Bacteriocin‐producing bacteria in aquaculture may prevent spreading of potentially pathogenic microorganisms, and could be used as an alternative to the empirical use of antimicrobial drugs, especially for prophylaxis. Bacteriocinogenic bacteria inoculated as probiotics should not carry undesirable traits, such as antibiotic resistance. This study aimed to find potentially bacteriocinogenic bacteria in an aquaculture system and evaluate their antimicrobial susceptibility patterns. Selective cultures for enterobacteria, non‐fermenting Gram‐negative rods and Gram‐positive cocci were obtained from water samples before bacterial isolation and biochemical identification. Overall, 160 representative strains were recovered and for 57 antagonism was observed against selected strains such as Staphylococcus aureus, with antagonism being expressed better on Brain Heart Infusion medium. After exclusion of interfering factors, bacteriocin or bacteriocin‐like substances were suggested to be related to the antagonism observed. Higher drug‐resistant rates were observed among potentially bacteriocinogenic bacteria for different antimicrobials of clinical relevance. Although antibiotic resistance is a global health problem and bacteriocins are attractive alternatives to classical antibiotic even to multiresistant bacteria, the data obtained suggest that bacteriocin‐producing bacteria may harbour resistance genes available for transference in different environments. From the ecological and biotechnological perspective, antimicrobial susceptibility tests must be always performed when prospecting potentially bacteriocinogenic bacteria as probiotic candidates in the environment.

Techniques to decrease losses from bacterial diseases are always important to improve the fish production. The use of antagonistic substances (bacteriocins) has been proven to be a viable option. The aim of this study was to evaluate different methods of purification for bacteriocin like inhibitory substances (BLIS). For the purification process, we isolated and used two Gram-positive bacilli that produce antagonistic substances for pathogens in aquaculture. Tests for detection of interfering factors were also performed. After the confirmation that the antagonistic action was due the BLIS activity, we carried out the purification methods. The methods tested were: cell free supernatant, acid extraction and ammonium sulfate precipitation at two concentrations (20 and 50%). Salmonella Tiphy CFP/IAL1472 and Aeromonas hydrophila (isolated in a tilapia production environment) were used as indicators of the efficiency of extracts in controlling pathogenic potentials. Ammonium sulfate precipitation at 50% was the most appropriate for purifying the antagonistic substance for both indicators. The extracts of the two isolates remained active for 22 days at 25ºC. These are promising results regarding the water and fish health without the use of antibiotics, in this manner being a safer environmental practice.