INFLUENCE OF PRESERVATIVE ON STAINING TECHNIQUE FOR

BLASTOCYSTIS SPP.

Nithyamathi,K., Suresh, K. And Sivanandam,S.

Department of Parasitology, Faculty of Medicine

University Malaya.

Abstract :. The influence or effect of three types of preservative for blastocystis such as
10% Formalin, 70% Alcohol and Polyvinyl Alcohol (PVA) on three different staining
technique which are Field stain, Giemsa stain and Trichome stain was studied. Monkey stool
sample which is positive for blastocystis is collected and preserved in three different
preservatives and kept at room temperature and 4⁰C. An amount of blastocystis from in vitro
culture were added into the preservatives as well (for both cyst and vacuolar form). Stool
specimen kept in preservative to preserve morphology of blastocystis and to prevent the
continued development of cyst. Preserved sample was stained to observe morphology, clarity
of nuclear and cytoplasmic details, overall color differences and ease or difficulty detecting
blastocystis in fecal sample at different time intervals (1,7,14 and 30 days). The results
revealed that 10% formalin is the most effective preservative whereby for 70% alcohol, fecal
debris was clumped together and a number of blastocystis were seen and some of its
morphology likely lysed and has spike structure on surface. PVA fixative is not
recommended and Trichome staining is preferable to be used for unpreserved sample. Further
work is needed to complete the experiment to determine stability of blastocystis for 14 and 30
days in preservative. Hence, this study recommended that 70% alcohol can be used as
preservative instead of 10% formalin for permanent staining.