Topical Delivery of Lactobacillus Culture Supernatant Increases Survival and Wound Resolution in Traumatic Acinetobacter
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Topical Delivery of Lactobacillus Culture Supernatant Increases Survival and Wound Resolution in Traumatic Acinetobacter baumannii Infections Josh Stanbro 1 & Ju Me Park 1 & Matthew Bond 1 & Michael G. Stockelman 1 & Mark P. Simons 1 & Chase Watters 1
# This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2019
Abstract Species of Lactobacillus have been proposed as potential candidates for treating wound infections due to their ability to lower pH, decrease inflammation, and release antimicrobial compounds. This study investigated the impact of lactobacilli (Lactobacillus acidophilus ATCC 4356, Lactobacillus casei ATCC 393, Lactobacillus reuteri ATCC 23272) secreted products on wound pathogens in vitro and in a murine wound infection model. Evaluation of 1–5 day lactobacilli conditioned media (CM) revealed maximal inhibition against wound pathogens using the 5-day CM. The minimum inhibitory concentration (MIC) of 5-day Lactobacillus CMs was tested by diluting CM in Mueller-Hinton (MH) broth from 0 to 25% and was found to be 12.5% for A. baumannii. Concentrating the CM to 10× with a 3 kDa centrifuge filter decreased the CM MIC to 6.25–12.5% for A. baumannii planktonic cells. Minimal impact of 5-day CMs was observed against bacterial biofilms. No toxicity was observed when these Lactobacillus CMs were injected into Galleria melonella waxworms. For the murine A. baumannii wound infection studies, improved survival was observed following topical treatment with L. acidophilus ATCC 4356 or L. reuteri ATCC 23272, while L. reuteri ATCC 23272 treatment alone improved wound resolution. Overall, this study suggests that the topical application of certain Lactobacillus species byproducts could be effective against gram-negative multi-drug resistant (MDR) wound pathogens, such as A. baumannii. Keywords Lactobacillus . Acinetobacter baumannii . Wound infections . Lactic acid bacteria
Introduction Battlefield-associated wounds are often traumatic and heavily contaminated with foreign soil bacteria, and thus prone to develop debilitating bacterial infections. Multi-drug resistant (MDR) pathogens are one of the leading causes of infections in US-deployed military personnel [1]. The annual cost of MDR infections ranks in the billions of dollars in the USA, and is estimated to be $15–36 million for the US Army alone [2]. In recent surveys of combat-injured personnel, the isolated causes of wound infections in 46–51% of the patients were Staphylococcus aureus, Acinetobacter baumannii, Electronic supplementary material The online version of this article (https://doi.org/10.1007/s12602-019-09603-z) contains supplementary material, which is available to authorized users. * Chase Watters [email protected] 1
Wound Infections Department, Naval Medical Research Center, 503 Robert Grant Avenue, Silver Spring, MD 20910, USA
Enterococcus faecium, and Klebsiella spp. [3, 4]. In addition to carrying genes that confer antibiotic resistance, MDR pathogens commonly exhibit
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