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R esearch
(Published online:
11-12-2015)
2.
Flow cytometry based profiling of
leukocytes: A new method for diagnosis of tropical theileriosis
in crossbred cattle -
Ramesh B. Jagtap, Amit Gupta and Sushama R. Chaphalkar
Veterinary World, 8(12): 1379-1385
doi:
10.14202/vetworld.2015.1379-1385
Ramesh B. Jagtap:
Department of Virology and Immunology, Vidya Pratishthan’s
School of Biotechnology, Baramati, Pune, Maharashtra,
India; docrameshjagtap@gmail.com
Amit Gupta:
Department of Virology and Immunology, Vidya Pratishthan’s
School of Biotechnology, Baramati, Pune, Maharashtra,
India; amitvsbt@gmail.com
Sushama R. Chaphalkar:
Director, Vidya Pratishthan’s School of Biotechnology, Baramati,
Pune, Maharashtra, India;
director.vsbt@gmail.com
Received: 13-06-2015, Revised: 14-10-2015, Accepted: 25-10-2015,
Published online:
11-12-2015
Corresponding author:
Ramesh B. Jagtap, e-mail: docrameshjagtap@gmail.com
Citation:
Jagtap RB, Gupta A, Chaphalkar SR (2015) Flow cytometry based
profiling of leukocytes: A new method for diagnosis of tropical
theileriosis in crossbred cattle,
Veterinary World
8(12):
1379-1385
Abstract
Aim:
In India, dairy industries are important for the livelihood of
small scale farmers and dairy owners. Tropical theileriosis,
mostly affecting dairy cattle and buffaloes is a major threat to
dairy and related industries. Tropical theileriosis is caused by
Theileria annulata,
a hemoprotozoan parasite transmitted by Ixodid ticks of
Hyalomma
spp.
In the present study, we examined the clinical signs,
hematological parameters and flow cytometric profile of whole
blood in 30 theileriosis affected crossbred cattle. The aim of
our study is to analyze, in comparison with clinical and
hematological diagnosis, whether flow cytometry based profiling
of leukocytes could be used as better, quick and alternative
method for diagnosis and screening of bovine tropical
theileriosis in crossbred cattle.
Materials and Methods:
In this study, we screened parasites in 30 peripheral blood
samples from clinical cases of theileriosis by Giemsa’s staining
technique in crossbred cattle. Hematological analysis was done
to estimate hemoglobin (Hb) content, total red blood cell (RBC)
count, total leukocyte count and differential leukocyte count.
Further, flow cytometric analysis of whole blood was carried out
to study leukocytes profile in affected cattle.
Results:
Microscopic examination of stained blood films revealed the
presence of piroplasms in erythrocytes and schizonts in
lymphocytes. Hematological examination revealed significant
(p<0.05) decrease of Hb percent (Hb %), reduced total RBC and
total leukocytes, lymphocytosis, eosinopenia, and neutropenia
compared to that of apparently healthy cattle. Flow cytometric
profiling of leukocytes revealed the severe effect on shape,
size, and granularity of leukocytes, marked decrease in
granulocytes and 3-5 fold increase in lymphocytes count compared
to clinically healthy cattle. Thus, in both methods, namely
conventional and flow cytometric analysis, marked lymphocytosis
and decrease in other blood cell counts were observed compared
to clinically healthy cattle.
Conclusions:
From results, it can be concluded that though conventional
staining techniques and hematology are efficient in diagnosis of
theileriosis, leukocytes profiling based on flow cytometry
combined with clinical examination could be a quick, novel and
alternative method for diagnosis and screening of clinical
tropical theileriosis in crossbred cattle. Thus, there is
potential to offer a flow cytometry based diagnostic service for
tropical theileriosis in crossbred cattle.
Keywords:
clinical examination, flow cytometry, hematology, lymphocytosis,
Theileria annulata.
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