Bovine mastitis is one of the most important diseases in the dairy industry and has detrimental impact on the economy and welfare of the animals. Further, treatment failure results in increased antibiotic use in the dairy industry, as some of these mastitis cases for unknown reasons are not resolved despite standard antibiotic treatment. Chronic biofilm infections are notoriously known to be difficult to eradicate with antibiotics and biofilm formation could be a possible explanation for mastitis cases that are not resolved by standard treatment. This paper reviews the current literature on biofilm in bovine mastitis research to evaluate the status and methods used in the literature. Focus of the current research has been on isolates from milk samples and investigation of their biofilm forming properties in vitro. However, in vitro observations of biofilm formation are not easily comparable with the in vivo situation inside the udder. Only two papers investigate the location and distribution of bacterial biofilms inside udders of dairy cows with mastitis. Based on the current knowledge, the role of biofilm in bovine mastitis is still unclear and more in vivo investigations are needed to uncover the actual role of biofilm formation in the pathogenesis of bovine mastitis.

In chronic wounds in humans, biofilm formation and wound chronicity are linked, as biofilms contribute to chronic inflammation and delayed healing. Biofilms are aggregates of bacteria, and living as biofilms is the default mode of bacterial life; within these aggregates, the bacteria are protected from both antimicrobial substances and the immune response of the host. In horses, delayed healing is more commonly seen in limb wounds than body wounds. Chronic inflammation and hypoxia are the main characteristics of delayed wound healing in equine limbs, and biofilms might also contribute to this healing pattern in horses. However, biofilm formation in equine wounds has been studied to a very limited degree. Biofilms have been detected in equine traumatic wounds, and recent experimental models have shown that biofilms protract the healing of equine limb wounds. Detection of biofilms within wounds necessitates advanced techniques that are not available in routine diagnostic yet. However, infections with biofilm should be suspected in equine limb wounds not healing as expected, as they are in human wounds. Treatment should be based on repeated debridement and application of topical antimicrobial therapy.

Objective: Biofilm infections in chronic wounds are common and pose a significant clinical challenge. This challenge was addressed by developing the SoftOx Biofilm Eradicator (SBE) composed of hypochlorous acid (HOCl) and acetic acid with strong broad-spectrum antimicrobial activity. Approach: First-in-human study investigating the safety and tolerability as primary endpoints and wound size effect and antimicrobial efficacy as secondary endpoints of SBE treatment in chronic leg wound patients. The study was divided into two as follows: a randomized, double-blinded, Single Ascending Dose (SAD) phase ( n = 16 SBE; n = 4 placebo), where patients were treated with SBE or saline (placebo) only once, followed by an open-label, Multiple Ascending Dose (MAD) phase ( n = 8), where patients were treated with SBE once daily or twice daily over five days. Reporting is according to CONSORT guidelines. Results: SBE was safe and well-tolerated in chronic leg wound patients. There were no significant differences in pain during and after treatment with SBE or the placebo. The SBE treatment reduced bioburden in wounds compared to baseline, with 98% and 49% median reduction after SBE or placebo treatment, respectively. A dose-dependent trend in absolute wound size reduction was observed in the MAD groups with a median (min, max) change of −2.99 (−14.25, −1.5) cm ² in the once-daily and −10.48 (−17.95, −0.38) cm ² in the twice-daily group, respectively. Innovation and Conclusion: This study demonstrated the safe use of HOCl-based SBE in chronic leg wounds with promising trends of immediate antimicrobial action and beneficial effect on wound healing.