Bronchoalveolar lavage cytology in cynomolgus monkeys and identification of cytologic alterations following sequential saline lavage

Total and differential cell counts were determined on cytolytic specimens obtained by fiberoptic bronchoscopy and bronchoalveolar lavage (BAL) of five normal cynomolgus monkeys. Total nucleated cell counts ranged from 100 to 430 cells/microliters. Macrophages were approximately 91% of total nucleated cells, while lymphocytes were 3%, neutrophils 4%, and eosinophils 2% of the initial BAL from each monkey. Less than 1% of the cells were mast cells and ciliated or nonciliated epithelial cells. The effects of repeated saline BAL on pulmonary cell populations were evaluated. Saline lavage of individual lung lobes resulted in a marked rise in circulating blood neutrophils at 4 hr after BAL; there was a similar rise in neutrophils in lavage fluids 24 hr after the initial lavage. Differential and total cell counts of both blood and lavage fluid returned to normal if subsequent lavages were spaced at 48-hr intervals. Lymphocytes were not present in saline-lavaged lung lobes, and protein levels of lavage fluids did not rise significantly. BAL produced a transient, reversible, intra-alveolar influx of neutrophils which was preceded by mobilization of bone marrow-stored neutrophils. Neutrophilia in the lavage fluid and blood was not detectable if lavage and blood sampling procedures were done at 48-hr intervals (which did not alter Ia antigen expression among BAL cells). These observations indicate that BAL is a valid method for sampling and assessing pulmonary cellular and fluid constituents if the procedures are done at intervals of at least 48 hr.     

Effects of lung site and fluid volume on results of bronchoalveolar lavage fluid analysis in horses.

Bronchoalveolar lavage (BAL) fluid was analyzed in healthy horses, using different lavage fluid volumes and lung sites. The only significant difference in the cellular composition of BAL fluid between the right and left lungs was the mast cell numbers, which were significantly higher in the left lung. Total cell count ranged from 34 to 330 cells/microliter for the right lung and 43 to 330 cells/microliter for the left lung. Percentage of neutrophils ranged from 1 to 7% in the right lung and 1 to 5% in the left lung. The small-volume (50 ml) lavage had a greater percentage of neutrophils and a lesser percentage of mast cells in the large-volume (350 ml) lavage. Statistical difference in the composition of BAL fluid recovered was not detected between the 3 sequential 100-ml lavages and a single 300-ml lavage, except that macrophages were significantly higher in the 3 sequential 100-ml lavages. Values for BAL fluid analysis in healthy horses have varied considerably and this variation is from a failure to adhere to any standard technique for volume of fluid infused.     

Effect of repetitive bronchoalveolar lavage on cytologic findings in healthy dogs

OBJECTIVE: To determine reference values for cytologic examination results of bronchoalveolar lavage fluid (BALF) and to investigate effects of repeated lavages on pulmonary health and on results of cytologic examination of BALF in dogs. ANIMALS: 16 healthy adult Beagles. PROCEDURE: All dogs underwent pulmonary lavage to obtain BALF. Eleven dogs were repeatedly lavaged 6 times at 5- to 7-week intervals. Analyses for total and differential cell counts and for viability of cells before and after cell processing were performed. Arterial blood gas analysis before and after bronchoalveolar lavage was used to study the safety of the lavage procedure. Histologic and radiologic examinations were used to study effects of repeated lavages on pulmonary health. RESULTS: Mean (+/- SD) cell count was 104 +/- 69 cells/microl, comprising 75 +/- 7% alveolar macrophages, 13 +/- 6% lymphocytes, 5 +/- 4% neutrophils, 4 +/- 5% eosinophils, 2 +/- 2% mast cells, 0.6 +/- 0.7% epithelial cells, and 0.3 +/- 0.4% plasma cells. Centrifugation of samples and washing of cells caused significant cell loss (59 +/- 13%). Repeated lavages did not cause significant variations in cell counts of BALF or results of arterial blood gas analysis, thoracic radiography, or histologic examination of pulmonary specimens. Only a moderate, although significant, decrease in arterial oxygen content was observed after bronchoalveolar lavage. CONCLUSIONS AND CLINICAL RELEVANCE: Analysis indicated that several lavages performed at 5- to 7-week intervals can safely and reliably be used to study the kinetics of pathologic processes in pulmonary tissues or for evaluation of therapeutic efficacy.     

Cytological analysis of equine bronchoalveolar lavage fluid. Part 1: Comparison of sequential and pooled aliquots

The aim of this study was to investigate whether initial equine bronchoalveolar lavage fluid (BALF) aliquots were more representative of bronchial cytology that bronchiolar and alveolar cytology. Cell viability and total nucleated (TCC), differential (DCC) and absolute cell counts of cytocentrifuged preparations of 3 sequentially collected BALF aliquots (Aliquots 1-3) were compared with those of pooled BALF (Aliquot 4) to assess whether all aliquots were representative of the lavaged lung segment. BALF samples (n = 21) were collected from control horses (n = 5) or heaves-affected horses (n = 5). There were nonsignificant trends of increasing TCC and absolute macrophage count from Aliquot 1 to Aliquot 3 and significant differences in macrophage (P<0.05) and lymphocyte (P<0.01) DCC among aliquots of all horses; however, no linear trend in this DCC data was observed. There was a significant decrease in mast cell DCC (P<0.01) from Aliquot 1 to Aliquot 3 in control horses. Cell viability did not differ significantly among aliquots. There was no diagnostically significant difference in TCC, DCC, absolute cell counts or cell viability, among sequential and pooled BALF aliquots and, therefore, all aliquots can be considered to represent the cytology of the lavaged lung segment. This indicates that even if BALF recoveries are very low, cytological analysis of samples will be of diagnostic value.     

Cellular content of secretions obtained by lavage from different levels of the equine respiratory tract

Lavage procedures were used to obtain samples of respiratory secretions from the nasal cavity, trachea, bronchi and bronchoalveolar level of apparently normal conscious horses, and the cellular composition of the lavage fluids was assessed. There was a progressive increase in total cell count of the secretions obtained from the upper to the lower respiratory tracts. Nasal lavage fluid was composed chiefly of epithelial cells, whereas tracheal, bronchial and bronchoalveolar fluids contained higher proportions of macrophages, lymphocytes and neutrophils. Eosinophils and mast cells were identified in small numbers. The results of the differential cell counts were compared with previous published reports, which revealed some major differences between studies. It is suggested that these differences may relate to variations in the collection technique, the cytological interpretation of cell types, the presence of subclinical airway disease, and the degree of exposure to airborne environmental contaminants.     

Evaluation of peritoneal fluid following intestinal resection and anastomosis in horses.

Postoperative abdominal fluid changes were compared in 2 groups of horses; those undergoing double small-colon resection and anastomosis (n = 10) and those undergoing exploratory celiotomy alone (n = 5). Peritoneal fluid was collected before surgery and on postoperative days 1, 3, 5, and 7. Total and differential nucleated cell counts, RBC numbers, and total protein and fibrinogen concentrations were evaluated. In both groups, all values were significantly higher than normal on the first postoperative day (after small-colon resection and anastomoses, WBC = 130,350 +/- 23,310 cells/microliters, RBC = 7,389,000 +/- 6,234,000 cells/microliters, total protein = 3.63 +/- 0.16 g/dl; after exploratory celiotomy alone, WBC = 166,620 +/- 34,340 cells/microliters, RBC = 295,000 +/- 86,070 cells/microliters, total protein 4.38 +/- 0.54 g/dl). The number of total peritoneal nucleated cells and RBC significantly decreased after the first postoperative day, whereas total protein and fibrinogen concentrations, percent neutrophils, and percent mononuclear cells remained unchanged. None of the values had returned to normal by postoperative day 7 (after small-colon resection and anastomoses, WBC = 45,600 +/- 8,765 cells/microliters, RBC = 95,390 +/- 53,380 cells/microliters, total protein = 4.39 +/- 0.23 g/dl; after exploratory celiotomy alone, WBC = 43,340 +/- 7,746 cells/microliters, RBC = 12,860 +/- 11,790 cells/microliters, total protein = 3.92 +/- 2.20 g/dl.) The resection and anastomosis group had a significantly lower total protein concentration on the first postoperative day and a significantly higher mean total RBC count over the entire 7-day postoperative evaluation than did horses that underwent celiotomy alone. Other values in the 2 groups of horses did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)     

Volume-controlled bronchopulmonary lavage of normal and pneumonic calves

Saline bronchopulmonary lavage of the right lung of 16 anesthetized calves was performed using a single-lumen cuffed endotracheal tube. The initial volume of saline introduced was based on the functional residual capacity (FRC) of the right lung lobes as determined from the proportional weights of the right (58% of total FRC) and left (42% of total FRC) lung lobes. Calves were divided into "pneumonic" and "normal" groups based on clinical signs. Five sequential washes were done on each calf. There was no difference in the percentage of total lavage fluid volume recoverable between normal (83.8 +/- 4.2%) and pneumonic (81.1 +/- 8.2%) calves. Cell yield in the initial wash was consistently greater than in subsequent washes for both normal (12.7 +/- 6.6 X 10(6) cells/kg body weight) and pneumonic (58.1 +/- 37.6 X 10(6) cells/kg body weight) calves, and constituted 62.0% (normal) and 75.4% (pneumonic) of the total recoverable cell yield. Total cell yields were higher (P less than 0.05) in pneumonic calves, primarily due to neutrophil leukocytes (PMN). Neutrophils constituted 53.7 +/- 25% of the total cell yield in the pneumonic calves, but only 12.3 +/- 9.5% in the normal calves. The pulmonary alveolar macrophage (PAM) was the major recoverable cell in normal calves (85.7 +/- 8.7% of total lavage cells). Macrophages constituted a smaller (42.9 +/- 23.5) percentage of the total lavage cells in the pneumonic group due to increased PMN numbers. Viability of recovered cells from the pneumonic calves (91.5 +/- 4.8%) was lower than for the normal calves (94.1 +/- 2.5%), but the difference was not significant.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cytologic, microbiologic, and biochemical analysis of bronchoalveolar lavage fluid obtained from 24 healthy cats

Twenty-four healthy cats underwent bronchoscopy and bronchoalveolar lavage to determine the normal cytologic environment of the lower respiratory tract of cats. Initial screening to ensure the health of the study population included complete histories, physical examinations, thoracic radiography, CBC, serologic tests for feline leukemia virus, feline immunodeficiency virus, and occult heartworm, and sugar and Baermann fecal flotation. In 18 cats, protected catheter brush samples of airway secretions from the lavaged lung segment were taken for culture of aerobic and anaerobic bacteria and mycoplasma. Bronchial lavage fluid (5 sequential 10-ml aliquots of normal saline solution) was pooled and filtered with cotton gauze. The unspun sample was used for determination of a total nucleated cell count. Lavage fluid was cytocentrifuged and 500 cells/slide were scored for determination of the cellular differential. Activity of lactate dehydrogenase and concentrations of total protein and IgG within the supernatant were measured, and assays were performed to detect the presence of IgA and IgM. Complete histologic evaluation of the lavaged lung of each of 6 random-source cats was performed after differential cell counting revealed 18% eosinophils within bronchoalveolar lavage fluid recovered from this group. Alveolar macrophages were the predominant cells encountered; however, a quarter of all cells recovered were eosinophils. A significant relationship was not found between the abundance of eosinophils in the lavage fluid, and either isolation of aerobic bacteria, high total nucleated cell counts, total protein concentrations, or activity of lactate dehydrogenase. Histologic evaluation of the lungs of 5 of 6 random-source cats revealed normal lungs in 2 cats, and minimal abnormal change in 3 others. Evaluation of the lungs from 1 random source cat revealed acute, mild eosinophilic bronchiolitis. We conclude that large numbers of eosinophils may be retrieved from the bronchoalveolar lavage fluid of healthy cats.     

Using urea dilution to standardise cellular and non-cellular components of pleural and bronchoalveolar lavage (BAL) fluids in the cat

A technique to standardise the analysis of cellular and non-cellular components in epithelial lining fluid (ELF) collected during saline lavage of pulmonary and pleural cavities was developed using the urea dilution method. Bronchoalveolar lavage (BAL) and pleural lavage (PL) fluids were collected from 12 clinically healthy cats. Total and differential cell counts in BAL fluid were within normal ranges for the cat, while cell counts in PL fluid were assumed to be normal based on clinical health during examination, auscultation and lactate dehydrogenase (LDH) activities being comparable with other species. The major clinical implication of this study was that nucleated cell counts within feline ELF could not be predicted from analysis of lavage fluid which suggests that calculation of the proportion of ELF in lavage fluid by the urea dilution method may be necessary to avoid misdiagnosis of health or disease in pulmonary or pleural cavities.     

Role of platelet-activating factor in alveolar septal injury associated with experimentally induced pneumonic pasteurellosis in calves

OBJECTIVE: To determine whether platelet-activating factor (PAF) is involved in acute lung microvascular injury associated with pneumonic pasteurellosis in calves. ANIMALS: 15 healthy 2- to 4-week-old male Holstein calves. PROCEDURE: Calves were anesthetized and inoculated intrabronchially with saline (0.9% NaCl) solution (n = 5) or 1x10(9) Pasteurella haemolytica organisms (n = 10). Of the 10 calves inoculated with P haemolytica, 5 also were treated with WEB 2086, a potent inhibitor of PAF, and 5 were treated with vehicle. Blood and bronchoalveolar lavage samples were collected before and 1, 2, 4, and 6 hours after inoculation of P. haemolytica. Blood samples were analyzed to evaluate total number and differential counts of leukocytes, dilute whole-blood leukocyte deformability, size of neutrophils, and neutrophil CD11b expression. Bronchoalveolar lavage samples were analyzed for total number and differential counts of nucleated cells, total protein concentration, and hemoglobin concentration. Size and gross and histologic appearance of lung lesions also was determined. RESULTS: Treatment of calves with WEB 2086 reduced size of lung lesions, attenuated the increase in microvascular permeability, and reduced neutrophil infiltration in the first 4 hours after inoculation. Treatment with WEB 2086 also attenuated a decrease in leukocyte deformability, increase in size of neutrophils, and CD11b expression by circulating neutrophils. CONCLUSIONS AND CLINICAL RELEVANCE: It appears that PAF is a major mediator for altered lung microvascular permeability and activation of circulating neutrophils in the first 4 hours after onset of pneumonic pasteurellosis in calves.     

Diagnostic value of cytology of bronchoalveolar fluid for lung diseases of sheep

Bronchoalveolar lavage fluid was collected postmortem from the lungs of 113 sheep, and total and differential cell counts were analysed in relation to the presence of gross and microscopic lung pathology. The diffuse lung diseases, maedi and adenomatosis, were both characterised by an increase in overall cellularity and by increases in the percentages of lymphocytes and neutrophils, respectively. Focal parasitic lung disease was characterised by an increase in the percentage of eosinophils and mast cells. Consolidated lung lesions were characterised by a slight increase in cellularity but no change in the differential cell profile. In regions of parasitised and consolidated lungs without lesions the differential cell profile was consistent with focal lung pathology, although the slight increase in cellularity observed in the consolidated regions was not observed in the regions without lesions. A decision tree was developed to facilitate the interpretation and indicate the likely predictive capacity of the differential cytology of the fluid.     

Light microscopic and ultrastructural characterization of cells recovered by respiratory-tract lavage of 2- and 6-week-old chickens

This study characterized the cell population recovered by respiratory-tract lavage of 57 two-week-old and 59 six-week-old specific-pathogen-free chickens as a prerequisite to study the response of the avian respiratory tract to infectious agents. The respiratory tract of each bird was lavaged through the trachea with a series of three lavages of 10 ml of room-temperature, neutral phosphate-buffered saline per lavage. The three lavages per bird were pooled for analysis. Total recovery volumes were measured, lavage fluid cellularity was determined, and a 200-cell differential count of non-erythrocyte cells was performed. Lavage fluid recovery was greater from 2-week-old birds (91.3 percent) than from 6-week-old birds (86.3 percent). Total cells recovered were greater for 6-week-old chickens (6.79 x 10(5)) than for 2-week-old chickens (5.03 x 10(5)). Cells of epithelial origin included squamous cells, goblet cells, and both ciliated and non-ciliated columnar epithelial cells. Cells of non-epithelial origin consisted of heterophils, lymphocytes, macrophages, eosinophils, basophils, and erythrocytes. Cells of epithelial origin were the predominant cell type recovered from the 2-week-old chickens, followed by heterophils. In 6-week-old chickens, heterophils were the predominant cell type recovered, followed by cells of epithelial origin. In descending order of prevalence, the remainder of cell types recovered from chickens of both ages were lymphocytes, macrophages, eosinophils, and basophils.     

Histamine bronchoprovocation does not affect bronchoalveolar lavage fluid cytology, gene expression and protein concentrations of IL-4, IL-8 and IFN-gamma

In diagnosing inflammatory airway disease (IAD) in performance horses, a histamine bronchoprovocation (HBP) test is often performed. In previously published studies, HBP is usually undertaken prior to cytological examination of the bronchoalveolar lavage (BAL) cells. The purpose of this study was to determine if HBP alters (1) the total nucleated cell numbers and distribution in BAL fluid (BALF) and (2) the mRNA and protein concentrations of selected cytokines in BAL cells and BALF, respectively. BALF was initially collected endoscopically from the right middle or diaphragmatic lung lobe in eight healthy young Standardbred horses. Five to six days later, HBP was performed by aerosolization of histamine (8mg) over a 2min period. BALF was again collected within 2-4h of the HBP from the left middle or diaphragmatic lung lobe. In both samples, total and differential WBC counts were obtained. The gene expressions of interleukin-4 (IL-4), IL-8, interferon-gamma (IFN-gamma) and beta-actin in BAL cells were measured using real-time RT-PCR. The cytokine protein concentrations were measured in the BALF using ELISA. HBP was not associated with either a change in the total BAL cell number or in the distribution of the BAL cells. BAL cell expression of IL-4, IL-8 and IFN-gamma, detected in all samples with the exception of IL-4 in one horse (post-HBP), was not altered as a result of HBP. HBP was not associated with a significant change in IL-8 or IFN-gamma concentrations in the BALF. IL-4 protein was undetectable in BALF either prior to or following HBP. We conclude that HBP can precede BALF collection performed within 2-4h of the former without affecting selected parameters analysed in the BAL cells or BALF.     

Bronchoalveolar lavage in ponies with heaves during disease remission

Bronchoalveolar ravage (BAL) fluid was collected from control ponies and from affected ponies with heaves which were in clinical remission. Significant differences were not detected between the two groups in total nucleated cell count or relative percentages (relative cell counts) of small lymphocytes, large mononuclear cells, eosinophils, or mast cells. The mean relative neutrophil count of BAL fluid from the affected group was statistically greater than that of controls; however, the difference between the two groups was not sufficient to be of diagnostic significance.     

Concentrations of gentamicin in serum and bronchial lavage fluid after once-daily aerosol administration to horses for seven days

OBJECTIVE: To assess gentamicin concentrations in serum and bronchial lavage fluid (BLF) of horses during a 24-hour period after once-daily aerosol administration of gentamicin (GAER) for 7 days and the pattern and degree of bronchial tree inflammation associated with repeated GAER. ANIMALS: 13 healthy adult horses (9 geldings and 4 mares). PROCEDURE: The treatment group comprised 8 horses, and 5 horses were untreated control animals. Gentamicin (20 mL of gentamicin [50 mg/mL]) was administered via aerosol once daily for 7 days. Samples of serum and BLF were obtained from all horses before GAER and 0.5, 4, 8, and 24 hours after the final day of GAER. Gentamicin concentrations were determined for all samples from treated horses, and cytologic examinations were performed on all BLF samples. RESULTS: Peak median BLF gentamicin concentration detected at 0.5 hours was 2.50 microg/mL. Median serum gentamicin concentration was < 0.50 microg/mL at all time points. Significant differences were not observed in total nucleated cell counts or differential cell counts in BLF between groups at any time point. Neutrophil count in BLF for all horses was increased over baseline at 4 and 24 hours. CONCLUSIONS AND CLINICAL RELEVANCE: We did not detect evidence of gentamicin accumulation or respiratory inflammation after once-daily GAER for 7 days. This protocol appears unlikely to result in local or systemic toxicosis. Repeated daily GAER to horses appears to be a safe procedure and may have clinical use in the treatment of horses with bacterial infections of the airways.     

Bronchoscopic findings in 48 cats with spontaneous lower respiratory tract disease (2002-2009)

Background: Diagnosis of lower respiratory disease requires collection of airway samples to confirm the etiology of disease. Bronchoscopic evaluation is commonly performed in dogs but less information is available in cats. Hypothesis: The presence and number of bronchoscopic abnormalities visualized during bronchoscopic evaluation of cats with lower respiratory disease will correlate with the type of disease and total and differential cell counts in bronchoalveolar lavage (BAL) fluid. Animals: Forty‐eight cats prospectively evaluated by a single bronchoscopist. Methods: Bronchoscopy was performed during clinical evaluation of cats presenting with cough, respiratory distress, or both. Cats were evaluated for airway hyperemia, stenosis, or collapse, mucus accumulation, bronchiectasis, and epithelial irregularities. Cats were placed into groups of bronchitis/“asthma,” pneumonia, or neoplasia based on BAL findings, histopathology, and response to appropriate medical therapy. Summation of bronchial abnormalities and total and differential cell counts were compared among groups. Results: Endobronchial abnormalities were common in cats with feline bronchitis/asthma, pneumonia, and neoplasia and no differentiating features were found. Excessive mucus accumulation was common (83%), followed by stenosis of bronchial openings and nodular epithelial irregularities (56%), airway hyperemia (54%), airway collapse (48%), and bronchiectasis (27%). Total bronchoscopic score and total cell count did not differ among groups, although differential cell counts were significantly different. A weak correlation (R²= 0.16, P= .006) between age and total bronchoscopic score was noted. Conclusions and Clinical Relevance: Bronchoscopic abnormalities are common in cats with lower respiratory tract disease, and visualization of the airways provides additional nonspecific clinical information in cats.     

Effect of sex, age, number of bronchoalveolar lavages and quantitation methods on the bronchoalveolar cell counts in rats.

This study was conducted to investigate the effect of sex, age, number of bronchoalveolar lavages and method of quantitation on the number of bronchoalveolar cells in rats. Forty Long Evans rats were divided into four age-sex subgroups of ten animals each. Nine consecutive bronchoalveolar lavages were done in every rat and the number of bronchoalveolar cells/mL in lavages 1-3 (L1), 4-6 (L2) and 7-9 (L3) were determined by hemocytometer and electronic cell counts (Coulter Counter). The sex or age of the rats did not show a significant effect (p less than 0.05) in the number of bronchoalveolar cells recovered from the lungs; however, there was a significant difference (p less than 0.05) in the number of cells/mL among lavages 1-3, 4-6 and 7-9 (L1 approximately equal to L2 greater than L3). A discrepancy of approximately 8% in the counts of bronchoalveolar cells was found between the hemocytometer and the Coulter Counter; however, these two methods of cell quantitation showed a significant (p less than 0.01) positive correlation (r = 0.89). No significant (p greater than 0.05) differences were found in the percentage of fluid recovery (overall mean = 94.5%) among lavages L1, L2 and L3. It was concluded that the electronic cell counting of bronchoalveolar cells is as reliable as manual counting. Although sex or age did not significantly affect the number of cells recovered from the lung, caution should be used in the number of lavages done per rat since this variable may significantly affect the results.     

Bronchoalveolar lavage of cranial and caudal lung regions in selected normal calves: cellular, microbiological, immunoglobulin, serological and histological variables.

Of a group of 30 clinically normal male Holstein calves two to eight weeks of age, six two week old and six four week old calves met various radiographical and clinicopathological criteria for normality. Bronchoalveolar lavage was performed by fiberoptic bronchoscopy on cranial and caudal lung regions in all 30 calves and samples analyzed for free cells, microorganisms, and immunoglobulins. Lateral chest radiographs and lung biopsies were also conducted on each calf. Calves were euthanized and necropsied ten days after bronchoalveolar lavage was conducted. Reported in this paper are results from the 12 normal calves. Microorganisms were present in small numbers in the lower respiratory tract of some normal calves. There were no differences in the above parameters between cranial and caudal lobes. There were statistically significant changes in bronchoalveolar lavage cell proportions with age although there were no detectable differences in clinical signs. Four week old calves had a lower percentage of macrophages and a higher percentage of epithelial cells than two week old animals (p less than 0.05). There was also a trend toward an increased percentage of neutrophils in older calves but this was not significant (p greater than 0.05). Total bronchoalveolar lavage protein also appeared to increase with age (p less than 0.05). In both groups a higher proportion of IgG2 in bronchoalveolar lavage compared to serum was found, suggesting the presence of a local selective transfer mechanism into respiratory secretions.     

Detection of annexin I and IV and haptoglobin in bronchoalveolar lavage fluid from calves experimentally inoculated with Pasteurella haemolytica

OBJECTIVE: To determine whether annexins or haptoglobin could be detected in bronchoalveolar lavage (BAL) fluid specimens obtained from calves experimentally inoculated with Pasteurella haemolytica. ANIMALS: Twelve 2- to 3-month-old male Holstein calves. PROCEDURE: Pasteurella haemolytica was inoculated into the right lung lobes of each of 6 calves. Six other calves received vehicle alone and were used as control calves. Specimens of BAL fluid were obtained from 3 control and 3 inoculated calves 1 day after inoculation and from the other calves 2 days after inoculation. The amount of annexins I, II, IV, and VI, and haptoglobin in BAL fluid specimens was examined by use of immunoblot analysis. RESULTS: Annexins I and IV were detected in BAL fluid specimens obtained from the right lung lobes of each of the inoculated calves, but annexins II and VI were not. Annexin I also was found in BAL fluid specimens obtained from the left lung lobes of each inoculated calf and from left and right lung lobes of the control calves. By comparison, detection of annexin IV was essentially limited to the right lung lobes of inoculated calves. Haptoglobin was detected in some, but not all, BAL fluid specimens from the right lung lobes of inoculated calves, and its detection in BAL fluid was associated with serum proteins such as albumin. CONCLUSIONS AND CLINICAL RELEVANCE: Annexin IV was detected most specifically in response to inoculation of P haemolytica. This protein could be used as a marker for inflammatory pulmonary disease caused by P haemolytica.     

Peritoneal fluid values and collection technique in young,normal calves

Peritoneal fluid from 10 healthy young male Holstein calves was analyzed three times (2 to 3 days, 12 to 15 days and 27 to 30 days) during the first month of life. A new technique for collection of peritoneal fluid from calves positioned in left lateral recumbency was developed. The technique was found to be reliable and without noticeable complications. Mean peritoneal fluid nucleated cell counts, red blood cell counts, and absolute counts for mononuclear cells, lymphocytes and eosinophils did not change significantly (P 相似文献