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Topical Delivery of Lactobacillus Culture Supernatant Increases Survival and Wound Resolution in Traumatic Acinetobacter baumannii Infections

  • Josh Stanbro
  • Ju Me Park
  • Matthew Bond
  • Michael G. Stockelman
  • Mark P. Simons
  • Chase WattersEmail author
Article

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 

Notes

Acknowledgments

Special thanks to the Army Wound Infections department at WRAIR for their support and assistance with the animal studies, specifically Yonas Alamneh, Rania Abu-Taleb, Jonathan Shearer, Samandra Demons, Anna Jacobs, Kathleen Umayam, Natalie-Makenna Gingras, Daniel Zurawski, and Yuanzheng Si.

Funding Information

The support for this work was provided by the Office of Naval Research under work unit number A1601.

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no conflicts of interest.

Ethical Approval

All procedures were performed in accordance with protocols approved by the Walter Reed Army Institute of Research (WRAIR)/Naval Medical Research Center (NMRC) Institutional Animal Care and Use Committee in compliance with all applicable Federal regulations governing the protection of animals in research.

Disclaimers

The views expressed are those of the authors and do not necessarily reflect the official policy or position of the Department of the Navy, Army, Department of Defense, nor the US Government. Some authors are service members of the US Government. I am a military service member or federal/contracted employee of the US government. This work was prepared as part of my official duties. Title 17 U.S.C. 105 provides that “copyright protection under this title is not available for any work of the United States Government.” Title 17 U.S.C. 101 defines a US Government work as work prepared by a military service member or employee of the US Government as part of that person’s official duties.

Supplementary material

12602_2019_9603_MOESM1_ESM.docx (192 kb)
Fig. S1 (DOCX 49 kb)

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Copyright information

© This is a U.S. government work and not under copyright protection in the U.S.; foreign copyright protection may apply 2019

Authors and Affiliations

  1. 1.Wound Infections DepartmentNaval Medical Research CenterSilver SpringUSA

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