PMC9364985_01
Pulmonary imaging manifestations of the patient. (A) Irregular nodules in the left upper lung. (B) Ground glass opacities and small nodules in the upper lobe of the right lung. (C) Diffuse ground glass opacities in both lungs. Lesions mentioned above were pointed by the green arrows.
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Pulmonary CT showed irregular nodules in the left upper lung (Figure 1A). Lung CT showed ground glass opacities and small nodules in the upper lobe of the right lung (Figure 1B). Lung CT showed diffuse ground glass opacities in his lungs and suspected alveolar haemorrhage (Figure 1C).
The morphology of R. pusillus. Hyphae forms of R. pusillus in sputum stained with Gram stain (A) Original magnification X 1000) and fluorescent stain (B), Original magnification X 400). R. pusillus colony grown in 28 C PDA medium for 7 days. (C) Lactophenol cotton blue (LPCB) staining of the R. pusillus colony. (D), Original magnification X 400).
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Microscopic examination of the sputum by fluorescence staining and Gram staining showed fungal hyphae (Figure 2A-B). The colony showed a brownish and woolly appearance (Figure 2C). Fungal morphology on staining with lactophenol cotton blue (LPCB) showed some stolon mycelia and globose sporangium terminating a branch (Figure 2D).
Comparisons of the R. pusillus isolate and reference sequences. (A) All 18S rRNA reference sequences and genomes were obtained from the NCBI database. Muscle (V3.8.31) was used for multisequence comparison of 18S rRNA sequences. IQTree (V 1.6.12) was used to draw the evolutionary tree. (B) All reference genome sequences used were obtained from the NCBI database. Nucmer (V 3.1) was used to compare the whole-genome sequences, Mummer (V 3.23) was used to compare the large-fragment sequences in the genome, and FastTree (V 2.1.11) was used to draw the whole-genome phylogenetic tree after data integration. Finally, ITOL was used to visualize the figure.
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WGS identified R. pusillus on the basis of 18S rRNA phylogenetic tree analysis performed using CLCbio (QIAGEN, https://www.qiagen.com), which showed that the microorganism clustered closely with strains R. pusillus (Figure 3A) and very far from other Mucor species. FCH5.7 (GenBank accession no. FWWN00000000) according to Geneious (BioMatters, https://www.geneious.com) (Figure 3B), indicating that this rare fungus is R. pusillus.