Blood Safety and Clinical Technology Guidelines on Standard Operating Procedures for MICROBIOLOGY Chapter 20-Parasitological Examination of Faeces

The examination of faeces for parasitological diagnosis is done to detect: Adult worms Segments of tapeworms Ova and cysts Larvae Trophozoites Cellular exudates such as WBCs, RBCs, macrophages and Charcot-Leyden (CL) c rystals For this, the sample should be properly collected and preserved. Collection of faecal sample Ask the patient to pass the stool sample directly into a waxed cardboard or a plastic cup with a tight fitting lid. Collection of sample in a match box or on plant leaves is not a satisfactory method. About 20-40 grams of well-formed stool or 5-6 table spoonfuls of watery sto ol will suffice for a routine examination. Ingestion of some medicines prior to collection of faecal sample may interf ere with the detection of parasites. These include tetracyclines, sulfonamides, antiprotozoal agents, laxatives, antacids, castor oil, magnesium hydroxide, bari um sulphate, bismuth kaolin compounds and hypertonic salts etc. These should not be taken 1-2 weeks before the examination of stool sample. All specimens must be properly labelled with patient s name, age, sex, and da te of collection. The specimen must reach the laboratory within 30 minutes of passing of the stool, since amoebic trophozoites die and become unrecognizable after that. Note Do not keep the specimen at warm temperatures. Try to keep it in cool, shad y places. Prevent the drying of the specimen. Prevent contamination with urine or dirt particles. Multiple stool examinations are required before the presence of parasitic i nfections is ruled out. Stool should not be collected from bed-pans containing disinfectants. Transportation of samples If looking for trophozoites, stool specimen must be transported very rapidly to the laboratory to avoid disintegration of trophozoites. Stool samples should be examined within 30 minutes of collection of the specimen and not receipt of the specimen in the laboratory. Stool specimens should never be frozen and thawed or placed in an incubator because parasitic forms deteriorate very rapidly. For permanent fixation of the stool specimen, 10% formol-saline (prepared by add ing 100 ml formaldehyde to 900 ml of 0.85% sodium chloride) is a well known pres ervative. Polyvinyl alcohol (PVA) is a widely used preservative because the perf ormance of concentration procedures and preparation of permanent stained smears are both possible with this. Macroscopic examination Various points to be noted are: Consistency: The consistency of the stool could be formed, soft, loose or w

atery. The cysts are found maximum in the formed stool while trophozoites are mo st abundant in watery stool. Presence of blood and mucus. Presence of round worms, thread worms or tapeworm proglottids. Colour and smell of the stool. Microscopic examination (temporary wet mounts) It is the simplest and easiest technique. A wet mount can be prepared directly f rom faecal material or from the concentrated specimens. The basic types of wet m ounts that should be made from each sample include: a) Saline wet mount: It is used to detect worm eggs or larvae, protozoan tr ophozoites and cysts. In addition it can reveal the presence of RBCs and WBCs. b) Iodine wet mount: It is used to stain glycogen and nuclei of the cysts. Procedure Place a drop of saline on left half of the slide and one drop of iodine on the right half. With an applicator stick pickup a small portion of the specimen (equivalent to the size of a match head) and mix with saline drop. Similarly pickup similar amount and mix with a drop of iodine. Put the cover slip separately on both and examine under the microscope. Ova, cysts, trophozoites and adult worms can be identified as per their cha racteristic features (Fig 1 and Table 1). Iodine wet mount is examined for amoebic and flagellar cysts.