KATO THICK SMEAR

Background

 The cellophane-covered thick smear was originally described by Kato and Miura (1954)
for use in field surveys for helminthic infections;
 A very useful technique for helminth eggs, but not for helminth larvae or protozoa; not
recommended for stool specimens with large amounts of fiber or gas;
 A significant advantage of the procedure is the large quantity of eggs (50-60 mgs)
examined directly without employing other time-consuming concentration procedures.
Cellophane strips immersed in glycerine and malachite green mixtures are utilized.

Materials

 Glycerol, distilled water, 3% malachite green solution

 Transparent wettable cellophane
 Slide and coverslip
 Fecal sample

Procedure

 Cut wettable cellophane into 22 x 30 mm strips, soak for 24 hours or longer in a mixture
of 100 parts glycerol, 100 parts distilled water, and 1 part 3% malachite green solution;
 Place 50 to 60 milligrams fresh feces on a clean standard slide and cover with one of the
cellophane strips. Invert preparation above paper towels, press to spread fecal material
uniformly to edges of the cellophane strips;
 Reverse slide and let stand at +40C for 30 minutes, or at room temperature for 1 hour;
 Examine slide under LPO; confirm identification using HPO.

Comment

Since the technique employs cellophane strips immersed in a glycerine-malachite green solution,
this leads to glare reduction and improved visibility of helminth eggs due to the clearing action
of the glycerine.

Examination of a large amount of feces also greatly increases the sensitivity of this diagnostic
method in detecting helminthic eggs.

A major disadvantage of the Kato Thick smear technique has been the restriction of its
application to fresh specimens only or to those refrigerated for a relatively short period of time.

The “incubation period” in the glycerol-malachite green solution is crucial. If specimens do not
clear for a sufficient period of time, the eggs will be obscured. Likewise, for smears which are
allowed to clear for too long, some eggs (e.g., those of hookworm and schistosomes) will
collapse and thus they become too difficult to see or recognize.
In most routine diagnostic laboratories, the direct fecal smear and Kato Thick smear preparations
should be standard procedures for fecal examinations.

MODIFIED KATO-KATZ EGG COUNTING TECHNIQUE

Background

 A quantitative diagnostic technique developed by Katz et. al. (1972) for schistosomiasis
mansoni; involves measuring a known amount of feces by a piece of cardboard with a
small hole and examining it using the Kato cellophane thick smear technique;
 Very useful for mass surveys on helminthiasis due to its reliability, low cost, and easy
performance;
 Especially useful for epidemiological research since worm burden or intensity can be
estimated, the level of endemicity can be determined; also valuable in the evaluation of
eradication and/or control programs

Materials

 Cellophane coverslips used in Kato thick smear technique

 Wire (60 or 80 mesh) or nylon net (105 mesh)
 Rectangular cards (3 cm x 4 cm x 1.37 mm) with a central hole (6 mm in diameter)
 Absorbent paper and toothpicks

Procedure

1. Place fecal sample on absorbent paper and press upper part of sample with the wire net;
2. Withdraw the feces that is strained through the net and fill up the hole of the card which
lies over the glass slide. After filling up the central hole, carefully withdraw card, thus
leaving feces on the glass slide;
3. Cover the feces with cellophane coverslip, invert the slide, press against a sheet of
absorbent paper to evenly spread the fecal sample;
4. Examine microscopically after a few minutes. The number of eggs present in the fecal
material multiplied by 23 gives the number of eggs per grams (EPG) of feces.

Reading

Parasite Very Light Light Moderate Heavy Very

Heavy
Ascaris 100 – 9,999 10,000 – 50,000 – 100,000 –
lumbricoides EPG 49,000 EPG 99,999 EPG 299,999 EPG
Hookworm 100 – 699 700 – 2,599 2,600 – 12,599 12,600 – 25,100 and
EPG EPG EPG 25,099 EPG above
EPG
Trichuris 100 – 999 1,000 – 4,999 5,000 – 10,000 and
trichuria EPG EPG 9,000 EPG above
EPG
WHO classification of intensity of infections with soil-transmitted helminths and schistosomiasis
Parasite Light Intensity Moderate Intensity Heavy Intensity
Ascaris lumbricoides 1 – 4,999 EPG 5,000 – 49,999 EPG 50,000 EPG and
above
Hookworm 1 – 1,999 EPG 2,000 – 3,999 EPG 4,000 EPG and above
Schistosoma 1 – 99 EPG 100 – 399 EPG 400 EPG and above
japonicum
Trichuris trichuria 1 – 999 EPG 1,000 – 9,999 EPG 10,000 EPG and
above

Comment

On the average, the amount of feces produced when the cardboard hole is full is approximately
43.4 milligrams. Thus, 23 is the “standard factor” to estimate EPG.

If using templates of different sizes, note the standard factor to estimate EPG using the
following:
Hole Diameter (mm) Thickness (mm) Sample Capacity (mg) Factor
9 1 50 20
6 1.5 41.7 24
6.5 0.5 20 50

Quantitative diagnosis of helminthic infections is usually made –

a. To evaluate quantitatively the effect of parasite control;

b. To evaluate the efficacy of antihelminthics

Density of helminth eggs may be classified into four categories according to the number of eggs
found in the entire microscopic field – 1-9: +, 10-99: ++, 100-999: +++, and over 1,000: ++++.

The disadvantage of this method is that counting of eggs can be laborious if stool is derived from
heavily infected subjects. However, there is high sensitivity and the chances of missing light
infection is less.