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Chapter 32 : Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis

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Abstract:

Pancreatic digests of casein are major ingredients of media used in the axenic cultivation of lumen-dwelling parasitic protozoa. Unfortunately, the digest used almost exclusively in the development of these media has not been available since the early 1980s. Many digest products have been tried in the interim with marginal results in supporting the growth of . Specimens would include cerebrospinal fluid (CSF), biopsy tissue, and autopsy tissue of the brain; for spp., corneal scrapings or biopsy material, contact lenses and contact lens paraphernalia such as lens cases and solutions, skin abscess material, ear discharge, or feces can also be used. Three continuous cell lines (HeLa, LLC, and Vero) and three cell culture methods (culture in conventional flasks, culture in membrane-based flasks, and an automated culture system) were investigated. Most routine clinical laboratories do not have the animal care facilities necessary to provide animal inoculation capabilities for the diagnosis of parasitic infections. Host specificity for many animal parasite species is a well-known fact which limits the types of animals available for these procedures. In certain suspect infections, animal inoculation may be requested and can be very helpful in making the diagnosis, although animal inoculation certainly does not take the place of other, more routine procedures. Xenodiagnosis is a technique that uses the arthropod host as an indicator of infection. Uninfected reduviid bugs are allowed to feed on the blood of a patient who is suspected of having Chagas’ disease ( infection).

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Figures

Image of Figure 32.1
Figure 32.1

Protozoa from culture systems. (Upper left) trophozoite from liquid medium containing rice starch (note that there are no definitive erythrocytes within the cytoplasm, so that it is not possible to differentiate the true pathogen, , from the nonpathogen, ). (Upper right) trophozoite from nonnutrient agar culture with bacterial overlay (note that this trophozoite has been stained). (Lower left) spp. trophozoite from nonnutrient agar culture with bacterial overlay (note the spiky acanthapodia). (Lower right) spp. cysts from nonnutrient agar culture with bacterial overlay (note the double hexagonal wall appearance.

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.2
Figure 32.2

flagellate stage. When placed in distilled water (enflagellation test), , the causal agent of primary amebic meningoencephalitis, undergoes transformation to a pear-shaped flagellate, usually with two flagella but occasionally with three or four flagella; the flagellate stage is a temporary nonfeeding stage and usually reverts to the trophozoite stage.

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.3
Figure 32.3

Illustration of the InPouch TV culture system for From top to bottom: (1) introduction of the specimen into the upper chamber containing a small amount of medium; (2) application of a plastic holder for microscope viewing prior to expressing medium into the lower chamber (optional); (3) transfer of a small amount of medium in the upper chamber to the lower chamber; (4) rolling down the upper chamber and sealing it with the tape; (5) plastic viewing frame used to immobilize the medium in the pouch for examination under the microscope. Specific photographs can also be seen in Figure 5.22. (Courtesy of BIOMED Diagnostics.)

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.4
Figure 32.4

InPouch TV culture system for Note the pouch, swabs, and plastic pouch holder for microscopic examination of the pouch contents. (Courtesy of BIOMED Diagnostics.)

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.5
Figure 32.5

Illustration of a tube of NNN medium, used for the culture and recovery of spp. (A) Fluid condensate and tissue culture medium overlay containing the developing organisms. (B) Blood agar medium. (Illustration by Sharon Belkin.)

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.6
Figure 32.6

spp. from culture systems. (Upper left) Wet mount of culture fluid sediment showing promastigotes of spp. (Upper right) Stained smear of culture fluid sediment showing promastigotes of spp. (Lower left) Stained spp. promastigotes. (Lower right) Stained promastigotes (note the round nucleus, bar-shaped kinetoplast, and flagella).

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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Image of Figure 32.7
Figure 32.7

Illustration of the process of xenodiagnosis used for the diagnosis of Chagas’ disease. (Illustration by Sharon Belkin.)

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
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References

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1. Arrowood, M. J. 2002. In vitro cultivation of Cryptosporidium species. Clin. Microbiol. Rev. 15:390400.
2. Asami, K.,, Y. Nodake, and, T. Ueno. 1955. Cultivation of Trichomonas vaginalis on solid medium. Exp. Parasitol. 4:3439.
3. Beal, C.,, R. Goldsmith,, M. Kotby,, M. Sherif,, A. El-Tagi,, A. Farid,, S. Zakaria, and, J. Eapen. 1992. The plastic envelope method, a simplified technique for culture diagnosis of trichomoniasis. J. Clin. Microbiol. 30:22652268.
4. Borchardt, K. A.,, M. Z. Zhang,, H. Shing, and, K. Flink. 1997. A comparison of the sensitivity of the InPouch TV, Diamond’s and Trichosel media for detection of Trichomonas vaginalis. Genitourin. Med. 73:297298.
5. Bowman, E. K.,, A. A. Vass,, R. Mackowski,, B. A. Owen, and, R. L. Tyndall. 1996. Quantitation of free-living amoebae and bacterial populations in eyewash stations relative to flushing frequency. Am. Ind. Hyg. Assoc. J. 57:626633.
6. Chang, T. H.,, S. Y. Tsing, and, S. Tzang. 1986. Monoclonal antibodies against Trichomonas vaginalis. Hybridoma 5:4351.
7. Clark, C. G.,, and L. S. Diamond. 2002. Methods for cultivation of luminal parasitic protists of clinical importance. Clin. Microbiol. Rev. 15:329341.
8. Cote, R. (ed.) 1984. American Type Culture Collection. American Type Culture Collection, Rockville, Md.
9. Dedet, J. P.,, F. Pratlong,, R. Pradinaud, and, B. Moreau. 1999. Delayed culture of Leishmania in skin biopsies. Trans. R. Soc. Trop. Med. Hyg. 93:673674.
10. Desjardins, R.,, and J. Bowdre. 1981. In vitro cultivation of malaria parasites. Clin. Microbiol. Newsl. 3:5253.
11. Diamond, L. S. 1983. Lumen dwelling protozoa: Entamoeba, Trichomonads, and Giardia, p. 65–109. In J. B. Jensen (ed.), In Vitro Cultivation of Protozoan Parasites. CRC Press, Inc., Boca Raton, Fla.
12. Diamond, L. S. 1987. Entamoeba, Giardia and Trichomonas, p. 1–28. In A. E. R. Taylor and, J. R. Baker (ed.), In Vitro Methods for Parasite Cultivation. Academic Press, Inc., Orlando, Fla.
13. Diamond, L. S.,, C. G. Clark, and, C. C. Cunnick. 1995. YI-S, a casein-free medium for axenic cultivation of Entamoeba histolytica, related Entamoeba, Giardia intestinalis, and Trichomonas vaginalis. J. Eukaryot. Microbiol. 42:277278.
14. Dolkart, R.,, and B. Halpern. 1958. A new monophasic medium for the cultivation of Entamoeba histolytica. Am. J. Trop. Med. Hyg. 7:595596.
15. Druilhe, P.,, D. Mazier,, O. Brandicourt, and, M. Gentilini. 1983. One-step Plasmodium falciparum cultivation—application to in-vitro drug testing. Trop. Med. Parasitol. 34:233234.
16. Evans, D. A. 1978. Kinetoplastida, p. 55–58. In A. E. R. Taylor and, J. R. Baker (ed.), Methods of Culturing Parasites In Vitro. Academic Press, Inc., New York, N.Y.
17. Evans, D. A. 1987. Leishmania, p. 52–75. In A. E. R. Taylor and, J. R. Baker (ed.), In Vitro Methods for Parasite Cultivation. Academic Press, Inc., Orlando, Fla.
18. Evans, R.,, J. M. Chatterton,, D. Ashburn,, A. W. Joss, and, D. O. Ho-Yen. 1999. Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. Eur. J. Clin. Microbiol. Infect. Dis. 18:879884.
19. Fairlamb, A. H.,, D. C. Warhurst, and, W. Peters. 1985. An improved technique for the cultivation of Plasmodium falciparum in vitro without daily medium change. Ann. Trop. Med. Parasitol. 75:717.
20. Garcia, L. S. 1999. Practical Guide to Diagnostic Parasitology. ASM Press, Washington, D.C.
21. Gorlin, A. I.,, M. M. Gabriel,, L. A. Wilson, and, D. G. Ahearn. 1996. Effect of adhered bacteria on the binding of Acanthamoeba to hydrogel lenses. Arch. Ophthalmol. 114:576580.
22. Hendricks, L. D.,, D. E. Wood, and, M. E. Hajduk. 1978. Hemoflagellates: commercially available liquid media for rapid cultivation. Parasitology 76:309316.
23. Hollander, D. H. 1976. Colonial morphology of Trichomonas vaginalis in agar. J. Parasitol. 62:826828.
24. Isenberg, H. D. (ed.). 2004. Clinical Microbiology Procedures Handbook, 2nd ed., p. 7.0.17.10.8.2. ASM Press, Washington, D.C.
25. Jensen, J. B. 2005. Reflections on the continuous cultivation of Plasmodium falciparum. J. Parasitol. 91:487491.
26. Jensen, J. B.,, W. Trager, and, J. Doherty. 1979. Plasmodium falciparum: continuous cultivation in a semiautomated apparatus. Exp. Parasitol. 48:3641.
27. Krogstad, D. A.,, G. S. Visvesvara,, K. W. Walls, and, J. W. Smith. 1985. Blood and tissue protozoa, p. 612–630. In E. H. Lennette,, A. Balows,, W. J. Hausler, Jr., and, H. J. Shadomy (ed.), Manual of Clinical Microbiology, 4th ed. American Society for Microbiology, Washington, D.C.
28. Kulda, J.,, S. M. Honigberg,, J. K. Frost, and, H. D. Hollander. 1970. Pathogenicity of Trichomonas vaginalis. A clinical and biologic study. Am. J. Obstet. Gynecol. 108:908918.
29. Kuroki, T.,, S. Sata,, S. Yamai,, K. Yagita,, Y. Katsube, and, T. Endo. 1998. Occurrence of free-living amoebae and Legionella in whirlpool bathes. Kansenshogaku Zasshi 72:10561063.
30. Landers, P.,, K. G. Kerr,, T. J. Rowbotham,, J. L. Tipper,, P. M. Keig,, E. Ingham, and, M. Denton. 2000. Survival and growth of Burkholderia cepacia within the free-living amoeba Acanthamoeba polyphaga. Eur. J. Clin. Microbiol. Infect. Dis. 19:121123.
31. Lennette, E. H.,, A. Balows,, W. J. Hausler, Jr., and, H. J. Shadomy (ed.). 1985. Manual of Clinical Microbiology, 4th ed. American Society for Microbiology, Washington, D.C.
32. Linstead, D. 1990. Cultivation, p. 91–111. In B. M. Honigberg (ed.), Trichomonads Parasitic in Man. Springer-Verlag, New York, N.Y.
33. Ma, P.,, G. S. Visvesvara,, A. J. Martinez,, F. H. Theodore,, P.-M. Daggett, and, T. K. Sawyer. 1990. Naegleria and Acanthamoeba infections: review. Rev. Infect. Dis. 12:490513.
34. Maekelt, G. 1964. A modified procedure of xenodiagnosis of Chagas’ disease. Am. J. Trop. Med. Hyg. 13:1115.
35. Marciano-Cabral, F.,, and G. Cabral. 2003. Acanthamoeba spp. as agents of disease in humans. Clin. Microbiol. Rev. 16:273307.
36. Markell, E.,, and M. Voge. 1976. Medical Parasitology. The W. B. Saunders Co., Philadelphia, Pa.
37. Martinez, A. J. 1985. Free-Living Amebas: Natural History, Prevention, Diagnosis, Pathology, and Treatment of the Disease. CRC Press, Inc., Boca Raton, Fla.
38. Mata-Cardenas, B. D.,, J. Vargas-Villarreal,, L. Navarro-Marmolejo, and, S. Said-Fernandez. 1998. Axenic cultivation of Trichomonas vaginalis in a serum-free medium. J. Parasitol. 84:638639.
39. Mazur, T.,, E. Hadas, and, I. Iwanicka. 1995. The duration of the cyst stage and the viability and virulence of Acanthamoeba isolates. Trop. Med. Parasitol. 46:106108.
40. McMillan, A. 1990. Laboratory diagnostic methods and cryopreservation of trichomonads, p. 297–310. In B. M. Honigberg (ed.), Trichomonads Parasitic in Man. Springer-Verlag, New York, N.Y.
41. Michel, R.,, and M. Borneff. 1989. The significance of amoebae and other protozoa in water conduit systems in dental units. Zentbl. Bakteriol. Mickobiol. Hyg. Ser. B 187:312323.
42. Michel, R.,, H. Burghardt, and, H. Bergmann. 1995. Acanthamoeba, naturally intracellularly infected with Pseudomonas aeruginosa, after their isolation from a micro-biologically contaminated drinking water system in a hospital. Zentbl. Hyg. Umweltmed. 196:532544.
43. Newsome, A. L.,, T. M. Scott,, R. F. Benson, and, B. S. Fields. 1998. Isolation of an amoeba naturally harboring a distinctive Legionella species. Appl. Environ. Microbiol. 64:16881693.
44. Novy, F. G.,, and W. J. McNeal. 1903. The cultivation of Trypanosoma brucei. A preliminary note. JAMA 41:12661268.
45. Novy, F. G.,, and W. J. McNeal. 1904. On the cultivation of Trypanosoma brucei. J. Infect. Dis. 1:130.
46. Ohlemeyer, C. L.,, L. L. Hornberger,, D. A. Lynch, and, E. M. Swierkosz. 1998. Diagnosis of Trichomonas vaginalis in adolescent females: InPouch TV culture versus wet-mount microscopy. J. Adolesc. Health 22:205208.
47. Pabst, U.,, J. Demuth,, T. Gebel, and, H. Dunkelberg. 1997. Establishment of a method for determining the association between Legionella sp. and Amoeba sp. using polymerase chain reaction. Zentbl. Hyg. Umweltmed. 199:568577.
48. Page, F. C. 1988. A New Key to Fresh Water and Soil Gymnamoebae. Fresh Water Biological Association, Ambleside, England.
49. Penland, R. L.,, and K. R. Wilhelmus. 1997. Comparison of axenic and monoxenic media for isolation of Acanthamoeba. J. Clin. Microbiol. 35:915922.
50. Penland, R. L.,, and K. R. Wilhelmus. 1998. Laboratory diagnosis of Acanthamoeba keratitis using buffered charcoal-yeast extract agar. Am. J. Ophthalmol. 126:590592.
51. Rondanelli, E. G. 1987. Amphizoic Amoebae: Human Pathology. Piccin Nuova Libraria, Padua, Italy.
52. Schuster, F. L. 2002. Cultivation of Babesia and Babesia-like blood parasites: agents of an emerging zoonotic disease. Clin. Microbiol. Rev. 15:365373.
53. Schuster, F. L. 2002. Cultivation of pathogenic and opportunistic free-living amebas. Clin. Microbiol. Rev. 15:342354.
54. Schuster, F. L. 2002. Cultivation of Plasmodium spp. Clin. Microbiol. Rev. 15:355364.
55. Schuster, F. L.,, and J. J. Sullivan. 2002. Cultivation of clinically significant hemoflagellates. Clin. Microbiol. Rev. 15:374389.
56. Shepp, D.,, R. Hackman,, F. Conley,, J. Anderson, and, J. Meyers. 1985. Toxoplasma gondii reactivation identified by detection of parasitemia in tissue culture. J. Intern. Med. 103:218221.
57. Smith, R. F. 1986. Detection of Trichomonas vaginalis in vaginal specimens by direct immunofluorescence assay. J. Clin. Microbiol. 24:11071108.
58. Taylor, A. E. R.,, and J. R. Baker. 1968. The Cultivation of Parasites In Vitro. Blackwell Scientific Publications Ltd., Oxford, United Kingdom.
59. Trager, W. 1976. Prolonged cultivation of malaria parasites (Plasmodium coatneyi and P. falciparum), p. 427–434. In H. Van den Bossche (ed.), Biochemistry of Parasites and Host-Parasite Relationships. Elsevier-North Holland Biomedical Press, Amsterdam, The Netherlands.
60. Trager, W. 1979. Plasmodium falciparum in culture: improved continuous flow method. J. Protozool. 26:125129.
61. Trager, W.,, and J. B. Jensen. 1976. Human malaria parasites in continuous culture. Science 193:673675.
62. Trager, W.,, and J. B. Jensen. 2005. Human malaria parasites in continuous culture. 1976. J. Parasitol. 91:484486.
63. Visvesvara, G. S. 1995. Pathogenic and opportunistic free-living amebae, p. 1196–1203. In P. R. Murray,, E. J. Baron,, M. A. Pfaller,, F. C. Tenover, and, R. H. Yolken (ed.), Manual of Clinical Microbiology, 6th ed. American Society for Microbiology, Washington, D.C.
64. Visvesvara, G. S. 2002. In vitro cultivation of microsporidia of clinical importance. Clin. Microbiol. Rev. 15:401413.
65. Visvesvara, G. S.,, and L. S. Garcia. 2002. Culture of protozoan parasites. Clin. Microbiol. Rev. 15:327328.
66. Walochnik, J.,, O. Picher,, C. Aspock,, M. Ullmann,, R. Sommer, and, H. Aspock. 1998. Interactions of “Limax amoebae” and gram-negative bacteria: experimental studies and review of current problems. Tokai J. Exp. Clin. Med. 23:273278.
67. Walton, B. C.,, J. J. Shaw, and, R. Lainson. 1977. Observations on the in vitro cultivation of Leishmania braziliensis. J. Parasitol. 63:11181119.
68. Winiecka-Krusnell, J.,, and E. Linder. 1999. Free-living amoebae protecting Legionella in water: the tip of an iceberg? Scand. J. Infect. Dis. 31:383385.

Tables

Generic image for table
Table 32.1

Approaches to isolation of free-living amebae

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32
Generic image for table
Table 32.2

Issues related to free-living amebae and symbiotic bacteria

Citation: Garcia L. 2007. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 910-935. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch32

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