DETECTION OF HAEMOPARASITES AND HAEMOPLASMAS IN DOMESTIC CATS OF KERALA
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Date
2019-09-19
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES, POOKODE WAYANAD
Abstract
The present study was conducted with the objective of detection of
haemoparasites and haemoplasmas of domestic cats using microscopy and
polymerase chain reaction (PCR). A total of 111 cat blood samples were collected
from four different districts of Kerala viz., Wayanad, Kozhikode, Ernakulam and
Thiruvananthapuram. Peripheral blood smears were prepared and stained with
Giemsa’s stain and were microscopically examined for the presence of any
organisms. The examination of blood smears could not detect any parasitemia in
any of the smears. The whole blood samples collected in EDTA vials were
processed for the isolation of genomic DNA for PCR amplification. The genomic
DNA were used as template for all PCR reactions. The piroplasmid primers
targeting 18S rRNA gene with a product size of 358 bp showed amplification in
13 samples. Sequence analysis and BLAST results of the PCR products revealed
10 samples infected with Hepatozoon felis (9.01 per cent), and three infected with
Cytauxzoon felis (2.70 per cent). The phylogenetic tree constructed for
Hepatozoon felis based on 18S rRNA gene revealed the isolates of Kerala being
diverged into two clades. Nine of the isolates grouped with H. felis sequences of
cats from Hyderabad, India. One isolate of Kerala clustered with sequences of
domestic cats from Japan and Austria and also with wild felids. This indicated the
existence of two genetically different populations of H. felis infecting the
domestic cats of Kerala. The phylogenetic tree based on 18S rRNA gene plotted
for C. felis showed that the three sequences of C. felis from Kerala formed a
separate clade and do not cluster with any other isolates of pathogenic C. felis, C.
manul and undetermined Cytauxzoon sp. of Europe, proving the existence of a
non pathogenic population of C. felis existing in the apparently healthy cats of
Kerala. Similarly, PCR amplification of 16S rRNA gene specific for Mycoplasma
spp. yielded ~600 bp product in 10 samples. NCBI-BLAST of nucleotide
sequences revealed the existence of three different populations of Mycoplasma
spp. (9.01 per cent) circulating in the blood of infected cat viz., Mycoplasma
haemofelis in two cats, Candidatus M. haemominutum in seven cats and
Candidatus M. turicensis in one cat. The phylogenetic analysis of feline
85 Mycoplasma spp. isolates from Kerala using 16S rRNA gene revealed two
sequences of Mycoplasma haemofelis clustered with M. haemofelis isolates from
South Africa, USA, UK and Australia, seven sequences of Candidatus M.
haemominutum belonged to the same clade with Candidatus M. haemominutum
isolates of other countries and one sequence of Candidatus M. turicensis grouping
in one clade with Candidatus M. turicensis isolates from other countries. Thus,
the present study established the prevalence of H. felis, C. felis and three
population of Mycoplasma spp. viz., Mycoplasma haemofelis (1.80 per cent),
Candidatus M. haemominutum (6.31 per cent) and Candidatus M. turicensis (0.90
per cent) in domestic cats of Kerala.