Routine diagnosis of pulmonary disorders in bovines by physical examination, hematology and radiography is not sufficient to establish confirmatory diagnosis. Tracheo-bronchial aspirates (TBA) can be used as a reliable sample for the diagnosis of lower respiratory tract affections in cattle. Percutaneous tracheo-bronchial aspirate sampling was performed in 173 standing conscious cattle using two types of catheters. Fifty four cows were sampled with Large Animal transtracheal wash Kit (MILA International, Inc. USA) and 119 with infant feeding tube. Cytologic analysis of TBA smears was performed to assess the diagnostic utility of samples. Diagnostic samples were obtained in 87.8 per cent cows (152/173). Sampling using commercial catheter and infant feeding tube retrieved diagnostic samples in 90.7 per cent and 86.6 per cent of cows, whereas repeated infusions were required in 61.2 per cent and 78.7 per cent of cases, respectively. None of the cows exhibited complications post sampling. Infant feeding tube (7 Fr) with 10 G steel introduction catheter was effectively used in the collection of TBA and was comparable with commercial catheter. Present communication reports first systematic study on collection of TBA by percutaneous method in adult pneumonic cattle.