flame cells of the miracidium within the shell can be seen to move in a rapid flickering motion.
2. The eggs within the tissue can be removed (carefully tease the tissue apart in saline solution) and subjected to a hatching procedure. If the miracidia are released from the egg and swim to the top of the hatching flask, this movement is also proof of viability. (The procedure can be found in chapter 4.)
Digestion Procedure in Diagnosis of Schistosomiasis
Small pieces of tissue may be digested in 4% potassium hydroxide (4% sodium hydroxide may be used) for 2 to 3 h at 60 to 80°C. The material may then be concentrated by sedimentation or centrifugation and examined under the microscope for eggs (low power with low-intensity light).
References
1. Schwartz DA, Bryan RT, Hewanlowe KO, Visvesvara GS, Weber R, Cali A, Angritt P. 1992. Disseminated microsporidiosis (Encephalitozoon hellem) and acquired immunodeficiency syndrome—autopsy evidence for respiratory acquisition. Arch Pathol Lab Med 116:660–668. PMID 1616428
2. Current WL, Garcia LS. 1991. Cryptosporidiosis. Clin Microbiol Rev 4:325–358. PMID 1889046
3. Garcia LS. 1999. Practical Guide to Diagnostic Parasitology. ASM Press, Washington, DC.
4. Garcia LS, Shimizu RY. 1997. Evaluation of nine immunoassay kits (enzyme immunoassay and direct fluorescence) for detection of Giardia lamblia and Cryptosporidium parvum in human fecal specimens. J Clin Microbiol 35:1526–1529. PMID 9163474
5. Siddons CA, Chapman PA, Rush BA. 1992. Evaluation of an enzyme immunoassay kit for detecting Cryptosporidium in faeces and environmental samples. J Clin Pathol 45:479–482. PMID 1624594
6. Weber R, Bryan RT, Owen RL, Wilcox CM, Gorelkin L, Visvesvara GS, The Enteric Opportunistic Infections Working Group. 1992. Improved light-microscopical detection of microsporidia spores in stool and duodenal aspirates. N Engl J Med 326:161–166. PMID 1370122
7. Ryan NJ, Sutherland G, Coughlan K, Globan M, Doultree J, Marshall J, Baird RW, Pedersen J, Dwyer B. 1993. A new trichrome-blue stain for detection of microsporidial species in urine, stool, and nasopharyngeal specimens. J Clin Microbiol 31:3264–3269. PMID 7508457
8. Morris A, Norris KA. 2012. Colonization by Pneumocystis jirovecii and its role in disease. Clin Microbiol Rev 25:297–317. PMID 22491773
9. Botterel F, Cabaret O, Foulet F, Cordonnier C, Costa JM, Bretagne S. 2012. Clinical significance of quantifying Pneumocystis jirovecii DNA by using real-time PCR in bronchoalveolar lavage fluid from immunocompromised patients. J Clin Microbiol 50:227–231. PMID 22162560
10. Kovacs JA, Masur H. 2009. Evolving health effects of Pneumocystis: one hundred years of progress in diagnosis and treatment. JAMA 301:2578–2585. PMID 19549975
11. Brinn NT. 1983. Rapid metallic histological staining using the microwave oven. J Histotechnol 6:125–129.
12. Pintozzi RL. 1978. Technical methods: modified Grocott’s methenamine silver nitrate method for quick staining of Pneumocystis carinii. J Clin Pathol 31:803–805. PMID 80411
13. Arasteh KN, Simon V, Musch R, Weiss RO, Przytarski K, Futh UM, Pleuger F, Huhn D, L’age MP. 1998. Sensitivity and specificity of indirect immunofluorescence and Grocott-technique in comparison with immunocytology (alkaline phosphatase anti alkaline phosphatase = APAAP) for the diagnosis of Pneumocystis carinii in broncho-alveolar lavage (BAL). Eur J Med Res 3:559–563. PMID 9889176
14. Magee JG, McDade KJ, Cunningham J, Harrison V. 1991. Pneumocystis carinii pneumonia—detection of parasites by immunofluorescence based on a monoclonal antibody. Med Lab Sci 48:235–237. PMID 1787784
15. Midgley J, Parsons PA, Shanson DC, Husain OAN, Francis N. 1991. Monoclonal immunofluorescence compared with silver stain for investigating Pneumocystis carinii pneumonia. J Clin Pathol 44:75–76. PMID 1705263
16. Stratton N, Hryniewicki J, Aarnaes SL, Tan G, de la Maza LM, Peterson EM. 1991. Comparison of monoclonal antibody and calcofluor white stains for the detection of Pneumocystis carinii from respiratory specimens. J Clin Microbiol 29:645–647. PMID 1709951
17. Wolfson JS, Waldron MA, Sierra LS. 1990. Blinded comparison of a direct immunofluorescent monoclonal antibody staining method and a Giemsa staining method for identification of Pneumocystis carinii in induced sputum and bronchoalveolar lavage specimens of patients infected with human immunodeficiency virus. J Clin Microbiol 28:2136–2138. PMID 1699970
18. Homer KS, Wiley EL, Smith AL, McCollough L, Clark D, Nightingale SD, Vuitch F. 1992. Monoclonal antibody to Pneumocystis carinii. Comparison with silver stain in bronchial lavage specimens. Am J Clin Pathol 97:619–624. PMID 1374213
19. Rafanan AL, Klevjer-Anderson P, Metersky ML. 1998. Pneumocystis carinii pneumonia diagnosed by non-induced sputum stained with a direct fluorescent antibody. Ann Clin Lab Sci 28:99–103. PMID 9558448
20. Tuncer S, Erguven S, Kocagoz S, Unal S. 1998. Comparison of cytochemical staining, immunofluorescence and PCR for diagnosis of Pneumocystis carinii on sputum samples. Scand J Infect Dis 30:125–128. PMID 9730296
21. Khan MA, Farrag N, Butcher P. 1999. Diagnosis of Pneumocystis carinii pneumonia: immunofluorescence staining, simple PCR or nPCR. J Infect 39:77–80. PMID 10468133
22. Reid AB, Chen SC, Worth LJ. 2011. Pneumocystis jirovecii pneumonia in non-HIV-infected patients: new risks and diagnostic tools. Curr Opin Infect Dis 24:534–544. PMID 21986616
23. Musto L, Flanigan M, Elbadawi A. 1982. Ten-minute silver stain for Pneumocystis carinii and fungi in tissue sections. Arch Pathol Lab Med 106:292–294. PMID 6178384
24. Settnes OS, Larsen P. 1979. Inhibition of toluidine blue O stain for Pneumocystis carinii by additives in the diethyl ether. Am J Clin Pathol 72:493–494. PMID 89811
25. Bottone EJ. 1991. Diagnosis of acute pulmonary toxoplasmosis by visualization of invasive and intracellular tachyzoites in Giemsa-stained smears of bronchoalveolar lavage fluid. J Clin Microbiol 29:2626–2627. PMID 1774274
26. Blanshard C, Hollister WS, Peacock CS, Tovey DG, Ellis DS,