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Detection of a Lophomonas, a rare pathogen in Bronchoalveolar lavage.

ABSTRACT: Lophomonas infection is an emerging parasitic disease-causing respiratory infection. Although common in immunocompromised patient, it has been observed also in some immunocompetent cases. We report the case of a 45-year-old male who presented with productive cough, fever, and chest pain, with marked eosinophilia and cavitary lesion in the X-ray chest. KOH preparation and acid-fast bacilli microscopy of bronchoalveolar lavage (BAL) were negative. Direct microscopic examination of BAL accidentally showed a large number of living Lophomonas species with the movement of flagella. Methylene blue and Giemsa staining showed the plume of flagella and the nucleus. The patient was managed conservatively with metronidazole and get cured. It was concluded that the patient presented with signs and symptoms of pneumonia must be evaluated for rare events also if the patient was not responding with typical management of pneumonia. We reported the first case of this rare entity in Chhattisgarh state in an immunocompetent young Indian male.

SUBMITTER: Keche A

PROVIDER: S-EPMC9832492 | biostudies-literature | 2022 Jul-Dec

REPOSITORIES: biostudies-literature

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Detection of a <i>Lophomonas</i>, a rare pathogen in Bronchoalveolar lavage.

Keche Archana A Khatoon Shagufta S Sahu Dibakar D

Tropical parasitology 20220701 2

<i>Lophomonas</i> infection is an emerging parasitic disease-causing respiratory infection. Although common in immunocompromised patient, it has been observed also in some immunocompetent cases. We report the case of a 45-year-old male who presented with productive cough, fever, and chest pain, with marked eosinophilia and cavitary lesion in the X-ray chest. KOH preparation and acid-fast bacilli microscopy of bronchoalveolar lavage (BAL) were negative. Direct microscopic examination of BAL accid ...[more]

PMID: 36643984

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Project description:Pneumonia is the leading cause of morbidity and mortality in children and needs rapid and accurate pathogenic diagnosis. The aim of this study was to evaluate the diagnostic value of bronchoalveolar lavage fluid (BALF) metagenomic next-generation sequencing (mNGS) and conventional microbiological tests (CMTs) for pathogen detection in children with severe or refractory pneumonia. In this retrospective study, the clinical data of 127 children with severe or refractory pneumonia admitted to the respiratory department from June 2021 to March 2022 were analyzed. BALF mNGS and CMTs were utilized for pathogen diagnosis and comparison of their detection performance for different pathogens. The pathogenic diagnosis rate was 95.28% (121/127) by combining mNGS and CMTs. mNGS had significantly higher overall (96.06% vs 72.44%, P < 0.001), bacterial (69.29% vs 12.60%, P < 0.001), and fungal (11.81% vs 3.15%, P = 0.009) detection rates than CMTs. However, there was no significant difference of detection rates between them for respiratory viruses (33.86% vs 33.75%, P = 0.99) and Mycoplasma pneumoniae (48.03% vs 45.67%, P = 0.71). The sensitivities of mNGS for total pathogens, bacteria, and fungi were 99.17%, 100%, and 87.50%, respectively, which were higher than those of CMTs. CMTs for M. pneumoniae had the highest sensitivity (91.23%) compared with mNGS (89.47%) and multiplex PCR (88.57%). For respiratory viruses, mNGS and mPCR had similar sensitivities (97.67% vs 96.43%). mNGS was superior to CMTs in bacterial and fungal detection, while it was comparable to multiplex PCR for the detection of M. pneumoniae and respiratory viruses. Different detection methods should be rationalized for different pathogens.ImportanceThis study on 127 patients with severe and refractory pneumonia showed that mNGS was significantly superior to CMTs in terms of bacterial and fungal detection. We also found that multiplex PCR assay was comparable to mNGS for the detection of Mycoplasma pneumoniae and respiratory viruses and may have greater application advantages in combination with CMTs, such as M. pneumoniae IgM. For severe and refractory pneumonia, or when empiric treatment is not effective, collecting BALF for mNGS can help to quickly identify the causative organisms at an early stage. It is also important to choose more appropriate methods or combinations for different pathogens.

Project description:Three small antimicrobial anionic peptides (AP) were originally isolated from an ovine pulmonary surfactant. However, their presence in bronchoalveolar lavage (BAL) fluid and tissues of the respiratory tract is unknown. In this study, we made affinity-purified rabbit polyclonal and mouse monoclonal antibodies to synthetic H-DDDDDDD-OH. Antibody specificity was assessed by a competitive enzyme-linked immunosorbent assay (ELISA), and the exact epitope binding sites were determined with analog peptides synthesized on derivatized cellulose. These antibodies were used to detect AP in BAL fluid by ELISA and in respiratory tissues by Western blot analysis and immunocytochemistry. BAL fluid from 25 sheep contained 0.83 +/- 0.33 mM AP (mean +/- standard deviation; range, 0.10 to 1.59 mM) and was antimicrobial. The presence of AP in BAL fluid was confirmed by reverse-phase high-pressure liquid chromatography fractionation followed by matrix-assisted laser desorption/ionization (MALDI) mass spectrometry on those fractions which were positive by competitive ELISA and demonstrated antimicrobial activity. In Western blots, polyclonal antibody PAB96-1 and monoclonal antibody 1G9-1C2 (5.0 micrograms/ml) detected four bands in solubilized turbinate and tracheal epithelial cells (53.7, 31.2, 28.0, and 25.7 kDa) and five bands in lung homogenates (53.5, 37.1, 31.2, 28.0, and 25.7 kDa). Only a single band was seen in solubilized liver and small-intestine homogenates, and no bands were seen in blots containing BAL fluid, albumin, or kidney or spleen homogenates. In pulmonary-tissue sections, both antibodies PAB96-1 and 1G9-1C2 identified accumulated protein in the apical cytoplasm of the bronchial and bronchiolar epithelia, in the cytoplasm of pulmonary endothelial cells, and in an occasional alveolar macrophage. As a first step in identifying a candidate AP precursor gene(s), degenerate oligonucleotides representing all possible coding combinations for H-GADDDDD-OH and H-DDDDDDD-OH were synthesized and used to probe Southern blots of sheep genomic DNA. Following low-stringency washes and a 2-day exposure, strongly hybridizing bands could be identified. One degenerate oligonucleotide, SH87, was used as a hybridization probe to screen a sheep phage genomic library. Two independent phage contained the H-GADDDDD-OH coding sequence as part of a larger predicted protein. AP may originate as part of an intracellular precursor protein, with multistep processing leading to the release of the heptapeptide into mucosal secretions. There it may interact with other innate pulmonary defenses to prevent microbial infection.

Dataset Information

Detection of a Lophomonas, a rare pathogen in Bronchoalveolar lavage.

Publications

Detection of a <i>Lophomonas</i>, a rare pathogen in Bronchoalveolar lavage.

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