5.3.2 Ectopic liver infestation of Ascaris lumbricoides

Ectopic infestation of Ascaris lumbricoides in the hepatobiliary tract has been reported [74]. A farmer housewife aged 40's complained of abdominal pain. Liver abscess was indicated by diagnostic imaging, and partial hepatectomy specimen revealed multiple yellow-colored xanthogranulomatous nodules at the liver hilus. Microscopically, a small number of dead and calcified ova were distributed in necrotic substance, while the parasite body had been absorbed [75]. The content of the ova was immunoreactive with the diluted patient's serum [7, 20, 21, 23, 75]. Ouchterlony's diffusion-in-gel test using the extract of varied helminths identified a

clear precipitation line against A. lumbricoides (Figure 46). The precipitation line was abolished by preabsorbing the serum with the roundworm extract [75]. The final diagnosis was ectopic ascariasis, caused by infestation of a young (immature) female worm. The serum was unreactive with ova of Schistosoma japonicum and Paragonimus miyazakii seen in paraffin sections. The inner surface of the cell wall of

Hepatic ascariasis (left upper, cut surface of the resected liver; right upper, Ouchterlony's diffusion-in-gel test using the extract of varied helminths; left lower, H&E; right lower, reactivity with patient's own serum). Yellow-colored xanthogranulomatous nodules are seen at the liver hilus. Microscopically, a small number of dead and calcified ova are distributed in necrotic substance. The content of the ova is immunoreactive with the patient's serum. In Ouchterlony's test, a precipitation line against Ascaris lumbricoides (As) has been absorbed by the Ascaris extract, as shown on the right hand (Em, Echinococcus multilocularis; Fh, Fasciola hepatica;

Low-Specificity and High-Sensitivity Immunostaining for Demonstrating Pathogens…

DOI: http://dx.doi.org/10.5772/intechopen.85055

Figure 47 demonstrates a splenic lesion with ectopic migration of A.

lumbricoides. Necrotizing granulomas were formed beneath the splenic capsule, and dead parasite body fragments were seen in the necrotic area. It was impossible to determine the species of nematode under microscopy. The abovementioned serum

Splenic ascariasis (left, cut surface of the formalin-fixed spleen; center, H&E; right, reactivity with the serum of patient with hepatic ascariasis). Ectopic migration of A. lumbricoides has provoked necrotizing granulomas beneath the splenic capsule. Dead parasite body fragments are seen in the necrotic area. The abovementioned serum of patient with hepatic ascariasis reacts with the dead worm body, confirming the diagnosis of ectopic

A. simplex larva was weakly stained with the serum [21].

Tc, Toxocara canis; EV, Enterobius vermicularis; CRP, C-reactive protein).

Figure 46.

Figure 47.

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splenic ascariasis.

#### Figure 45.

Propionibacterium acnes folliculitis (upper panels, dilated hair follicle with histiocytic reaction in the perifollicular dermis; lower panels, higher powered view of the infiltrating macrophages; left, H&E; right, reactivity with patient's own serum; inset, immunostaining for PAC3 antigen specific to P. acnes). Macrophages clustered around the dilated hair follicle contain numerous granular-looking microbes. The patient's own serum clearly demonstrates the pathogen not only in macrophages but also in commensal bacterial colonies in the hair follicle (arrow). Immunostaining for PAC3 antigen confirms the diagnosis of P. acnes-induced folliculitis.

Low-Specificity and High-Sensitivity Immunostaining for Demonstrating Pathogens… DOI: http://dx.doi.org/10.5772/intechopen.85055

#### Figure 46.

clinical information and histopathological appearance with the morphology of the pathogen. Of no doubt, this gives us the most powerful and useful situation in

Invasive opportunistic infection of Propionibacterium acnes, a commensal in the hair follicle, has been described [72]. An aged afebrile Japanese male complained of a skin nodule on the chest. Biopsy revealed hair follicle-centered inflammation, and macrophages clustered around the hair follicle contained numerous granularlooking microbes. Microscopically, the possibility of yeast-form mycosis,

rickettsiosis, and protozoan infection was suspected. Grocott positivity was faint. Immunostaining using B. cereus antiserum revealed positivity (see Figure 22), while antisera to BCG,T. pallidum, and E. coli did not. The patient's own serum diluted at 1:500 clearly demonstrated the pathogen not only in macrophages but also in commensal bacterial colonies in the hair follicle [23]. Finally, PAC3 antigen specific to P. acnes was positive (through the courtesy of Prof. Yoshinobu Eguchi, Department of Pathology, Tokyo Medical and Dental University, Tokyo) [73], and the diagnosis

Ectopic infestation of Ascaris lumbricoides in the hepatobiliary tract has been reported [74]. A farmer housewife aged 40's complained of abdominal pain. Liver abscess was indicated by diagnostic imaging, and partial hepatectomy specimen revealed multiple yellow-colored xanthogranulomatous nodules at the liver hilus. Microscopically, a small number of dead and calcified ova were distributed in necrotic substance, while the parasite body had been absorbed [75]. The content of the ova was immunoreactive with the diluted patient's serum [7, 20, 21, 23, 75]. Ouchterlony's diffusion-in-gel test using the extract of varied helminths identified a

Propionibacterium acnes folliculitis (upper panels, dilated hair follicle with histiocytic reaction in the perifollicular dermis; lower panels, higher powered view of the infiltrating macrophages; left, H&E; right, reactivity with patient's own serum; inset, immunostaining for PAC3 antigen specific to P. acnes). Macrophages clustered around the dilated hair follicle contain numerous granular-looking microbes. The patient's own serum clearly demonstrates the pathogen not only in macrophages but also in commensal bacterial colonies in the hair follicle (arrow). Immunostaining for PAC3 antigen confirms the diagnosis of P. acnes-induced folliculitis.

of P. acnes-induced folliculitis was thus confirmed (Figure 45).

5.3.2 Ectopic liver infestation of Ascaris lumbricoides

Figure 45.

104

applying immunostaining using the patient serum.

5.3.1 Propionibacterium acnes folliculitis

Immunohistochemistry - The Ageless Biotechnology

Hepatic ascariasis (left upper, cut surface of the resected liver; right upper, Ouchterlony's diffusion-in-gel test using the extract of varied helminths; left lower, H&E; right lower, reactivity with patient's own serum). Yellow-colored xanthogranulomatous nodules are seen at the liver hilus. Microscopically, a small number of dead and calcified ova are distributed in necrotic substance. The content of the ova is immunoreactive with the patient's serum. In Ouchterlony's test, a precipitation line against Ascaris lumbricoides (As) has been absorbed by the Ascaris extract, as shown on the right hand (Em, Echinococcus multilocularis; Fh, Fasciola hepatica; Tc, Toxocara canis; EV, Enterobius vermicularis; CRP, C-reactive protein).

clear precipitation line against A. lumbricoides (Figure 46). The precipitation line was abolished by preabsorbing the serum with the roundworm extract [75]. The final diagnosis was ectopic ascariasis, caused by infestation of a young (immature) female worm. The serum was unreactive with ova of Schistosoma japonicum and Paragonimus miyazakii seen in paraffin sections. The inner surface of the cell wall of A. simplex larva was weakly stained with the serum [21].

Figure 47 demonstrates a splenic lesion with ectopic migration of A. lumbricoides. Necrotizing granulomas were formed beneath the splenic capsule, and dead parasite body fragments were seen in the necrotic area. It was impossible to determine the species of nematode under microscopy. The abovementioned serum

#### Figure 47.

Splenic ascariasis (left, cut surface of the formalin-fixed spleen; center, H&E; right, reactivity with the serum of patient with hepatic ascariasis). Ectopic migration of A. lumbricoides has provoked necrotizing granulomas beneath the splenic capsule. Dead parasite body fragments are seen in the necrotic area. The abovementioned serum of patient with hepatic ascariasis reacts with the dead worm body, confirming the diagnosis of ectopic splenic ascariasis.

of patient with hepatic ascariasis reacted with the dead worm body, confirming the diagnosis of ectopic splenic ascariasis. The serum functioned as an immunohistochemical probe specific to A. lumbricoides [21].

aged 60's suffering from alcoholic liver cirrhosis manifested left hemiparesis [79]. Computed tomography disclosed multifocal low densities in his right hemisphere. Herpetic encephalitis was clinically suspected. HIV antibody was negative. Progressive intracranial edema necessitated decompressive craniotomy with brain biopsy. The brain tissue microscopically showed perivascular chronic active inflammation, with ameba-like cells somewhat resembling macrophages being scattered. The 1:500 diluted patient's serum clearly reacted with the protozoan bodies, and mouse antiserum to Acanthamoeba culbertsoni (a gift from Prof. Yuji Tachibana, Department of Infectious Diseases, Tokai University School of Medicine, Isehara) also gave positivity (Figure 49) [7, 8, 21, 23, 77]. No reactivities for A. polyphaga and A. castellanii were noted. High immunofluorescence titer against A. culbertsoni was serologically confirmed in the patient's serum. Detailed microscopic observation of H&E-stained preparations disclosed the presence of acanthamebic trophozoites and cysts in the brain tissue. The final diagnosis was opportunistic acanthamebic meningoencephalitis associated with liver cirrhosis. Balamuthia mandrillaris may cause amebic meningoencephalitis in both immunocompromised and immunocompetent individuals [80]. An amoeba-induced skin nodule may be formed before the onset of meningoencephalitis [81]. A healthy Japanese farmer housewife aged 50's suddenly complained of progressive consciousness disturbance and seizure. She daily grew vegetables. Two weeks after onset, the patient expired. At autopsy, the basal side of the brain grossly revealed hemorrhagic meningoencephalitis. Microscopically, large-sized, basophilic amebic trophozoites were clustered mainly in Virchow-Robin's spaces. Smaller-sized cysts

Low-Specificity and High-Sensitivity Immunostaining for Demonstrating Pathogens…

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were focally observed. PCR analysis disclosed infection of B. mandrillaris.

Figure 49.

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Acanthamoeba subspecies.

Immunostaining using both the patient's own serum and the patient's serum of the abovementioned acanthamebic meningoencephalitis gave distinct positivity (Figure 50) [23]. Balamuthia-infected skin nodule seen in another patient gave clear positivity of the microbe using these two patients'sera. Cross-reactivity of the acanthamebic antigens to Balamuthia species was thus confirmed, but the serum

Acanthamebic meningoencephalitis (left upper, low-powered H&E; right upper, high-powered H&E; inset, an amebic cyst; left lower, reactivity with patient's own serum; right lower, reactivity with mouse antiserum to Acanthamoeba culbertsoni). An HIV-negative Japanese male suffering from alcoholic liver cirrhosis manifested left hemiparesis. Progressive intracranial edema necessitated decompressive craniotomy with brain biopsy. The brain tissue microscopically shows perivascular chronic active inflammation, and amebic trophozoites and cysts (inset) are scattered. The diluted patient's serum clearly reacts with the amebic bodies.

A. culbertsoni infection has been confirmed by using a panel of mouse antisera against different
