Tick born infections are most common problem in dairy animals of tropical region. This infection causes heavy economical losses in terms reduced milk yield and meet production. Keeping in view the economic importance of tick born infection i.e Theileriosis, present study was organized to resolve the consequences of Theileriosis on naturally infected cattle and buffaloes. The objective of the study was occurrence of disease and correlate exposure of ticks and tick-borne Theileriosis at the peri-urban and urban areas of Hyderabad, Sindh, Pakistan. Data was collected from the selected farmers in study areas through survey questionnaire. Ticks and blood samples of 2400 buffaloes and 300 cows were collected from 20 dairy farms of urban and Peri-urban areas of Hyderabad, during November 2013 to October 2014. Ticks were identified on the basis of their physical characteristics. Thin blood smears were prepared and stained for identification of Theileria spp. DNA was extracted from blood samples of non infected (control) and infected cows and buffaloes. Extracted DNA was amplified by specific genes of Theileria spp. through Polymerase Chain Reaction (PCR) to detect the Theilerial infection in both animal species. Microscopic examination of blood from suspected cows revealed 73 % and 76% prevalance of thielarial infection in urban and peri-urban areas respectively. Buffaloes showed 65% and 75% prevalnce in urban and peri-urban areas respectively. Overall prevlance of thielariosis in cows was higher i.e 75% than buffaloes which is 70%. Cows and bufflaoes in urabn areas showed less prevalance of Theilariosis than peri-urban areas. PCR results given overall infection rate in cows was calculated as 79% that is higher than that in buffaloes which is 76%. Comparative results indicated higher infection rate on PCR than by staining method. Analytical investigations of data revealed significant difference (P<0.05) between prevalance percentage between both species as well as between urban and perii-urban areas of Hyderabad. Identification of ticks indicated that Hyalomma antolicum anatolicum was the highly prevalent tick in both peri-urban and urban areas i.e. 44.32 %, and 44.57% respectively. High tick infestation in cows were in external genitalia, udder and perineum (45%) followed by dewlap (20%), inner thighs (15%).Where as in buffaloes external genitalia, udder and perineum (55%) followed by dewlap (23%). Clinical examination of infected cows indicted anorexia (88%), fever (87%), pale mucus membrane (75%) and enlargement of lymph nodes (47%). Whereas, infected buffaloes had anorexia 95%, fever (91%), pale mucous membrane (80%) and enlarged lymph nodes (49%). Nasal discharge, skin lesions and diarrhea were the minor signs in both species. Analysis of associated risk factors affecting prevalence of tick infestation and Theilerial infection in cows and buffaloes determined the type of house and housing, environmental conditions, gender and age of significantly affects on prevalence of theileriosis. Prevalence was high in semi-intensive house with kacha floor, during months of July to October in adult females. Complete Blood count investigations showed statistically a significant decrease in mean Hgb, PCV, MCH, RBCs, WBCs (P<0.001),whereas significant increase was noted LY count corpuscular volume (P<0.001). It was concluded from the current study that Haylomma anatolicum anatolicum is a vector of Theileria annulata. Load of vector tick and risk of infection is greatly influenced by housing system, climatic conditions. Sensitivity to infection also varies in male, female and also different age groups; adult females are more sensitive to tick infestation as compare to male and young animals. Molecular detection is more sensitive and specific method for detection of Theileria. Theilerial infection influences the blood, causing blood loss anemia resulting in ill health of cows and buffaloes. Chemical analysis of milk of infected cows and buffaloes revealed that protein, ash and lactose were significantly declined (P<0.05) whereas, fat and total solids were significantly increased (P<0.05) in Theileria infected cows and buffaloes, but pH and acidity was unaffected. Financial losses occur in terms of reduced quality and quantity of milk and meat. The total financial fall of RS- 0.17 million / animal was recorded in current study. Theileriosis is one of main causes of financial fall for livestock holders.