Rusli, Nurul Ain
(2014)
Detection of intestinal parasites and
toxoplasma gondii in Kota Bharu pet cats via
microscopy and molecular techniques.
Project Report.
Universiti Sains Malaysia.
(Submitted)
Abstract
Toxoplasma gondii is a zoonotic parasitic protozoon that causes toxoplasmosis. Cat is a definitive host for T. gondii and a reservoir for transmission of other intestinal parasites.
Thus, this study was embarked upon to examine and identify intestinal parasites in pet
cats and developed an in-house duplex polymerase chain reaction (PCR) assay for collected from different areas in Kota Bharu, Kelantan. All stool samples were screened
using conventional saline wet mount and examined by direct microscopy. Subsequently,
the samples were concentrated by formalin ether sedimentation techniques and stained
with permanent modified acid-fast stain. For specific DNA detection, an in-house
duplex PCR assay was developed to detect an internal transcribed spacers (ITS 1) region
of 18S rRNA gene of T. gondii. Plasmodium falciparum gene was incorporated into the
PCR assay as the internal control to rule out false negative results. Result showed that
58% were positive with one or more parasites. For direct wet mount, eggs of Toxocara
cati, Toxascaris leonine, Ancylostoma sp., Trichuris vulpis, Uncinaria stenocephala,
Trichuris trichiura, Ascaris sp. and Capillaria sp. were observed in 5(14%), 5(14%),
3(8%), 1(3%) ,1(3%), 1(3%), 1(3%) and 1(3%) stool sample, respectively. In the
concentration technique, 8(22%) of stool samples were positive for T. cati', 6(17%) for
T. leonine, 3(8%) for Ancylostoma sp., 2(6%) for U. stenocephala, 1(3%) for T. vulpis,
T. trichiura, Ascaris sp. and Capillaria sp.. Interestingly, neither oocysts nor DNA of
T. gondii was detected by microscopy and PCR assay. The latter assay revealed that the
results were all true negatives because each PCR assay produced the expected internal control amplicon of 150 bp. In conclusion, none of the pet cats was infected with T.
gondii but they carried other intestinal parasites, in which T. cati was the most common.
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