Background South Africa has a high prevalence of people living with human immunodeficiency virus (HIV; PLWH) who have shown to affect the prevalence and severity of infection and sepsis particularly gallbladder disease. Empirical Antimicrobial (EA) therapy for acute cholecystitis (AC) is based largely on bacteria colonisation of bile (bacteriobilia) and antimicrobial susceptibility patterns (antibiograms) obtained from the developed world where the prevalence of PLWH is very low. In an ever-emerging era of increasing antimicrobial resistance, monitoring and updating local antibiograms is underscored. Objective Due to the paucity of data available locally to guide treatment we found it pertinent to examine gallbladder bile for bacteriobilia and antibiograms in a setting with a high prevalence of PLWH to determine if this may demand a review of our local antimicrobial policies for gallbladder infections for both EA and pre-operative antimicrobial prophylaxis (PAP) for laparoscopic cholecystectomies (LC). Methodology A retrospective observational descriptive study was undertaken at King Edward VIII Hospital, Durban, KwaZulu-Natal, South Africa. Hospital records were reviewed for all patients undergoing cholecystectomy over a 3-year period. Gallbladder bacteriobilia and antibiograms were assessed and compared between PLWH and HIV uninfected (HIV-U). Pre-operative age, ERCP, PCT, CRP and NLR were used as predictors for bacteriobilia. Statistical analyses were performed using R Project and p values of less than 0.05 were considered as statistically significant. Results There were no differences in bacteriobilia or antibiograms between PLWH and HIV-U. There was >30% resistance to amoxicillin/clavulanate and cephalosporins. Aminoglycoside-based therapy, had good susceptibility patterns whilst carbapenem-based therapy demonstrated the lowest resistance levels. ERCP and age were predictors of bacteriobilia (p<0.001 and 0.002 respectively). PCT, CRP and NLR were not. Conclusion PLWH should follow the same PAP and EA recommendations as HIV-U. For EA, we recommend, a combination of amoxicillin/clavulanate with aminoglycoside-based therapy (amikacin or gentamycin) or piperacillin/tazobactam as monotherapy. Carbapenem-based therapy should be reserved for drug resistant species. For PAP, we recommend the routine use in older patients and patients with history of ERCP undergoing LC.