Background: Information is lacking on the prevalence and susceptibility patterns of bacterial isolates in dogs and cats with suspected hepatobiliary disease.Objectives: To characterize the prevalence, identity, and antimicrobial susceptibility of common hepatobiliary isolates from such patients.Animals: Dogs and cats presented to the University of Wisconsin-Madison Veterinary Medical Teaching Hospital for which samples of bile, gallbladder, or liver were submitted for culture from 1998 to 2003, including 190 dogs (192 culture episodes) and 58 cats (61 culture episodes).Methods: Cases were identified from the microbiology laboratory database. Data from patient medical records were extracted, including the history of antimicrobial administration, the presence of fever, the results of CBC and serum biochemistry, the presence of biliary obstruction or hepatobiliary inflammation, and the results of aerobic and anaerobic bacterial cultures and aerobic antimicrobial susceptibilities.Results: Biliary cultures yielded a significantly higher percentage of positive results overall (30% [18 of 60]) than did hepatic cultures (7% [15 of 215]). In patients with cholecystitis, 62% (8 of 13) had positive biliary cultures. In patients with hepatic inflammation, 23% (7 of 30) had positive bile cultures, whereas only 6% (6 of 103) had positive hepatic cultures. Escherichia coli, Enterococcus spp., Bacteroides spp., Streptococcus spp., and Clostridium spp. were the most common true-positive isolates. More than 80% of Enterobacteriaceae were susceptible to ciprofloxacin or aminoglycosides, with only 30-67% susceptible to first-generation aminopenicillins and cephalosporins. Liver samples obtained by surgery or laparoscopy were more likely to yield positive cultures than those obtained by percutaneous needle biopsy.