Question

Write down the grading, result interpretation and ways of documentation of acid fast stain microscopy
for intestinal coccidian.

Answer 1

Ziehl – neelsen staining is used to staining the both mycobacterium and protozoa. Because both are acid fast stain. This staining helps to identify the pathological intestinal coccidian protozoa in the stool.

Intestinal coccidian comprises of cysts of cryptosporidium pavum, isopora belli, cyclospora cayetanesis are stained in acid fast and recorded its result after staining. If these are colored with in a particular stain, these species retain it red color. Its red color will not be decolorized with alcohol and acid. Here methylene blue is being served as counter stain and able to identify red colored species of protozoa in a sample

Items to be taken for procedure are

Ziehl Neelsen A contains fuschin base, Ziehl Neelsen B contains decolorize substance, Ziehl –Neelsen C contains mehylene blue substance, staining table, water, microscopic slide for staining, burner, stool sample for testing

Procedure

Initially the sample of test is smeared on the microscopic slide and it should be dry before testing and then the slide should be crossed over the low flame in order to fix the smear. After that it can be kept on staining rack and fuchsin base is mixed with smear slide over a 4 minute. Then it is washed in flowing water. Then it is washed lightly after it counterstained with mehylene blue. Then it is dried. It is examined with light microscope with 400× and 1000× magnification. Oil immersion can be used for the 1000× magnification

Interpretation of results based on staining results

It normally bright red to intestive purple, red, bright, or slightly curved rods, bean shaped. It is differ depend on the protozoa

If it is cryptosporidium pavum, it will be spherical form, and size of oocytes 4-6µm and it Is stained bright pinkish red.

If it is cyclospora cayetanesis, it will be round spherical form, size of oocytes 8-10µ m and it will different in color between unstained to pink to deep red

If it is isopora belli, ellipsoidal shape, it size will be 20-30 µm long and 10 -20µm wide, two round spherocytes may be seen in staining

Documentation of test

The result of the test is recorded in a register and health personnel records the rest if it is completed and records the reason if it is not completed and reason for conducting test

Then the health care personnel should record the result of test either positive or negative based on result of test

Then the health care personnel should record the species of protozoa based on identification

Then the health care personnel should the record the number of protozoa based on identification

Grading of acid fast of intestinal coccidian can be done on the number of protozoa

Number of protozoa	Grading
No parasites	Negative
If it contains Lower than 1 parasites/10 observation field of 1000× magnification	+
If it contains 1-10 parasites/10 observation field of 1000× magnification	++
If it contains 1-10 parasites /observation field of 1000×magnification	+++
If it contains higher than 10 parasites /observation field of 1000×magnification	++++