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Bovine anaplasmosis (BA) is tick-borne disease of cattle caused by Anaplasma marginale and it remains an economically important disease in the United States (U.S.). We have anecdotal information that Veterinary Feed Directive prescriptions in Kentucky (KY) are written most often for treatment and prevention of BA. However, there are no recent prevalence estimates of this disease in KY. Thus, this study was aimed at determining the seroprevalence of and factors associated with BA in KY. Data were obtained from an active slaughter survey (n = 232) performed between May and July 2013 as well as from reviewing The University of Kentucky Veterinary Diagnostic Laboratory (UKVDL) records of specimens submitted for BA testing from 2002-2012 (n = 2,573). With competitive ELISA, the apparent prevalence of BA in KY was 10.78% (95% CI: 7.41-15.42%) and 11.58% (95% CI: 10.31-12.98%) for the slaughter survey and laboratory records, respectively. Whereas the estimated true prevalence was 9.44% (95% CI: 5.65-14.48%) and 10.3% (95% CI: 8.92-11.8%), respectively. From the laboratory records, factors associated with positive BA results were age, breed, whether specimens were submitted singularly or as a group, year and quarter of the year the specimens were submitted. The odds of the outcome were 5 times as high when cattle were adults (vs juvenile) and almost 4 times as high when specimens were submitted singularly (vs group). In comparison to Holstein breed, the odds of the outcome were 3.5 and 2.5 times higher in Angus and mixed breeds, respectively. The odds of a diagnosis of BA varied in an undulating pattern by year of sample submission. When compared to 2011, the odds of a diagnosis of BA was approximately 3 times as high in 2005, 2008, and 2009 and approximately 5 times as high in 2004, 2006, and 2012. In comparison to the duration from January to March, the odds of the outcome were almost 20 times as high from July to September but 10 times as high from October to December durations. Counties with specimen submissions for BA testing had a significantly greater cattle population and number of cattle farms than counties without specimen submissions. Future prevention and control measures for BA should target these factors and should be weighted more on counties with higher cattle population. Furthermore, current records from the UKVDL appear sufficient for the surveillance of BA in KY., (Published by Elsevier B.V.)