1887

Chapter 33 : Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods

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

Preview this chapter:
Zoom in
Zoomout

Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, Page 1 of 2

| /docserver/preview/fulltext/10.1128/9781555816018/9781555813802_Chap33-1.gif /docserver/preview/fulltext/10.1128/9781555816018/9781555813802_Chap33-2.gif

Abstract:

Adequate fixation of parasites is important not only for diagnostic procedures but also as a means of preserving positive material for personnel and student training. There are many fixatives and preservatives available; however, only the more common ones are presented in this chapter. To optimize molecular analyses of fixed tissues, morphologic and RNA integrity in rat liver was assessed when sections were fixed in 70% neutralbuffered formalin, modified Davidson’s II medium, 70% ethanol, UMFIX, modified Carnoy’s solution, modified methacarn, Bouin’s medium, phosphate-buffered saline, or 30% sucrose. Each sample was treated with standard or microwave fixation and standard or microwave processing, and sections were evaluated microscopically. RNA was extracted and assessed for preservation of quality and quantity. Definitive morphology needed for identification of protozoan trophozoites and cysts is best seen on the permanent stained smear, which can be prepared from fresh fecal specimens or from stool that has been submitted to the laboratory in fixative. Nematodes can be killed with hot water (60 to 63°C) and then transferred to a preservative, such as alcohol-glycerin or alcohol-formalin-acetic acid (AFA). A solution of alcohol, formalin, and acetic acid can be routinely used for nematodes, trematodes, and cestodes. The fixative should be used hot (60 to 63°C); after fixation for 24 h, parasites can be stored in the alcohol-glycerin mixture. Whole fecal specimens can be mixed directly with an appropriate fixative, although it is recommended that the organisms be concentrated before fixation. The use of formalin-saline is recommended for helminth eggs and larvae.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33

Key Concept Ranking

Taenia solium
0.61516184
Ascaris lumbricoides
0.6076598
0.61516184
Highlighted Text: Show | Hide
Loading full text...

Full text loading...

Figures

Image of Figure 33.1
Figure 33.1

(Left) cyst (unstained). (Right) cyst (unstained).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.2
Figure 33.2

Adult nematode (male worm).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.3
Figure 33.3

Adult nematode (note the “whiplike” appearance; this is a much smaller adult worm than ).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.4
Figure 33.4

fertilized eggs. Note the fully developed larva within each egg; these eggs were viable when photographed. In some cases, the moving larva can be seen, even in formalin-preserved specimens.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.5
Figure 33.5

Adult trematodes. (Upper) Preserved but unstained fluke; (lower) stained fluke (note the morphologic details that can be seen after staining).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.6
Figure 33.6

unstained proglottid. Note that morphologic details cannot be seen in this preserved but unstained preparation.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.7
Figure 33.7

(Left) spp. egg (note the striated shell and hooklets seen on the oncosphere). (Right) egg (note the six-hooked embryo and the polar filaments that are found between the oncosphere and the shell).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.8
Figure 33.8

Semipermanent mounts of small nematodes. (Left) Small nematode from soil sample. (Right) Female (pinworm) nematode.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.9
Figure 33.9

Stained trematodes. (Left) sp. (Right) sp. Note that the morphologic details are clearly visible after staining.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.10
Figure 33.10

gravid proglottids after India ink injection of the uterine branches. (Left) (Right)

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.11
Figure 33.11

itch mites in wet mounts. These specimens could be seen using the high dry objective (magnification, ×400).

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.12
Figure 33.12

(Left) Body louse, (Right) Flea,

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.13
Figure 33.13

Ticks. (Left) Example of a hard tick, (Right) Example of a soft tick, sp.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint
Image of Figure 33.14
Figure 33.14

Example of a large hard-body insect, which is a triatomid bug. These large insects can be pinned, labeled, and stored in boxes containing naphthalene flakes or paradichlorobenzene.

Citation: Garcia L. 2007. Fixation and Special Preparation of Fecal Parasite Specimens and Arthropods, p 936-946. In Diagnostic Medical Parasitology, Fifth Edition. ASM Press, Washington, DC. doi: 10.1128/9781555816018.ch33
Permissions and Reprints Request Permissions
Download as Powerpoint

References

/content/book/10.1128/9781555816018.ch33
1. Ash, L. R.,, and T. C. Orihel. 1987. Parasites: a Guide to Laboratory Procedures and Identification. ASCP Press, Chicago, Ill.
2. Berlin, O. G.,, and M. J. Miller. 1980. Euparal as a permanent mounting medium for helminth eggs and proglottids. J. Clin. Microbiol. 12:700703.
3. Bostwick, D. G.,, N. al Annouf, and, C. Choi. 1994. Establishment of the formalin-free surgical pathology laboratory. Utility of an alcohol-based fixative. Arch. Pathol. Lab. Med. 118:298302.
4. Cox, M. L.,, C. L. Schray,, C. N. Luster,, Z. S. Stewart,, P. J. Korytko,, K. N. Khan,, J. D. Paulauskis, and, R. W. Dunstan. 2006. Assessment of fixatives, fixation, and tissue processing on morphology and RNA integrity. Exp. Mol. Pathol. 80:183191.
5. Dillon, N.,, A. D. Austin, and, E. Bartowsky. 1996. Comparison of preservation techniques for DNA extraction from hymenopterous insects. Insect Mol. Biol. 5:2124.
6. Faust, E. C.,, P. C. Beaver, and, R. C. Jung. 1975. Animal Agents and Vectors of Human Disease, 4th ed. Lea & Febiger, Philadelphia, Pa.
7. France, S. C.,, and T. D. Kocher. 1996. DNA sequencing of formalin-fixed crustaceans from archival research collections. Mol. Mar. Biol. Biotechnol. 5:304313.
8. Fritsche, T. R. 1999. Arthropods of medical importance, p. 1449–1466. In P. R. Murray,, E. J. Baron,, M. A. Pfaller,, F. C. Tenover, and, R. H. Yolken (ed.), Manual of Clinical Microbiology, 7th ed. ASM Press, Washington, D.C.
9. Garcia, L. S.,, and L. R. Ash. 1979. Diagnostic Parasitology: Clinical Laboratory Manual, 2nd ed. The C. V. Mosby Co., St. Louis, Mo.
10. Goddard, J. 2003. Physician’s Guide to Arthropods of Medical Importance, 4th ed. CRC Press, Inc., Boca Raton, Fla.
11. Horsefall, W. R. 1962. Medical Entomology. The Ronald Press Co., New York, N.Y.
12. Hunter, G. W.,, J. C. Swartzwelder, and, D. F. Clyde. 1976. Tropical Medicine, 5th ed. The W. B. Saunders Co., Philadelphia, Pa.
13. Ishiwata, K.,, A. Shinohara,, K. Yagi,, Y. Horii,, K. Tsuchiya, and, Y. Nawa. 2004. Identification of tissue-embedded ascarid larvae by ribosomal DNA sequencing. Parasitol. Res. 92:5052.
14. James, M. T.,, and R. F. Harwood. 1969. Herms’ Medical Entomology, 6th ed. The Macmillan Co., New York, N.Y.
15. Kettle, D. S. 1995. Medical and Veterinary Entomology, 2nd ed. CAB International, Wallingford, United Kingdom.
16. Lane, R. P.,, and R. W. Crosskey. 1993. Medical Insects and Arthropods. Chapman & Hall, Ltd., London, United Kingdom.
17. Malek, E. T. A. 1951. Menthol relaxation of helminths before fixation. J. Parasitol. 37:321.
18. Matheson, R. 1950. Medical Entomology, 2nd ed. Comstock Publishing Co., Ithaca, N.Y.
19. Miething, F.,, S. Hering,, B. Hanschke, and, J. Dressler. 2006. Effect of fixation to the degradation of nuclear and mitochondrial DNA in different tissues. J. Histochem. Cytochem. 54:371374.
20. Nace, E. K.,, F. J. Steurer, and, M. L. Eberhard. 1999. Evaluation of Streck tissue fixative, a nonformalin fixative for preservation of stool samples and subsequent parasitologic examination. J. Clin. Microbiol. 37:41134119.
21. Palmer-Toy, D. E.,, B. Krastins,, D. A. Sarracino,, J. B. Nadol, Jr., and, S. N. Merchant. 2005. Efficient method for the proteomic analysis of fixed and embedded tissues. J. Proteome Res. 4:24042411.
22. Ramos, F.,, R. Zurabian,, P. Moran,, M. Ramiro,, A. Gomez,, C. G. Clark,, E. I. Melendro,, G. Garcia, and, C. Ximenez. 1999. The effect of formalin fixation on the polymerase chain reaction characterization of Entamoeba histolytica. Trans. R. Soc. Trop. Med. Hyg. 93:335336.
23. Reese, N. E.,, W. M. Boyce,, I. A. Gardner, and, D. M. Nelson. 1996. Fixation affects morphometric characters of Psoroptes cuniculi mites (Acari: Psoroptidae). J. Med. Entomol. 33:835838.
24. Scholten, T. H.,, and J. Yang. 1974. Evaluation of unpreserved and preserved stools for the detection and identification of intestinal parasites. Am. J. Clin. Pathol. 62:563567.
25. Steyskal, G. C.,, W. L. Murphy, and, E. M. Hoover. 1987. Insects and Mites: Techniques for Collection and Preservation. USDA miscellaneous publication no. 1443. U.S. Department of Agriculture, Washington, D.C.
26. Vince, D. G.,, A. Tbakhi,, A. Gaddipati,, R. M. Cothren,, J. F. Cornhill and, R. R. Tubbs. 1997. Quantitative comparison of immunohistochemical staining intensity in tissues fixed in formalin and Histochoice. Anal. Cell Pathol. 15:119129.

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