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Abstract:

Once known collectively by the single genus and species, "," it is now understood that distinct species of infect different mammalian hosts. A recent report of the ability of spp. to form biofilms outside the lung holds promise for development of an in vitro system. Almost every mammal examined to date appears to harbor at least one species of that is not found in any other mammal. Human- and rat-derived species have been the most extensively studied of the species in this group, and discussion of the life cycle in this chapter is based on these findings. The chapter discusses morphological criteria for recognition of stained by various procedures. Recent studies of humans reported the ability to detect -specific DNA in nasopharyngeal aspirates and oropharyngeal washes after amplification by PCR with -specific primers. There is currently only one recognized species of that causes pneumonia in humans, . It may become desirable in the future to track the emergence of organisms that are resistant to trimethoprim-sulfamethoxazole (TMP-SMX), or evaluate the potential for therapeutic response. The microscopic demonstration of in tissue and fluids by staining with Gomori methenamine silver (GMS) or a rapid variant of the Wright-Giemsa stain, by immunofluorescent assay (IFA), or by other stains such as Papanicolaou should be considered sufficient for diagnosis.

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116

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Image of FIGURE 3
FIGURE 3

Morphology and tinctorial characteristics of in clinical samples stained with various stains (magnification, ∼×960 unless stated otherwise). (A) Calcofluor stain of BALF. Cyst walls with internal thickenings (double comma) are highly fluorescent (color varies with barrier filter used). (B) Rapid Giemsa-like (Diff-Quik) stain of BALF. A thick cluster of mostly trophic forms (2 to 3 μm) with small reddish-purple nuclei and light blue to red-violet cytoplasm is shown. Boundaries of trophic forms are rarely discernible with this stain. Trophic forms overlay each other to produce darker-staining blue cytoplasm. Large dark purple host nuclei are admixed in the cluster. (C) Gomori's methenamine silver stain (Grocott) of organisms from BALF. Cyst walls and thickenings (double comma) can be observed as well as collapsed cup shapes and a crinkled raisin-like appearance. Note the lack of budding. Trophic forms are not stained with silver-based stains. (D) Papanicolaou's stain of BALF. Note the distinctive alveolar cast morphology. Magnification, ∼×380. (E) Toluidine blue O stain of in BALF. Cyst walls are stained light purple. The crinkled appearances of the cysts are illustrated with this stain as well as the darker-staining central body. Note the lack of budding with this and other cyst wall stains. Trophic forms are not stained. (F) Direct fluorescent antibody stain of an organism cluster from BALF. Note the apple green fluorescence distributed unevenly over the cluster, with accumulation on a cyst wall (lower left of cluster). Structures within the cysts are unstained and appear black.

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116
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Image of FIGURE 1
FIGURE 1

Major developmental forms of (A) Single trophic form; (B) sporocyte (precyst); (C) cyst with 4 visible spores (intracystic bodies); (D) cyst with 3 intracystic spores and a spore that has apparently excysted (arrow); (E) cyst with localized thickening of the cell wall (arrow) and 8 visible spores. Nomarski interference contrast microscopy; magnification, ×1,000.

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116
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Image of FIGURE 2
FIGURE 2

Proposed life cycle of (A) The primary site of infection is the lung alveoli. Three clusters of alveoli are illustrated. An expanded schematic of an alveolus (box) is shown in panel B. (B) Single alveolus with organisms depicted as the shape with an arrow attached to the cells lining the alveoli in some areas (type I pneumocytes) and unattached to other alveolar cells (type II pneumocytes). (C) Putative sexual cycle of in the lung alveoli: ( ) opposite mating types fuse and undergo karyogamy, resulting in a diploid zygote; ( ) the zygote then undergoes meiosis, resulting in four nuclei; ( ) additional postmeiotic mitosis increases the number of nuclei to eight; ( ) the nuclei and mitochondria (not shown) are compartmentalized by invagination of the inner plasma membrane, resulting in eight spores. Spores are released from the ascus (cyst) and presumably enter into the vegetative phase of the cycle. (D) Asexual replication cycle of Trophic forms undergo binary fission after mitotic replication of the nucleus. (Drawn with SmartDraw Suite, version 7.3.)

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116
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Tables

Generic image for table
TABLE 1

Comparison of stains used to detect

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116
Generic image for table
TABLE 2

Suggested primers and conditions for PCR detection of

From reference 101.

From reference 99.

Dihydropteroate synthase gene; from references 4 and 55.

Citation: Cushion M. 2011. , p 1822-1835. In Versalovic J, Carroll K, Funke G, Jorgensen J, Landry M, Warnock D (ed), Manual of Clinical Microbiology, 10th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816728.ch116

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