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      Safety of transtympanic application of probiotics in a chinchilla animal model

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          Abstract

          Background

          Chronic suppurative otitis media can be recalcitrant and difficult to treat, particularly with the increasing occurrence of antibiotic resistance. Lactobacillus plantarum is a probiotic that has been shown to decrease S. aureus and P. aeruginosa growth in wounds, making it a good candidate for the treatment of chronic suppurative otitis media. However, before it can be applied in the ear, its ototoxicity potential must be evaluated.

          Methods

          A prospective controlled trial was conducted in a chinchilla animal model at the Animal care research facilities of the Montreal Children’s Hospital Research Institute to determine whether Lactobacillus plantarum is ototoxic when applied transtympanically. Ten chinchillas each had one ear randomly assigned to receive 10 9 CFU/mL of Lactobacillus plantarum solution, while the contralateral ear received saline. Auditory brainstem responses were measured bilaterally at 8, 20, 25 kHz before, at 7–10 days after application, and at 28 days after application of probiotic or saline. Facial nerve and vestibular function were assessed clinically.

          Results

          There were no statistically significant differences in hearing thresholds between control and experimental ears at 28 days after application. A difference of 11 dB was noted in the 25 kHz range at day 7–10, but resolved by day 28. No animals receiving probiotics developed vestibular nerve dysfunction. There was no histologic evidence of auditory hair cell damaged evidenced by scanning electron microscopy.

          Conclusion

          Our study suggests that a single application of Lactobacillus plantarum at 10 9 CFU/mL does not cause ototoxicity in a chinchilla animal model. These preliminary safety evaluations and the pathogen inhibitory effects of L. plantarum demonstrated by previous studies present this probiotic as a candidate of interest for further investigation.

          Electronic supplementary material

          The online version of this article (10.1186/s40463-017-0242-y) contains supplementary material, which is available to authorized users.

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          Most cited references28

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          Probiotics: definition, sources, selection, and uses.

          Interest in probiotics is at an all-time high in the United States, driven in part by new products emerging in the market, by US researchers eager to evaluate efficacy claims rigorously, and by consumers interested in potential therapeutic and preventive health benefits. The US marketplace is a mixed bag of products, some well-defined and properly evaluated in controlled clinical studies and others with unsubstantiated claims of efficacy. Validation of probiotic contents in commercial products is needed to ensure consumer confidence. The term "probiotic" should be used only for products that meet the scientific criteria for this term-namely, products that contain an adequate dose of live microbes that have been documented in target-host studies to confer a health benefit. Probiotics must be identified to the level of strain, must be characterized for the specific health target, and must be formulated into products using strains and doses shown to be efficacious. Several characteristics commonly presumed to be essential to probiotics, such as human origin and the ability to improve the balance of the intestinal microbiota, are discussed.
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            Interference of Lactobacillus plantarum with Pseudomonas aeruginosa in vitro and in infected burns: the potential use of probiotics in wound treatment.

            This study evaluated the ability of the probiotic organism Lactobacillus plantarum to inhibit the pathogenic activity of Pseudomonas aeruginosa, both in vitro and in vivo, and investigated the mechanisms involved in such protection. L. plantarum whole cultures, culture filtrates (acid filtrate and neutralised acid filtrate) and isolated, washed cells were tested in vitro for their effects on the production of the P. aeruginosa quorum-sensing signal molecules, acyl-homoserine-lactones (AHLs), and two virulence factors controlled by these signal molecules, elastase and biofilm. All were inhibited by L. plantarum cultures and filtrates, but not by isolated, washed cells. The acid L. plantarum growth medium itself had some inhibitory activity, but the greatest activity was exerted by the whole culture. To test the in-vivo activity of L. plantarum, a burned-mouse model was used in which burns infected with P. aeruginosa were treated with L. plantarum at 3, 4, 5, 7 and 9 days post-infection. Samples from skin, liver and spleen taken after 5, 10 and 15 days demonstrated inhibition of P. aeruginosa colonisation by L. plantarum. There was also an improvement in tissue repair, enhanced phagocytosis of P. aeruginosa by tissue phagocytes, and a decrease in apoptosis at 10 days. These results indicate that L. plantarum and/or its by-products are potential therapeutic agents for the local treatment of P. aeruginosa burn infections.
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              Bacteriotherapy with Lactobacillus plantarum in burns.

              Bacterial colonisation and infection remain the major causes of delayed healing and graft rejection following burns. Topical treatment is necessary to reduce the incidence of burn wound infection. Silver sulphadiazine (SD-Ag) is an often used microbicidal agent. However, this treatment produces adverse reactions and side-effects. On the basis of experimental data and clinical application of lactobacilli as probiotics, we performed this exploratory study to establish the effectiveness of bacteriotherapy with topical application of the innocuous bacteria Lactobacillus plantarum cultured in De Man, Rogosa and Sharpe medium to provide an alternative method for burn treatment using SD-Ag as a reference. These innocuous bacteria would compete with other bacteria that are wound pathogens and would modify the wound environment and promote tissue repair. Eighty burned patients from the Plastic Surgery and Burns Unit were grouped into infected (delayed) second- and third-degree and non infected (early) third-degree burns and treated with L. plantarum or SD-Ag. The proportion of patients with delayed second-degree burns was 0.71 for L. plantarum and 0.73 for SD-Ag (relative rate: -2.72%) with respect to the decrease in bacterial load (<10(5) bacteria/g of tissue), promotion of granulating tissue wound bed and healing. In early third-degree burns, the values were 0.75 for L. plantarum and 0.84 for SD-Ag (relative rate: -1.07%) in preventing wound infection and promotion of granulation tissue, 0.90 in graft taking for both treatments (relative rate: 0%) and 0.75 for L. plantarum and 0.77 for SD-Ag (relative rate: -2.60%) in healing. In delayed third-degree burns, values were 0.83 for L. plantarum and 0.71 for SD-Ag (relative rate: +16.90%) with respect to the decrease in the bacterial load (<10(5) bacteria/g of tissue) and providing a granulating tissue wound bed, 0.90 in graft taking for both treatments (relative rate: 0%) and 0.75 for L. plantarum and 0.64 for SD-Ag (relative rate: + 17.19%) in healing. Although the number of patients (between 12 and 15 per group) did not enable the application of a power statistical test, these results suggest that the L. plantarum treatment should be studied in greater depth and could be used as a valid alternative for the topical treatment of burns.
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                Author and article information

                Contributors
                carol.nhan@gmail.com
                aren.bezdjian@gmail.com
                shyamali.saha2@gmail.com
                satya.prakash@mcgill.ca
                lily.hp.nguyen@gmail.com
                514 412-4400 , sam.daniel@mcgill.ca
                Journal
                J Otolaryngol Head Neck Surg
                J Otolaryngol Head Neck Surg
                Journal of Otolaryngology - Head & Neck Surgery
                BioMed Central (London )
                1916-0208
                1916-0216
                22 November 2017
                22 November 2017
                2017
                : 46
                : 63
                Affiliations
                [1 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Department of Otolaryngology – Head & Neck Surgery and McGill Auditory Sciences Laboratory, , The Montréal Children’s Hospital, McGill University, ; 1001 Boulevard Décarie, Montréal, Québec, H4A 3J1 Canada
                [2 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Department of Experimental Surgery and McGill Auditory Sciences Laboratory, , The Montréal Children’s Hospital, McGill University, ; Montréal, Québec, Canada
                [3 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Departments of Biomedical Engineering, Physiology, and Artificial Cells and Organs Research Center, Biomedical Technology and Cell Therapy Research Laboratory, , McGill University, ; Montréal, Québec, Canada
                [4 ]ISNI 0000 0004 1936 8649, GRID grid.14709.3b, Department of Otolaryngology – Head & Neck Surgery and Center for Medical Education, , McGill University, ; Montreal, Québec, Canada
                Article
                242
                10.1186/s40463-017-0242-y
                5700520
                29166927
                2ec3a388-5a30-45a1-9e97-401a0184cc3a
                © The Author(s). 2017

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 10 April 2017
                : 8 November 2017
                Categories
                Original Research Article
                Custom metadata
                © The Author(s) 2017

                probiotic,otitis,ototoxicity,transtympanic,chinchilla
                probiotic, otitis, ototoxicity, transtympanic, chinchilla

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