The present study describes the frequency of Foot and Mouth Disease (FMD) virus serotypes (O,A and Asia-1) in major regions (all provinces) of Pakistan using Indirect Sandwich ELISA.Also,spatial distribution of various FMD serotypes and their comparison is discussed.A total of 590 samples (Epithelial tissue) have been analyzed during a period of five years (2005-2009).Out of 590 samples,180 were found positive,giving an overall confirmation of FMDV about 33.2 ％.Of the prevalent serotypes,FMDV ‘O’ serotype caused most outbreaks (20.7 ％),followed by serotype A (6.6 ％) and serotype Asia-1 (4.6 ％) while there was no positive case oftype ‘C’.The study clearly showed that the disease was more frequent in the agro-climatic zones than in hilly areas.Based on the data of 590 samples (＞50 outbreaks),the overall prevalence of FMDV in cattle and buffaloes in Pakistan was 33.2 ％,while in cattle alone,it was 37.1％,higher than in buffalo (28.7 ％).There were eight cases of mixed serotypes infection,indicating the presence of endemic state of disease.Another significant feature was the change over time.In phase-I (2005-2007),there was an overall prevalence of 29.4 ％,while the occurrence of the serotype O,A and Asia-1 was 20.4 ％,2.9 ％ and 4.7 ％,respectively.During phase-II (2008-2009),the overall prevalence was 59.21％,while those of serotype O,A and Asia-1 were 22.4 ％,31.6 ％ and 4.0 ％,respectively.This clearly indicated a shift from serotype O to A,which may help to explain the occurrence of more severe outbreaks,despite vaccination.