| 1. |
- Butler, Éile, et al.
(författare)
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A pilot study investigating lactic acid bacterial symbionts from the honeybee in inhibiting human chronic wound pathogens
- 2016
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Ingår i: International Wound Journal. - Oxford : Blackwell Publishing. - 1742-4801 .- 1742-481X. ; 13:5, s. 729-37
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Tidskriftsartikel (refereegranskat)abstract
- Treatment and management of chronic wounds is a large burden on the health sector and causes substantial suffering for the patients. We believe that 13 lactic acid bacteria (LAB) symbionts isolated from the honey crop of the honeybee are important players in the antimicrobial action of honey, by producing antimicrobial substances and can be used in combination with heather honey as an effective treatment in wound management. A total of 22 patients with chronic ulcers were included; culture-dependent and molecular-based (MALDI-MS and 16S rRNA gene sequencing) techniques were used to identify bacteria from chronic wounds. These clinical isolates were used for in vitro antimicrobial testing with standardised viable LAB and sterilised heather honey mixture. Twenty of the patients' wounds were polymicrobial and 42 different species were isolated. Patient isolates that were tested in vitro were inhibited by the LAB and honey combination with inhibitory zones comparable with different antibiotics. LAB and heather honey in combination presents a new topical option in chronic wound management because of the healing properties of honey, antimicrobial metabolite production from the LAB and their bactericidal effect on common chronic wound pathogens. This new treatment may be a stepping stone towards an alternative solution to antibiotics.
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| 2. |
- Butler, Éile
(författare)
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Lactic acid bacterial symbionts from the honeybee: Characteristics and applications in wound management
- 2015
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Antibiotic resistance is now a large threat with worldwide agreement that we will fall into a post-antibiotic era unless there is suitable treatment alternatives discovered. In recent years, chronic wounds have become a major burden on society and treatment is complicated. Honey has been widely used as a topical treatment for wounds yet there is much discourse between studies examining its effect and still some of its antimicrobial effect has not been characterized. In this thesis we investigate some characteristics of 13 novel Lactic acid bacterial (LAB) symbionts originating from the honey stomach of the western honeybee. The LABs involvement in honey production and their antimicrobial action against many environmental bacteria and yeasts hypothesizes that these LAB symbionts may be a good alternative to antibiotics for the topical treatment of chronic wounds. First we investigated the effect of stress on the extracellular protein production of all 13 LAB separately. We revealed that this extracellular production varied between species and genera, and that different microbial stressors had varying effects. These putative proteins may have links with the LAB survival in their niche possibly in attachment or as antimicrobials. Secondly, we investigated some characteristics of the LAB and discovered a link between the therapeutic effect of honey and these symbionts. We identified a myriad of bioactive substances that the LAB produce and had broad-spectrum antimicrobial activity against human pathogens including antibiotic resistant species. The effect varied between species, but all 13 LAB combined were the most effective. Finally we investigated the antimicrobial effect the LAB symbionts had together in a standardized concentration with heather honey, in vitro against human and animal pathogens, and in vivo in two pilot studies. We showed once again the LAB formulation had broad-spectrum activity when combined. When applied to chronic wounds on horses and finally on humans, we saw in both cases remarkable results towards wound healing. Bacterial diversity was also investigated and we observed that the wounds were polymicrobial in nature and bacterial diversity varied between subjects, but there were a number of genera that are readily identified throughout the majority of samples. This thesis studies the hypothesis of a novel LAB microbiota as an alternative tool in wound management and provides knowledge about these symbionts’ therapeutic and antimicrobial characteristics in vitro and in vivo. It contributes to an understanding of how interdisciplinary research can proceed starting from basic knowledge to medical applications. In conclusion, further work, including more in vitro experiments and controlled clinical trials, could in a future perspective establish these LAB symbionts as an alternative to antibiotics in wound treatment, and possibly other infections.
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| 3. |
- Butler, Eile, et al.
(författare)
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Lactobacillus reuteri DSM 17938 as a Novel Topical Cosmetic Ingredient : A Proof of Concept Clinical Study in Adults with Atopic Dermatitis
- 2020
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Ingår i: Microorganisms. - : MDPI. - 2076-2607. ; 8:7
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Tidskriftsartikel (refereegranskat)abstract
- Atopic Dermatitis (AD) is a chronically relapsing skin condition characterized by dry, itchy, and inflamed skin where sufferers can frequently be subject to infections. Probiotics are known to be potent immune-modulators, and liveLactobacillus reuteriDSM 17938 has shown to be anti-inflammatory but also to possess antimicrobial and barrier function properties. This study aimed to investigate and compare two investigational ointment products (topical probiotic and control) for cutaneous acceptability, safety, and efficacy under normal conditions of use, in adult subjects with atopic dermatitis. The products were applied twice daily for 8 weeks, and cutaneous acceptability, SCORAD index, local SCORAD, and adverse events were evaluated after 4 and 8 weeks of treatment. At the end of the observations, it was demonstrated that both the probiotic-containing and probiotic-free ointments were both cutaneously acceptable and safe. It importantly showed a statistically and clinically significant improvement of the SCORAD index and local SCORAD in adult subjects with AD after 4 and 8 weeks of continuous use. In conclusion, we show evidence that the probiotic product, containing liveL. reuteriDSM 17938 as an extra ingredient, is safe and promising as a novel topical cosmetic ointment and with further testing could be a standard topical product for the management of atopic dermatitis or other disorders associated with the skin.
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| 4. |
- Butler, Éile, et al.
(författare)
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Proteins of novel lactic acid bacteria from Apis mellifera mellifera: an insight into the production of known extra-cellular proteins during microbial stress
- 2013
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Ingår i: BMC Microbiology. - : Springer Science and Business Media LLC. - 1471-2180. ; 13
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Tidskriftsartikel (refereegranskat)abstract
- Background: Lactic acid bacteria (LAB) has been considered a beneficial bacterial group, found as part of the microbiota of diverse hosts, including humans and various animals. However, the mechanisms of how hosts and LAB interact are still poorly understood. Previous work demonstrates that 13 species of Lactobacillus and Bifidobacterium from the honey crop in bees function symbiotically with the honeybee. They protect each other, their hosts, and the surrounding environment against severe bee pathogens, bacteria, and yeasts. Therefore, we hypothesized that these LAB under stress, i.e. in their natural niche in the honey crop, are likely to produce bioactive substances with antimicrobial activity. Results: The genomic analysis of the LAB demonstrated varying genome sizes ranging from 1.5 to 2.2 mega-base pairs (Mbps) which points out a clear difference within the protein gene content, as well as specialized functions in the honeybee microbiota and their adaptation to their host. We demonstrate a clear variation between the secreted proteins of the symbiotic LAB when subjected to microbial stressors. We have identified that 10 of the 13 LAB produced extra-cellular proteins of known or unknown function in which some are arranged in interesting putative operons that may be involved in antimicrobial action, host interaction, or biofilm formation. The most common known extra-cellular proteins secreted were enzymes, DNA chaperones, S-layer proteins, bacteriocins, and lysozymes. A new bacteriocin may have been identified in one of the LAB symbionts while many proteins with unknown functions were produced which must be investigated further. Conclusions: The 13 LAB symbionts likely play different roles in their natural environment defending their niche and their host and participating in the honeybee's food production. These roles are partly played through producing extracellular proteins on exposure to microbial stressors widely found in natural occurring flowers. Many of these secreted proteins may have a putative antimicrobial function. In the future, understanding these processes in this complicated environment may lead to novel applications of honey crop LAB proteins.
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| 5. |
- Hernández, Aura Rocio, et al.
(författare)
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New concepts for transdermal delivery of oxygen based on catalase biochemical reactions studied by oxygen electrode amperometry
- 2019
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Ingår i: Journal of Controlled Release. - : Elsevier. - 0168-3659 .- 1873-4995. ; 306, s. 121-129
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Tidskriftsartikel (refereegranskat)abstract
- The development of formulation concepts for improved skin tissue oxygenation, including methods for measuring oxygen (O) transport across biological barriers, are important research topics with respect to all processes that are affected by the O concentration, such as radiation therapy in oncology treatments, wound healing, and the general health status of skin. In this work we approach this topic by a novel strategy based on the antioxidative enzyme catalase, which is naturally present in the skin organ where it enables conversion of the reactive oxygen species hydrogen peroxide (HO) into O. We introduce various applications of the skin covered oxygen electrode (SCOE) as an in-vitro tool for studies of catalase activity and function. The SCOE is constructed by placing an excised skin membrane directly on an O electrode and the methodology is based on measurements of the electrical current generated by reduction of O as a function of time (i.e. chronoamperometry). The results confirm that a high amount of native catalase is present in the skin organ, even in the outermost stratum corneum (SC) barrier, and we conclude that excised pig skin (irrespective of freeze-thaw treatment) represents a valid model for ex vivo human skin for studying catalase function by the SCOE setup. The activity of native catalase in skin is sufficient to generate considerable amounts of O by conversion from HO and proof-of-concept is presented for catalase-based transdermal O delivery from topical formulations containing HO. In addition, we show that this concept can be further improved by topical application of external catalase on the skin surface, which enables transdermal O delivery from 50 times lower concentrations of HO. These important results are promising for development of novel topical or transdermal formulations containing low and safe concentrations of HO for skin tissue oxygenation. Further, our results indicate that the O production by catalase, derived from topically applied S. epidermidis (a simple model for skin microbiota) is relatively low as compared to the O produced by the catalase naturally present in skin. Still, the catalase activity derived from S. epidermidis is measurable. Taken together, this work illustrates the benefits and versatility of the SCOE as an in vitro skin research tool and introduces new and promising strategies for transdermal oxygen delivery, with simultaneous detoxification of HO, based on native or topically applied catalase.
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| 6. |
- Khmaladze, Ia, et al.
(författare)
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Lactobacillus reuteri DSM 17938 : A comparative study on the effect of probiotics and lysates on human skin.
- 2019
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Ingår i: Experimental dermatology. - : John Wiley & Sons. - 0906-6705 .- 1600-0625. ; 28:7, s. 822-828
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Tidskriftsartikel (refereegranskat)abstract
- Human skin microbiota might play an important role in maintaining skin health and potentially prevent premature skin ageing. The use of probiotics in therapeutic skin applications is an attractive idea, as it could offer an alternative option for certain inflammatory skin disorders and dry or sensitive skin. Here, we investigated for the first time, a comparative study of live and the lysate products of probiotic strain Lactobacillus reuteri DSM 17938 in skin topical applications using ex vivo skin models focusing on anti-inflammatory and skin barrier function and in vitro assays for antimicrobial activity. Our results in ultraviolet B radiation (UVB-R)-induced inflammation model demonstrated that both live bacteria and the lysate of L. reuteri DSM 17938 reduced proinflammatory IL-6 and IL-8, illustrated in both reconstructed human epidermis (RHE) and native skin models. Live L reuteri DSM 17938 significantly increased aquaporin 3 (AQP3) gene expression, while the lysate enhanced laminin A/B levels in a healthy (unstimulated) state of RHE, suggesting a positive impact on skin barrier. In addition, live L. reuteri DSM 17938 had antimicrobial action against pathogenic skin bacteria (Staphylococcus aureus, Streptococcus pyogenes M1, Cutibacterium acnes AS12, Pseudomonas aeruginosa), whereas the lysate did not have such an effect. Therefore, it is hypothesized that L. reuteri DSM 17938 could be beneficial for general skin health, to avoid the UVB-R-mediated inflammatory cascade and/or prevent photoageing, improve barrier function or in the management of unhealthy skin prone to inflammatory conditions due to its antimicrobial, anti-inflammatory and skin barrier enhancing functions.
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| 7. |
- Olofsson, Tobias C., et al.
(författare)
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Fighting Off Wound Pathogens in Horses with Honeybee Lactic Acid Bacteria.
- 2016
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Ingår i: Current Microbiology. - : Springer Science and Business Media LLC. - 0343-8651 .- 1432-0991. ; 73:4, s. 463-73
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Tidskriftsartikel (refereegranskat)abstract
- In the global perspective of antibiotic resistance, it is urgent to find potent topical antibiotics for the use in human and animal infection. Healing of equine wounds, particularly in the limbs, is difficult due to hydrostatic factors and exposure to environmental contaminants, which can lead to heavy bio-burden/biofilm formation and sometimes to infection. Therefore, antibiotics are often prescribed. Recent studies have shown that honeybee-specific lactic acid bacteria (LAB), involved in honey production, and inhibit human wound pathogens. The aim of this pilot study was to investigate the effects on the healing of hard-to-heal equine wounds after treatment with these LAB symbionts viable in a heather honey formulation. For this, we included ten horses with wound duration of >1 year, investigated the wound microbiota, and treated wounds with the novel honeybee LAB formulation. We identified the microbiota using MALDI-TOF mass spectrometry and DNA sequencing. In addition, the antimicrobial properties of the honeybee LAB formulation were tested against all wound isolates in vitro. Our results indicate a diverse wound microbiota including fifty-three bacterial species that showed 90 % colonization by at least one species of Staphylococcus. Treatment with the formulation promoted wound healing in all cases already after the first application and the wounds were either completely healed (n = 3) in less than 20 days or healing was in progress. Furthermore, the honeybee LAB formulation inhibited all pathogens when tested in vitro. Consequently, this new treatment option presents as a powerful candidate for the topical treatment of hard-to-heal wounds in horses.
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| 8. |
- Olofsson, Tobias, et al.
(författare)
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Lactic acid bacterial symbionts in honeybees - an unknown key to honey's antimicrobial and therapeutic activities
- 2016
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Ingår i: International Wound Journal. - : Wiley. - 1742-4801 .- 1742-481X. ; 13:5, s. 668-79
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Tidskriftsartikel (refereegranskat)abstract
- Could honeybees' most valuable contribution to mankind besides pollination services be alternative tools against infections? Today, due to the emerging antibiotic-resistant pathogens, we are facing a new era of searching for alternative tools against infections. Natural products such as honey have been applied against human's infections for millennia without sufficient scientific evidence. A unique lactic acid bacterial (LAB) microbiota was discovered by us, which is in symbiosis with honeybees and present in large amounts in fresh honey across the world. This work investigates if the LAB symbionts are the source to the unknown factors contributing to honey's properties. Hence, we tested the LAB against severe wound pathogens such as methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas aeruginosa and vancomycin-resistant Enterococcus (VRE) among others. We demonstrate a strong antimicrobial activity from each symbiont and a synergistic effect, which counteracted all the tested pathogens. The mechanisms of action are partly shown by elucidating the production of active compounds such as proteins, fatty acids, anaesthetics, organic acids, volatiles and hydrogen peroxide. We show that the symbionts produce a myriad of active compounds that remain in variable amounts in mature honey. Further studies are now required to investigate if these symbionts have a potential in clinical applications as alternative tools against topical human and animal infections.
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| 9. |
- Olofsson, Tobias, et al.
(författare)
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Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov., and Lactobacillus kullabergensis sp. nov., isolated from the honey stomach of the honeybee Apis mellifera.
- 2014
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Ingår i: International Journal of Systematic and Evolutionary Microbiology. - : Microbiology Society. - 1466-5026 .- 1466-5034. ; 64, s. 3109-3119
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Tidskriftsartikel (refereegranskat)abstract
- We discovered a symbiotic lactic acid bacterial (LAB) microbiota in the honey stomach of the honeybee Apis mellifera. The microbiota was composed of several phylotypes of Bifidobacterium and Lactobacillus. 16S ribosomal ribonucleic acid (rRNA) gene analyses and phenotypic and genetic characteristics revealed that the Lactobacillus phylotypes isolated represent seven novel species. One is grouped with Lactobacillus kunkeei and the others belong to the Lactobacillus buchneri and Lactobacillus delbrueckii subgroups of Lactobacillus. We propose the names Lactobacillus apinorum sp. nov., Lactobacillus mellifer sp. nov., Lactobacillus mellis sp. nov., Lactobacillus melliventris sp. nov., Lactobacillus kimbladii sp. nov., Lactobacillus helsingborgensis sp. nov., and Lactobacillus kullabergensis sp. nov., with the respective type strains being Fhon13NT ( = DSM 26257T = CCUG 63287T), Bin4NT ( = DSM 26254T = CCUG 63291T), Hon2NT ( = DSM 26255T = CCUG 63289T), Hma8NT ( = DSM 26256T = CCUG 63629T), Hma2NT ( = DSM 26263T = CCUG 63633T), Bma5NT ( = DSM 26265T = CCUG 63301T) and Biut2NT ( = DSM 26262T = CCUG 63631T).
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