American Journal of Microbiological Research, 2020, Vol. 8, No. 1, 7-18 Available online at http://pubs.sciepub.com/ajmr/8/1/2 Published by Science and Education Publishing DOI:10.12691/ajmr-8-1-2 Molecular Characterization of Bacillus-Genus Bacteria with Fibrinolytic Potential Isolated from Squashes «NTETE»in Brazzaville in the Republic of Congo Faly Armel SolokaMabika 1,2 , Etienne Nguimbi 1,2,3,* , AiméChristian Kayath 1,3 , Gabriel Ahombo 1,2 1 Laboratoire de Biologie Cellulaire et Moléculaire, Facultédes Sciences et Techniques, UniversitéMarien Ngouabi 2 Unitéde Biologie moléculaire et Bioinformatique, Facultédes Sciences et Techniques, UniversitéMarien Ngouabi 3 Institut de Recherche en Sciences Naturelles et Exactes (IRSEN) *Corresponding author: etienne.ng1612@gmail.com Received December 20, 2019; Revised February 03, 2020; Accepted February 20, 2020 Abstract Thromboses appearing in the blood and causing cardiovascular disease by the presence of fibrin remain a problem of concern worldwide. The fibrinolytic enzymes developed by bacteria of the Bacillus-genus are today an interesting and promising alternative to chemicals with multiple consequences. It is in this perspective that three (3) samples of cooked squash wrapped in sheets and consumed in Brazzaville and collected and in three Brazzaville markets were explored. The estimated count of the total flora of each sample for each of the three markets, namely Total, Moukondo and Tsiéméwas carried out on PCA and represents respectively (5.2 ±0.12) 107 CFU / g, (1.73 ± 0 , 16) 107 CFU / g, and (9.43 ± 1.06) 107 CFU / g while bacteria of the genus Bacillus are estimated in Mossel respectively at (3.5 ±1.16) 106 CFU / g, (4, 01 ±0.85) 106 CFU / g (8.96 ±0.60) 106 CFU / g. Sixty-six isolates of bacteria of the genus Bacillus isolated from squash by conventional microbiology techniques have been phenotypically characterized. The morphological types characterized are essentially the bacillary form and the spherical form. The ability to produce fibrinolytic enzymes correlated with growth was assessed. The growth in terms of optical density varies from 0.800 to 0.97 and the enzymatic production in all the isolates tested varies from 12 to 21 mm. After DNA extraction from 36 isolates, PCR amplification of the rR16S gene revealed fragments of approximately 1500bp by electrophoresis on Agarose Gel. The sequencing of thirty-four (34) fragments made it possible to obtain fifteen (15) sequences having a strong similarity between them and also with the homologs of the databases (97% to 100%) and therefore the molecular identification of: Bacillus sp 40%, Bacillus amyloliquefaciens 6.66%, Bacillus subtilis 33.33%, Bacillus Pumilus 6.66%, Bacillus megaterum 6.66%, Bacillus velezensis 6.66%. Five (5) of these sequences have been submitted to GenBank and the accession numbers are successively: MK193815.1 (Bacillus subtilis strain ASM1), MK207434.1 (Bacillus subtilis strain ASM3), MK207435.1 (Bacillus pumilus strain ASM5) , MK207436.1 (Bacillus subtilis strain ASM4), MK207437.1 (Bacillus megaterium strain ASM2).The multiple alignment of sequences obtained shows a high conservation of this gene in bacteria of the genus Bacillus. The phylogenetic classification clearly shows this monophyletic class of bacteria of the genus Bacillus with very short distances(less than 3%). Keywords: molecular characterization, fibrinolytic potential, Bacillus, squash, Brazzaville Cite This Article: Faly Armel SolokaMabika, Etienne Nguimbi, Aimé Christian Kayath, and Gabriel Ahombo, “Molecular Characterization of Bacillus-Genus Bacteria with Fibrinolytic Potential Isolated from Squashes «NTETE»in Brazzaville in the Republic of Congo.” American Journal of Microbiological Research, vol. 8, no. 1 (2020): 7-18. doi: 10.12691/ajmr-8-1-2. 1. Introduction In recent years the idea of a direct relationship between an optimal diet and a healthy lifestyle has gained ground among the general public. Thus, healthy food markets have developed strongly in order to meet the demand of consumers who increasingly seek to control their quality of life and their state of health through better nutrition [1]. These foods are either subjected to transformation processes under the influence of microorganisms in the case of fermented foods or undergo no transformation in the case of non-fermented foods. Ntete is a food product obtained from the seeds of squash (cucurbitaceae). Squash is an ornamental and vegetable plant, with a long climbing stem and large orange flowers belonging to the cucurbitaceae family, of which there are roughly 100 genera and 750 species listed worldwide. This family is known for its great genetic diversity and its great adaptation to tropical, subtropical regions.They are particularly cultivated in the departments of Pool, Bouenza and Plateaux for their seeds and leaves for food and medicinal purposes (In traditional pharmacopoeia). In