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Culture of Protozoan Parasites

Article in Clinical Microbiology Reviews · August 2002

DOI: 10.1128/CMR.15.3.327-328.2002 · Source: PubMed

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CLINICAL MICROBIOLOGY REVIEWS, July 2002, p. 327–328 Vol. 15, No. 3
0893-8512/02/$04.00⫹0 DOI: 10.1128/CMR.15.3.327–328.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

GUEST COMMENTARY

Culture of Protozoan Parasites

Govinda S. Visvesvara1* and Lynne S. Garcia2
Centers for Disease Control and Prevention, Atlanta, Georgia 30341,1 and LSG & Associates,
Santa Monica, California 904022

There are a number of issues involved in the culture of disease is invaluable, as it provides not only information on the
protozoan parasites that make these procedures highly com- development of the parasite but also avenues for new ap-
plex and subject to many variables, some of which are known proaches to the containment and/or eradication of the parasite.
and some of which are still undefined. Certainly protozoan In vitro cultivation is important for a number of reasons, as
parasites have complex life cycles. They may have different follows: (i) as an important adjunct to diagnosis; (ii) to produce
morphological stages within the life cycle and may have both antigens used to prepare monoclonal and polyclonal antibod-
cold-blooded and warm-blooded animals as intermediate or ies against the organisms for use in immunologic tests; (iii) to
definitive hosts within the life cycle. In vitro culture of organ- identify specific proteins that may enhance the invasive prop-
isms at any one of these stages within the life cycle involves a erties of the parasite and in turn the development of mono-
tremendous number of variables, including parasite stage, host clonal antibodies that will help neutralize parasitic invasion;
site, host temperature, host immune responses, parasite spe- (iv) to assess functional antibodies and cell-mediated protec-
cies and/or strain, and parasite-protective mechanisms. To sim- tive systems against the parasites, assessments that can only be
ulate the host environment in an in vitro culture system can be made in a cost-effective manner using in vitro culture; (v) to
extremely demanding, assuming one can actually determine all screen drugs, in vitro, in order to identify potential therapeutic
the relevant variables (2, 3, 10). agents; (vi) to differentiate susceptible from resistant isolates
Often the organisms are very fastidious in their growth require- so that advances in chemotherapy can be made; (vii) to differ-
ments, and different lots of media and/or medium components entiate clinical isolates using techniques such as isoenzyme
may be toxic. Something as simple as the type of glass used for the electrophoresis, monoclonal antibody techniques, and/or DNA
culture container can have tremendous influence over the success probe techniques; (viii) to elucidate isolate and strain differ-
or failure of culture trials. Many culture medium components ences which will be a useful tool for molecular epidemiology;
require filter sterilization and have relatively short shelf lives. and (ix) to produce vaccines, as relatively large numbers of
Another requirement for many medium formulations includes parasites at specific stages can be produced in culture. (x) In
the addition of human or animal sera, which are expensive and addition, continuous culture over long periods of time may
highly variable and may contain factors that are detrimental to cause attenuation of strains, and therefore, attenuated strains
parasite growth. Much research has been devoted to the devel- have potential in the development of suitable vaccines. (xi) In
opment of defined medium formulations, although, even with the vitro cultivation also provides a system to assess vaccine effi-
elimination of serum, various other components may not have cacy, since it can only be done by using intact parasites that can
been totally defined. In some cases, growth factors have been be obtained in large quantities and without the contaminating
identified and substituted for serum or serum components. influences of host components. Finally, culture can be used
At the present time, are there any advantages in the in vitro (xii) to provide the parasite inoculum used for experimental
culture of protozoan parasites, especially when revolutionary animal disease models; (xiii) to study the biochemistry, physi-
developments are going on in the field of molecular biology? ology, and metabolism of the parasites as well as determine
One can argue that many types of research, including the their nutritional requirements; (xiv) to understand the ultra-
diagnosis of etiologic agents, can be accomplished even when structural organization of the parasite; and (xv) to provide a
very small numbers of organisms are available. However, many system suitable for the assay of lymphokines and other cyto-
areas of research can be undertaken only when large numbers kines that may block invasion of the parasite.
of parasites with no contaminating bacteria or host materials Individuals working in this field have indicated that “black
are available. One of the great advantages of in vitro culture, magic” may be required for culture success; therefore, clinical
especially axenic culture, is that obtaining a continuous supply laboratories have been somewhat hesitant to undertake these
of pure organisms without any bacterial and/or fungal contam- types of diagnostic procedures on a routine basis. However,
ination is possible (4, 9). with proper culture controls in addition to patient specimen
In vitro cultivation of parasitic protozoa that cause human cultures, clinically relevant information can be obtained from
these cultures when other diagnostic methods may be unsuc-
cessful (1, 5, 6, 8). This is particularly true for cultures of some
* Corresponding author. Mailing address: Division of Parasitic Dis-
eases, MS F36, Centers for Disease Control and Prevention, 4770
of the free-living amebae and the hemoflagellates (1, 7, 10).
Buford Highway NE, Atlanta, GA 30341-3724. Phone: (770) 488-4417. Another potential problem is associated with earlier research
Fax: (770) 488-4253. E-mail: [email protected]. publications in which various media are mentioned but not

327
328 GUEST COMMENTARY CLIN. MICROBIOL. REV.

thoroughly described; the same medium is often referred to by A. E. R. Taylor and J. R. Baker (ed.), In vitro methods for parasite cultiva-
tion. Academic Press, Orlando, Fla.
different names.
3. Evans, D. A. 1987. Leishmania, p. 52–75. In A. E. R. Taylor and J. R. Baker
In vitro cultivation of the various parasites discussed in this (ed.), In vitro methods for parasite cultivation. Academic Press, Orlando,
issue will continue to be a challenging diagnostic option but, Fla.
nevertheless, a fruitful area of research. It is anticipated that 4. 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
further research on the emerging and reemerging opportunis- tachyzoites. Eur. J. Clin. Microbiol. Infect. Dis. 18:879–884.
tic parasites will likely include surface antigens that may be 5. Garcia, L. S. 2001. Diagnostic medical parasitology, 4th ed., p. 850–872.
involved in the invasion of the host cell. ASM Press, Washington, D.C.
6. Garcia, L. S. (ed.). 1992. Section 7. Parasitology, p. 7.0.1–7.10.8.2. In H. D.
Following is an excellent, comprehensive collection of works Isenberg (ed.), Clinical microbiology procedures handbook. American Soci-
on the current status of protozoan culture systems. This infor- ety for Microbiology, Washington, D.C.
mation should be helpful to all microbiologists in understand- 7. Novy, F. G., and W. J. McNeal. 1904. On the cultivation of Trypanosoma
brucei. J. Infect. Dis. 1:1–30.
ing the role that in vitro culture plays, not only in the research 8. Taylor, A. E. R., and J. R. Baker. 1968. The cultivation of parasites in vitro.
setting but also in the clinical arena. Blackwell Scientific Publications, Ltd., Oxford, United Kingdom.
9. Trager, W., and J. Jensen. 1976. Human malaria parasites in continuous
REFERENCES culture. Science 193:673–675.
1. Dedet, J. P., F. Pratlong, R. Pradinaud, and B. Moreau. 1999. Delayed 10. Visvesvara, G. S. 1999. Pathogenic and opportunistic free-living amebae, p.
culture of Leishmania in skin biopsies. Trans. R. Soc. Trop. Med. Hyg. 1383–1390. In P. R. Murray, E. J. Baron, M. A. Pfaller, F. C. Tenover, and
93:673–674. R. H. Yolken (ed.), Manual of clinical microbiology, 7th ed. ASM Press,
2. Diamond, L. S. 1987. Entamoeba, Giardia and Trichomonas, p. 1–28. In Washington, D.C.

The views expressed in this Commentary do not necessarily reflect the views of the journal or of ASM.

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