Unguided bronchoalveolar lavage techniques and residual effects in dogs

Objectives To investigate the use of unguided bronchoalveolar lavage techniques in dogs without fibreoptic bronchoscopy, using an adapted single vascular catheter and a double-lumen catheter made from two single vascular catheters.
Animals Sixty-nine dogs were examined with the single-catheter technique and 110 dogs with the double-catheter technique.
Design A prospective study.
Procedure Sixty-nine and 220 samples, collected with the single catheter and the double catheter respectively, were examined cytologically Lungs of 69 dogs were examined grossly and histologically. Radiographic examination was performed on 11 dogs.
Results The double-catheter technique produced samples with significantly higher cellularity (P < 0.01) and fewer red blood cells (P < 0.01) than the single-catheter technique. Repeat samples collected with a double catheter were not significantly different (P > 0.01) in any value. A reference range for nucleated cell counts of 62 to 1210 – 10⁶/L was calculated from 57 clinically and histologically normal dogs. The major residual effects of the technique were localised pulmonary oedema, and alveolar distension with collapse and congestion of distant parenchyma. Thoracic radiographs revealed increased lung opacity for at least up to 7 h after the procedure.
Conclusions The cellularity of the bronchoalveolar lavage fluid obtained was adequate and sufficient fluid was retrieved when the single catheter was located in a proper position. However, the double catheter obtained better samples more quickly and easily, with less damage to the respiratory tract.     

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.     

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.     

Collection of bronchoalveolar lavage fluid in cats, using an endotracheal tube

Bronchoalveolar lavage fluid was collected from 12 anesthetized cats by use of an endotracheal tube and syringe adapter. The safety of the technique was evaluated by monitoring mucous membrane color, capillary refill time, pulse rate, respiratory rate, ECG, and arterial blood gas tensions and by necropsy findings. Group A consisted of 3 cats that were administered (by lavage) 4 aliquots of 20 ml of saline solution during anesthesia for placement of femoral artery catheters. Group B consisted of 4 cats that were administered a smaller total volume of saline solution (3 aliquots of 5 ml/kg of body weight) during a separate anesthetic period, other than the one for placement of catheters. Group C consisted of 5 cats administered 3 aliquots (5 ml/kg) of saline solution during a separate anesthetic period and administered supplemental oxygen for 5 to 10 minutes before and for 20 minutes after the lavage procedure. Group-A cats had a prolonged recovery period that was attributed to the lengthy anesthetic period required for placement of femoral catheters. The effect was eliminated in the cats of the other groups in which the lavage procedure itself accounted for only 5 to 10 minutes of anesthetic time. Evaluation of mucous membrane color, capillary refill time, ECG, pulse, and respiratory rate revealed no persistent abnormalities. Transient increase in pulse and respiratory rate was seen in some cats. Blood gas analysis revealed noticeable decrease in arterial oxygen pressures (Pao2) after the lavage procedure. In group-C cats, oxygen supplementation allowed the maintenance of normal or above normal Pao2.(ABSTRACT TRUNCATED AT 250 WORDS)     

Quantitative bacterial cultures and cytological examination of bronchoalveolar lavage specimens in dogs

Cytology and quantitative bacterial cultures of lower respiratory tract secretions are widely used in human medicine to differentiate airway infection from simple bacterial colonization. A retrospective study was conducted to determine the usefulness of quantitative aerobic cultures and Gram stain intracellular bacteria counts from bronchoalveolar lavage (BAL) specimens in dogs in diagnosing lower respiratory tract infection (LRTI) and to determine whether chronic bronchitis is associated with marked bacterial growth in dogs. The threshold determined to define clinically relevant bacterial growth was 1.7 x 10(3) colony-forming units per milliliter of BAL fluid. We used this threshold and found that diagnostic sensitivity and specificity were 86% and 100%, respectively. With a threshold for infection of >2 intracellular bacteria observed in any of 50 fields, microscopic examination of Gram stain BAL preparations had a sensitivity of 71% and a specificity of 97% in establishing LRTI. There was a high correlation between bacterial morphology on BAL Gram stain and bacterial cultures. Combining the results of intracellular bacteria counts from the BAL Gram stain with those from the quantitative cultures, the sensitivity in diagnosing LRTI was 87% and the specificity was 97%. BAL quantitative cultures as well as quantitating intracellular bacteria on Gram stain BAL cytology were revealed to be useful in identifying LRTI in dogs. Chronic bronchitis does not appear to be associated with marked bacterial growth in dogs.     

Cytological and parasitological analysis of bronchoalveolar lavage fluid for the diagnosis of Angiostrongylus vasorum infection in dogs

Bronchoalveolar lavage (BAL) is a procedure that retrieves cells and other elements from the lungs for evaluation, which helps in the diagnosis of many pulmonary diseases. The aims of this work were to perform this procedure in dogs in the acute and chronic phases of an Angiostrongylus vasorum infection for cytological analysis and to evaluate the potential of this technique as a diagnostic method for this lung-heart worm. The BAL procedure was performed through the use of an endotracheal tube on seven A. vasorum infected dogs and on five non-infected dogs lined as a control group. Sixty days post-infection (dpi) active and live larvae were retrieved from the bronchoalveolar fluid (BALF) of all infected dogs. Furthermore, in one animal it was possible to retrieve larvae in its BALF before the pre-patent period. This work reports that the A. vasorum infection resulted in an increase of relative neutrophils and eosinophils counts. In contrast, there was a significant decrease in the alveolar macrophage relative count in infected animals from 60 to 330 dpi. This study shows that the BAL is an accurate technique for the diagnosis of canine angiostrongylosis. Moreover, the technique allows us to retrieve cells and other elements that line the lung surface for cytological evaluation, which provides information about inflammatory diseases, and the diagnosis and prognosis of pulmonary parasites such as A. vasorum.     

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.     

Use of a jugular vein graft in a modified Blalock-Taussig procedure in dogs

OBJECTIVE: To determine whether autologous jugular veins provide functional grafts with high 30-day patency rates in an experimental model of systemic-to-pulmonary shunting performed with a modified Blalock-Taussig procedure. ANIMALS: 15 healthy Beagles. PROCEDURE: A segment of the left jugular vein was implanted between the left subclavian and pulmonary arteries. Echocardiograms were obtained prior to surgery, at day 4 to 7, and at day 30 after surgery. Selective angiograms were performed immediately after surgery and on day 30. Oximetric shunt calculations were made via terminal angiography prior to euthanasia. Gross and histologic evaluations of the grafts were conducted. RESULTS: Grafts were patent in 12 of 15 dogs 30 days after surgery as assessed via auscultation, color Doppler ultrasonography, angiography, and histologic examination. Echocardiographic analysis revealed compensatory eccentric left ventricular hypertrophy. Mean pulmonary-to-systemic flow ratio was 1.5:1. Histologic evidence of endothelialization of the anastomotic sites and vein graft arterialization was detectable at 30 days. CONCLUSIONS AND CLINICAL RELEVANCE: Autologous jugular vein grafts were effectively used to create a systemic-to-pulmonary shunt by use of a modified Blalock-Taussig procedure. High patency, ready accessibility, low cost, and theoretical adaptative remodeling during patient growth make autologous jugular vein grafts a valuable alternative to synthetic materials.     

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.     

Diagnosis of naturally occurring toxoplasmosis by bronchoalveolar lavage in a cat

A 5-year-old cat presented with a history of fever and respiratory distress of 7 days' duration. The cat did not respond to initial symptomatic treatment. Through use of a bronchoalveolar lavage (BAL) technique, the diagnosis of toxoplasmosis was obtained rapidly without evidence of adverse effects to the cat. Antemortem diagnosis of naturally occurring active Toxoplasma gondii infection can be difficult in cats. Identification of organisms obtained by BAL may be beneficial in the diagnosis of feline toxoplasmosis.     

Alveolar lavage in dogs

Alveolar lavage was performed in 10 healthy dogs. After tracheal intubation was done, a sterile fiberoptic bronchoscope was wedged in a distant bronchus and the lungs were lavaged with sterile saline solution. An average of 140 ml of saline solution was flushed into the lungs of each dog, and an average of 79% of the solution was recovered. Examination of the recovered fluid revealed average total cell counts of 6.4 X 10(6) cells/dog. Average differential cell counts were as follows: macrophages, 50.5%; lymphocytes, 46.0%; and neutrophils, 3.5%. Results of bacterial culture of the recovered fluid were negative in 8 dogs and positive in 2; Bordetella bronchiseptica was isolated in 1 dog and Klebsiella pneumoniae was isolated in the other.     

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.     

Elevated levels of fragmented laminin-5 gamma2-chain in bronchoalveolar lavage fluid from dogs with pulmonary eosinophilia

Inflammation causes epithelial cell sloughing and basement membrane (BM) exposure in canine pulmonary eosinophilia (PE), leading to degradation of the epithelial cell attachment component, laminin-5 gamma2-chain, into small molecular weight fragments. The subsidence of inflammation after treatment down-regulates degradation. Laminin-5 gamma2-chain levels and molecular forms in bronchoalveolar lavage fluid (BALF) were analysed semiquantitatively by Western immunoblotting to compare PE affected (n=20) and healthy dogs (n=16) as well as PE dogs (n=6) before and after corticosteroid treatment. PE dogs expressed significantly elevated levels of total (P<0.01), 36 kDa (P<0.05) and 53 kDa (P<0.05) laminin-5 gamma2-fragments. The 36 Da fragment decreased significantly (P<0.05) after treatment. The laminin-5 gamma2-chain degradation products may be linked to epithelial cell sloughing and BM exposure or healing.     

Respiratory disease markers in porcine bronchoalveolar lavage fluid

In bronchoalveolar lavage fluid (BALF) of pigs originating from different herds bacteria, cells and the antibacterial peptide PR-39 were examined to gain information about the lung health status. In a high health nucleus herd 56% and in low health herds 20-100% of the examined pigs were found positive for potentially pathogenic bacteria. Based on these findings, a novel definition for bacterial respiratory tract disease was established using an 8% cut-off for the relative number of neutrophils in bronchoscopic and a 40% cut-off in transtracheal BALF in combination with the occurrence of potentially pathogenic microorganisms. The antibacterial peptide PR-39 was highly correlated to this definition of respiratory disease. An assessment of the bacteriological respiratory health status appears to be possibly based on the determination of PR-39 concentrations in BALF using different cut-off values according to the lavage method (2.5 nM for bronchoscopic and 5 nM for transtracheal BALF).     

Association between quantitative bacterial culture of bronchoalveolar lavage fluid and antibiotic requirement in dogs with lower respiratory tract signs

BackgroundHistorically, positive bacterial cultures from the lower respiratory tract (LRT) have been considered clinically relevant when quantitative bacterial cultures of bronchoalveolar lavage fluid (BALF) were >1700 colony forming units (cfu)/mL. However, this threshold might not accurately predict a requirement for antibiotics.ObjectivesTo study whether quantitative BALF bacterial culture results were predictive of antibiotic requirement in dogs with LRT signs.AnimalsThirty‐three client‐owned dogs.MethodsCross‐sectional study. Dogs with positive quantitative bacterial culture of BALF were included. Dogs were divided into 2 groups, depending on whether they had a LRT infection requiring antibiotics (LRTI‐RA) or LRT disease not requiring antibiotics (LRTD‐NRA), based on thoracic imaging features, presence of intracellular bacteria on BALF cytology, and response to treatment. Predictive effect of cfu/mL and BALF total nucleated cell count (TNCC) on antibiotic requirement, adjusting for ongoing or prior antibiotic therapy and age, were studied using logistic regression.ResultsTwenty‐two and 11 dogs were included in the LRTI‐RA and LRTD‐NRA groups, respectively. The cfu/mL was not significantly predictive of antibiotic requirement, independent of ongoing or prior antibiotic treatment and age (LRTI‐RA: median, 10 000 cfu/mL; range, 10‐3 × 10⁸; LRTD‐NRA: median, 10  000 cfu/mL; range, 250‐1.3 × 10⁹; P = .27). The TNCC was not significantly predictive of antibiotic requirement when only dogs with bronchial disease were considered (LRTI‐RA: median, 470 cells/μL; range, 240‐2260; LRTD‐NRA: median, 455 cells/μL; range, 80‐4990; P = .57).Conclusion and Clinical ImportanceThe cfu/mL is an inappropriate measure for determining whether antibiotics are of benefit in dogs with LRT signs.     

Severe bronchoconstriction after bronchoalveolar lavage in a dog with eosinophilic airway disease

Bronchoalveolar lavage (BAL) is considered to be a safe procedure in humans and other animals. However, in some instances, potentially life-threatening complications can arise. In this clinical report, a dog with eosinophilic airway disease that underwent a BAL and subsequently became profoundly dyspneic during the recovery period is described. The dyspnea was severe enough to warrant mechanical ventilation for almost 24 hours. Several anti-inflammatory and bronchodilatory medications were also used. The dog was successfully weaned off the ventilator and made a full recovery. On the basis of radiographic findings and clinical response to treatment, we believe the dog had acute exacerbation of eosinophilic airway disease and severe bronchoconstriction secondary to the BAL. Caution should be exercised when performing a BAL if there is suspicion of a reactive airway disease.     

Cellular composition of bronchial brushings obtained from healthy dogs and dogs with chronic cough and cytologic composition of bronchoalveolar lavage fluid obtained from dogs with chronic cough

OBJECTIVE: To determine whether bronchial brushings from dogs with chronic cough have increased numbers of goblet cells and WBCs, compared with numbers for healthy dogs, or have differing WBC populations, compared with populations in bronchoalveolar lavage (BAL) fluid obtained from dogs with chronic cough. ANIMALS: 9 healthy dogs and 10 dogs with chronic cough. PROCEDURE: Specimens were collected by use of bronchoscopy. Cellular composition was determined for brushings, and results from dogs with chronic cough were compared with those from healthy dogs. Cellular composition of brushings was compared with composition of BAL obtained from dogs with chronic cough. RESULTS: Brushings from healthy dogs contained a median of 2.9 x 10(6) epithelial cells, comprising 100% epithelial cells (96% ciliated, 3% goblet, and 1% other) and no WBCs. Brushings from dogs with chronic cough had 4.5 x 10(6) epithelial cells, comprising 93% epithelial cells (86% ciliated, 2% goblet, and 12% other). Dogs with chronic cough had significantly greater percentages of WBCs (7%) and neutrophils (6%), compared with values for healthy dogs. Five dogs with chronic cough had no neutrophilic inflammation evident in BAL, but 4 of these had evidence of neutrophilic inflammation in brushings. CONCLUSIONS AND CLINICAL RELEVANCE: Neutrophils, but not goblet cells, were increased in brushings from dogs with chronic cough. Analysis of bronchial brushings provides information about airway inflammation that differs from that found by examination of BAL in some dogs with chronic cough and is a more sensitive indicator of airway inflammation than cytologic examination of BAL in these dogs.