1887

Chapter 8 : Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis

MyBook is a cheap paperback edition of the original book and will be sold at uniform, low price.

Ebook: Choose a downloadable PDF or ePub file. Chapter is a downloadable PDF file. File must be downloaded within 48 hours of purchase

Buy this Chapter
Digital (?) $30.00

Preview this chapter:
Zoom in
Zoomout

Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555819002/9781555819002_ch8-1.gif /docserver/preview/fulltext/10.1128/9781555819002/9781555819002_ch8-2.gif

Abstract:

  • Culture methods
    • Intestinal protozoa
    • Pathogenic free-living amebae
    • spp. ()
    • Pathogenic flagellates
    • Flagellates of blood and tissue
    • and spp.
    • spp.
    • Microsporidia
  • Animal inoculation
    • spp.
    • spp.
  • Xenodiagnosis

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Image of Figure 8.1
Figure 8.1

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. doi:10.1128/9781555819002.ch8.f1

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.2
Figure 8.2

flagellate stage. (Upper) 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. (Lower) flagellate stage (microtubules are highlighted in green, basal bodies in red, and DNA is stained blue). (Courtesy of Lillian Fritz-Laylin; from http://genome.jgi-psf.org/Naegr1/Naegr1.home.html.) doi:10.1128/9781555819002.ch8.f2

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.3
Figure 8.3

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. doi:10.1128/9781555819002.ch8.f3

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.4
Figure 8.4

InPouch TV culture system for . Note the pouch, swabs, and plastic pouch holder for microscopic examination of the pouch contents. doi:10.1128/9781555819002.ch8.f4

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.5
Figure 8.5

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.) doi:10.1128/9781555819002.ch8.f5

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.6
Figure 8.6

spp. from culture systems. (Upper left) Stained smear of culture fluid sediment [U3] showing promastigotes of sp. (Upper right) Stained smear of culture fluid sediment showing promastigotes of sp. (higher magnification). (Lower left) Stained promastigote. (Lower right) promastigotes during infection of primary fibroblast culture. Cells are stained with antitubulin (green) and antiactin (red). (Courtesy of the Pasteur Institute, Molecular Parasitology and Signaling Image Bank). doi:10.1128/9781555819002.ch8.f6

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.7
Figure 8.7

RH tachyzoites replicated in Vero cell cultures. A rosette of many tachyzoites is seen at the left and several parasite pairs are at right (arrows). (Courtesy of I. Canedo-Solares, Abstract 42.009, 15 International Congress on Infectious Diseases, Bangkok, Thailand, 2012). doi:10.1128/9781555819002.ch8.f7

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.8
Figure 8.8

(A) An E6 cell infected with . Magnification, ×1,200. Note the well-defined multiple parasitophorous vacuoles (PV); N, host cell nucleus. (B) An HLF cell culture completely destroyed by . Note the spores with everted polar tubules (at arrows), Magnification, ×600. All three species of destroy the cell culture, and often the cell cultures are completely covered by spores that either are intact or have discharged their polar tubules. Note It is very unusual to see the extruded polar tubules in routine clinical specimens such as urine or stool. (Courtesy of Govinda Visvesvara, from Visvesvara G, 401–413, 2002). doi:10.1128/9781555819002.ch8.f8

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.9
Figure 8.9

(A) A host cell infected with () . Note the arrangement of spores around the host (E6 cell) nucleus (N). A single spore is probably in the process of infecting an adjacent cell (arrowhead). Magnification, ×1,200. (B) A spore with an everted polar tubule. Magnification, ×1,200. (Courtesy of Govinda Visvesvara, from Visvesvara G, 401–413, 2002). doi:10.1128/9781555819002.ch8.f9

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 8.10
Figure 8.10

Illustration of the process of xenodiagnosis used for the diagnosis of Chagas' disease. (Illustration by Sharon Belkin.)doi:10.1128/9781555819002.ch8.f10

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555819002.garcia.ch08
1. Arrowood MJ. 2002. In vitro cultivation of Cryptosporidium species. Clin Microbiol Rev 15:390400.
2. Clark CG,, Diamond LS. 2002. Methods for cultivation of luminal parasitic protists of clinical importance. Clin Microbiol Rev 15:329341.
3. Isenberg HD (ed). 2004. Clinical Microbiology Procedures Handbook, 2nd ed., p. 7.0.17.10.8.2. ASM Press, Washington, DC.
4. Garcia LG (ed). 2010. Clinical Microbiology Procedures Handbook, 3rd ed., ASM Press, Washington, DC.
5. Schuster FL. 2002. Cultivation of Babesia and Babesia-like blood parasites: agents of an emerging zoonotic disease. Clin Microbiol Rev 15:365373.
6. Schuster FL. 2002. Cultivation of pathogenic and opportunistic free-living amebas. Clin Microbiol Rev 15:342354.
7. Schuster FL. 2002. Cultivation of Plasmodium spp. Clin Microbiol Rev 15:355364.
8. Schuster FL,, Sullivan JJ. 2002. Cultivation of clinically significant hemoflagellates. Clin Microbiol Rev 15:374389.
9. Taylor AER,, Baker JR. 1968. The Cultivation of Parasites In Vitro. Blackwell Scientific Publications Ltd., Oxford, United Kingdom.
10. Trager W,, Jensen JB. 2005. Human malaria parasites in continuous culture. 1976. J Parasitol 91:484486.
11. Visvesvara GS. 2002. In vitro cultivation of microsporidia of clinical importance. Clin Microbiol Rev 15:401413.
12. Roberts T,, Barratt J,, Harkness J,, Ellis J,, Stark D. 2011. Comparison of microscopy, culture, and conventional polymerase chain reaction for detection of Blastocystis sp. in clinical stool samples. Am J Trop Med Hyg. 84:308312.
13. Visvesvara GS,, Garcia LS. 2002. Culture of protozoan parasites. Clin Microbiol Rev 15:327328.
14. Diamond LS,, Clark CG,, Cunnick CC. 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.
15. Garcia LS. 1999. Practical Guide to Diagnostic Parasitology. ASM Press, Washington, DC.
16. Dolkart R,, Halpern B. 1958. A new monophasic medium for the cultivation of Entamoeba histolytica. Am J Trop Med Hyg 7:595596.
17. Cote R (ed). 1984. American Type Culture Collection. American Type Culture Collection, Rockville, MD.
18. Ma P,, Visvesvara GS,, Martinez AJ,, Theodore FH,, Daggett P-M,, Sawyer TK. 1990. Naegleria and Acanthamoeba infections: review. Rev Infect Dis 12:490513.
19. Martinez AJ. 1985. Free-Living Amebas: Natural History, Prevention, Diagnosis, Pathology, and Treatment of the Disease. CRC Press, Inc., Boca Raton, FL.
20. Page FC. 1988. A New Key to Fresh Water and Soil Gymnamoebae. Fresh Water Biological Association, Ambleside, United Kingdom.
21. Visvesvara GS,. 1995. Pathogenic and opportunistic free-living amebae, p 11961203. In Murray PR,, Baron EJ,, Pfaller MA,, Tenover FC,, Yolken RH. (ed), Manual of Clinical Microbiology, 6th ed. American Society for Microbiology, Washington, DC.
22. Marciano-Cabral F,, Cabral G. 2003. Acanthamoeba spp. as agents of disease in humans. Clin Microbiol Rev 16:273307.
23. Diamond LS,. 1987. Entamoeba, Giardia and Trichomonas, p 128. In Taylor AER,, Baker JR (ed), In Vitro Methods for Parasite Cultivation. Academic Press, Inc., Orlando, FL.
24. Diamond LS,. 1983. Lumen dwelling protozoa: Entamoeba, Trichomonads, and Giardia, p 65109. In Jensen JB (ed), In Vitro Cultivation of Protozoan Parasites. CRC Press, Inc., Boca Raton, FL.
25. Linstead D,. 1990. Cultivation, p 91111. In Honigberg BM (ed), Trichomonads Parasitic in Man. Springer-Verlag, New York, NY.
26. McMillan A,. 1990. Laboratory diagnostic methods and cryopreservation of trichomonads, p 297310. In Honigberg BM (ed), Trichomonads Parasitic in Man. Springer-Verlag, New York, NY.
27. Borchardt KA,, Zhang MZ,, Shing H,, Flink K. 1997. A comparison of the sensitivity of the InPouch TV, Diamond's and Trichosel media for detection of Trichomonas vaginalis. Genitourin Med 73:297298.
28. Ohlemeyer CL,, Hornberger LL,, Lynch DA,, Swierkosz EM. 1998. Diagnosis of Trichomonas vaginalis in adolescent females: InPouch TV culture versus wet-mount microscopy. J Adolesc Health 22:205208.
29. Mata-Cardenas BD,, Vargas-Villarreal J,, Navarro-Marmolejo L,, Said-Fernandez S. 1998. Axenic cultivation of Trichomonas vaginalis in a serum-free medium. J Parasitol 84:638639.
30. Barratt JL,, Banik GR,, Harkness J,, Marriott D,, Ellis JT,, Stark D. 2010. Newly defined conditions for the in vitro cultivation and cryopreservation of Dientamoeba fragilis: new techniques set to fast track molecular studies on this organism. Parasitology 137:18671878.
31. Munasinghe VS,, Stark D,, Ellis JT. 2012. New advances in the in-vitro culture of Dientamoeba fragilis. Parasitology 139:864869.
32. Dedet JP,, Pratlong F,, Pradinaud R,, Moreau B. 1999. Delayed culture of Leishmania in skin biopsies. Trans R Soc Trop Med Hyg 93:673674.
33. Hendricks LD,, Wood DE,, Hajduk ME. 1978. Hemoflagellates: commercially available liquid media for rapid cultivation. Parasitology 76:309316.
34. Novy FG,, McNeal WJ. 1903. The cultivation of Trypanosoma brucei. A preliminary note. JAMA 41:12661268.
35. Novy FG,, McNeal WJ. 1904. On the cultivation of Trypanosoma brucei. J Infect Dis 1:130.
36. Walton BC,, Shaw JJ,, Lainson R. 1977. Observations on the in vitro cultivation of Leishmania braziliensis. J Parasitol 63:11181119.
37. Evans DA,. 1978. Kinetoplastida, p 5558. In Taylor AER,, Baker JR (ed), Methods of Culturing Parasites In Vitro. Academic Press, Inc., New York, NY.
38. Evans DA,. 1987. Leishmania, p 5275. In Taylor AER,, Baker JR (ed), In Vitro Methods for Parasite Cultivation. Academic Press, Inc., Orlando, FL.
39. Shepp D,, Hackman R,, Conley F,, Anderson J,, Meyers J. 1985. Toxoplasma gondii reactivation identified by detection of parasitemia in tissue culture. J Intern Med 103:218221.
40. Evans R,, Chatterton JM,, Ashburn D,, Joss AW,, Ho-Yen DO. 1999. Cell-culture system for continuous production of Toxoplasma gondii tachyzoites. Eur J Clin Microbiol Infect Dis 18:879884.
41. Cuellar JA,, Hernandez A,, Villegas E,, Gomez JE. 2012. Efficiency of in vitro culture of Toxoplasma gondii in THP1 and Vero cell lines. Biomedica 32:461466.
42. Trager W,, Jensen JB. 1976. Human malaria parasites in continuous culture. Science 193:673675.
43. Trager W. 1979. Plasmodium falciparum in culture: improved continuous flow method. J Protozool 26:125129.
44. Jensen JB,, Trager W,, Doherty J. 1979. Plasmodium falciparum: continuous cultivation in a semiautomated apparatus. Exp Parasitol 48:3641.
45. Castellanos-Gonzalez A,, Cabada MM,, Nichols J,, Gomez G,, White ACJr. 2013. Human primary intestinal epithelial cells as an improved in vitro model for Cryptosporidium parvum infection. Infect Immun 81:19962001.
46. Monaghan SR,, Rumney RL,, Vo NTY,, Bois NC,, Lee LE. 2011. In vitro growth of microsporidia Anncaliia algerae in cell lines from warm water fish. In Vitro Cell Dev Biol Anim 47:104113.
47. Maekelt G. 1964. A modified procedure of xenodiagnosis of Chagas' disease. Am J Trop Med Hyg 13:1115.

Tables

Generic image for table
TABLE 8.1

Approaches to isolation of free-living amebae

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8
Generic image for table
TABLE 8.2

Cell lines, media, and supplements that have been used to cultivate microsporidia causing human infections (adapted from and )

Citation: Garcia L. 2016. Parasite Recovery: Culture Methods, Animal Inoculation, and Xenodiagnosis, p 156-182. In Diagnostic Medical Parasitology, Sixth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555819002.ch8

This is a required field
Please enter a valid email address
Please check the format of the address you have entered.
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error