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Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food

Received: 17 August 2022     Accepted: 16 September 2022     Published: 29 September 2022
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Abstract

Congo has a wide range of traditional foods including Mbala pinda. The objective of this work was to explore the microbiological quality of this food by identifying bacteria of the genus Bacillus capable of producing antibacterial substances isolated from this food. Samples of Mbala pinda were collected in six localities namely Pointe-Noire, Dolisie, Makabana, Loudima, Nkayi and Madingou. Exposed to the open air, a quantification of the bacteria was hold and showed that the microorganisms only appear on the 5th day of exposure. The quantity of microorganisms varies from one sample to another. The total mesophilic microorganisms are slightly more numerous in the Dolisie samples. This quantity is between 44·103 UFC/g and 52·103 UFC/g. Enterobacteriaceae are between 0.441·103 and 0.142·103 and more represented in Makabana. Bacillus genus bacteria are between 4.51·103 and 2.24·103 and are more represented in Dolisie. All isolates were generally elongated in shape with two particular morphotypes with whitish or light yellow and isolated colonies. The isolates were all catalase positive, oxidase positive, gram positive, motile, assigned to the genus Bacillus. The analysis of the 16S rRNA gene of five isolates has been hold, with PCR, Agarose Gel electrophoresis, sequencing and bioinformatics analysis. All PCR fragments were about 1500pb od size. Five strains have been identified, which 16SrRNA sequences genes were put in GenBank. New accession numbers were given as the following strains: Bacillus safensis MPRN8 (MT107116), Bacillus megaterium MPRN5 (MT107117), Bacillus amyloliquefaciens MPRN2 (MT107118), Bacillus subtilis MPRN7 (MT107119), Bacillus velezensis MPRN1 (MT107120). All these strains belonging to the genus Bacillus are phylogenetically very similar forming a group within this genus keeping the same taxonomic level. Moreover, identified Bacillus strains were explored for their ability to produce antibacterial substances. It appears that the five strains inhibit differently the growth of three pathogenic strains namely E coli, Pseudomonas aeruginosa and Staphylococcus aureus IMa1 (Bacillus velezensis MPRN1), IL1 (Bacillus megaterieum MPRN5) and ID1 (Bacillus subtilis MPRN7) were those which inhibited the growth of the three pathogens with larger diameters, thus testifying the production of antibacterial substances by the identified strains. Correlations between identified strains and their inhibition according to each pathogen show the importance of these strains in the search for antibacterial substances and therefore the importance of promoting this food at a lower cost and adapted to our country.

Published in International Journal of Microbiology and Biotechnology (Volume 7, Issue 3)
DOI 10.11648/j.ijmb.20220703.15
Page(s) 143-158
Creative Commons

This is an Open Access article, 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 or format, provided the original work is properly cited.

Copyright

Copyright © The Author(s), 2022. Published by Science Publishing Group

Keywords

Mbala Pinda, Antibacterial, Sequencing, Bacillus, BLASTn

References
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    Priscillia Renée Nguimbi-Tsati, Saturnin Nicaise Mokemiabeka, Christian Aimé Kayath, Thantique Moutali Lingouangou, Yannich Okouakoua, et al. (2022). Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food. International Journal of Microbiology and Biotechnology, 7(3), 143-158. https://doi.org/10.11648/j.ijmb.20220703.15

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    Priscillia Renée Nguimbi-Tsati; Saturnin Nicaise Mokemiabeka; Christian Aimé Kayath; Thantique Moutali Lingouangou; Yannich Okouakoua, et al. Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food. Int. J. Microbiol. Biotechnol. 2022, 7(3), 143-158. doi: 10.11648/j.ijmb.20220703.15

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    Priscillia Renée Nguimbi-Tsati, Saturnin Nicaise Mokemiabeka, Christian Aimé Kayath, Thantique Moutali Lingouangou, Yannich Okouakoua, et al. Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food. Int J Microbiol Biotechnol. 2022;7(3):143-158. doi: 10.11648/j.ijmb.20220703.15

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  • @article{10.11648/j.ijmb.20220703.15,
      author = {Priscillia Renée Nguimbi-Tsati and Saturnin Nicaise Mokemiabeka and Christian Aimé Kayath and Thantique Moutali Lingouangou and Yannich Okouakoua and Gabriel Ahombo},
      title = {Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food},
      journal = {International Journal of Microbiology and Biotechnology},
      volume = {7},
      number = {3},
      pages = {143-158},
      doi = {10.11648/j.ijmb.20220703.15},
      url = {https://doi.org/10.11648/j.ijmb.20220703.15},
      eprint = {https://article.sciencepublishinggroup.com/pdf/10.11648.j.ijmb.20220703.15},
      abstract = {Congo has a wide range of traditional foods including Mbala pinda. The objective of this work was to explore the microbiological quality of this food by identifying bacteria of the genus Bacillus capable of producing antibacterial substances isolated from this food. Samples of Mbala pinda were collected in six localities namely Pointe-Noire, Dolisie, Makabana, Loudima, Nkayi and Madingou. Exposed to the open air, a quantification of the bacteria was hold and showed that the microorganisms only appear on the 5th day of exposure. The quantity of microorganisms varies from one sample to another. The total mesophilic microorganisms are slightly more numerous in the Dolisie samples. This quantity is between 44·103 UFC/g and 52·103 UFC/g. Enterobacteriaceae are between 0.441·103 and 0.142·103 and more represented in Makabana. Bacillus genus bacteria are between 4.51·103 and 2.24·103 and are more represented in Dolisie. All isolates were generally elongated in shape with two particular morphotypes with whitish or light yellow and isolated colonies. The isolates were all catalase positive, oxidase positive, gram positive, motile, assigned to the genus Bacillus. The analysis of the 16S rRNA gene of five isolates has been hold, with PCR, Agarose Gel electrophoresis, sequencing and bioinformatics analysis. All PCR fragments were about 1500pb od size. Five strains have been identified, which 16SrRNA sequences genes were put in GenBank. New accession numbers were given as the following strains: Bacillus safensis MPRN8 (MT107116), Bacillus megaterium MPRN5 (MT107117), Bacillus amyloliquefaciens MPRN2 (MT107118), Bacillus subtilis MPRN7 (MT107119), Bacillus velezensis MPRN1 (MT107120). All these strains belonging to the genus Bacillus are phylogenetically very similar forming a group within this genus keeping the same taxonomic level. Moreover, identified Bacillus strains were explored for their ability to produce antibacterial substances. It appears that the five strains inhibit differently the growth of three pathogenic strains namely E coli, Pseudomonas aeruginosa and Staphylococcus aureus IMa1 (Bacillus velezensis MPRN1), IL1 (Bacillus megaterieum MPRN5) and ID1 (Bacillus subtilis MPRN7) were those which inhibited the growth of the three pathogens with larger diameters, thus testifying the production of antibacterial substances by the identified strains. Correlations between identified strains and their inhibition according to each pathogen show the importance of these strains in the search for antibacterial substances and therefore the importance of promoting this food at a lower cost and adapted to our country.},
     year = {2022}
    }
    

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  • TY  - JOUR
    T1  - Molecular Identification and Phylogenetic Classification of Producing-Antibacterial Substances Bacteria Isolated from Mbala Pinda, a Congolese Traditional Food
    AU  - Priscillia Renée Nguimbi-Tsati
    AU  - Saturnin Nicaise Mokemiabeka
    AU  - Christian Aimé Kayath
    AU  - Thantique Moutali Lingouangou
    AU  - Yannich Okouakoua
    AU  - Gabriel Ahombo
    Y1  - 2022/09/29
    PY  - 2022
    N1  - https://doi.org/10.11648/j.ijmb.20220703.15
    DO  - 10.11648/j.ijmb.20220703.15
    T2  - International Journal of Microbiology and Biotechnology
    JF  - International Journal of Microbiology and Biotechnology
    JO  - International Journal of Microbiology and Biotechnology
    SP  - 143
    EP  - 158
    PB  - Science Publishing Group
    SN  - 2578-9686
    UR  - https://doi.org/10.11648/j.ijmb.20220703.15
    AB  - Congo has a wide range of traditional foods including Mbala pinda. The objective of this work was to explore the microbiological quality of this food by identifying bacteria of the genus Bacillus capable of producing antibacterial substances isolated from this food. Samples of Mbala pinda were collected in six localities namely Pointe-Noire, Dolisie, Makabana, Loudima, Nkayi and Madingou. Exposed to the open air, a quantification of the bacteria was hold and showed that the microorganisms only appear on the 5th day of exposure. The quantity of microorganisms varies from one sample to another. The total mesophilic microorganisms are slightly more numerous in the Dolisie samples. This quantity is between 44·103 UFC/g and 52·103 UFC/g. Enterobacteriaceae are between 0.441·103 and 0.142·103 and more represented in Makabana. Bacillus genus bacteria are between 4.51·103 and 2.24·103 and are more represented in Dolisie. All isolates were generally elongated in shape with two particular morphotypes with whitish or light yellow and isolated colonies. The isolates were all catalase positive, oxidase positive, gram positive, motile, assigned to the genus Bacillus. The analysis of the 16S rRNA gene of five isolates has been hold, with PCR, Agarose Gel electrophoresis, sequencing and bioinformatics analysis. All PCR fragments were about 1500pb od size. Five strains have been identified, which 16SrRNA sequences genes were put in GenBank. New accession numbers were given as the following strains: Bacillus safensis MPRN8 (MT107116), Bacillus megaterium MPRN5 (MT107117), Bacillus amyloliquefaciens MPRN2 (MT107118), Bacillus subtilis MPRN7 (MT107119), Bacillus velezensis MPRN1 (MT107120). All these strains belonging to the genus Bacillus are phylogenetically very similar forming a group within this genus keeping the same taxonomic level. Moreover, identified Bacillus strains were explored for their ability to produce antibacterial substances. It appears that the five strains inhibit differently the growth of three pathogenic strains namely E coli, Pseudomonas aeruginosa and Staphylococcus aureus IMa1 (Bacillus velezensis MPRN1), IL1 (Bacillus megaterieum MPRN5) and ID1 (Bacillus subtilis MPRN7) were those which inhibited the growth of the three pathogens with larger diameters, thus testifying the production of antibacterial substances by the identified strains. Correlations between identified strains and their inhibition according to each pathogen show the importance of these strains in the search for antibacterial substances and therefore the importance of promoting this food at a lower cost and adapted to our country.
    VL  - 7
    IS  - 3
    ER  - 

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Author Information
  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

  • Cellular and Molecular Biology Laboratory, Faculty of Science and Techniques, Marien Ngouabi University, Brazzaville, Congo

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