Isolation and purification of Giardia trophozoites from rat intestine

D. E. Feely, S. L. Erlandsen

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

A method for the isolation of Giarda trophozoites based on their ability to attach to warm surfaces has been developed. Mucosal scrapings were obtained from the small intestine of rats infected with Giardia and suspended in Hanks' balanced salt solution (HBSS). Trophozoites were concentrated by centrifugation and allowed to attach to the surfaces of polystyrene petri dishes incubated at 37 C. Incubation temperature significantly affected the recovery of trophozoites. After attachment at 37 C, trophozoites were separated from contaminating intestinal debris by incubation at cold temperature. The trophozoites detach at 4 C, whereas the intestinal debris remain adherent. Then the detached Trophozoites were isolated by reattachment at 37 C. Examination by scanning electron microscopy revealed a marked reduction in contamination of attached trophozoites and dish surfaces after the use of cold temperature detachment and reattachment at 37 C. Viability of trophozoites, as measured by erythrosin-B dye exclusion, remained above 90% up to 120 min after isolation. This method of isolation facilitates the recovery of this protozoan directly from small intestine for morphological and experimental study.

Original languageEnglish (US)
Pages (from-to)59-64
Number of pages6
JournalJournal of Parasitology
Volume67
Issue number1
DOIs
StatePublished - Jan 1 1981

Fingerprint

Giardia
Trophozoites
trophozoites
Intestines
purification
intestines
rats
incubation
temperature
dye
isolation techniques
viability
experimental study
scanning electron microscopy
Small Intestine
salt
small intestine
Erythrosine
erythrosine
Polystyrenes

ASJC Scopus subject areas

  • Parasitology
  • Ecology, Evolution, Behavior and Systematics

Cite this

Isolation and purification of Giardia trophozoites from rat intestine. / Feely, D. E.; Erlandsen, S. L.

In: Journal of Parasitology, Vol. 67, No. 1, 01.01.1981, p. 59-64.

Research output: Contribution to journalArticle

@article{93e5d5a369de4495a6b0b5dd55a63457,
title = "Isolation and purification of Giardia trophozoites from rat intestine",
abstract = "A method for the isolation of Giarda trophozoites based on their ability to attach to warm surfaces has been developed. Mucosal scrapings were obtained from the small intestine of rats infected with Giardia and suspended in Hanks' balanced salt solution (HBSS). Trophozoites were concentrated by centrifugation and allowed to attach to the surfaces of polystyrene petri dishes incubated at 37 C. Incubation temperature significantly affected the recovery of trophozoites. After attachment at 37 C, trophozoites were separated from contaminating intestinal debris by incubation at cold temperature. The trophozoites detach at 4 C, whereas the intestinal debris remain adherent. Then the detached Trophozoites were isolated by reattachment at 37 C. Examination by scanning electron microscopy revealed a marked reduction in contamination of attached trophozoites and dish surfaces after the use of cold temperature detachment and reattachment at 37 C. Viability of trophozoites, as measured by erythrosin-B dye exclusion, remained above 90{\%} up to 120 min after isolation. This method of isolation facilitates the recovery of this protozoan directly from small intestine for morphological and experimental study.",
author = "Feely, {D. E.} and Erlandsen, {S. L.}",
year = "1981",
month = "1",
day = "1",
doi = "10.2307/3280779",
language = "English (US)",
volume = "67",
pages = "59--64",
journal = "Journal of Parasitology",
issn = "0022-3395",
publisher = "American Society of Parasitologists",
number = "1",

}

TY - JOUR

T1 - Isolation and purification of Giardia trophozoites from rat intestine

AU - Feely, D. E.

AU - Erlandsen, S. L.

PY - 1981/1/1

Y1 - 1981/1/1

N2 - A method for the isolation of Giarda trophozoites based on their ability to attach to warm surfaces has been developed. Mucosal scrapings were obtained from the small intestine of rats infected with Giardia and suspended in Hanks' balanced salt solution (HBSS). Trophozoites were concentrated by centrifugation and allowed to attach to the surfaces of polystyrene petri dishes incubated at 37 C. Incubation temperature significantly affected the recovery of trophozoites. After attachment at 37 C, trophozoites were separated from contaminating intestinal debris by incubation at cold temperature. The trophozoites detach at 4 C, whereas the intestinal debris remain adherent. Then the detached Trophozoites were isolated by reattachment at 37 C. Examination by scanning electron microscopy revealed a marked reduction in contamination of attached trophozoites and dish surfaces after the use of cold temperature detachment and reattachment at 37 C. Viability of trophozoites, as measured by erythrosin-B dye exclusion, remained above 90% up to 120 min after isolation. This method of isolation facilitates the recovery of this protozoan directly from small intestine for morphological and experimental study.

AB - A method for the isolation of Giarda trophozoites based on their ability to attach to warm surfaces has been developed. Mucosal scrapings were obtained from the small intestine of rats infected with Giardia and suspended in Hanks' balanced salt solution (HBSS). Trophozoites were concentrated by centrifugation and allowed to attach to the surfaces of polystyrene petri dishes incubated at 37 C. Incubation temperature significantly affected the recovery of trophozoites. After attachment at 37 C, trophozoites were separated from contaminating intestinal debris by incubation at cold temperature. The trophozoites detach at 4 C, whereas the intestinal debris remain adherent. Then the detached Trophozoites were isolated by reattachment at 37 C. Examination by scanning electron microscopy revealed a marked reduction in contamination of attached trophozoites and dish surfaces after the use of cold temperature detachment and reattachment at 37 C. Viability of trophozoites, as measured by erythrosin-B dye exclusion, remained above 90% up to 120 min after isolation. This method of isolation facilitates the recovery of this protozoan directly from small intestine for morphological and experimental study.

UR - http://www.scopus.com/inward/record.url?scp=0019467214&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0019467214&partnerID=8YFLogxK

U2 - 10.2307/3280779

DO - 10.2307/3280779

M3 - Article

C2 - 7229820

AN - SCOPUS:0019467214

VL - 67

SP - 59

EP - 64

JO - Journal of Parasitology

JF - Journal of Parasitology

SN - 0022-3395

IS - 1

ER -