Cytologic analysis of tracheal wash specimens and bronchoalveolar lavage fluid in the diagnosis of mycotic infections in dogs

Tracheal wash and bronchoalveolar lavage fluid analyses were performed in 9 dogs that had mycotic infections with pulmonary involvement. Characteristic organisms were identified in tracheal wash fluid in 3 of 7 dogs with blastomycosis. Organisms were identified in bronchoalveolar lavage fluid in 5 of 7 dogs with blastomycosis and in one dog with histoplasmosis. Organisms were not found in either fluid in one dog with coccidioidomycosis. These procedures should be considered for dogs with suspected mycotic infections that involve the lungs and that cannot be diagnosed by less invasive means.     

Specific enzyme activities in bronchoalveolar lavage fluid as an aid to diagnosis of tracheobronchitis and bronchopneumonia in dogs

Lactate dehydrogenase (LDH), alkaline phosphatase (ALP), alanine aminotransferase and gamma-glutamyl transferase enzyme activities, and total protein (TP), calcium, inorganic phosphate, urea nitrogen (UN) and creatinine concentrations in bronchoalveolar lavage fluid were investigated for their relative importance in the diagnosis of respiratory diseases in dogs. Bronchoalveolar lavage (BAL) fluid was obtained from 26 dogs (20 with respiratory diseases and six controls) following anaesthesia with sodium pentothal. Enzyme activities and biochemical parameters were measured in BAL fluid. LDH and ALP levels were significantly increased in 12 dogs with bronchopneumonia, but not in eight dogs with tracheobronchitis. Insignificant and variable levels of TP and UN concentrations were found in both groups. It was concluded that LDH and ALP enzyme activities could be considered as pointers to pulmonary inflammation and/or damage while TP and UN measurements in BAL fluid may have a place in the identification of changes in respiratory and vascular permeability.     

Use of a protected catheter brush for culture of the lower respiratory tract in horses with small airway disease.

A protected catheter brush introduced by fiberoptic bronchoscopy was used to sample the tracheai and bronchial mucosa in 28 horses with small airway disease. Tracheal and bronchial brushings were examined for the presence of fungi, aerobic and anaerobic bacteria, and a cytoiogical evaluation was also done on fluid collected by the bronchoalveolar lavage (BAL) technique. Microorganisms (bacteria and fungi) were isolated more often in tracheal brushings (53.6%) than in bronchial brushings (10.7%). Anaerobic bacteria were not isolated. Results of this study indicate that fiberoptic bronchoscopy using a protected catheter brush is an easy and practical technique to obtain minimally contaminated samples for isolation of microorganisms from the lower respiratory tract of horses. However, no association was observed between isolation of high numbers of microorganisms from the bronchi and severity of small airway disease.     

Use of a modified stomach tube for bronchoalveolar lavage in dogs

A technique that did not require use of a bronchoscope for bronchoalveolar lavage (BAL) in dogs was developed. An inexpensive, readily available 16-F Levin-type stomach tube was modified for the procedure. The technique was effective for collecting BAL fluid in 9 dogs that ranged from 9.3 to 26.2 kg (20.5 to 57.6 lb). Recovered fluid was consistent with fluid collected bronchoscopically. Mean recovery volume was 84/125 ml (67%), mean WBC counts were high (> 300 cells/microl), and > 70% of cells were macrophages. Complications from use of the technique were not detected on the basis of pulse oximetry, thoracic radiography, and clinical observation. This effective, simple, and safe technique for BAL can be readily performed in clinical settings that do not have bronchoscopic capabilities. It also provides a less costly alternative than bronchoscopic BAL.     

Influence of age on bronchoscopic findings in healthy beagle dogs

The aims of this study were to evaluate the effect of age on bronchoscopic features and bronchoalveolar lavage fluid (BALF) cellularity in dogs. Thirty healthy beagle dogs from three age groups were included: young dogs (10 months to 4.5 years of age; n = 8), middle-aged dogs (5–8 years old; n = 13) and older dogs (>8 years; n = 9). Haematology, thoracic radiography, bronchoscopy and bronchoalveolar lavage were performed; bronchoscopic findings were scored and BALF total and differential cell counts were determined. The total bronchoscopic score was higher in older dogs; these dogs had more irregular bronchial mucosa, more prominent mucosal vessels and bronchiectasis. Younger dogs had a higher percentage of neutrophils in BALF compared with middle-aged and old dogs and a higher percentage of lymphocytes in BALF compared with middle-aged dogs. The results show that age has an effect on bronchoscopic features of airways and the composition of BALF in the dog.     

Using urea dilution to standardise components of pleural and bronchoalveolar lavage fluids in the dog

AIM: To develop a technique to estimate the volume of epithelial lining fluid (ELF) obtained during bronchoalveolar lavage (BAL) and pleural lavage (PL) in the dog, using the urea dilution method. METHODS: BAL and PL fluids were obtained by saline lavage of pulmonary and pleural cavities of nine clinically healthy mixed-breed dogs immediately after euthanasia. Cell counts in the BAL and PL fluids were measured using standard techniques. The concentration of ELF in each lavage fluid was calculated from the relative concentration of urea in plasma and in each type of lavage fluid. Cell counts in ELF were then calculated. RESULTS: There were substantially higher cell counts in ELF compared to BAL or PF fluid. However, nucleated cell counts in ELF could not be predicted from cell counts in BAL or PL fluid. CONCLUSIONS AND CLINICAL RELEVANCE: These results suggest that accurate assessment of cellular or non-cellular components in lavage fluids should include a calculation of the proportion of ELF recovered, using a method such as urea dilution.     

Use of vitamin B12 in joint lavage for determination of dilution factors of canine synovial fluid

OBJECTIVE: To test a modified saline (0.9% NaCl) solution joint washing (lavage) technique that includes the use of vitamin B12 as an internal marker for the evaluation of synovial fluid dilution in lavage samples from canine joints. SAMPLE POPULATION: 9 plasma samples obtained from blood samples of 9 healthy dogs and 9 synovial fluid samples aspirated from stifle joints of 9 cadaveric dogs. PROCEDURE: Photometric absorbances of 25% vitamin B12 solution, canine synovial fluid, and canine plasma were measured in a spectrophotometer to establish an optimal wavelength for analysis. Canine synovial fluid and plasma samples were mixed with the 25% vitamin B12 solution to obtain 1%, 3%, 5%, 10%, 20%, and 50% solutions of synovial fluid or plasma. Diluted synovial fluid and plasma samples were used to simulate joint lavage samples and to examine the possible interference of these substances (synovial fluid or plasma) with the absorbance of the 25% vitamin B12 solution in photometric analysis. RESULTS: The optimal wavelength was found to be at 550 nm. Canine synovial fluid and plasma samples did not interfere with the absorbance measurements of the 25% vitamin B12 solution up to a 50% dilution of plasma or synovial fluid. CONCLUSIONS AND CLINICAL RELEVANCE: The modified saline solution joint lavage method with the use of a 25% vitamin B12 solution as an internal standard provides an accurate and reliable technique for the evaluation of synovial fluid dilution in lavage samples from canine joints.     

Immunohistochemical localisation of alpha 2-macroglobulin in the horse

A peroxidase antiperoxidase technique was used to identify alpha 2-macroglobulin in formalin-fixed sections of normal equine lung and liver and in tissue sections and bronchoalveolar lavage fluid from horses with various lung diseases. Equine peripheral blood leucocytes and bronchoalveolar lavage samples from clinically normal horses were negative for alpha 2-macroglobulin. It was concluded that liver and pulmonary macrophages may be potential sources of alpha 2-macroglobulin. Although alpha 2-macroglobulin may play a role in various chronic bronchointerstitial pneumonias of the horse, it is doubtful that it is of importance in equine chronic small airway disease.     

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.     

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 components of pleural and bronchoalveolar lavage fluids in the dog

AIM: To develop a technique to estimate the volume of epithelial lining fluid (ELF) obtained during bronchoalveolar lavage (BAL) and pleural lavage (PL) in the dog, using the urea dilution method.

METHODS: BAL and PL fluids were obtained by saline lavage of pulmonary and pleural cavities of nine clinically healthy mixed-breed dogs immediately after euthanasia. Cell counts in the BAL and PL fluids were measured using standard techniques. The concentration of ELF in each lavage fluid was calculated from the relative concentration of urea in plasma and in each type of lavage fluid. Cell counts in ELF were then calculated.

RESULTS: There were substantially higher cell counts in ELF compared to BAL or PF fluid. However, nucleated cell counts in ELF could not be predicted from cell counts in BAL or PL fluid.

CONCLUSIONS AND CLINICAL RELEVANCE: These results suggest that accurate assessment of cellular or non-cellular components in lavage fluids should include a calculation of the proportion of ELF recovered, using a method such as urea dilution.     

Cytological Analysis of Bronchoalveolar Lavage Fluid in the Diagnosis of Spontaneous Respiratory Tract Disease in Dogs: A Retrospective Study

Results of cytological analysis of bronchoalveolar lavage (BAD fluid were compared with clinical diagnoses in dogs that presented with signs of respiratory disease to referral hospitals. Of 68 dogs in which a clinical diagnosis was possible, BAL cytological findings were considered definitive for the diagnosis in 17 cases (25%), supportive of the diagnosis in 34 cases (50%), and not helpful in 17 cases (25%). Findings were most often considered supportive of or definitive for the clinical diagnosis in dogs with alveolar or bronchial radiographic patterns, or the presence of pulmonary masses. BAL results among lung lobes differed in 23 of 63 dogs (37%) with diffuse radiographic patterns. Tracheal wash cytology differed from BAL fluid cytology in 45 of 66 dogs (68%). Bronchoalveolar lavage was a clinically useful procedure for the diagnostic evaluation of dogs with signs of respiratory disease.     

Cellophane banding for the gradual attenuation of single extrahepatic portosystemic shunts in eleven dogs

Objective To evaluate the efficacy and short term effects of a cellophane banding technique for progressive attenuation of canine single extrahepatic portosystemic shunts.
Design A prospective trial of 11 dogs with single congenital extrahepatic shunts.
Procedure Rectal ammonia tolerance testing and routine biochemical tests were performed preoperatively on all dogs. In seven dogs, preoperative abdominal Doppler ultrasonography was also performed. Exploratory laparotomy revealed a single extrahepatic portocaval shunt in each animal, which was attenuated using a cellophane band with an internal diameter of 2 to 3 mm. The abdomen was closed routinely. Follow-up biochemical analysis and abdominal Doppler ultrasonography or splenoportography were performed postoperatively.
Results The shunt was not amenable to total ligation in 11 dogs, based upon reported criteria. All dogs recovered uneventfully from surgery without evidence of portal hypertension, and showed clinical improvement thereafter. Shunt occlusion was deemed to have occurred in 10 dogs based on resolution of biochemical and/or sonographic abnormalities. One dog continued to have sonographic evidence of portosystemic shunting when evaluated 3 weeks after surgery, despite normal ammonia tolerance, but was lost to subsequent follow-up. Two dogs, in which 3 mm cellophane bands were placed, experienced delayed shunt occlusion.
Conclusion Cellophane banding is simple to perform, and causes progressive attenuation of single extrahepatic shunts in dogs. Further work is needed to determine the maximum diameter of a cellophane band which will produce total attenuation, and the long-term safety and reliability of the treatment.     

Evaluation of circulating amino terminal-pro-B-type natriuretic peptide concentration in dogs with respiratory distress attributable to congestive heart failure or primary pulmonary disease

OBJECTIVE: To evaluate assessment of circulating amino terminal-pro-B-type natriuretic peptide (NT-proBNP) concentration as a means to discriminate between congestive heart failure and primary pulmonary disease in dogs. DESIGN: Prospective case series. ANIMALS: 46 dogs with signs of respiratory distress or coughing. PROCEDURES: All dogs underwent physical and thoracic radiographic examinations. Dogs with evidence of heart disease (eg, murmur, arrhythmia, or large cardiac silhouette detected by radiography) also underwent echocardiography. Dogs with no evidence of heart disease or failure were included if they underwent bronchoalveolar lavage (with cytologic examination and bacterial culture of the lavage fluid). Blood samples for NT-proBNP assay were obtained within 12 hours of the diagnosis of heart failure or prior to bronchoalveolar lavage in dogs with primary pulmonary disease. Circulating concentrations of NT-proBNP were compared between groups and correlated with radiographic and echocardiographic measures of cardiac size. RESULTS: Congestive heart failure and primary pulmonary disease were diagnosed in 25 and 21 dogs, respectively. Dogs with congestive heart failure had significantly higher median serum or plasma NT-proBNP concentration (2,554 pmol/L; interquartile [25% to 75%] range, 1,651.5 to 3,475.5 pmol/L) than dogs with primary pulmonary disease (357 pmol/L; interquartile range, 192.5 to 565.5 pmol/L). Radiographic vertebral heart score and echocardiographic left atrial-to-aortic diameter ratio were not correlated with NT-proBNP concentration. Left ventricular end-diastolic diameter (measured echocardiographically) and NT-proBNP concentration were weakly correlated. CONCLUSIONS AND CLINICAL RELEVANCE: Serum or plasma NT-proBNP concentration assessment may be useful for discrimination of congestive heart failure from primary pulmonary disease in dogs with respiratory distress or cough.     

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.     

Comparison of bronchoalveolar lavage cytospins and smears in dogs and cats

Differences in the cytological interpretation of bronchoalveolar lavage fluid (BALF) after cytospin preparation (CP) or manual smearing of pelleted cells preparation (MSP) were investigated in client-owned dogs and cats with inflammatory or infectious lower respiratory disease. Bronchoalveolar lavage fluid from healthy cats was also examined. With MSP, cell lysis was more frequently observed, and cellular distribution was more heterogeneous throughout the slide. When samples from healthy and diseased animals were considered together, a significantly greater percentage of neutrophils was seen on CP than on MSP slides (P<0.002). Cytospin preparations were considered of better quality in all individual comparisons. Cytospin preparation is advised in the evaluation of BALF with low total cell count. When only MSPs are evaluated, clinicians should be aware that differential neutrophil counts may underestimate the counts found on CP slides.     

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.     

Tracheobronchial Brush Cytology and Bronchoalveolar Lavage in Dogs and Cats with Chronic Cough: 45 Cases (2012–2014)

Background

Animals with chronic cough can have normal bronchoalveolar lavage fluid cytology when small airway disease is absent. Cytology of a tracheobronchial brushing can detect inflammation in larger airways; however, evaluation of this technique has been limited in veterinary medicine.

Objective

To compare airway brush cytology to bronchoalveolar lavage fluid analysis in dogs and cats with chronic cough.

Animals

Forty dogs and five cats undergoing bronchoscopic investigation of chronic cough.

Methods

Prospective study. Bronchoscopy and bronchoalveolar lavage were performed followed by tracheobronchial brushing of central airways. Results of cytologic assessment of BAL fluid and brush cytology were compared for the presence or absence of inflammation and concordance of inflammatory cell type.

Results

Brush cytology detected central airway inflammation in 34 of 40 (85%) dogs with inflammatory BAL fluid. However, the type of inflammation reported differed in 23 of 34 dogs. In five cats with inflammation in BAL fluid, brush cytology detected inflammation in four; the type of inflammation was discordant in all cats.

Conclusions and clinical relevance

Brush cytology has good agreement with BAL regarding the presence of inflammation, although the type of inflammation detected with the different sampling techniques commonly varies. Brush cytology can provide supplementary information to BAL, and additional studies will provide further information on the role of tracheobronchial brush cytology in the diagnosis and management of respiratory conditions.     

Hemodynamic Changes During Laparoscopic Radiofrequency Ablation of Normal Adrenal Tissue in Dogs

Objective— To determine the hemodynamic response to radiofrequency ablation (RFA) of normal adrenal tissue in dogs.
Study Design— Experimental study.
Animals— Healthy adult mixed-breed dogs (n=6).
Methods— During general anesthesia a Swan–Ganz thermodilution catheter was flow directed into the pulmonary artery and used to quantify cardiac output. An arterial catheter was used for direct blood pressure measurements. An RFA device was introduced into the left adrenal gland under observation through laparoscopic instrumentation. Blood samples were collected and hemodynamic variables studied after a stable surgical anesthetic depth was achieved (time 1), during CO₂ insufflation of the abdomen (time 2), during adrenal RFA (time 3), and after completed RFA (time 4). Catecholamine determinations were performed with a human enzyme immunoassay. Histopathology was performed to verify medullary necrosis.
Results— Arterial, pulmonary arterial and central venous pressure, and plasma norepinephrine increased more during RFA than during abdominal insufflation. Heart rate and cardiac index did not differ between time points. High baseline epinephrine was present and significant differences between time points were not detected. Systemic vascular resistance had very high individual variation and differences were not detected.
Conclusions— RFA of normal adrenal tissues is associated with severe hemodynamic alterations. Further studies of the optimal blockage of catecholamine-induced hypertension in dogs are warranted.
Clinical Relevance— Clinicians should prepare for potential hypertensive crisis during RFA of adrenal masses, especially if treating a margin of normal tissue.     

Pulmonary cryptococcosis and Capillaria aerophila infection in an FIV-positive cat

A 12-year-old, FIV-positive, domestic longhair cat was presented with a history of sneezing and coughing during the previous seven months. On thoracic radiographs, a prominent bronchial pattern and three focal, opacified nodules were seen. Cytology of bronchoalveolar lavage fluid demonstrated spherical, capsulate, narrow-necked, budding yeasts within macrophages. Culture of the fluid yielded a heavy growth of Cryptococcus neoformans var neoformans. The serum latex cryptococcal antigen agglutination test titre was 158. The cat was treated with itraconazole and the cough resolved over a 5-month period but then recurred. Repeat thoracic radiographs showed resolution of the pulmonary nodules but a persistent bronchial pattern. Adult nematodes and ova with morphology characteristic of Capillaria aerophila were seen in bronchoalveolar lavage fluid and no yeasts were cultured from the fluid. The cryptococcal titre was zero. The lungworm infection was treated successfully with abamectin and the cough resolved. Immunosuppression related to FIV infection may have predisposed this cat to sequential respiratory tract infections.