Contamination of wounds with fecal bacteria in immuno-suppressed mice.
Animals
Anti-Bacterial Agents
/ pharmacology
Coinfection
/ drug therapy
Disease Models, Animal
Feces
/ microbiology
Humans
Immunocompromised Host
/ drug effects
Immunosuppression Therapy
/ adverse effects
Mice
Wound Healing
/ drug effects
Wound Infection
/ drug therapy
Wounds and Injuries
/ drug therapy
Journal
Scientific reports
ISSN: 2045-2322
Titre abrégé: Sci Rep
Pays: England
ID NLM: 101563288
Informations de publication
Date de publication:
13 07 2020
13 07 2020
Historique:
received:
31
12
2019
accepted:
15
06
2020
entrez:
15
7
2020
pubmed:
15
7
2020
medline:
17
12
2020
Statut:
epublish
Résumé
Immunocompromised patients are predisposed to chronically infected wounds. Especially ulcers in the dorsal region often experience secondary polymicrobial infections. However, current wound infection models mostly use single-strain bacteria. To mimic clinically occurring infections caused by fecal contamination in immunocompromised/immobile patients, which differ significantly from single-strain infections, the present study aimed at the establishment of a new mouse model using infection by fecal bacteria. Dorsal circular excision wounds in immunosuppressed mice were infected with fecal slurry solution in several dilutions up to 1:8,000. Impact of immunosuppressor, bacterial load and timing on development of wound infections was investigated. Wounds were analyzed by scoring, 3D imaging and swab analyses. Autofluorescence imaging was not successful. Dose-finding of cyclophosphamide-induced immunosuppression was necessary for establishment of bacterial wound infections. Infection with fecal slurry diluted 1:166 to 1:400 induced significantly delayed wound healing (p < 0.05) without systemic reactions. Swab analyses post-infection matched the initial polymicrobial suspension. The customized wound score confirmed significant differences between the groups (p < 0.05). Here we report the establishment of a simple, new mouse model for clinically occurring wound infections by fecal bacteria and the evaluation of appropriate wound analysis methods. In the future, this model will provide a suitable tool for the investigation of complex microbiological interactions and evaluation of new therapeutic approaches.
Identifiants
pubmed: 32661287
doi: 10.1038/s41598-020-68323-5
pii: 10.1038/s41598-020-68323-5
pmc: PMC7359036
doi:
Substances chimiques
Anti-Bacterial Agents
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
11494Références
Report Global Wound Dressings Market 2018–2022, TechNavio, Infiniti Research Ltd., London, UK (2018)
Gottrup, F., Apelqvist, J., Price, P. & European Wound Management Association Patient Outcome, G. Outcomes in controlled and comparative studies on non-healing wounds: recommendations to improve the quality of evidence in wound management. J Wound Care 19, 237–268. https://doi.org/10.12968/jowc.2010.19.6.48471 (2010).
Sunderkotter, C. & Becker, K. Frequent bacterial skin and soft tissue infections: Diagnostic signs and treatment. J Dtsch Dermatol Ges 13, 501–524. https://doi.org/10.1111/ddg.12721 (2015) ((quiz 525–506)).
doi: 10.1111/ddg.12721
pubmed: 26018361
Organization, W. H. WHO | WHO Global Strategy for Containment of Antimicrobial Resistance. WHO (2016).
Neil, J. A. Perioperative care of the immunocompromised patient. AORN J 85, 544–560. https://doi.org/10.1016/S0001-2092(07)60126-4 (2007) ((quiz 561–544)).
doi: 10.1016/S0001-2092(07)60126-4
pubmed: 17352893
Livesley, N. J. & Chow, A. W. Infected pressure ulcers in elderly individuals. Clin. Infect. Dis. 35, 1390–1396. https://doi.org/10.1086/344059 (2002).
doi: 10.1086/344059
pubmed: 12439803
Pendleton, J. N., Gorman, S. P. & Gilmore, B. F. Clinical relevance of the ESKAPE pathogens. Expert Rev. Anti Infect. Ther. 11, 297–308. https://doi.org/10.1586/eri.13.12 (2013).
doi: 10.1586/eri.13.12
pubmed: 23458769
Santajit, S. & Indrawattana, N. Mechanisms of antimicrobial resistance in ESKAPE pathogens. Biomed. Res. Int. 2016, 2475067. https://doi.org/10.1155/2016/2475067 (2016).
doi: 10.1155/2016/2475067
pubmed: 27274985
pmcid: 4871955
Dai, T. et al. Animal models of external traumatic wound infections. Virulence 2, 296–315. https://doi.org/10.4161/viru.2.4.16840 (2011).
doi: 10.4161/viru.2.4.16840
pubmed: 21701256
pmcid: 3173676
Tatara, A. M., Shah, S. R., Livingston, C. E. & Mikos, A. G. Infected animal models for tissue engineering. Methods 84, 17–24. https://doi.org/10.1016/j.ymeth.2015.03.025 (2015).
doi: 10.1016/j.ymeth.2015.03.025
pubmed: 25843609
pmcid: 4526327
Dai, T., Tegos, G. P., Zhiyentayev, T., Mylonakis, E. & Hamblin, M. R. Photodynamic therapy for methicillin-resistant Staphylococcus aureus infection in a mouse skin abrasion model. Lasers Surg. Med. 42, 38–44. https://doi.org/10.1002/lsm.20887 (2010).
doi: 10.1002/lsm.20887
pubmed: 20077489
pmcid: 2820267
Zolfaghari, P. S. et al. In vivo killing of Staphylococcus aureus using a light-activated antimicrobial agent. BMC Microbiol. 9, 27. https://doi.org/10.1186/1471-2180-9-27 (2009).
doi: 10.1186/1471-2180-9-27
pubmed: 19193212
pmcid: 2642833
Kraft, W. G., Johnson, P. T., David, B. C. & Morgan, D. R. Cutaneous infection in normal and immunocompromised mice. Infect. Immun. 52, 707–713 (1986).
doi: 10.1128/IAI.52.3.707-713.1986
Kugelberg, E. et al. Establishment of a superficial skin infection model in mice by using Staphylococcus aureus and Streptococcus pyogenes. Antimicrob. Agents Chemother. 49, 3435–3441. https://doi.org/10.1128/AAC.49.8.3435-3441.2005 (2005).
doi: 10.1128/AAC.49.8.3435-3441.2005
pubmed: 16048958
pmcid: 1196267
Gaspari, A. A. et al. CD86 (B7–2), but not CD80 (B7–1), expression in the epidermis of transgenic mice enhances the immunogenicity of primary cutaneous Candida albicans infections. Infect. Immun. 66, 4440–4449 (1998).
doi: 10.1128/IAI.66.9.4440-4449.1998
Jeray, K. J. et al. Evaluation of standard surgical preparation performed on superficial dermal abrasions. J. Orthop. Trauma 14, 206–211 (2000).
doi: 10.1097/00005131-200003000-00011
Walker, H. L. & Mason, A. D. Jr. A standard animal burn. J. Trauma 8, 1049–1051. https://doi.org/10.1097/00005373-196811000-00006 (1968).
doi: 10.1097/00005373-196811000-00006
pubmed: 5722120
Stieritz, D. D. & Holder, I. A. Experimental studies of the pathogenesis of infections due to Pseudomonas aeruginosa: Description of a burned mouse model. J. Infect. Dis. 131, 688–691. https://doi.org/10.1093/infdis/131.6.688 (1975).
doi: 10.1093/infdis/131.6.688
pubmed: 805812
Katakura, T., Yoshida, T., Kobayashi, M., Herndon, D. N. & Suzuki, F. Immunological control of methicillin-resistant Staphylococcus aureus (MRSA) infection in an immunodeficient murine model of thermal injuries. Clin. Exp. Immunol. 142, 419–425. https://doi.org/10.1111/j.1365-2249.2005.02944.x (2005).
doi: 10.1111/j.1365-2249.2005.02944.x
pubmed: 16297152
pmcid: 1809536
Stevens, E. J. et al. A quantitative model of invasive Pseudomonas infection in burn injury. J. Burn Care Rehabil. 15, 232–235 (1994).
doi: 10.1097/00004630-199405000-00005
Manafi, A. et al. Active immunization using exotoxin A confers protection against Pseudomonas aeruginosa infection in a mouse burn model. BMC Microbiol. 9, 23. https://doi.org/10.1186/1471-2180-9-23 (2009).
doi: 10.1186/1471-2180-9-23
pubmed: 19183501
pmcid: 2644702
Hamblin, M. R., O’Donnell, D. A., Murthy, N., Contag, C. H. & Hasan, T. Rapid control of wound infections by targeted photodynamic therapy monitored by in vivo bioluminescence imaging. Photochem. Photobiol. 75, 51–57 (2002).
doi: 10.1562/0031-8655(2002)075<0051:RCOWIB>2.0.CO;2
Burkatovskaya, M., Castano, A. P., Demidova-Rice, T. N., Tegos, G. P. & Hamblin, M. R. Effect of chitosan acetate bandage on wound healing in infected and noninfected wounds in mice. Wound Repair Regen. 16, 425–431. https://doi.org/10.1111/j.1524-475X.2008.00382.x (2008).
doi: 10.1111/j.1524-475X.2008.00382.x
pubmed: 18471261
pmcid: 2805166
Simonetti, O. et al. RNAIII-inhibiting peptide enhances healing of wounds infected with methicillin-resistant Staphylococcus aureus. Antimicrob. Agents Chemother. 52, 2205–2211. https://doi.org/10.1128/AAC.01340-07 (2008).
doi: 10.1128/AAC.01340-07
pubmed: 18391046
pmcid: 2415788
Mahoney, E. et al. Bacterial colonization and the expression of inducible nitric oxide synthase in murine wounds. Am. J. Pathol. 161, 2143–2152. https://doi.org/10.1016/s0002-9440(10)64492-6 (2002).
doi: 10.1016/s0002-9440(10)64492-6
pubmed: 12466130
pmcid: 1850895
Bowler, P. G., Duerden, B. I. & Armstrong, D. G. Wound microbiology and associated approaches to wound management. Clin. Microbiol. Rev. 14, 244–269. https://doi.org/10.1128/CMR.14.2.244-269.2001 (2001).
doi: 10.1128/CMR.14.2.244-269.2001
pubmed: 11292638
pmcid: 88973
Klein, P. et al. A porcine model of skin wound infected with a polybacterial biofilm. Biofouling 34, 226–236. https://doi.org/10.1080/08927014.2018.1425684 (2018).
doi: 10.1080/08927014.2018.1425684
pubmed: 29405092
Mastropaolo, M. D. et al. Synergy in polymicrobial infections in a mouse model of type 2 diabetes. Infect. Immun. 73, 6055–6063. https://doi.org/10.1128/IAI.73.9.6055-6063.2005 (2005).
doi: 10.1128/IAI.73.9.6055-6063.2005
pubmed: 16113326
pmcid: 1231087
Dalton, T. et al. An in vivo polymicrobial biofilm wound infection model to study interspecies interactions. PLoS ONE 6, e27317. https://doi.org/10.1371/journal.pone.0027317 (2011).
doi: 10.1371/journal.pone.0027317
pubmed: 22076151
pmcid: 3208625
Starr, M. E. et al. A new cecal slurry preparation protocol with improved long-term reproducibility for animal models of sepsis. PLoS ONE 9, e115705. https://doi.org/10.1371/journal.pone.0115705 (2014).
doi: 10.1371/journal.pone.0115705
pubmed: 25531402
pmcid: 4274114
Zuluaga, A. F. et al. Neutropenia induced in outbred mice by a simplified low-dose cyclophosphamide regimen: Characterization and applicability to diverse experimental models of infectious diseases. BMC Infect. Dis. 6, 55. https://doi.org/10.1186/1471-2334-6-55 (2006).
doi: 10.1186/1471-2334-6-55
pubmed: 16545113
pmcid: 1434751
Piñar, G., Poyntner, C., Lopandic, K., Tafer, H. & Sterflinger, K. Rapid diagnosis of biological colonization in cultural artefacts using the MinION nanopore sequencing technology. Int. Biodeterior. Biodegrad. 148, 104908. https://doi.org/10.1016/j.ibiod.2020.104908 (2020).
doi: 10.1016/j.ibiod.2020.104908
Xiao, L. et al. A catalog of the mouse gut metagenome. Nat Biotechnol 33, 1103–1108. https://doi.org/10.1038/nbt.3353 (2015).
doi: 10.1038/nbt.3353
pubmed: 26414350
Wu, Y. C. et al. Autofluorescence imaging device for real-time detection and tracking of pathogenic bacteria in a mouse skin wound model: preclinical feasibility studies. J. Biomed. Opt. 19, 085002. https://doi.org/10.1117/1.JBO.19.8.085002 (2014).
doi: 10.1117/1.JBO.19.8.085002
pubmed: 25089944
Kline, K. A. & Bowdish, D. M. Infection in an aging population. Curr. Opin. Microbiol. 29, 63–67. https://doi.org/10.1016/j.mib.2015.11.003 (2016).
doi: 10.1016/j.mib.2015.11.003
pubmed: 26673958
Hamblin, M. R. et al. Polycationic photosensitizer conjugates: effects of chain length and Gram classification on the photodynamic inactivation of bacteria. J. Antimicrob. Chemother. 49, 941–951. https://doi.org/10.1093/jac/dkf053 (2002).
doi: 10.1093/jac/dkf053
pubmed: 12039886
Hamblin, M. R., Zahra, T., Contag, C. H., McManus, A. T. & Hasan, T. Optical monitoring and treatment of potentially lethal wound infections in vivo. J. Infect. Dis. 187, 1717–1725. https://doi.org/10.1086/375244 (2003).
doi: 10.1086/375244
pubmed: 12751029
pmcid: 3441051
Dai, T. et al. Photodynamic therapy for Acinetobacter baumannii burn infections in mice. Antimicrob. Agents Chemother. 53, 3929–3934. https://doi.org/10.1128/AAC.00027-09 (2009).
doi: 10.1128/AAC.00027-09
pubmed: 19564369
pmcid: 2737832
Wolcott, R. D. et al. Analysis of the chronic wound microbiota of 2,963 patients by 16S rDNA pyrosequencing. Wound Repair Regen. 24, 163–174. https://doi.org/10.1111/wrr.12370 (2016).
doi: 10.1111/wrr.12370
pubmed: 26463872
Kalan, L. R. & Brennan, M. B. The role of the microbiome in nonhealing diabetic wounds. Ann. N. Y. Acad. Sci. 1435, 79–92. https://doi.org/10.1111/nyas.13926 (2019).
doi: 10.1111/nyas.13926
pubmed: 30003536
Citron, D. M., Goldstein, E. J., Merriam, C. V., Lipsky, B. A. & Abramson, M. A. Bacteriology of moderate-to-severe diabetic foot infections and in vitro activity of antimicrobial agents. J. Clin. Microbiol. 45, 2819–2828. https://doi.org/10.1128/JCM.00551-07 (2007).
doi: 10.1128/JCM.00551-07
pubmed: 17609322
pmcid: 2045270
Tipton, C. D. et al. Chronic wound microbiome colonization on mouse model following cryogenic preservation. PLoS ONE 14, e0221565. https://doi.org/10.1371/journal.pone.0221565 (2019).
doi: 10.1371/journal.pone.0221565
pubmed: 31442275
pmcid: 6707584
Roine, E., Bjork, I. T. & Oyen, O. Targeting risk factors for impaired wound healing and wound complications after kidney transplantation. Transplant Proc. 42, 2542–2546. https://doi.org/10.1016/j.transproceed.2010.05.162 (2010).
doi: 10.1016/j.transproceed.2010.05.162
pubmed: 20832540
Abalo, A. et al. Risk factors for surgical wound infection in HIV-positive patients undergoing surgery for orthopaedic trauma. J. Orthop. Surg. (Hong Kong) 18, 224–227. https://doi.org/10.1177/230949901001800218 (2010).
doi: 10.1177/230949901001800218
Davis, P. A., Corless, D. J., Gazzard, B. G. & Wastell, C. Increased risk of wound complications and poor healing following laparotomy in HIV-seropositive and AIDS patients. Dig. Surg. 16, 60–67. https://doi.org/10.1159/000018695 (1999).
doi: 10.1159/000018695
pubmed: 9949269
Guo, S. & Dipietro, L. A. Factors affecting wound healing. J. Dent. Res. 89, 219–229. https://doi.org/10.1177/0022034509359125 (2010).
doi: 10.1177/0022034509359125
pubmed: 20139336
pmcid: 2903966
Su, Y. & Richmond, A. Chemokine regulation of neutrophil infiltration of skin wounds. Adv. Wound Care (New Rochelle) 4, 631–640. https://doi.org/10.1089/wound.2014.0559 (2015).
doi: 10.1089/wound.2014.0559
Gad, F., Zahra, T., Francis, K. P., Hasan, T. & Hamblin, M. R. Targeted photodynamic therapy of established soft-tissue infections in mice. Photochem. Photobiol. Sci. 3, 451–458. https://doi.org/10.1039/b311901g (2004).
doi: 10.1039/b311901g
pubmed: 15122362
pmcid: 3071693
Manepalli, S. et al. Characterization of a cyclophosphamide-induced murine model of immunosuppression to study Acinetobacter baumannii pathogenesis. J. Med. Microbiol. 62, 1747–1754. https://doi.org/10.1099/jmm.0.060004-0 (2013).
doi: 10.1099/jmm.0.060004-0
pubmed: 24000227
pmcid: 4083505
Bairy, L., Ganesh, S. B., Adiga, S. & Shalini, A. Impaired wound healing due to cyclophosphamide (CLP) alleviated by supplemental Ginkgo biloba (GB). J. Nat. Remed. 6, 31–34 (2006).
Wie, H., Bruaset, I. & Eckersberg, T. Effects of cyclophosphamide on open, granulating skin wounds in rats. Acta Pathol. Microbiol. Scand. A 87A, 185–192 (1979).
pubmed: 463564
Thompson, M. G. et al. Validation of a novel murine wound model of Acinetobacter baumannii infection. Antimicrob. Agents Chemother. 58, 1332–1342. https://doi.org/10.1128/AAC.01944-13 (2014).
doi: 10.1128/AAC.01944-13
pubmed: 24342634
pmcid: 3957858
Nair, N., Biswas, R., Gotz, F. & Biswas, L. Impact of Staphylococcus aureus on pathogenesis in polymicrobial infections. Infect. Immun. 82, 2162–2169. https://doi.org/10.1128/IAI.00059-14 (2014).
doi: 10.1128/IAI.00059-14
pubmed: 24643542
pmcid: 4019155
Leise, B. S. Topical wound medications. Vet. Clin. N. Am. Equine Pract. 34, 485–498. https://doi.org/10.1016/j.cveq.2018.07.006 (2018).
doi: 10.1016/j.cveq.2018.07.006
Barrett, S. Wound-bed preparation: A vital step in the healing process. Br. J. Nurs. 26, S24–S31. https://doi.org/10.12968/bjon.2017.26.12.S24 (2017).
doi: 10.12968/bjon.2017.26.12.S24
pubmed: 28640728
Garcez, A. S. et al. Effects of photodynamic therapy on Gram-positive and Gram-negative bacterial biofilms by bioluminescence imaging and scanning electron microscopic analysis. Photomed Laser Surg. 31, 519–525. https://doi.org/10.1089/pho.2012.3341 (2013).
doi: 10.1089/pho.2012.3341
pubmed: 23822168
pmcid: 3818004
Wang, Y. et al. In vivo investigation of antimicrobial blue light therapy for multidrug-resistant Acinetobacter baumannii burn infections using bioluminescence imaging. J. Vis. Exp. https://doi.org/10.3791/54997 (2017).
doi: 10.3791/54997
pubmed: 29364212
pmcid: 5908403
DaCosta, R. S. et al. Point-of-care autofluorescence imaging for real-time sampling and treatment guidance of bioburden in chronic wounds: First-in-human results. PLoS ONE 10, e0116623. https://doi.org/10.1371/journal.pone.0116623 (2015).
doi: 10.1371/journal.pone.0116623
pubmed: 25790480
pmcid: 4366392
Ottolino-Perry, K. et al. Improved detection of clinically relevant wound bacteria using autofluorescence image-guided sampling in diabetic foot ulcers. Int. Wound J 14, 833–841. https://doi.org/10.1111/iwj.12717 (2017).
doi: 10.1111/iwj.12717
pubmed: 28244218
Jamadagni, P. S. et al. Experimental and histopathological observation scoring methods for evaluation of wound healing properties of Jatyadi Ghrita. Ayu 37, 222–229. https://doi.org/10.4103/ayu.AYU_51_17 (2016).
doi: 10.4103/ayu.AYU_51_17
pubmed: 29491675
pmcid: 5822989
Panuncialman, J. & Falanga, V. The science of wound bed preparation. Clin. Plast. Surg. 34, 621–632. https://doi.org/10.1016/j.cps.2007.07.003 (2007).
doi: 10.1016/j.cps.2007.07.003
pubmed: 17967618