The diagnostics and treatment of tropical diseasesStitt, E. R. (Edward Rhodes)
Science
The diagnostics and treatment of tropical diseases
Stitt, E. R. (Edward Rhodes)
Tropical medicine
For bringing out the nuclear characteristics of human amoebae
Walker recommends fixation of thin moist smears in Giemsa’s
sublimate alcohol (absolute alcohol 1 part, sat. aq. sol.
bichloride 2 parts) for 10 to 15 minutes. These smears are then
well washed with water and stained with alum haematoxylin for five
minutes. The nuclear characteristics are noted under etiology. In
such staining the preparations, which are best made on cover-slips,
should never be allowed to become dry.
An excellent iron haematoxylin method is that of Rosenbusch:
Rapidly smear out with a toothpick a small particle of faeces or
other material containing protozoa and, while still moist, fix by
Giemsa’s method and, after getting rid of the mercury with iodine
solution followed by 95% alcohol, treat smears with a 3.5% solution
of iron-alum in distilled water for one-half hour or overnight,
then wash thoroughly in distilled water.
Then stain from five to twenty minutes in the following
haematoxylin stain: (1) 1% solution of haematoxylin in 95% alcohol.
It takes at least ten days to ripen. (2) A saturated solution of
lithium carbonate. Add to 10 cc. of the haematoxylin solution
5 to 6 drops of the lithium carbonate one. Next wash well and
differentiate with about 1% solution of the iron-alum. Again wash
in water, pass through alcohols to xylol and mount in balsam. With
vegetative amoebae I have obtained beautiful results with vital
staining which can best be done by tinging the faeces emulsion with
a 1% aqueous solution of neutral red. I have also had good results
by emulsifying the faeces in a drop of 1 or 2% formalin and then
adding a drop of 2% acetic acid. The mixture is then tinged with
either neutral red or methyl green.
For distinguishing the encysted form of _Entamoeba coli_ one can
obtain excellent results by emulsifying the faeces in Gram’s iodine
solution. Owing to the glycogenic reaction given by _E. coli_, the
round amoeba, with its 8 nuclei stands out very distinctly.
For diagnosing the 4-nucleated cyst of the pathogenic amoeba one
gets better results with haematoxylin as this brings out not only
the 4 nuclei but the chromidial bodies as well. It was formerly
customary to recommend the administration of salts prior to examining
for amoebae. Walker warns that such a procedure gives us amoebae
which are difficult to differentiate, the nuclear characteristics of
_E. coli_ and the tetragena nucleus of _E. histolytica_ being much
alike as they both contain much chromatin. In a dysenteric stool the
histolytica type of nucleus, containing but little chromatin, does
not resemble the nucleus of _E. coli_.
He prefers the examination of formed stools obtained without a
purgative.
DIFFERENTIATING CHARACTERISTICS OF PATHOGENIC AMOEBAE
(AFTER DOBELL AND O’CONNOR). MOTILE STAGE
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