title: Flow cytometry based profiling of leukocytes: A new method for diagnosis of tropical theileriosis in crossbred cattle.
abstract: OBJECTIVE
  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.
  
  
  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.
authors.affiliation: Department of Virology and Immunology, Vidya Pratishthan's School of Biotechnology, Baramati, Pune, Maharashtra, India.
authors.forename: Ramesh B
authors.initials: RB
authors.lastname: Jagtap
authors.affiliation: Department of Virology and Immunology, Vidya Pratishthan's School of Biotechnology, Baramati, Pune, Maharashtra, India.
authors.forename: Amit
authors.initials: A
authors.lastname: Gupta
authors.affiliation: Director, Vidya Pratishthan's School of Biotechnology, Baramati, Pune, Maharashtra, India.
authors.forename: Sushama R
authors.initials: SR
authors.lastname: Chaphalkar
keywords: Theileria annulata; clinical examination; flow cytometry; hematology; lymphocytosis