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ASCARIS SUUM EGG RECOVERY FROM SLUDGE SAMPLES AFTER PHASE EXTRACTION.

D Naidoo¹, C E Archer¹

¹Water, Sanitation and Hygiene Research and Development Centre, Howard College Campus, University of KwaZulu-Natal, Durban 4041, South Africa.

The Journal of Parasitology

|July 21, 2024

Summary

Phase extraction in helminth testing can lead to significant egg loss. Researchers recommend excluding this step to improve egg recovery from sanitation samples.

Keywords:

Ascaris suum Helminth eggs Hydrophilic Lipophilic Phase extraction Recovery Solvent

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Area of Science:

Environmental Science
Parasitology
Analytical Chemistry

Background:

Helminth testing in sanitation samples often involves phase extraction to reduce lipids and pellet size.
This method aims to concentrate helminth eggs for microscopy by creating two liquid phases and a debris plug.

Purpose of the Study:

To evaluate the impact of different hydrophilic and lipophilic solutions on helminth egg recovery during phase extraction.
To identify combinations that minimize egg loss and optimize sample preparation for microscopy.

Main Methods:

Tested combinations of hydrophilic solutions (10% formalin, acetoacetic buffer, acid alcohol) and lipophilic solvents (ethyl acetate, diethyl ether).
Assessed egg recovery from water, primary sludge, and fatty sludge samples.
Collected and examined both the sedimented pellet and the supernatant/debris plug to quantify egg loss.

Main Results:

Combinations of 10% formalin or acetoacetic buffer with ethyl acetate or diethyl ether resulted in less than 50% egg recovery.
Acid alcohol with ethyl acetate yielded high egg recovery (57.3–93.2%) but did not reduce pellet size.
Significant helminth egg loss was observed with certain phase extraction protocols.

Conclusions:

The phase extraction step in current helminth testing methods can cause substantial egg loss.
Excluding the phase extraction step is recommended to improve the accuracy of helminth egg detection in sanitation samples.
Further optimization of sample preparation methods is needed for reliable helminth egg quantification.