta 1-glycoprotein may be found free in horse serum or complexed with alpha-1-proteinase inhibitor to form pre-alpha 2-elastase inhibitor. There has been little information published concerning alpha 2-beta 1-glycoprotein and its possible tissue sources in horses. A peroxidase-antiperoxidase technique was used to identify alpha 2-beta 1-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 alpha 2-beta 1-glycoprotein. Alpha 2-beta 1-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, alpha 2-beta 1-glycoprotein was observed in some airway secretions and in macrophages.

Effect of transportation stress on bronchoalveolar lavage fluid analysis in female horses.

Four bronchoalveolar lavages were performed sequentially on 9 control and 8 transport-stressed female horses. Alterations in results of fluid cytologic analyses, microbial content, and phagocyte function of recovered pulmonary macrophages in all horses were determined. Seemingly, absolute and relative increase in the number of inflammatory cells detected in the second bronchoalveolar lavage fluid of control horses was the result of irritation of the first lavage. This increased response was not observed in transport-stressed horses until 5 days after transport (third lavage; 10 days after initial lavage). Seemingly, delayed inflammatory response was the result of the transport stress. Microbial content and macrophage function were not significantly different between the 2 groups (P greater than 0.05).

Effect of bovine herpesvirus-1 or parainfluenza-3 virus on immune receptor-mediated functions of bovine alveolar macrophages in the presence or absence of virus-specific serum or pulmonary lavage fluids collected after virus infection.

The immune receptor-mediated functions of bovine alveolar macrophages (AM) inoculated in vitro with bovine herpesvirus-1 (BHV-1) or parainfluenza-3 (PI-3) virus were tested in the presence or absence of virus-specific antiserum or pulmonary lavage fluids collected from calves 6 days after inoculation with BHV-1 or PI-3 virus. The Fc and C3b phagocytic indices of noninoculated AM, collected from 6- to 16-week-old calves, ranged from 75 to 87 and 59 to 64, respectively, and the binding indices ranged from 5 to 8 and 22 to 28, respectively. Infection of AM with either BHV-1 or PI-3 virus had no significant effect on receptor-mediated phagocytosis or binding, with the exception of a significant (P less than 0.05) decrease, from 64 to 46, of the C3b phagocytic index of PI-3 virus-infected AM. The addition of lavage fluids, collected after BHV-1 or PI-3 virus infection, to AM infected with the respective virus caused a significant (P less than 0.05) decrease in phagocytic indices with values for the Fc and C3b indices in BHV-1-infected AM decreasing from 81 to 49 and from 47 to 8, respectively, and those for the PI-3 virus-infected AM from 79 to 51 and from 46 to 15, respectively. The binding indices of virus-infected AM increased with the addition of viral lavage fluids, but the only significant (P less than 0.05) increase was for C3b binding in PI-3 virus-infected cells, which increased from 33 to 56.(ABSTRACT TRUNCATED AT 250 WORDS)

Prostaglandin and thromboxane concentrations in plasma and lung lavage fluids during sequential infection of vaccinated and nonvaccinated calves with bovine respiratory syncytial virus.

The potential action of immunologic reactions and mediators released during the course of bovine respiratory syncytial virus infection in pathogenesis of the ensuing disease process was examined in an experimental infection study. Prostaglandin (PG) E2, PGF2 alpha 6-keto-PGF1 alpha, and thromboxane B2 (TxB2) concentrations were quantitated in plasma and lung lavage fluid by radioimmunoassay at 3- to 4-day intervals during a primary and secondary virus infection of vaccinated, nonvaccinated, and control (mock-infected) calves. A significant increase in the plasma PGE2 concentration for the nonvaccinated calves was noticed on day 3 after primary infection and on day 7 after secondary infection. The PGF2 alpha plasma concentrations increased significantly for the nonvaccinated groups on day 10 after primary infection. Plasma 6-keto-PGF1 alpha concentrations increased for nonvaccinated and vaccinated calves 3 days after the secondary infection. Plasma TxB2 concentrations during the primary exposure did not vary significantly. However, 14 days after the secondary exposure, both experimental groups had concentrations significantly greater than did the control group. Lung lavage fluid concentrations of TxB2 had peaks of activity 7 days after primary and secondary viral infections for the nonvaccinated group. Increases in plasma PG concentrations corresponded variably with disease expression, whereas plasma TxB2 concentrations did not have any correlation with disease expression. However, there was a significant correlation between TxB2 concentration in lung lavage fluid of the nonvaccinated group with disease expression 7 days after primary and secondary virus infection.(ABSTRACT TRUNCATED AT 250 WORDS)

An improved method of bronchoalveolar lavage of lungs of small laboratory animals: short report.

This report describes a semi-automatic method for standardized bronchoalveolar lavage of small laboratory animals. In essence the method is constructed from a dispenser and a pressure chamber.

A method for bronchoalveolar lavage in live pigs.

In order to isolate porcine alveolar macrophages and to quantitatively study the components of recovered lung fluid, a bronchoalveolar lavage technique in living pigs was developed. Lung lavage was performed after introducing a catheter through the mouth via the trachea in the diaphragmatical lobe. Thirty ml of phosphate-buffered saline (PBS) was introduced into the lung and the fluid was aspirated after one minute. Following this, another 15 ml of PBS was introduced into the lung and aspirated after one minute. The recovered volume of the second lavage averaged 15 ml (+/- 0.4 S.E.M.). Cells thus obtained from specific-pathogen-free (SPF) pigs were composed of 98% macrophages. Lavage fluids from conventionally bred pigs contained 67% macrophages, 17% neutrophilic granulocytes and about 16% lymphocytes, demonstrated by their morphology and acid phosphatase activity. The viability of the recovered cells was over 98% in both SPF and conventionally bred pigs. The dilution of the aspirated lung liquid was determined by using methylene blue in the introduced fluid. The calculated dilution factor of the recovered lavage fluid was 0.58 (S.E.M. 0.02). No influence was noticed on the number or composition of cells nor on the dilution factor when lung lavage was done in SPF pigs twice a week during a four week period. The protein concentration in lavage fluid from SPF pigs was 142 (SD +/- 26) mcg/ml. In conventionally bred pigs, however, a wide variation (276 +/- 229 mcg/ml) in protein content was noted. Lavage fluid supernatant of some animals had a bactericidal effect on Haemophilus pleuropneumoniae strain 13261, whereas no bactericidal effect was noted in other lavage samples.

Bronchoalveolar lavage in the newborn foal.

Two bronchoalveolar lavages, 24 h apart, were performed on 15 foals, ranging in age from 1 to 21 days. In the first lavage, a numerical deficiency in alveolar macrophages was demonstrated in foals up to 2 weeks of age when compared with older (2-3 years of age) horses. Alveolar macrophages obtained from the lungs of 2-3-day-old foals also demonstrated significant impaired chemotactic function. In the second lavage, although an increase of alveolar macrophages was noted, a dramatic increase of polymorphonuclear neutrophil (PMN) mobilization occurred in the foals, thus providing a phagocytic back-up for the alveolar macrophage host defence mechanism. In contrast, PMN response to lower respiratory tract perturbation in older horses was negligible. The results of this study suggest that the numerical and chemotactic deficiency found in alveolar macrophages of the neonate may play a role in the foal's susceptibility to respiratory disease.

Pneumonia in lambs inoculated with Bordetella parapertussis: bronchoalveolar lavage and ultrastructural studies.

Eight colostrum-deprived lambs were inoculated intratracheally with ovine isolates of Bordetella parapertussis. Fluids obtained by bronchoalveolar lavage had a large increase in total cell counts 24 hours after inoculation; up to 93% of cells were neutrophils. From 3 days after inoculation, the number of alveolar macrophages in lavage samples was markedly increased. From 5 days onwards, many alveolar macrophages had moderate to severe cytoplasmic vacuolation. Topographically, tracheal and bronchial epithelium was covered by a large amount of inflammatory exudate 24 hours after inoculation. Later, the tracheobronchial epithelium showed focal extrusions from ciliated cells, which were occasionally associated with B. parapertussis organisms. Ultrastructurally, cytopathological changes associated with B. parapertussis infection were mild focal degeneration of airway epithelium with slight loss of cilia, moderate to severe degeneration of type I and type II alveolar epithelial cells, and focal inflammation in the lungs. These results suggest that the primary targets of B. parapertussis infection are alveolar macrophages and the epithelial cells of bronchioles and alveoli.

[Surfactant lipids of bovine lungs]. / Lipidy surfaktantu plucnego bydla.

Lung surfactant from bronchoalveolar lavage and post-lavaged lung tissue of calf and adult cows was isolated. In both species of the surfactant, the main fractions are: phospholipids and free fatty acids. Phosphatidylcholine was the main phospholipid fraction. Phosphatidylglycerol level is about 4% of total phospholipids. In both phosphatidylcholine and phosphatidylglycerol fractions, palmitic, oleic and linoleic acids are the main fatty acids. There are no differences between bronchoalveolar lavage and post-lavaged lung tissue surfactants and between calf and adult cow surfactants in the aspect of their lipid composition.

Bronchoalveolar eosinophilia in random-source versus purpose-bred dogs.

Eosinophils (EOS) have been implicated in changes in airway and vascular reactivity in a variety of disease states. Analysis of cells in bronchoalveolar lavage samples from chronic, heartworm-free random-source (RS) dogs indicated higher leukocyte counts with markedly higher percent and total numbers of EOS than were present in purpose-bred (PB) animals. Bronchoalveolar lavage fluid (BALF) obtained from RS dogs had a significantly elevated total nucleated cell count: 0.8 x 10(6) vs 0.4 x 10(6) for the PB dogs. RS dogs had 24% +/- 5% and PB dogs had 3% +/- 0.7% EOS. The RS animals with elevated EOS had similar percentages of neutrophils: 4% +/- 0.6% as the PB animals. Despite aggressive anthelminthic treatment, the abnormal BALF cellular profile of the RS animals persisted even though circulating levels of EOS in this group decreased. Analysis of BALF for thromboxane B2 (TxB2) and 6-keto-prostaglandin F1(1a) (6-keto-PGF1a) indicated that only the TxB2 levels were significantly different between groups. The RS BALF TxB2 levels were 73 +/- 14 pg/ml vs 23 +/- 3 pg/ml for the PB group (P < 0.05). Regression analysis of the relationship between increasing TxB2 levels and the absolute number of EOS per milliliter of BALF obtained from the RS dogs indicated a significant correlation (r = 0.83, P < 0.0001). No difference in plasma levels of these mediators was observed. Other physiologic parameters also differed between the two groups: the RS group had significantly increased heart rates and cardiac output under baseline conditions.(ABSTRACT TRUNCATED AT 250 WORDS)