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Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture

OBJECTIVES: We aimed to explore the bacterial community composition following ocean bacterial infection using an animal model. METHODS: This animal-based experiment was conducted from September 2019 to November 2019. Eighteen seawater filter membranes were collected from Changle City, Fujiian Provin...

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Autores principales: Wang, Du, Zheng, Qingcong, Lv, Qi, Zhang, Chaofan, Zheng, Yun, Chen, Huidong, Zhang, Wenming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8293552/
https://www.ncbi.nlm.nih.gov/pubmed/34289854
http://dx.doi.org/10.1186/s13018-021-02553-9
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author Wang, Du
Zheng, Qingcong
Lv, Qi
Zhang, Chaofan
Zheng, Yun
Chen, Huidong
Zhang, Wenming
author_facet Wang, Du
Zheng, Qingcong
Lv, Qi
Zhang, Chaofan
Zheng, Yun
Chen, Huidong
Zhang, Wenming
author_sort Wang, Du
collection PubMed
description OBJECTIVES: We aimed to explore the bacterial community composition following ocean bacterial infection using an animal model. METHODS: This animal-based experiment was conducted from September 2019 to November 2019. Eighteen seawater filter membranes were collected from Changle City, Fujiian Province, China, on September 8, 2019. Ten filter membranes were used for implantation. Eight filter membranes that were used in the bacterial culture for the exploration of seawater bacteria were assigned to the seawater group (SG). Fourteen healthy adult New Zealand rabbits were randomly divided into the experimental group (EG) and control group (CG). Seawater filter membranes and asepsis membranes were implanted into the tibia in the EG and CG, respectively. One week after surgery, tibial bone pathology tissues were collected and assessed using light microscopy and scanning electron microscopy (SEM). Medullary cavity tissues were collected for the performance of Illumina MiSeq sequencing and bacterial culture. The differences between EG and CG were assessed by pathological observation under light microscopy and SEM, high-throughput bacterial sequencing, and bacterial culture. RESULTS: Compared with the CG, the infection rate was 100%, and the mortality value was 20% after the implantation of the filter membranes in the EG. Both light microscopy and SEM showed that a large number of bacteria were distributed in the bone marrow cavity after ocean bacterial infection. No bacterial growth was found in the CG. Illumina MiSeq sequencing found that Firmicutes, Proteobacteria, Thermotogae, Fusobacteria, Bacteroidetes, and Actinobacteria were the dominant bacteria at the phylum level and Clostridium_sensu_stricto_7, Haloimpatiens, Clostridium_sensu_stricto_15, Clostridiaceae_1, Clostridium_sensu_stricto_18, and Oceanotoga were the dominant bacteria in genus level among the EG. In the bacterial culture of the medullary cavity tissues, Klebsiella pneumoniae, Shewanella algae, Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, and Vibrio vulnificus were the predominant infective species. Moreover, compared with the SG, the EG showed a higher detection rate of E. coli and S. aureus (P = 0.008 and P = 0.001, respectively). The detection rates of V. alginolyticus, V. parahaemolyticus, and V. fluvialis were higher in the SG than the EG (P = 0.007, P = 0.03, and P = 0.03, respectively). CONCLUSIONS: Our model, which was comprehensively evaluated using four techniques: histopathology and SEM observation, gene detection, and bacteria culture, provides a scientific basis for the clinical diagnosis and treatment of patients in such settings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13018-021-02553-9.
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spelling pubmed-82935522021-07-21 Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture Wang, Du Zheng, Qingcong Lv, Qi Zhang, Chaofan Zheng, Yun Chen, Huidong Zhang, Wenming J Orthop Surg Res Research Article OBJECTIVES: We aimed to explore the bacterial community composition following ocean bacterial infection using an animal model. METHODS: This animal-based experiment was conducted from September 2019 to November 2019. Eighteen seawater filter membranes were collected from Changle City, Fujiian Province, China, on September 8, 2019. Ten filter membranes were used for implantation. Eight filter membranes that were used in the bacterial culture for the exploration of seawater bacteria were assigned to the seawater group (SG). Fourteen healthy adult New Zealand rabbits were randomly divided into the experimental group (EG) and control group (CG). Seawater filter membranes and asepsis membranes were implanted into the tibia in the EG and CG, respectively. One week after surgery, tibial bone pathology tissues were collected and assessed using light microscopy and scanning electron microscopy (SEM). Medullary cavity tissues were collected for the performance of Illumina MiSeq sequencing and bacterial culture. The differences between EG and CG were assessed by pathological observation under light microscopy and SEM, high-throughput bacterial sequencing, and bacterial culture. RESULTS: Compared with the CG, the infection rate was 100%, and the mortality value was 20% after the implantation of the filter membranes in the EG. Both light microscopy and SEM showed that a large number of bacteria were distributed in the bone marrow cavity after ocean bacterial infection. No bacterial growth was found in the CG. Illumina MiSeq sequencing found that Firmicutes, Proteobacteria, Thermotogae, Fusobacteria, Bacteroidetes, and Actinobacteria were the dominant bacteria at the phylum level and Clostridium_sensu_stricto_7, Haloimpatiens, Clostridium_sensu_stricto_15, Clostridiaceae_1, Clostridium_sensu_stricto_18, and Oceanotoga were the dominant bacteria in genus level among the EG. In the bacterial culture of the medullary cavity tissues, Klebsiella pneumoniae, Shewanella algae, Staphylococcus aureus, Escherichia coli, Enterobacter cloacae, and Vibrio vulnificus were the predominant infective species. Moreover, compared with the SG, the EG showed a higher detection rate of E. coli and S. aureus (P = 0.008 and P = 0.001, respectively). The detection rates of V. alginolyticus, V. parahaemolyticus, and V. fluvialis were higher in the SG than the EG (P = 0.007, P = 0.03, and P = 0.03, respectively). CONCLUSIONS: Our model, which was comprehensively evaluated using four techniques: histopathology and SEM observation, gene detection, and bacteria culture, provides a scientific basis for the clinical diagnosis and treatment of patients in such settings. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s13018-021-02553-9. BioMed Central 2021-07-21 /pmc/articles/PMC8293552/ /pubmed/34289854 http://dx.doi.org/10.1186/s13018-021-02553-9 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research Article
Wang, Du
Zheng, Qingcong
Lv, Qi
Zhang, Chaofan
Zheng, Yun
Chen, Huidong
Zhang, Wenming
Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title_full Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title_fullStr Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title_full_unstemmed Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title_short Assessment of seawater bacterial infection in rabbit tibia by Illumina MiSeq sequencing and bacterial culture
title_sort assessment of seawater bacterial infection in rabbit tibia by illumina miseq sequencing and bacterial culture
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8293552/
https://www.ncbi.nlm.nih.gov/pubmed/34289854
http://dx.doi.org/10.1186/s13018-021-02553-9
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