This document presents a proposed study on the molecular diagnosis of babesiosis in cattle in southern Khyber Pakhtunkhwa, Pakistan. Babesiosis is caused by protozoan parasites that infect cattle and other animals, causing economic losses. The study aims to 1) detect babesia species in cattle early in infection and 2) develop a tool to investigate the epidemiology and control the disease. Blood samples will be collected from clinically infected cattle and examined microscopically and through PCR to identify babesia species. Data will be statistically analyzed to understand the epidemiology of babesiosis in the region. The study seeks to better understand and control an important disease affecting the cattle industry in southern Khyber Pak

1. COMPARATIVE STUDIES ON MOLECULER DIAGNOSIS OF
BABESIOSIS IN CATTLES OF SOUTHERN PART OF KHYBER
PAKHTUNKHWA
By
Hamid Ullah
(Reg. No. …………)
Supervisor -I Department of Zoology ------------
KUST, Kohat Signature
Supervisor –II Department of Microbiology ------------
KUST, Kohat Signature
Chairman Dr. Shahid Niaz Khan Department of Zoology ------------
KUST, Kohat Signature
Department of Zoology
Kohat University of Science & Technology, Kohat-2600
Khyber Pakhtunkhwa, Pakistan
(2019)

2. Introduction
Babesiosis is caused by intraerythrocytic protozoan parasites of the genus Babesia that infect a
wide range of domestic and wild animals and occasionally humans. Babesioses are characterized
by high fever, ataxia, anorexia, general circulatory shock, and sometimes also nervous signs as a
result of sequestration of infected erythrocytes in cerebral capillaries. Anaemia and
haemoglobinuria may appear later in the course of the disease [1, 2]. It is an acute hemolytic
feverous disease which is most common in domesticated animals and is transmitted by hard
tick’s bite. This disease is more Prevalence in tropical and subtropical regions and it cause severe
loss of sensitive animals. Mortality of this disease in absence of treatment in case of Babesia
bigemina is about 30% in cows and in case of Babesia bovis is about 70-80% in sheep [3]. The
greatest infection rate is seen in animals at 6-12 months of age. The average age at which calves
in enzootic areas become infected is 11 weeks, but at this early age clinical signs and
pathological changes are mild and short lived [4].
The disease has the potential to cause large economic and sanitary disruptions compromising the
livestock industry all over the world [5]. In several countries of the world including Asia,
Australia, Africa, South and Central America and United States the occurrence of Babesiosis in
cattle is about 1.2 billion [6, 7]
In Pakistan, the highest incidence of babesiosis was reported during the months of June, July,
August, September and November. These may also be considered as the tick active months [8].
In Pakistan its prevalence is so endemic in 2008, 5.5- 42.8% cattle’s was infected by Babesia
species in Pakistan [9]. Keeping in view, the importance of the disease due to economic losses to
the animal’s husbandry the present work is designed to study the epidemiology of Babesiosis in
cattle’s in Southern parts of Khyber Pakhtunkhwa.
Hypothesis
Babesia species may affect domestic cattle’s in Southern Khyber Pakhtunkhwa causing
economic losses to the dairy industries.
Objectives
1. To detect babesia species in cattle in the early phase of infection.
2. To develop a tool for epidemiological investigation and subsequent disease eradication.

3. Materials and Methods
Study location/area: This study will be conducted in the southern parts of Khyber Pakhtunkhwa
(KPK), Pakistan.
Sample collection: Blood samples will be collected from clinically suspected cattle`s jugular
vein in sterilized vacationer and will be kept in ice jar at -20°
C for further analysis.
Microscopic study & PCR: Slides will be prepared and stained with Giemsa stain according to
the prescribed protocol. Erythrocytes on the blood films will be examined by microscope for
intracellular piroplasms of the Babesia species. DNA from the samples will be separated and
polymerase chain reaction will be conducted.
Statistical Analysis: Data will be statistically analyzed through ANOVA and chi-square
test using SPSS software.
Time Frame
S. No Research Component Time Required
1 Literature’s review Two months
2 Blood samples collection and experimental work Five months
3 Data Interpretation Two months
4 Thesis writing Three months

4. References
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a bovine blood parasite of veterinary and zoonotic importance”, Clin Microbiol Rev, 4,
636.
[2] H. S. Hossini (2003), “Parasitic Infection of domesticated animals”, Tehran university
Press, Tehran, Iran, 65.
[3] J. Mosqueda, A. O. Ramirez, G. A. Tipacamu, G. J. Canto (2012), “Current advances in
detection and treatment of babesiosis”, Curr Med Chem, 10, 1518.
[4] O. Radostitis, M. C. Blood, C. C. Gay, K. W. Hinchcliff (2000), “Veterin ary Medicine”,
ed. 9th
, London: W. B. Saunders, 1296.
[5] R. A. Spickler (2008), “Emerging and Exotic Diseases of Animal”, ed. 3rd
, Ames, Iowa:
CFSPH Iowa State University, 135.
[6] C. Kim, H. Iseki, M. S. Herbas, N. Yokoyama, H. Suzuki, X. Xuan et al. (2007),
“Development of Taqman-Based Real-Time PCR Assays for Diagnostic Detection of
Babesia bovis and Babesia bigemina”, Am. J. Trop. Med. Hyg, 5, 841.
[7] S. Zulfiqar, S. Shahnawaz, M. Ali, A. M. Bhutta, S. Iqbal, S. Hayat et al. (2012),
“Detection of Babesia bovis in blood samples and its effect on the hematological and
serum biochemical profile in large ruminants from Southern Punjab”, .Asian Pacific
Journal of Tropical Biomedicine, 104-108.
[8] S. Gosh, G. C. Bansal, S. C. Gupta, D. Ray, M. Q. Khan, H. Irshad et al. (2007), “Status
of tick distribution in Bangladesh, India and Pakistan”, Parasitol Res, 101, 207-216.
[9] B. D. Perry, T. F. Randolph, J. J. McDermott, P. K. Thornton (2002), “Investing in
animal health research to alleviate poverty”, International Livestock Research Institute
(ILRI), Nairobi, Kenya.