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Chapter 10 : Laboratory Diagnosis of Microsporidiosis
Authors:
Rainer Weber1,
David A. Schwartz2,
Peter Deplazes3
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Affiliations:
1: Division of Infectious Diseases and Hospital Epidemiology, Department of Internal Medicine, University Hospital, CH-8091 Zurich, Switzerland;
2: Department of Pathology, Grady Memorial Hospital, 80 Butler Street SE, Atlanta, Georgia 30335;
3: Institute of Parasitology, University of Zurich, Winterthurerstrasse 266A, CH-8057 Zurich, Switzerland
Content Type: Monograph
Publication Year:
1999
Category: Fungi and Fungal Pathogenesis
Book DOI:
10.1128/9781555818227
Chapter DOI:
10.1128/9781555818227.ch10
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Laboratory Diagnosis of Microsporidiosis, Page 1 of 2
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Abstract:
The most robust and widely practicable technique for the diagnosis of microsporidial infection is light microscopic morphological demonstration of the organisms themselves. Evaluation of patients with suspected microsporidiosis should begin with light microscopic examination of stool specimens and urine or cytological examination of other body fluids. The most common clinical finding of ocular microsporidiosis is keratoconjunctivitis. In patients with suspected ocular microsporidiosis, urine and respiratory secretions also should be examined for microsporidia. In patients with ocular microsporidiosis, urine and respiratory secretions should also be examined for the presence of microsporidial spores. Serologic assays (including carbon immunoassay [CIA], IFAT, enzyme-linked immunosorbent assay [ELISA], and Western blot [WB]) have been applied in detecting specific IgG antibodies directed to Encephalitozoon intestinalis spores in humans and to Encephalitozoon cuniculi spores in humans and several animal species. Aside from studies on ocular microsporidiosis in presumably otherwise healthy persons, detailed histopathologic investigation of human microsporidial infection has been performed only in immunodeficient individuals. One clue that microsporidiosis may be present is based on the observation that infected epithelial cells containing mature spores are often shed intact into the lumen from the underlying basement membrane. Upper and lower respiratory tract infections due to microsporidia are associated almost exclusively with disseminated disease produced by all three members of the genus Encephalitozoon. Microsporidia have been found in almost every organ system. Molecular techniques have been used to compare microsporidial isolates obtained from humans and animals in order to study possible sources of infection and reservoir hosts.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
Figures
Laboratory Diagnosis of Microsporidiosis
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Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-1
FIGURE 1
Stool specimen from a patient with AIDS and chronic diarrhea, showing pinkish red-stained spores of Enterocytozoon bieneusi measuring 0.7 to 1.0 by 1.1 to 1.6 μm. Chromotrope stain was used. Magnification, ×1,000 (oil immersion).
10.1128/9781555818227/fig10-1_thmb.gif
10.1128/9781555818227/fig10-1.gif
FIGURE 1
Stool specimen from a patient with AIDS and chronic diarrhea, showing pinkish red-stained spores of Enterocytozoon bieneusi measuring 0.7 to 1.0 by 1.1 to 1.6 μm. Chromotrope stain was used. Magnification, ×1,000 (oil immersion).
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-2
FIGURE 2
Stool specimen from a patient with AIDS and chronic diarrhea, showing microsporidial spores stained with a chemofluorescent optical brightening agent (Fungi-Fluor Kit, Polysciences, Inc., Warrington, Pa.). Magnification, ×1,000 (oil immersion).
10.1128/9781555818227/fig10-2_thmb.gif
10.1128/9781555818227/fig10-2.gif
FIGURE 2
Stool specimen from a patient with AIDS and chronic diarrhea, showing microsporidial spores stained with a chemofluorescent optical brightening agent (Fungi-Fluor Kit, Polysciences, Inc., Warrington, Pa.). Magnification, ×1,000 (oil immersion).
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-3
FIGURE 3
Urine sediment from a patient with AIDS and disseminated Encephalitozoon cuniculi infection, showing pinkish red-stained nricrosporidial spores measuring 1.0 to 1.5 by 2.0 to 3.0 μm. Chromotrope stain was used. Magnification, ×1,000 (oil immersion).
10.1128/9781555818227/fig10-3_thmb.gif
10.1128/9781555818227/fig10-3.gif
FIGURE 3
Urine sediment from a patient with AIDS and disseminated Encephalitozoon cuniculi infection, showing pinkish red-stained nricrosporidial spores measuring 1.0 to 1.5 by 2.0 to 3.0 μm. Chromotrope stain was used. Magnification, ×1,000 (oil immersion).
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-4
FIGURE 4
Cytospin preparation of bronchoalveolar lavage fluid from a patient with AIDS and intestinal E. bieneusi infection, showing intracellular gram-positive microsporidial spores. Gram stain was used. Magnification ×1,000 (oil immersion).
10.1128/9781555818227/fig10-4_thmb.gif
10.1128/9781555818227/fig10-4.gif
FIGURE 4
Cytospin preparation of bronchoalveolar lavage fluid from a patient with AIDS and intestinal E. bieneusi infection, showing intracellular gram-positive microsporidial spores. Gram stain was used. Magnification ×1,000 (oil immersion).
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-5
FIGURE 5
Cytospin preparation of bronchoalveolar lavage fluid from a patient with AIDS and intestinal E. bieneusi infection, showing intracellular microsporidia. Giemsa stain was used. Magnification, ×1,000 (oil immersion).
10.1128/9781555818227/fig10-5_thmb.gif
10.1128/9781555818227/fig10-5.gif
FIGURE 5
Cytospin preparation of bronchoalveolar lavage fluid from a patient with AIDS and intestinal E. bieneusi infection, showing intracellular microsporidia. Giemsa stain was used. Magnification, ×1,000 (oil immersion).
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-6
FIGURE 6
Cytospin preparation of cerebrospinal fluid, showing intracellular clusters of pink-stained microsporidial spores measuring 2 to 3 µm. Chromotrope stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-6_thmb.gif
10.1128/9781555818227/fig10-6.gif
FIGURE 6
Cytospin preparation of cerebrospinal fluid, showing intracellular clusters of pink-stained microsporidial spores measuring 2 to 3 µm. Chromotrope stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-7
FIGURE 7
Immunofluorescence staining of sputum from a patient with disseminated E. cuniculi infection (Weber et al., 1997a). Polyclonal rabbit antibody to E. cuniculi was used. Spore walls and extruded polar tubes are seen. Magnification, ×600.
10.1128/9781555818227/fig10-7_thmb.gif
10.1128/9781555818227/fig10-7.gif
FIGURE 7
Immunofluorescence staining of sputum from a patient with disseminated E. cuniculi infection (Weber et al., 1997a). Polyclonal rabbit antibody to E. cuniculi was used. Spore walls and extruded polar tubes are seen. Magnification, ×600.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-8
FIGURE 8
Cytological preparation of conjunctival smear with superficial epithelial cells containing numerous gram-positive spores of Encephalitozoon hellem. Brown and Hopps stain was used. Magnification, ×1,000.
10.1128/9781555818227/fig10-8_thmb.gif
10.1128/9781555818227/fig10-8.gif
FIGURE 8
Cytological preparation of conjunctival smear with superficial epithelial cells containing numerous gram-positive spores of Encephalitozoon hellem. Brown and Hopps stain was used. Magnification, ×1,000.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-9
FIGURE 9
Stool specimen from a patient with AIDS and diarrhea, showing dark violet-stained spores of Encephalitozoon intestinalis measuring 1.0 to 1.2 by 2.0 to 2.5 μm. Quick hot Gram-chromotrope stain was used. Magnification, ×1,000 (oil immersion). (Reprinted with permission from G. S.Visvesvara, Centers for Disease Control, Atlanta, Ga.)
10.1128/9781555818227/fig10-9_thmb.gif
10.1128/9781555818227/fig10-9.gif
FIGURE 9
Stool specimen from a patient with AIDS and diarrhea, showing dark violet-stained spores of Encephalitozoon intestinalis measuring 1.0 to 1.2 by 2.0 to 2.5 μm. Quick hot Gram-chromotrope stain was used. Magnification, ×1,000 (oil immersion). (Reprinted with permission from G. S.Visvesvara, Centers for Disease Control, Atlanta, Ga.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-10
FIGURE 10
Stool specimen from an immunocompetent Mexican patient with diarrhea, showing microsporidial spores stained with a monoclonal antibody to E. cuniculi (Mab 3B6) ( Enriquez et al., 1997b ). The microsporidian was identified as E. intestinalis, as described by Bornay-Llinares et al. (1998). Immunofluorescence detection was used. (Reprinted with permission from J. Enriquez, University of Arizona, Tucson,Ariz.)
10.1128/9781555818227/fig10-10_thmb.gif
10.1128/9781555818227/fig10-10.gif
FIGURE 10
Stool specimen from an immunocompetent Mexican patient with diarrhea, showing microsporidial spores stained with a monoclonal antibody to E. cuniculi (Mab 3B6) ( Enriquez et al., 1997b ). The microsporidian was identified as E. intestinalis, as described by Bornay-Llinares et al. (1998). Immunofluorescence detection was used. (Reprinted with permission from J. Enriquez, University of Arizona, Tucson,Ariz.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-11
FIGURE 11
Section of duodenal villus tip, showing epithelial cells infected with E. bieneusi. Clusters of black-stained spores and plasmodia at various stages of development are shown. Modified Warthin-Starry stain was used. Magnification, ×600. (Reprinted with permission from A. S. Field, St.Vincent's Hospital, Darlinghurst, Australia.)
10.1128/9781555818227/fig10-11_thmb.gif
10.1128/9781555818227/fig10-11.gif
FIGURE 11
Section of duodenal villus tip, showing epithelial cells infected with E. bieneusi. Clusters of black-stained spores and plasmodia at various stages of development are shown. Modified Warthin-Starry stain was used. Magnification, ×600. (Reprinted with permission from A. S. Field, St.Vincent's Hospital, Darlinghurst, Australia.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-12
FIGURE 12
Section of duodenal villus tip, showing enterocytes and macrophages infected with blackstained clusters of E. intestinalis. Modified Warthin-Starry stain was used. Magnification, ×600. (Reprinted with permission from A. S. Field, St. Vincent's Hospital, Darlinghurst, Australia.)
10.1128/9781555818227/fig10-12_thmb.gif
10.1128/9781555818227/fig10-12.gif
FIGURE 12
Section of duodenal villus tip, showing enterocytes and macrophages infected with blackstained clusters of E. intestinalis. Modified Warthin-Starry stain was used. Magnification, ×600. (Reprinted with permission from A. S. Field, St. Vincent's Hospital, Darlinghurst, Australia.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-13
FIGURE 13
Transmission electron micrograph showing duodenal epithelium from an HIV-infected patient infected with E. bieneusi. The different developmental stages between the enterocyte nuclei and the microvillus border include a proliferative plasmodium (1), late sporogonial plasmodia (2), and mature spores (3). A sloughing enterocyte containing mature spores is also shown (arrow). Magnification, ×3,700. (Reprinted with permission from M. A. Spycher, University Hospital, Zurich, Switzerland.)
10.1128/9781555818227/fig10-13_thmb.gif
10.1128/9781555818227/fig10-13.gif
FIGURE 13
Transmission electron micrograph showing duodenal epithelium from an HIV-infected patient infected with E. bieneusi. The different developmental stages between the enterocyte nuclei and the microvillus border include a proliferative plasmodium (1), late sporogonial plasmodia (2), and mature spores (3). A sloughing enterocyte containing mature spores is also shown (arrow). Magnification, ×3,700. (Reprinted with permission from M. A. Spycher, University Hospital, Zurich, Switzerland.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-14
FIGURE 14
Transmission electron micrograph of a microsporidial spore.The polar tubes (arrow) lie in two rows characteristic of E. bieneusi. Bar, 0.5 μm. Magnification, ×51,000.
10.1128/9781555818227/fig10-14_thmb.gif
10.1128/9781555818227/fig10-14.gif
FIGURE 14
Transmission electron micrograph of a microsporidial spore.The polar tubes (arrow) lie in two rows characteristic of E. bieneusi. Bar, 0.5 μm. Magnification, ×51,000.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-15
FIGURE 15
Immature and mature spores of E. hellem within a parasitophorous vacuole in an infected renal tubular epithelial cell. Note the high concentration of spores within this vacuole. Original magnification, ×19,200.
10.1128/9781555818227/fig10-15_thmb.gif
10.1128/9781555818227/fig10-15.gif
FIGURE 15
Immature and mature spores of E. hellem within a parasitophorous vacuole in an infected renal tubular epithelial cell. Note the high concentration of spores within this vacuole. Original magnification, ×19,200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-16
FIGURE 16
Transmission electron micrograph of meronts and developing E. intestinalis spores within enterocytes separated by a fibrillar matrix. E. intestinalis develops within parasitophorous vacuoles. Magnification, ×5,300. (Reprinted with permission from M. A. Spycher, University Hospital, Zurich , Switzerland.)
10.1128/9781555818227/fig10-16_thmb.gif
10.1128/9781555818227/fig10-16.gif
FIGURE 16
Transmission electron micrograph of meronts and developing E. intestinalis spores within enterocytes separated by a fibrillar matrix. E. intestinalis develops within parasitophorous vacuoles. Magnification, ×5,300. (Reprinted with permission from M. A. Spycher, University Hospital, Zurich , Switzerland.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-17
FIGURE 17
Typical ultrastructural appearance of a mature spore of a Nosema sp. A diplokaryotic nucleus (with a dividing membrane) and 10 turns of the polar tubule are evident. Original magnification, ×38,000.
10.1128/9781555818227/fig10-17_thmb.gif
10.1128/9781555818227/fig10-17.gif
FIGURE 17
Typical ultrastructural appearance of a mature spore of a Nosema sp. A diplokaryotic nucleus (with a dividing membrane) and 10 turns of the polar tubule are evident. Original magnification, ×38,000.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-18
FIGURE 18
Mucosal biopsy specimen, showing duodenal epithelial cells infected with E. bieneusi. A cluster of gram-positive spores is located in the supranuclear portion of the cytoplasm. Brown and Brenn stain was used. Magnification, ×630.
10.1128/9781555818227/fig10-18_thmb.gif
10.1128/9781555818227/fig10-18.gif
FIGURE 18
Mucosal biopsy specimen, showing duodenal epithelial cells infected with E. bieneusi. A cluster of gram-positive spores is located in the supranuclear portion of the cytoplasm. Brown and Brenn stain was used. Magnification, ×630.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-19
FIGURE 19
E. intestinalis infection of the small intestine. The plastic-embedded semithin section shows spores larger than those of E. bieneusi. The parasites are not in direct contact with the host cell cytoplasm but instead develop within a parasitophorous vacuole. Toluidine blue stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-19_thmb.gif
10.1128/9781555818227/fig10-19.gif
FIGURE 19
E. intestinalis infection of the small intestine. The plastic-embedded semithin section shows spores larger than those of E. bieneusi. The parasites are not in direct contact with the host cell cytoplasm but instead develop within a parasitophorous vacuole. Toluidine blue stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-20
FIGURE 20
A solitary corneal epithelial cell infected with E. hellem from an enucleated globe obtained at autopsy. The spores are gram-positive. Brown and Hopps stain was used. Magnification, ×200.
10.1128/9781555818227/fig10-20_thmb.gif
10.1128/9781555818227/fig10-20.gif
FIGURE 20
A solitary corneal epithelial cell infected with E. hellem from an enucleated globe obtained at autopsy. The spores are gram-positive. Brown and Hopps stain was used. Magnification, ×200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-21
FIGURE 21
Conjunctival biopsy specimen, demonstrating shedding and necrotic conjunctival epithelial cells containing gram-positive microsporidial spores admixed with neutrophils and fibrin from a patient with AIDS and disseminated E. hellem infection. Brown and Hopps stain was used. Magnification, ×1,000.
10.1128/9781555818227/fig10-21_thmb.gif
10.1128/9781555818227/fig10-21.gif
FIGURE 21
Conjunctival biopsy specimen, demonstrating shedding and necrotic conjunctival epithelial cells containing gram-positive microsporidial spores admixed with neutrophils and fibrin from a patient with AIDS and disseminated E. hellem infection. Brown and Hopps stain was used. Magnification, ×1,000.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-22
FIGURE 22
Plastic embedded semithin section of a nasal mucosal biopsy specimen, showing epithelial infection with E. hellem spores. Toluidine blue stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-22_thmb.gif
10.1128/9781555818227/fig10-22.gif
FIGURE 22
Plastic embedded semithin section of a nasal mucosal biopsy specimen, showing epithelial infection with E. hellem spores. Toluidine blue stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-23
FIGURE 23
Microsporidial sinusitis due to an Encephalitozoon sp. Numerous infected epithelial cells can be seen to contain faintly gram-positive clusters of spores. Brown and Hopps stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-23_thmb.gif
10.1128/9781555818227/fig10-23.gif
FIGURE 23
Microsporidial sinusitis due to an Encephalitozoon sp. Numerous infected epithelial cells can be seen to contain faintly gram-positive clusters of spores. Brown and Hopps stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-24
FIGURE 24
Plastic-embedded semithin section of tracheal mucosa with numerous spores of E. hellem present in respiratory lining epithelium. Toluidine blue stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-24_thmb.gif
10.1128/9781555818227/fig10-24.gif
FIGURE 24
Plastic-embedded semithin section of tracheal mucosa with numerous spores of E. hellem present in respiratory lining epithelium. Toluidine blue stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-25
FIGURE 25
Open lung biopsy specimen, showing bronchiolitis and bronchopneumonia due to E. cuniculi. Microsporidia cannot be seen in this section, but the identity of this microsporidian was confirmed with both fluorescent antibody and PCR methods. Hematoxylin and eosin stain was used. Magnification, ×200.
10.1128/9781555818227/fig10-25_thmb.gif
10.1128/9781555818227/fig10-25.gif
FIGURE 25
Open lung biopsy specimen, showing bronchiolitis and bronchopneumonia due to E. cuniculi. Microsporidia cannot be seen in this section, but the identity of this microsporidian was confirmed with both fluorescent antibody and PCR methods. Hematoxylin and eosin stain was used. Magnification, ×200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-26
FIGURE 26
Alveolar macrophage containing phagocytosed spores of E. cuniculi. Brown and Hopps stain was used. Magification, ×1,000.
10.1128/9781555818227/fig10-26_thmb.gif
10.1128/9781555818227/fig10-26.gif
FIGURE 26
Alveolar macrophage containing phagocytosed spores of E. cuniculi. Brown and Hopps stain was used. Magification, ×1,000.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-27
FIGURE 27
Chronic interstitial nephritis in a patient with disseminated E. hellem infection. The glomeruli are not involved. Hematoxylin and eosin stain was used. Magnification, ×200.
10.1128/9781555818227/fig10-27_thmb.gif
10.1128/9781555818227/fig10-27.gif
FIGURE 27
Chronic interstitial nephritis in a patient with disseminated E. hellem infection. The glomeruli are not involved. Hematoxylin and eosin stain was used. Magnification, ×200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-28
FIGURE 28
Renal tubule with necrosis, infected with E. hellem. The granular material filling the lumen represents microsporidial spores and necrotic cellular debris. Hematoxylin and eosin stain was used. Magnification, ×400.
10.1128/9781555818227/fig10-28_thmb.gif
10.1128/9781555818227/fig10-28.gif
FIGURE 28
Renal tubule with necrosis, infected with E. hellem. The granular material filling the lumen represents microsporidial spores and necrotic cellular debris. Hematoxylin and eosin stain was used. Magnification, ×400.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-29
FIGURE 29
The lumen of this renal tubule is filled with E. hellem spores. Steiner stain was used. Magnification, ×200.
10.1128/9781555818227/fig10-29_thmb.gif
10.1128/9781555818227/fig10-29.gif
FIGURE 29
The lumen of this renal tubule is filled with E. hellem spores. Steiner stain was used. Magnification, ×200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-30
FIGURE 30
Microsporidial prostate abscess. The abscess material contained numerous spores which cannot be seen with routine hematoxylin and eosin staining. Hematoxylin and eosin stain was used. Magnification, ×200.
10.1128/9781555818227/fig10-30_thmb.gif
10.1128/9781555818227/fig10-30.gif
FIGURE 30
Microsporidial prostate abscess. The abscess material contained numerous spores which cannot be seen with routine hematoxylin and eosin staining. Hematoxylin and eosin stain was used. Magnification, ×200.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-31
FIGURE 31
Muscle fiber with a sporont of Pleistophora containing several sporoblasts. The sporont has a thick, electron dense pansporoblastic membrane (large arrows). Sporoblasts have a nucleus (N), electron-dense exospore (arrowheads), and a coiled polar tubule (small arrows) with 9 to 12 turns. (Reprinted with permission from L. S. Adelman and from J. Alroy, Department of Pathology, Tufts University School of Medicine, Boston, Mass.)
10.1128/9781555818227/fig10-31_thmb.gif
10.1128/9781555818227/fig10-31.gif
FIGURE 31
Muscle fiber with a sporont of Pleistophora containing several sporoblasts. The sporont has a thick, electron dense pansporoblastic membrane (large arrows). Sporoblasts have a nucleus (N), electron-dense exospore (arrowheads), and a coiled polar tubule (small arrows) with 9 to 12 turns. (Reprinted with permission from L. S. Adelman and from J. Alroy, Department of Pathology, Tufts University School of Medicine, Boston, Mass.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-32
FIGURE 32
Section of brain tissue from a rabbit with torticollis, showing a microgranuloma with a central focus of necrosis but without detectable spores. Hematoxylin and eosin stain was used. Magnification, ×250. (Reprinted with permission from F. Guscetti, University of Zurich, Zurich, Switzerland.)
10.1128/9781555818227/fig10-32_thmb.gif
10.1128/9781555818227/fig10-32.gif
FIGURE 32
Section of brain tissue from a rabbit with torticollis, showing a microgranuloma with a central focus of necrosis but without detectable spores. Hematoxylin and eosin stain was used. Magnification, ×250. (Reprinted with permission from F. Guscetti, University of Zurich, Zurich, Switzerland.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.fig10-33
FIGURE 33
Section of brain tissue from an emperor tamarin with lethal neonatal encephalitozoonosis. Immunohistochemical detection of E. cuniculi spores in a granuloma was done with a polyclonal rabbit antibody to E. cuniculi. Visualization was by the labeled streptavidin-biotin method. Magnification, ×250. (Reprinted with permission from F. Guscetti, University of Zurich, Zurich, Switzerland.)
10.1128/9781555818227/fig10-33_thmb.gif
10.1128/9781555818227/fig10-33.gif
FIGURE 33
Section of brain tissue from an emperor tamarin with lethal neonatal encephalitozoonosis. Immunohistochemical detection of E. cuniculi spores in a granuloma was done with a polyclonal rabbit antibody to E. cuniculi. Visualization was by the labeled streptavidin-biotin method. Magnification, ×250. (Reprinted with permission from F. Guscetti, University of Zurich, Zurich, Switzerland.)
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
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Tables
Laboratory Diagnosis of Microsporidiosis
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Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.tab10-1
TABLE 1
Microsporidial species pathogenic in humans and their clinical manifestations
a Species not yet classified.
b Microsporidium is a collective generic name for microsporidia that cannot be classified because available information is not sufficient.
10.1128/9781555818227/tab10-1_thmb.gif
10.1128/9781555818227/tab10-1.gif
TABLE 1
Microsporidial species pathogenic in humans and their clinical manifestations
a Species not yet classified.
b Microsporidium is a collective generic name for microsporidia that cannot be classified because available information is not sufficient.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.tab10-2
TABLE 2
Sites of detection of microsporidia a
a −, not reported; +, case report(s) of detection; ++, consistently reliable specimens for detection of microsporidia.
b Pituitary gland.
10.1128/9781555818227/tab10-2_thmb.gif
10.1128/9781555818227/tab10-2.gif
TABLE 2
Sites of detection of microsporidia a
a −, not reported; +, case report(s) of detection; ++, consistently reliable specimens for detection of microsporidia.
b Pituitary gland.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.tab10-3
TABLE 3
Diagnostic techniques and identification of microsporidia a
a The table has been modified from Weber et al. (1994a).
b +, recommended;(+), useful, but not widely available (research technique or commercially not available); −, not recommended.
10.1128/9781555818227/tab10-3_thmb.gif
10.1128/9781555818227/tab10-3.gif
TABLE 3
Diagnostic techniques and identification of microsporidia a
a The table has been modified from Weber et al. (1994a).
b +, recommended;(+), useful, but not widely available (research technique or commercially not available); −, not recommended.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.tab10-4
TABLE 4
Chromotrope stains and their modifications a
a Laboratory procedures are described in the appendix to this chapter.
b FEA, formalin-ethyl acetate; SAF, sodium acetate-acetic acid-formalin.
10.1128/9781555818227/tab10-4_thmb.gif
10.1128/9781555818227/tab10-4.gif
TABLE 4
Chromotrope stains and their modifications a
a Laboratory procedures are described in the appendix to this chapter.
b FEA, formalin-ethyl acetate; SAF, sodium acetate-acetic acid-formalin.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10
/content/10.1128/9781555818227.chap10.tab10-5
TABLE 5
Murine monoclonal antibodies with diagnostic potential for the detection of microsporidia in humans
a MAbs, monoclonal antibodies.
10.1128/9781555818227/tab10-5_thmb.gif
10.1128/9781555818227/tab10-5.gif
TABLE 5
Murine monoclonal antibodies with diagnostic potential for the detection of microsporidia in humans
a MAbs, monoclonal antibodies.
Citation:
Weber R, Schwartz D, Deplazes P.
1999.
Laboratory Diagnosis of Microsporidiosis,
p 315-362.
In
Wittner M, Weiss L (ed),
The Microsporidia and Microsporidiosis.
ASM Press, Washington, DC.
doi: 10.1128/9781555818227.ch10