This study aimed to the molecular identification of isolated bacteria from the soils of Likouala (Congo-Brazzaville) peat bog area. Counting and isolation were carried out on Mossel and TSB media enriched with petroleum and vegetable oil; the amplification and sequencing of 16S RNA genes by PCR and Artic Oxford Nanopore Technology. The results showed bacterial loads of (5.81 ± 1.08) × 104; (6.64 ± 1.94) × 104 et (8.56 ± 1.19) × 103 CFU/g on Mossel respectively for samples 1, 2 and 3 against (2.12 ± 4.1) × 108 et (8.15 ± 10.1) × 107 CFU/g respectively on TSB enriched with petroleum and vegetable oil exclusively with sample 2. The analysis of the 16S rRNA of the isolates gene made it possible, after PCR, agarose gel electrophoresis, sequencing and bioinformatics analysis, to identify eight (08) strains with similarities of 99 to 100% whose sequences genes placed in GenBank have made it possible to obtain accession numbers corresponding to: Bacillus thuringiensis strain (ON303633); Bacillus cereus strain (ON350770); Bacillus thuringiensis strain (ON350771); Bacillus thuringiensis strain (ON738723); Priestia megaterium strain (ON738719); Bacillus anthracis strain (ON738720); Bacillus subtilis (ON738721); Enterobacter sp (ON738722). The phylogenetic classification of strains was done and revealed two genuses which are Bacillus and Enterobacteriaceae.
References
[1]
Fatima, L.D. and Francis, M. (2008) Les tourbières et leur role de stockage de carbone face aux changements climatiques. Zones Humides Info, 59-60, 22-24.
https://hal-insu.archives-ouvertes.fr/insu-00321655
[2]
Poliakova, A.V., Chernov, I. and Panikoc, N.S. (2001) Yeast Biodiversity in Hydromorphic Soils with Reference to Grass-Sphagnum Swamp in Western Siberia and the Hammocky Tundra Region. Mikrobiologia, 70, 714-720.
[3]
Brown-Elliott, B. and Wallace, J. (2002) Clinical and Taxonomic Status of Pathogenic Nonpigmented or Late-Pigmenting Rapidly Growing Mycobacteria. Clinical Microbiology Reviews, 15, 716-746. https://doi.org/10.1128/CMR.15.4.716-746.2002
[4]
Joosten, H., Tapio-Bistrom, M.L. and Tol, S. (2012) Peatlands: Guidance for Climate Change Mitigation through Conservation, Rehabilitation and Sustainable Use. 2nd Edition, Mitigation of Climate Change in Agriculture Series 5, FAO & Wetlands International, 101.
[5]
Dorrepaal, E., Toet, S., van Loegtestijn, R.S.P., Swart, E., van de Weg, M.J., Callaghan, T.V. and Aerts, R. (2009) Carbon Respiration from Subsurface Peat Accelerated by Climate Warming in the Subarctic. Nature, 460, 616-619.
https://doi.org/10.1038/nature08216
[6]
Jonas, M.C., Viennechie, G.E., Cecile, M.K.I.M., Samuel, O.I., Averti, I.S. and Etienne, N. (2022) Characterization of Isolated Bacteria from Soils in the Likouala Peat Bog Area (Republic of Congo). American Journal of Microbiological Research, 10, 59-70.
[7]
Coulibaly, K. (2005) Etude de la qualité Physico-Chimique et bactériologique des puits de certains quartiers du district de Bamako. Ph.D. Thesis, Université de Bamako, Bamako.
[8]
Soloka, M.F.A., Nguimbi, E., Kayath, A.C. and Ahombo, G. (2020) Molecular Charactrisation of Bacillus-Genus Bacteria with Fibrinolytic Potential Isolated from Squashes « NTETE » Brazzaville in the Republic of Congo. Amicain Journal of Microbiolocal Research, 8, 7-18. https://doi.org/10.12691/ajmr-8-1-2
[9]
Ngo, I., Nguimbi, E., Kayath, C. and Ampa, R. (2019) Molecular Identification and Phylogenetic Classification and Proteolytic Capacity of cultivable Bacteria Isolated from Soils in Brazzaville, Republic of Congo. Journal of Biochemestry, Microbiology and Biotechnology, 7, 1-7.
[10]
De Vos, P., Garrity, G.M., Jones, D., Krieg, N.R., Ludwig, W., Rainey, F. and Whitman, W.B. (2009) Bergey’s Manuel of Systematic Bacteriology, 2nd Edition, Vol. 3, The Firmicutes. Springer, New York.
[11]
Dahou, A., Homrani, A. and Medjahed, F.B.M. (2015) La microflore lactique d’un fromage traditionnel Algérien « type j’ ben »: Connaissance des écosystèmes microbiens laitiers locaux et de leurs roles dans la fabrication des fromages. Afrique Science, 11, 1-13.
[12]
Weisberg, W.G., Barns, S.M., Pelletier, D.A. and Lane, D.J. (1991) 16S Ribosomal DNA Amplification for Phylogenetic Study. Journal of Bacteriology, 173, 697-703.
https://doi.org/10.1128/jb.173.2.697-703.1991
[13]
Jay, A.G. (1995) Validity of Nucleic Acid Purities Monitored by 260nm/280nm Absorbance Ratios. Bio Techniques, 18, 62-63.
[14]
Onyankouang, I., Morabandza, C.J., Kimbatsa, I.M.C.M., Mabika, F.A.S., Ngô, I., Lingouangou, T.M., Moyen, R. and Nguimbi, E. (2022) Diversity and Phylogenetic Relationships of Proteolytic Bacteria Isolated from Fermented Pepper and Soil in Brazzaville, Republic of Congo. International Journal of Microbiology and Biotechnology, 7, 124-134.
[15]
Bravo, A., Gómez, I., Porta, H., García-Gómez, I.B., Rodriguez-Almazan, C., Pardo, L. and Soberón, M. (2013) Evolution of Bacillus thuringiensis Cry Toxins Insecticidal Activity. Microbial Biotechnology, 6, 17-26.
https://doi.org/10.1111/j.1751-7915.2012.00342.x
[16]
Fanfani, G. (2014) Les bactéries du groupe de Bacillus cereus dans les conserves à pH peu acide (petit pois); détection, caractérisation par méthode probabiliste numérisée, moléculaire et anti-bio résistance. Master’s Thesis, université badji Mokhtar Annaba, Annaba.
[17]
Etienne, N., Armel, S.M.F., Valentin, D., Eckzechel, N.S.A., Christian, K.A. and Rachel, M. (2021) New Phylogenetic Molecular Markers in Bacteria of the Genus Bacillus: Fibrinolytic Proteases. International Journal of Microbiology and Biotechnology, 6, 86-94. https://doi.org/10.11648/j.ijmb.20210603.14
[18]
Helgason, E., Caugant, D.A., Lecadet, M.M., Chen, Y., Mahillon, J., Lovgreen, A., Hegna, I., Kvaloy, K. and Kolsto, A.B. (1998) Genetic Diversity of Bacillus cereus/Bacillus thuringiensis Isolates from Natural Sources. Current Microbiology, 37, 80-87. https://doi.org/10.1007/s002849900343
[19]
Chen, M.L. and Tsen, H.Y. (2002) Discrimination of Bacillus cereus and Bacillus thuringiensis with 16S rRNA and gyrB Gene Based PCR Primers and Sequencing of Their Annealing Sites. Journal of Applied Microbiology, 92, 912-919.
https://doi.org/10.1046/j.1365-2672.2002.01606.x
