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The objective of this study was to determine whether a chlamydial isolate recovered from nasal swabs from swine with pneumonia could cause pneumonia and rhinitis in gnotobiotic pigs. The identity of the isolate currently is unknown, but it shares characteristics with Chlamydia trachomatis. After propagation in Vero cells and preparation of the inoculum (2.5 x 1010 inclusion-forming units/ml), Chlamydiae were instilled into nostrils (1.0 ml/nostril) and lungs (2.0 ml intralaryngeally) of 15 anesthetized 3-day-old gnotobiotic piglets. Five age-matched gnotobiotic piglets were anesthetized and sham infected with uninfected cell culture lysates. Two treated piglets were moribund and 2 were severely dyspneic prior to necropsy 7 days postinfection (DPI), whereas remaining treated piglets showed mild dyspnea upon exertion throughout the study. All treated piglets developed diarrhea. All treated piglets necropsied 7-21 DPI had extensive consolidation in cranial, middle, and accessory lung lobes; a majority of these piglets also had extensive consolidation in the caudal lobes. Treated piglets necropsied 28 and 35 DPI had a lobular pattern of consolidation in all lung lobes. Histologically, lesions in lungs from treated piglets necropsied 7, 14, and 21 DPI were characterized by bronchointerstitial pneumonia with foci of type II pneumocyte hypertrophy and hyperplasia; pneumocytes and bronchial and bronchiolar epithelial cells were markedly vacuolated. Alveolar macrophages, peribronchitis, peribronchiolitis, and perivasculitis were seen in lungs from treated piglets necropsied 28 and 35 DPI; those necropsied 28 DPI also had foci of lymphohistiocytic and plasmacytic infiltrates. Turbinate lesions in all treated piglets were characterized by mild multifocal lymphoplasmacytic and occasionally neutrophilic rhinitis. Immunohistochemistry detected chlamydial antigen in bronchial and bronchiolar epithelial cells, pneumocytes, and inflammatory cells in treated piglets necropsied 7, 14, and 21 DPI. Positive staining was limited to alveolar macrophages in treated piglets necropsied 28 and 35 DPI. Chlamydial antigen was detected in turbinate epithelial cells at all necropsy intervals. Ultrastructurally, chlamydiae were seen with glycogen particles in vacuoles or free in the cytoplasm of bronchial and bronchiolar epithelial cells and pneumocytes.