Bronchoalveolar Lavage Cytology Research Articles

Pathology and bronchoalveolar lavage fluid cellular and oxidative stress changes in canine pneumonia in Nigeria

Studies on canine respiratory diseases are scant in the literature especially in Africa, unlike the situation on ruminants. This study is on the pathology and bronchoalveolar cellular and oxidative stress changes in naturally occurring canine pneumonia in Nigeria. The dogs used were collected from cases submitted for necropsy in the institutional Department of Veterinary Pathology. Gross examination, bronchoalveolar lavage (BAL) cytology, oxidative biochemical assay of BAL fluid (BALf) supernatant and histology of sections from all the lung lobes were done using standard procedures. Grossly, 22 (31%) were normal while 48 (69%) were congested, haemorrhagic and/or pneumonic out of the 70 canine lungs examined. Microscopically, there were pulmonary congestion (4%), pulmonary oedema (20%), pulmonary haemorrhages (7%), atelectasis (4%), interstitial pneumonia (10%), bronchointerstitial pneumonia (14%) and bronchopneumonia (9%). The BALf cytology of pneumonic lungs showed increases in neutrophils, alveolar macrophages, lymphocytes and haemosiderophages including free and phagocytosed bacteria, and fungal hyphae. Glutathione peroxidase (GPx) and glutathione transferase (GST) activities increased in congested and bronchointerstitial pneumonic lungs but were reduced in other pneumonic lungs. The superoxide dismutase (SOD) activity was significantly reduced in the pneumonic canine lungs. Hydrogen peroxide generation increased in congestion and oedema, bronchopneumonia and bronchointerstitial pneumonia. Malondialdehyde (MDA) level increased in other congested and pneumonic lungs. There was also significant positive correlation of BALf pro-oxidant assay to the type and severity of pneumonia (+ 0.75, P < 0.05). Pneumonias account for 57.2% of the respiratory conditions of which bronchointerstitial and interstitial pneumonias are the prevalent types. The study also showed that oxidative stress markers in BALf may serve as predictors of pneumonia especially in the early stages. Since treatment recommendations for pneumonia often centre primarily on supportive care and anti-oxidant and antibiotic administration, further studies on causal agents, treatment and prevention strategies for canine pneumonia in Nigeria are warranted.

Reliability of cytological evaluation of mast cells from bronchoalveolar lavage fluid in horses: Intraobserver agreement and mast cell identification

SummaryEndoscopy, including bronchoalveolar lavage (BAL) fluid cytology, are invaluable tools for equine lower airway diagnostics. Objectives of the study were to evaluate the reliability of two staining methods, May–Grünwald–Giemsa (MGG) and toluidine blue (TB), for the evaluation of mast cells in cytospin smears from BAL fluid samples and the reliability of a 500‐cell differential cell count using both staining methods. Endoscopies and BAL fluid sampling were performed and BAL cytology was evaluated from cytospin smears. BAL cytology slides from 78 horses were evaluated. Three experienced observers evaluated the BAL cytology cytospin smears for the mast cell comparison study. The intra‐ and interobserver agreements were calculated using intraclass correlation coefficients (ICC). BAL cytology mast cell counts were significantly higher with the TB staining method compared with the MGG staining method. Observer agreements showed below acceptable agreement between observers for mast cell counts for both staining methods, even with fixed coordinates. Based on results from this study recommendation for BAL cytology staining would be to use both the MGG and the TB staining method, relying on the TB for the enumeration of mast cells.

Bronchoalveolar Lavage Cytology Characteristics and Seasonal Changes in a Herd of Pastured Teaching Horses.

Equine asthma syndrome (EAS) is a common problem that affects horses of any age. Severe EAS is reported to affect 10–20% of adult horses in the northern hemisphere, while mild/moderate EAS is reported to affect 60–100% of adult horses, depending on the population and geographic region. For both severe and mild/moderate EAS, the presence of lower airway inflammation is attributed to airborne “triggers” such as dust, mold, and bacterial components that horses encounter in hay and stable-environments; and treatment recommendations for horses with EAS often include full-time pasture turnout. The caveat to this recommendation is horses with summer-pasture associated EAS (SP-EAS), who experience allergic lower airway inflammation when exposed to summer pasture. The prevalence of EAS in horses on pasture that do not have SP-EAS has not been reported. The purpose of this study was to use bronchoalveolar lavage (BAL) cytology to determine the prevalence of EAS in a herd of pastured, adult research horses with no history of respiratory disease. The horses were members of a teaching animal herd housed on pasture in the southeastern United States and fed round-bale Bermuda-grass hay. BAL fluid (BALF) cytology was analyzed in both summer (May–August 2017) and winter (November 2017–February 2018). Similar to previous reports, the prevalence of severe EAS in our study population was 10% in summer and 4.3% in winter. The prevalence of mild/moderate EAS was 60% in summer and 87% in winter. The high prevalence of mild/moderate EAS in this population was unexpected, given the 24-h, year-round pasture environment and the lack of history of respiratory disease. Additionally, 61.1% of horses with both summer and winter data had a different BALF cytology profile between the two seasons. To the authors' knowledge, this is the first study to use BAL cytology to diagnose, and monitor changes in, EAS phenotype in pastured adult horses. These results help to inform discussions regarding prevalence of EAS in pastured, adult horses in the southeastern region of North America.

Bronchoalveolar lavage hemosiderosis in dogs and cats with respiratory disease.

Hemosiderophages can be found in bronchoalveolar lavage samples and have been reported in association with a wide range of respiratory and cardiovascular disorders in cats and humans. This study aimed to retrospectively evaluate the presence of hemosiderin in canine and feline bronchoalveolar lavage (BAL) samples. It also aimed to examine the association of BAL hemosiderin with signalment, clinical signs, and historical disease prior to BAL, with prior transthoracic fine-needle aspiration (FNA), with bronchoalveolar lavage duration, and with cytologic interpretation. The medical records of dogs and cats with respiratory disease that underwent BAL between 2007 and 2016 were reviewed. Appropriate medical information and BAL results were available from 171 dogs and 34 cats. Cases were assigned to four disease categories based on BAL cytologic findings: pneumonia, inflammatory disease, neoplasia, or normal airways. The degree of hemosiderosis was classified based on a semi-quantitative scoring scale. Exact logistic regression analysis was used to evaluate the relationship between risk factors and the presence of BAL hemosiderosis on cytology. Hemosiderin was identified in 13/171 (7.6%) canine samples and 18/34 (52.9%) feline samples. Cats were 13.33 times more likely to have pulmonary hemosiderosis on bronchoalveolar lavage cytology compared with dogs (P<0.001). Increased respiratory rates, prolonged bronchoalveolar lavage times, concurrent transthoracic FNAs, and cytologic diagnoses were associated with an increased risk of hemosiderosis in dogs. No specific risk factors associated with pulmonary hemosiderosis in cats were identified. Hemosiderosis is more common in BAL samples from cats than from dogs and is associated with a diverse range of disease conditions.

Fungi in respiratory samples of horses with inflammatory airway disease.

BackgroundFungi contribute to the inflammatory response of lungs in horses with recurrent airway obstruction and in some forms of asthma in humans. The role of fungi in inflammatory airway disease (IAD) has not been assessed.ObjectivesEvaluate the prevalence of fungi in the respiratory samples of horses diagnosed with IAD, describe clinical signs associated with the presence of fungi in respiratory samples, and assess the risk factors associated with IAD and with the presence of fungi in the airways.AnimalsSeven‐hundred thirty‐one active horses referred to a specialized ambulatory practice for signs of respiratory disease or poor performance.MethodsA prospective observational study was performed, collecting clinical data, environmental conditions, and results of a tracheal wash (TW; cytology, fungal culture, and bacterial culture), and bronchoalveolar lavage (cytology).ResultsA positive fungal culture was obtained in 55% (402/731) of horses. Horses with fungal elements observed on the TW cytology had 2 times greater chance of having IAD than horses without fungi (odds ratio [OR] = 2.1; 95% CI 1.08‐3.33; P = .0003). Risks of being diagnosed with IAD and likelihood of fungi in TW were higher when horses were bedded on straw (OR = 2.0; 95% CI 1.2‐3.2 and OR = 1.9; 95% CI 1.3‐2.6, respectively) or fed dry hay (OR = 2.7; 95% CI 1.7‐4.4 and OR = 2.6; 95% CI 1.6‐3.4, respectively).Conclusions and Clinical ImportanceHorses inhaling aerosolized fungal particles are at a significantly higher risk of developing IAD. The type of bedding and forage represent significant risk factors for IAD and fungal contamination of equine airways.

356. Bronchoalveolar Lavage Fluid Cytology by GMS Stain for the Diagnosis of Invasive Pulmonary Aspergillosis in Patients With Hematologic Malignancies: Analysis of 67 Episodes

BackgroundThe yield of direct fungal visualization by GMS (Gomori–methenamine–silver) stain in bronchoalveolar lavage (BAL) cytology has rarely been studied in the diagnosis of invasive pulmonary aspergillosis (IPA) in patients with hematological malignancies (HM).MethodsWe analysed a series of patients with proven or probable culture-documented IPA (EORTC/MSG criteria) in HM patients (1999–2015). All patients had BAL cultures that were positive for Aspergillus spp. and had concurrently obtained BAL cytology GMS available for analysis.ResultsWe identified 67 such patients. BAL cytology based on GMS showed hyalohyphomycetes consistent with Aspergillus in 28/67 (41.8%) patients, whereas only in 2/67 (3.6%) direct smear Calcofluor White stain was positive. Based on BAL GMS cytology, co-infections were identified in six patients: two Pneumocystis and five viral infections with cytopathic changes (one had both). The yield of cytology was not different in patients with IPA caused by non-fumigatus Aspergillus, although patients with IPA and >1 Aspergillus in BAL culture had more often positive cytology GMS (100% vs. 0%, p 0.027). Cytology was also more often positive when obtained from a lesion-targeted BAL as compared with non-targeted bronchial washings (60.7% vs. 7.1%, p 0.038). Patients with IPA and cavitary lesions (32.1% vs. 5.1%, P = 0.006), history of SCT (64.3% vs. 33%, P = 0.015) or prior exposure to itraconazole (75% vs. 41%, P = 0.007) had positive cytology GMS results more often than did patients without these characteristics. In the multivariate analysis, only cavitary lesions were significantly associated with positive BAL GMS cytology.ConclusionGMS stain in cytology of BAL in patients with HM and culture-documented IPA had a sensitivity of 41.8% and was more often positive in patients with cavitary lesions. Although there were no differences in the proportion of GMS-positive cytology rates among differing Aspergillus spp. causing IPA, mixed Aspergillus spp. IPA was associated with an increase in positive cytology. BAL cytology was diagnostic for co-infections in more than 10% of patients. BAL cytology should be part of the diagnostic wok up in HM patients with suspected IPA.Disclosures All authors: No reported disclosures.

DIAGNOSING MILLIARY MYCOBACTERIUM BOVIS FROM THE PROSTATE OF AN IMMUNOCOMPETENT HOST

SESSION TITLE: Chest Infections 1 SESSION TYPE: Med Student/Res Case Report PRESENTED ON: 10/09/2018 03:45 PM - 04:45 PM INTRODUCTION: Zoonotic infection from Mycobacterium Bovis remains an important consideration for tuberculous disease in the developing world. We present a challenging case of hematogenous Mycobacterial dissemination from an uncommon culprit. CASE PRESENTATION: A 53 year old Bacillus-Calmette-Guerin (BCG) vaccinated and otherwise healthy Mexican male presented with complaints of fever, productive cough, arthralgias and weight loss over the preceding 21 days. He had already completed two separate outpatient courses of oral antibiotics without improvement. He denied any travel outside of the continental United States over the last 13 years and denied any recent sick contacts. He also owned 2 pet parrots. His physical exam and initial laboratory work up was unremarkable. A Chest X-ray (Fig. 1) revealed diffuse, bilateral, micronodular opacities, warranting further evaluation with Computed Tomography (CT) (Fig. 2). Initial acid-fast bacilli (AFB) sputum stains, bronchoalveolar lavage stains, cytology brushings and trans-bronchial biopsies were all negative. Hypersensitivity pneumonitis and opportunistic fungal panels were obtained which were unremarkable. Interferon gamma release assay was positive however equivocal given patient's BCG vaccination status. CT Chest, Abdomen and Pelvis (Fig. 3) was performed, which incidentally revealed an enlarged, heterogeneous prostate with fluid collections. Ultrasound guided aspiration of the prostate yielded pus and a strongly positive AFB smear. Eventually, the AFB cultures from the initial BAL came back positive. The patient was started on RIPE therapy with symptomatic improvement. Final speciation returned as M. Bovis and accordingly, pyrazinamide was stopped. DISCUSSION: M. bovis disease was common in the industrialized world before the practice of pasteurization became widely implemented in the 1940s. Current estimates in the USA and Europe identify between 1.3%-1.6% of tuberculosis cases attributable to M. Bovis. Infections are typically contracted via ingestion of unpasteurized dairy products from infected livestock and may remain dormant for many years, as was likely the situation in this case. Given the oral inoculation route, most cases of M.bovis manifest primarily with extrapulmonary findings. Of note, M. Bovis has documented intrinsic resistance to pyrazinamide which necessitates modification of typical RIPE treatment strategy. CONCLUSIONS: Diagnosis of disseminated TB remains a challenge. Regarding M. Bovis, the mean time to diagnosis has increased among practitioners given the diminishing incidence of infection. Vigilance of the disseminated and often extra-pulmonary form of tuberculosis caused by M. Bovis continues to be warranted. Patients at particular risk for M.bovis are those from developing countries, with frequent exposure to livestock and/or the practice of unpasteurized dairy product consumption. Reference #1: Scott. C. Human Tuberculosis Caused by Mycobacterium bovis in the United States, 2006-2013. Clin Infect Dis. 2016 Sep 1;63(5):594-601. Reference #2: Sherman LF. Patient and health care system delays in the diagnosis and treatment of tuberculosis. Int J Tuberc Lung Dis. 1999 Dec;3(12):1088-95. Reference #3: Majoor, C.J. Epidemiology of Mycobacterium bovis disease in humans, The Netherlands, 1993-2007. Emerg Infect Dis. 2011 Mar;17(3):457-63. DISCLOSURES: no disclosure on file for Christopher Barrios; No relevant relationships by Armin Krvavac, source=Web Response No relevant relationships by Scott Maughan, source=Web Response No relevant relationships by Pujan Patel, source=Web Response

Seasonal Variation in Tracheal Mucous and Bronchoalveolar Lavage Cytology for Adult Clinically Healthy Stabled Horses

Seasonal changes affect the concentration of dust particles and aeroallergens in the equine stable environment and are hypothesized to change airway cytology in healthy stabled horses. The objective of the study was to evaluate if seasonal changes from May to November influenced the lower airway health cytological findings in clinically healthy stabled horses. Endoscopic examination, including tracheal aspiration (TA) and bronchoalveolar lavage (BAL) cytology was performed in November and May on 63 clinically healthy stabled Danish Warmblood geldings, aged 12.9 (±4.6) years. Tracheal aspiration and BAL cytology were evaluated via May-Grünwald-Giemsa stain after cytocentrifugation of samples. Five-hundred cells were identified microscopically as macrophages, lymphocytes, neutrophils, eosinophils, or mast cells. A significant difference in climatic conditions with lower temperatures (P < .001), fewer sunlight hours (P = .004), less precipitation (P = .006), and higher humidity (P < .001) was found in November compared to May. A significantly higher percentage of neutrophils (Median 6.10%, Quartiles 4.00–8.80, P < .001) and a higher tracheal mucus score (Median 3.50%, Quartiles 1.95–5.20, P = .010) were found during November compared to May period. Furthermore, a positive correlation between TA and BAL neutrophils was found for both sampling times. Mild winter weather conditions with lower temperatures, fewer minutes of sunlight, and a higher percent humidity were associated with significantly higher BAL neutrophil percentage and significantly higher mucus score in stabled clinical healthy horses. The clear seasonal influence on BAL neutrophilia and tracheal mucous findings in clinically normal horse needs to be included in the working diagnosis of asthma in horses.

Identifying aerosolized cyanobacteria in the human respiratory tract: A proposed mechanism for cyanotoxin-associated diseases

Cyanobacteria produce harmful toxins that have been associated with several acute conditions and chronic human diseases, like gastroenteritis, non-alcoholic liver disease, and amyotrophic lateral sclerosis. Aerosol from waterbodies appears to be a likely mechanism for exposure. We conducted a study of human biospecimens focused on the cyanobacterial aerosilization process by evaluating the extent to which cyanobacteria can invade the human respiratory tract. Our study suggests that humans routinely inhale aerosolized cyanobacteria, which can be harbored in the nostrils and the lungs. Using PCR, cyanobacteria were found at high frequencies in the upper respiratory tract (92.20%) and central airway (79.31%) of our study subjects. Nasal swabs were not predictive of bronchoalveolar lavage (BAL) when detecting inhaled cyanobacteria. Interestingly, we found no evidence that time of year was a significant factor for cyanobacteria positivity (BAL cytology p = 1.0 and PCR p = 1.0); (nasal swab cytology p = 0.051 and PCR p = 0.65). Additionally, we found that proximity to a waterbody was not a significant factor for cyanobacteria positivity in BAL and nasal swabs collected during cyanobacteria bloom season [May–October] (p = 0.46 and p = 0.38). These data suggest that cyanobacteria exposure may be a prevalent and chronic phenomenon not necessarily restricted to waterbodies alone. Sources of indoor exposure warrant future investigation. Given the widespread prevalence of cyanobacterial exposure in the airway, investigation of the aerosol spread of cyanotoxins, more specifically, is warranted. Our findings are consistent with the hypothesis that aerosol is a significant route for cyanobacteria exposure, and thus a likely route of transmission for cyanotoxin-associated human diseases.

Comparative Study of efficacy of Broncho-alveolar lavage, bronchial brush and bronchial biopsy in diagnosis of lung tumours

Background: Lung carcinoma is oneof the most common malignancies in the industrialized countries and now a leading cause of death in developing countries like India. There are different methods to diagnose lung carcinoma. Broncho-alveolar lavage cytology, bronchial brush cytology and bronchial biopsy are three important techniques which are becoming more popular. The aimof the study is to compare the efficacy of these three methods. Materials and methods: This is a retrospective study in known patients with lung cancer, during a period of 18 months from July 2016 to December 2017. The materials obtained from Broncho-alveolar lavage, bronchial brush and bronchial lung biopsy are studied and their efficacy in diagnosing lung malignancy is compared. Results: The present study showed that sensitivity of Broncho-alveolar lavage, Bronchial brush and bronchial biopsy are 44%, 74% and 84% respectively. Conclusion: Bronchial biopsy and bronchial brush cytology are more sensitive than broncho-alveolar lavage cytology. However, combination of these three techniques helps to increase the rate of positive diagnosis.

Eosinophilic Bronchopneumopathy in Dogs

Background: The eosinophilic bronchopneumopathy (EBP) is characterized by pulmonary infiltration with eosinophils. The etiology of canine EBP remains unclear, although hypersensitivity to aeroallergens is suspected. Dogs affected are usually young. The persistent cough is the most common clinical sign, often associated with respiratory difficulty and exercise intolerance. The diagnosis is based on signalment, radiographic and bronchoscopic findings, and the tissue eosiphilic infiltration demonstrated by cytology of bronchoalveolar lavage (BAL). No reports were found in Brazil. The aim of this paper is report four cases of canine BPE, with emphasis on clinical aspects, diagnosis and therapy.Cases: Case 1. A 18-month-old female dog with 12 kg of body weight was presented for consultation with a 60-day history of cough, inappetence and weariness. It presented cough and tachypnea. The complementary exams demonstrated peripheral eosinophilia, bronchointersticial pulmonary pattern on radiography, moderate amount of mucus on bronchoscopy andtissue eosinophilis infiltration on the BAL. The treatment was based on steroids, with improvement of clinical signs. Case 2. A 24-month-old female dog with 16 kg of body weight was presented for consultation with one-year history of cough, inappetence and lethargy. It presented only cough, and in complementary exams showed transitory peripheral eosinophilia, bronchointersticial pattern on radiography and predominantly eosinophilic inflammation on citology of BAL. The therapy was based on steroids, with improvement followed by worsening of signs, with need of readjustment of doses. Case 3. A 8-year-old male dog with 6.2 kg of body weight was presented for consultation with 3-weeks history of productive cough,vomiting and weight loss. The complementary exams showed peripheral eosinophilia, bronchointersticial pulmonary patternon radiography and eosinophils infiltration on cytology of BAL. It was used steroids as therapy, with improvement of signs. Case 4. A 2-month-old female dog with 2.8 kg of body weight was presented for consultation with 3-days history of cough, nasal discharge and inappetence. It presented severe expiratory dyspnoea, tachypnea, harsh cough, lung crackles on auscultation and lethargy. The complementary exams demonstrated leukocytosis by peripheral eosinophilia and neutrophilia, bronchointersticial and alveolar pulmonar radiographic pattern. The therapy was started with steroids, febendazole and antibiotics. Tracheal swab was obtained and it showed eosinophilic inflammation on cytology. After 20-days treatment, the signs improved. Maintenance therapy was performed with steroids.Discussion: The EBP is usually reported in young patients, as described in three cases. The cough showed predominant clinically, associated with lethargy and inappetence in half of cases, as well as with dyspnea and weariness. The peripheral eosinophilia was observed in all patients, although transient in one of them, not being mandatory for the diagnosis.Despite there are not a specific radiographic pattern to the diagnosis, the bronchointersticial pattern was common to the four animals, with worsening of the radiographic findings possibly related to worsening clinical findings. The definitive diagnosis made through identification of eosinophilic inflammatory infiltration by bronchoalveolar lavage (BAL). Steroidtherapy proved to be effective in the four cases.Keywords: eosinophils, bronchus, lung parenchyma, dog.

Safety, tolerance, and effects on alveolar macrophages of a nebulized TLR agonist combination in foals

Abstract The intracellular pathogen R. equi causes pneumonia in foals and people that closely resembles that caused by Mycobacterium tuberculosis (Mtb). Development of resistance to antimicrobials by R. equi and Mtb represents a major health problem for veterinary and human medicine. There is a need for control methods and treatment alternative to the use of antibiotics. Our alternative approach is to use a TLR2/6 and TLR9 agonist combination known as PUL-042 to stimulate innate immunity in the lungs. Our objectives were to evaluate safety and ex vivo efficacy of PUL-042 (Pulmotect Inc, Houston, TX) in newborn foals. Ten newborn foals were divided in 2 groups (PUL-042 and saline control) and submitted to a bronchoalveolar lavage (BAL) on day 2 of age, 6 nebulizations in a 2-week interval and a second BAL 24 h after the last nebulization. BAL cells were infected with a virulent strain of R. equi, and then lysed at either 0 h (T0) and after 48 h of culture (T48). Supernatants were saved for determination of INF-γ, IL-1β, and IL-10 concentration by ELISA. PUL-042 did not induce either adverse effects or airway inflammation in foals, as assessed by clinical examination and BAL cytology. At the tested doses, nebulization of PUL-042 did not significantly (P &amp;gt; 0.05) affect ex vivo killing capacity of R. equi by AMs of foals at 2 and 21 days of age relative to controls. Results of cytokines are pending. Repeat nebulization of newborn foals with PUL-042 did not induce any adverse effects nor an inflammatory response in the lungs. At the dose tested, PUL-042 did not affect the killing capacity of R. equi by AMs, but its effects on other immune cells, and at other doses, need further evaluation.

Comparison of Tracheal Wash and Bronchoalveolar Lavage Cytology in 154 Horses With and Without Respiratory Signs in a Referral Hospital Over 2009-2015.

Most equine lower respiratory diseases present as increased airway neutrophilia, which can be detected in tracheal wash (TW) or bronchoalveolar lavage fluid (BALF) cytology samples. The aim was to compare the TW and BALF results in a population of client-owned horses with and without clinical respiratory disease signs. A secondary aim was to determine the sensitivity (Se) and specificity (Sp) of TW and BALF neutrophilia in detecting respiratory disease. The cutoff values for neutrophils were also evaluated. Retrospective data from 154 horses of various breeds that had been subject to both TW and bronchoalveolar lavage (BAL) sampling at rest during 2009−2015 were used. The horses were divided into three groups based on the presenting signs, physical examination, and endoscopy mucus score. Neutrophil counts of >20% in TW and >5% in BAL were considered abnormal. Cytology results between groups, correlations between TW and BALF cell types, and tracheal mucus score were analyzed. Two graph receiving operating characteristic (ROC) curves of the neutrophil percentage values of TW and BALF were created to determine the optimal cutoff values and to calculate the diagnostic Se and Sp for diagnosing airway inflammation in horses with and without clinical respiratory signs. The Se and Sp of TW and BALF neutrophil percentages were further estimated using a two-test one-population Bayesian latent class model. The two tests showed substantial agreement, and only 17.5% of the horses were classified differently (healthy vs. diseased). The neutrophil percentage was found to correlate between TW and BALF. The Se and Sp of TW were generally higher than for BAL when estimated with area under the curve or Bayesian model. Cutoff values of 17.7% for TW and 7% for BALF were indicated by the ROCs. We conclude that TW is a more sensitive and specific method in our patient population. We suggest that the current neutrophil cutoff values of 20% for TW and 5% for BALF would still be appropriate to use in clinical diagnosis of airway inflammation. However, further studies with other cell types and in other populations are warranted to determine the best sampling method for individual horses.

The progression of heartworm associated respiratory disease (HARD) in SPF cats 18\xa0months after Dirofilaria immitis infection

BackgroundHeartworm-associated respiratory disease (HARD) in cats is induced by the arrival and death of immature adult Dirofilaria immitis in the pulmonary system and is indistinguishable from mature adult heartworm infection.MethodsA controlled, blind research study investigated the long-term (18 months post infection, PI) consequences of the inflammatory response associated with the death of immature adult heartworms in cats.Three groups of cats, 10 per group, were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated with selamectin (Revolution®; Zoetis) per label directions at 28 days PI and once monthly for 17 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg) at 70 days PI, then every 2 weeks for 15 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, 240, 309, 380, and 505 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected.ResultsThe selamectin-treated cats (Group A) and ivermectin-treated cats (Group B) were free of heartworms or heartworm fragments at necropsy. All cats became heartworm antibody positive at some time point in the study except for one cat in Group A. Only cats in Group C (all with adult heartworms) were heartworm antigen positive. The heartworm antibody titer for Group B was highest on Days 110 to 168 and then decreased over time and 50% were serologically antibody negative on Day 240. Eosinophilic bronchoalveolar lavage (BAL) cytology and peripheral eosinophilia were most pronounced on Day 110 in all cats. Randomly distributed myofibrocytes in the lungs of some Group A cats suggest that precardiac larval stages were affecting the lungs. Radiographs in Group B cats demonstrated partial resolution of the initial HARD reaction but chronic myofibrocyte proliferation was histologically evident 18 months after infection.ConclusionHARD was induced by immature adult worm infection with progressive improvement starting 6 to 8 months after infection but histologic lesions were evident in some cats 18 months after infection. The serologic antibody assay was negative in 50% of cats at 8 months and 100% of cats at 18 months post infection. Abnormal radiographic lung patterns continued in a subset of Group B cats for months after heartworm antibody serology and BAL cytology returned to normal.

Heartworm-associated respiratory disease (HARD) induced by immature adult Dirofilaria immitis in cats

BackgroundA controlled, blind research study was conducted to define the initial inflammatory response and lung damage associated with the death of immature adult Dirofilaria immitis in cats as compared with cats developing adult heartworm infections and cats on preventive medication.MethodsThree groups of cats were utilized, 10 per group. All cats were infected with 100 third-stage (L3) larvae by subcutaneous injection. Group A cats were treated topically with selamectin (Revolution®; Zoetis) per label directions at 28 days post infection (PI) and once monthly for 8 months. Group B cats were treated orally with ivermectin (Ivomec®; Merial) at 150 μg/kg at 70 days PI, then every 2 weeks for 5 months. Group C cats were untreated PI. At baseline (Day 0) and on Days 70, 110, 168, and 240 PI, peripheral blood, serum, bronchial lavage, and thoracic radiographic images were collected on all cats. Upon completion of the study (Day 245), cats were euthanized and necropsies were conducted.ResultsResults were analyzed statistically between groups by ANOVA and by paired sample T testing for changes within the group over time. The selamectin-treated cats (Group A) did not develop radiographically evident changes throughout the study and were free of adult heartworms or worm fragments at necropsy. The heartworm life cycle was abbreviated with oral doses of ivermectin (Group B), shown by the absence of adult heartworms or worm fragments at necropsy. The early stage of immature adult worm in Group B cats, however, did induce severe pulmonary airway, interstitial, and arterial lung lesions, revealing that the abbreviated infection is a significant cause of respiratory pathology in cats. Cats in Groups B and C could not be differentiated based on radiographic changes, serologic antibody titers, complete blood count, or bronchoalveolar lavage cytology at any time point throughout the study. Eighty percent of cats in Group A and 100% of cats in Groups B and C became heartworm antibody positive at some time point post infection.ConclusionsThe clinical implications of this study are that cats that become infected with immature adult heartworms may not develop fully mature heartworms and are only transiently heartworm antibody positive, but do develop Heartworm-Associated Respiratory Disease (HARD).

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OBJECTIVES/SPECIFIC AIMS: Allergic asthma is a chronic lung disease driven by inappropriate inflammatory responses against inhaled allergens. Neuropilin-2 (NRP2) is a pleiotropic transmembrane receptor expressed in the lung, but its role in allergic airway inflammation is unknown. Here, we characterized NRP2 expression in lung immune cells and investigated the effects of NRP2 deficiency on airway inflammation. METHODS/STUDY POPULATION: NRP2 expression by lung immune cells from NRP2 reporter mice was determined by flow cytometry. NRP2 expression by human alveolar macrophages (AM) from healthy individuals was determined by mRNA analysis and flow cytometry. Airway inflammation in NRP2-deficient mice was assessed by bronchoalveolar lavage (BAL) cytology and inflammatory gene expression in lung tissue. RESULTS/ANTICIPATED RESULTS: NRP2 expression in lung immune cells was negligible under steady-state conditions. In contrast, inhalational exposure to lipopolysaccharide (LPS) adjuvant dramatically induced NRP2 expression in AM, as 63.3% of AM from LPS-treated mice were NRP2+ compared with 1.5% of AM from control mice. Ex vivo treatment of human AM with LPS resulted in a 1.5-fold and 2.6-fold increase in NRP2 mRNA and surface protein expression, respectively. Compared to littermate controls, NRP2-deficient mice had greater numbers of BAL leukocytes and increased lung expression of the T helper type 2 cytokines IL-4 and IL-5. Furthermore, NRP2 deficiency resulted in stochastic development of allergic airway inflammation, as spontaneous airway eosinophilia was detected in 25% (2/8) of NRP2-deficient mice compared with 0% (0/8) of littermate controls. DISCUSSION/SIGNIFICANCE OF IMPACT: NRP2 is expressed by activated human and murine AM and suppresses the spontaneous development of allergic airway inflammation. These findings suggest that NRP2 may play a key role in allergic asthma pathogenesis, and could prove to be an important therapeutic target in patients with asthma and other allergic diseases.

Comparative analyses of thoracic radiographs and bronchoalveolar lavage of dogs and cats with chronic bronchial diseases

There is a high incidence of bronchitis and asthma cases in veterinary medicine. Thoracic radiographs and bronchoalveolar lavage (BAL) are commonly performed for definitive diagnosis in dogs and cats with suspected bronchitis and asthma. It is believed that a combination of diagnostic tools is the best choice to achieve a diagnosis. The aim of this study was to evaluate the efficacy of thoracic radiographs and BAL in the diagnosis of chronic bronchial disease (CBD) in dogs and cats and whether there is any specific radiographic finding that could influence the indication for bronchoalveolar lavage. It was performed a cross-sectional, prospective, observational study including forty client-owned dogs and cats with lower respiratory tract signs and positive radiographic opacities that were evaluated with BAL followed by cytology and culture. The radiographic results compared with BAL culture showed a sensitivity of 38%, specificity of 95% and accuracy of 65% in detecting patients with pneumonia associated with chronic bronchial disease. Thoracic radiographs were effective in diagnosing 65% of the patients, radiographs plus BAL cytology diagnosed 75% of patients and the combination of radiographs, BAL cytology and culture diagnosed 95% of the patients with chronic bronchial disease. In conclusion, the combination of radiographic examination with BAL followed by cytological and microbiological analyses increases diagnostic success in CBD.

Diagnostic Efficacy of Bronchial Brush and Bronchoalveolar Lavage Cytology in Bronchopulmonary Lesions

Diagnostic Efficacy of Bronchial Brush and Bronchoalveolar Lavage Cytology in Bronchopulmonary Lesions

Shifted focus of bronchoalveolar lavage in patients with suspected thoracic malignancy: an analysis of 224 patients.

Bronchoscopies are extensively adopted for diagnosing and staging thoracic malignancies, but studies are missing as how to keep the process streamlined and more efficient. To evaluate current role of bronchoalveolar lavage (BAL) for cancer and possible infection diagnosis when practicing comprehensive bronchoscopy for patients suspected with thoracic malignancy, and provide foundation for possible practice modification. We retrospectively analyzed a prospectively kept database of immunocompetent patients undergoing bronchoscopy for suspected non-hematologic malignancies. Clinical, radiographic data, bronchoscopic sampling techniques and diagnostic results were recorded. Initially undiagnostic patients were followed up for 2 years for a definitive diagnosis. Of 224 patients included, 179 (79.9%) were confirmed with active thoracic malignancies. BAL diagnostic yield of cancer based on different radiographic characters of target lesion are as follow: isolated lymphadenopathies 0%, central lesions 45.5%, peripheral masses (diameter ≥3 cm) 21.4%, peripheral large nodules (2≤ diameter <3 cm) 15.8%, and peripheral small nodules (diameter <2 cm) 7.1%, while composite bronchoscopy achieved diagnostic yield of 93.3%, 95.5%, 91.7%, 76.9%, and 66.7% in corresponding lesion types. No cancer was diagnosed solely by BAL-cytology. Proportions of patients with positive BAL culture did not differ significantly between patients with and without pre-test suspicion for infections (P=0.199). In multivariable analysis, infections were associated with age ≥75 (OR 3.0; 95% CI: 1.29-7.06), chronic obstructive pulmonary disease (COPD) (OR 2.7; 95% CI: 1.14-6.26) and diabetes mellitus (DM) (OR 4.5; 95% CI: 1.90-10.44). Omitting BAL cytology in settings of comprehensive bronchoscopy may not compromise cancer diagnosis. For patients primarily suspected with thoracic malignancy, performing BAL culture only based on clinical suspicion could miss important infectious etiology.

Usefulness of Cyclophosphamide Pulse Therapy in Interstitial Lung Diseases

Background: Interstitial lung diseases (ILDs) are a group of disorders characterised by progressive lung function decline. Stabilisation of lung function under intermittent i.v. cyclophosphamide was shown in patients suffering from systemic sclerosis, yet data in ILD patients are scarce. Objectives: To retrospectively evaluate the usefulness of cyclophosphamide pulse therapy in ILD. Methods: We retrospectively analysed all patients who received i.v. cyclophosphamide in our centre from 2002 to 2012. Lung function, survival status, and bronchoalveolar lavage cytology were recorded during a follow-up period of 18 months. Results: Twenty-six patients with idiopathic pulmonary fibrosis, 6 with lymphocytic interstitial pneumonia (LIP), 8 with idiopathic non-specific interstitial pneumonia (NSIP), 7 with rheumatoid arthritis-associated ILD, and 7 with perinuclear anti-neutrophil cytoplasmic antibody-positive ILD (pANCA+ ILD) were included. Patients with LIP and NSIP had the best survival outcome, those with pANCA+ ILD the worst. In the total cohort, we found a significantly higher total lung capacity decline in the year before treatment compared to the year after treatment. Conclusions: This retrospective analysis of cyclophosphamide treatment shows a stabilisation of lung function in most patients with fibrotic ILDs, yet prospective studies in clearly defined diagnoses are urgently needed.