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Numeration and identification of thermotolerant endospore-forming Bacillus from two fermented condiments Bikalga and Soumbala.


Bikalga and Soumbala are condiment products of traditional uncontrolled alkaline fermentation of Hibiscus sabdariffa and Parkia biglobosa seeds respectively. These food additives are used and produced as major condiments in many African countries including Burkina Faso, Cameroon, Mali, Niger, Senegal, Sierra Leone and Sudan among others. Bikalga and Soumbala are produced by women and constitute economical resource. Soumbala is known under different appellations depending of the country: dawadawa or iru in Nigeria and North of Ghana [26,37,11], netetou in Senegal [23], afitin in Benin [4], kinda (Sierra Leone), natto in Japan and kinema in Nepal [5,38]. Bikalga is also called Dawadawa botso in Niger [34], Datou in Mali [34], Furundu in Sudan [40], Mbuja in Cameroon [22]. They are excellent sources of proteins with essential amino acids also containing lipids, carbohydrates, essential fatty acids and vitamins [6,33,40]. Many families in west Africa often used Soumbala and Bikalga as low-cost meat substitute. These condiments improve nutritional values of foods as well as sensory properties as taste enhancer; contain antioxidant and neutraceuticals that provide health.

Several works were dedicated to the characterization of the microflora of these fermented products [11,4,32,17,21,29]. These studies demonstrated the prevalence of Bacillus species (B. subtilis, B. coagulans, B. amyloliquefaciens, B. pumilus, B. cereus, B. thuringiensis, B. brevis and B. licheniformis) in the fermentation process and their role in the bioconversion of the products. Ouoba et al. [32] in their recent studies sowhed that Bacillus subtilis, Bacillus licheniformis, B. pumilus, B. cereus, B. badius, B.sphaericus and B. fusiformis are microorganisms present and involved in Bikalga fermentation. Parkouda et al. [34] were showed that these condiments contained several Bacillus strains which play a key role during fermentation for the manufacture of the product. Micro-organims involves or responsible for the fermentation of Soumbala have been identified as Bacillus spp. with B. subtilis as the predominant species [26,2,25,12,24].

The initial boiling of Parkii biglobosa and Hibiscus sabdariffa seeds during their traditional preparation process of these two Condiments may be way of thermotorant microorganisms selection and destroyed non-Bacillus species; this could explain the dominance of Bacillus species in Soumbala and Bikalga. However there is no information or few data on numeration of the thermotorant flora of these two condiments well consumed in Burkina Faso. This work deals with numeration and identification of thermotorant bacteria from Soumbala and Bikalga.

Materials and methods

Collection of Samples:

Bikalga and Soumbala samples were bought from various small markets (Table1) of Ouagadougou and Gaoua. Samples were transported at the microbiology laboratory immediately after collection and analysed or stored under refrigeration until analysis.

Microbial Analysis:

10 g of each sample was placed in a sterile stomacher bag containing 90 ml of peptone saline water and subsequent serial dilution up to [10.sup.-10] of each sample was made. Before analysis each sample (10 g plus 90ml peptone saline water) was heated at 90[degrees]C, 100[degrees]C, 105[degrees]C for 15 minutes. 0.2 ml of each appropriate dilution was inoculated triplicate in Plate Count Agar and incubated at 37[degrees]C for 24 H for viable colonies count. After incubation developed colonies on agar were counted for each sample.

Micro-organisms Primary Characterization:

To confirm that the isolated colonies are belonging to Bacillus species different tests were made. Sixten bacteria colonies from Soumbala and Bikalga were selected (5 from Soumbala and 11 from Bikalga), isolated and characterized according to methods described by Harrigan and McCance [13], ICSMF [15]; Collins and Lyne [10]. After colony counting different colonies were picked at random from petri dish and identified by cell morphology and motility, gram reaction, glucose metabolism [14], lecithinase, VP (Voges-Proskauer test), urease, catalase, starch hydrolysis, casein hydrolysis, sporulation, NaCl.

Selected Strains Characterization by PCR using Bacillus genus Specific Primer:

DNA Extraction and Preparation:

5ml of an overnight culture were used, cells were harvested by centrifugation and washed twice with Nacl 0.9%. Isolation of DNA was conducted with PROMEGA kit and protocol technics (Promega Corporation, Madison, WI 53711-5399 USA).

PCR Reaction and Electrophoresis:

Primers (B--K1/R: 5'-TCACCAAGGCRACGATGCG-3' and B-K1/F: 5'-CGTATTCACCGCGGCATG-3') were used for the identification of Bacillus strains [39]. PCR mixture consisted of 1.5 [micro]l of each primer (20 [micro]M), 12.5 [micro]l (1.2 [micro]M) of Master Mix (Fermentas GMBH, St-Leon, Rot, Germany), 8 [micro]l of [H.sub.2]O, 1.5 [micro]l of DNA (10-30 ng. [micro][l.sup.-1]) in a final volume of 25 [micro]l. Thermal cycling was carried out using a Eppendorf AG (Hamburg, German) Mastercycler gradient as follows: initial denaturation at 94[degrees]C for 3 min followed by 30 cycles of denaturation at 94[degrees]C for 1min, primer annealing at 43[degrees]C for 30 s primer extension at 72[degrees]C for 45 s and final extension at 72[degrees]C for 10min. The expected fragment size is 1114bp. 10[micro]l of the amplified products of PCR were analysed by electrophoresis in 1% (w/v) agarose gels stained with ethidium bromide (0.5 mg [m.sup.-1]). The gels were visualised with an Ultraviolet Illuminator and Digitally recorder (GelDOC Bio-Rad, Hercule, USA).

Dermination of Proteolytic Activity:

Proteolytic activity of the isolates was determined using skim milk agar. The culture supernatant were spotted on the surface of the skim milk agar plates and incubated at 37[degrees]C for 24h. The development of a clear zone was considered as proteolytic activity [8].

Amylolytic Activity:

For determination of amylolytic activity starch agar was used and incubated at 37[degrees]C for 48h. Enzymatic activity was indicated as clearing zones on the plates. For observation of amylolytic activity the agar plates were flooded with iodine solution.

Results and Discussion


The count of thermotolerant bacteria in Soumbala (Table 2) at 90[degrees]C, 95[degrees]C, 100[degrees]C and 105[degrees]C ranged from 1.35 x [10.sup.9] to 3 x [10.sup.9] cfu/g, 1.24 x [10.sup.9] to 2.78 x [10.sup.9] cfu/g, 0.54 x [10.sup.9] to 2.27 x [10.sup.9] cfu/g, 3.5 x [10.sup.7] to 1.54 x [10.sup.9] cfu/g respectively. Thermotolerant bacteria in Bikalga (Table 3) at 90[degrees]C, 95[degrees]C, 100[degrees]C and 105[degrees]C ranged from 1.9x[10.sup.7] to 12.8x [10.sup.7] cfu/g, 2x[10.sup.6] to 11.2 x [10.sup.7] cfu/g, 0 to 9.6 x [10.sup.7] cfu/g, 0 to 7.6 x [10.sup.7] cfu/g. These results indicated that Soumaba contained more themotolerant bacteria than Bikalaga. In all samples samples of Bikalga and Soumabala we numbered thermotolerant bacteria at 90[degrees]C, 95[degrees]C, 100[degrees]C and 105[degrees]C, except in of Bikalga samples B2 at 10[degrees]5C and B3 at 100[degrees]C and 105[degrees]C we didn't numbered no themotolerant . This analysis of heat treatment microbial flora in Bikalga and Soumbala revealed the occurring of both Bacillus genera. Colonies morphology on nutrient agar clearly indicates the presence of more than one type of isolate, confirming thus the diversity of flora.

-Primary Characterization, Proteolytic and Amylolytic Activities:

Five (5) representative strains from Soumbala and eleven (11) representative strains from Bikalga were selected for morphological and biochemical characterization (Table 4). All selected strains are Gram-positive, endospore-forming catalase positive; these selected strains are identified according to Biochemical, morphology and PCR characteristics (Table 4): five (5) are Bacillus coagulans, four (4) are Bacillus cereus, three (3) are Bacillus pumilis, two (2) are Bacillus firmus, one (1) is Bacillus subtilis and one (1) is Bacillus licheniformis. Bacillus subtilis, Bacillus cereus, Bacillus pumilus were identified in Soumbala and Bacillus licheniformis, Bacillus cereus, Bacillus pumilus, Bacillus coagulans, Bacillus firmus were identified in Bikalga (Table 4). Further examination (Table 4) showed that the selected strains hydrolysed casein (15 strains) and starch (6 strains), grew in medium with 5 % to 8% NaCl (10 strains for 5% NaCl, 6 strains for 6% NaCl, 5 strains for 7% NaCl and 3 strains for 8% NaCl). The heating has destroyed the non-Bacillus species and others microorganisms in volved in Soumbala and Bikalga process. Ouoba et al. [33] reported extra and intracellular protease enzymes pronounced activity in Bacillus species. Bacillus counts in soybean dawadawa by Ogbadu and Okagbue [25], Opai-Tetteh [30] and Omafuvbe et al. (2000) were estimated in order of [10.sup.10] to [10.sup.11] cfu/g.

PCR Characterization:

The amplification with PCR using Bacillus genus primers gave expected fragments 1114 bp for the sixteen selected strains (Figure 1). These results confirm that the selected strains are effectively belonging to the Bacillus genus.


The count of bacteria was dominated by Grampositive, catalase-positive, rods, endospore-forming all these characteristics are those of Bacillus group , this fact is due to heating during the numeration at 90[degrees]C ,95[degrees]C, 100[degrees]C and 105[degrees]C for 15 minutes. The initial boiling of Parkii biglobosa and Hibiscus sabdariffa seeds during their traditional preparation process of these two condiments and the heating during the numeration of microorganisms in our samples may be way of thermotorant microorganism selection and destroyed non-Bacillus species; this could explain the dominance of Bacillus species in Soumbala and Bikalga. Also The pH of these two condiments are alkaline this fact can explain why we get otptimum spore heat resistance and high number of Bacillus counted.

Among Sixteen (16) selected strains 15 have proteolytic activity this result indicate that Bikalga and Soumbala are rich in proteolytic Bacillus spp.


Bacillus spp. was found to be dominant in iru and sonru [3] in dawadawa [25] in Soumbala [12,5,33,31] in Meju and others fermented soybean foods [19,9,20,18]. Lipolysis and proteolysis are important processus observed in fermented condiments [1,33]. Bacillus subtilis was encountered and used as starter culture for production of locust bean daddawa [16,27].

B. subtilis was encountered and used as starter culture for production of locust bean daddawa [16,27].

Proteins degradation in fermented condiment contributes to the development of the texture, flour and organoleptic quality. Lipolysis and proteolysis are important processus observed in fermented condiments [1,39,33]. Proteolytic activity of Bacillus can lead to the liberation of bioactive compounds which could give somes interesting role and characteristics of fermented condiments. The production of several proteolytic enzymes enhance the growth of Bacillus species in fermented food [21]. The hydrolysis of protein during Bikalga and Soumbala fermentation leads to the alkalinity due to the protease and deaminase enzymes produced by the Bacillus group. Amylase activity has been reported to be the second most important activity during condiment production. Strain with protease activity will be relevant for flavor development and release of amino acids during condiment process. Strains with good protease and amylase will be more relevant as starter culture.

The Bacillus spp play important role in Maari fermentation [35] such as development of aroma and flavor. Bacillus subtilis was also present in others similar alkaline fermented condiments such as afitin [4] dawadawa [11] iru [26], Soumbala [31] kinema [36] mbuja [22]. Bacillus subtilis and Bacillus licheniformis members of Bacillus group have same ability to degrade locust bean proteins. Proteins degradation is likely to produce essential amino acids and bioactive peptides useful for human nutrition.

Fermented soybean foods have several functional properties like antioxidant, antimutagenesis, immunomodulatory, antithrombosis and fibrinolytic [7,9].


This work indicates that Bacillus group are endospore-forming thermotolerant bacteria involved in Soumbala and Bikalaga fermentation and contribute to their sensor and nutritional characteristics. Enumeration result shows that the thermoteroleant bacteria are important and dominant in these two condiments. These result can be explain by the fact that Bikalga and Soumbala are condiments product of traditional uncontrolled alkaline fermentation of Hibiscus sabdariffa and Parkii biglobosa seeds respectively after long time cooking process.


[1.] Aderibigbe, E.Y. and S.A. Odunfa, 1988. Purification and characterization of extracellular proteinases excreted by a strain of Bacillus subtilis BS2, isolated from fermented African locust bean "iru". Journal of Applied Bacteriology, 65: 361-369.

[2.] Antai, S.P. and M.H. Ibrahim, 1986. Microorganisms associated with African locust bean (Parkia filicoidea welw) fermentation for dawadawa production. Journal of Applied Bacteriology, 61: 145-148.

[3.] Azokpota, P., D.J. Hounhouigan and M.C. Nago, 2006. Microbiological and chemical changes during the fermentation of African locust bean (Parkia biglobosa) to produce afitin, iru and sonru, three traditional condiments produced in Benin. International Journal Food Microbiology, 107: 304-309.

[4.] Azokpota, P., P.L. Moller, J.D. Houghouigan and M. Jakobsen, 2008. Biodiversity of predominant Bacillus isolated from afitin, iru and sonru at different fermentation time. International Journal of Biological and Chemical Sciences, 1: 211-222.

[5.] Beaumont, M., 2002. Flavouring composition prepared by fermentation with Bacillus spp. International Journal Food Microbiology, 75: 189-196.

[6.] Bengaly, M.D., 2001. Etude microbiologique et valeur nutritionnelle d'un condiment traditionnel riche en proteines, obtenu par fermentation naturelle des graines d'Hibiscus sabdariffa. Memoire de Doctorat, UFR-SVT, Universite de Ouagadougou.116 pp.

[7.] Candela, T. and A. Fouet, 2006. Poly-gamma-glutamate in bacteria. Mol Microbiol., 60: 1091-1098

[8.] Chantawannakul, P., A. Oncharoen, K. Klanbut, E. Chukeatirote and S. Lumyong, 2002. Characterization of proteases of Bacillus subtilis strain 38 isolated from traditionally fermented soybean in northern Thailand. Sci Asia, 28: 241-245.

[9.] Cho, S.J., S.H. Oh, R.D. Pridmore, M.A. Juillerat and C.H. Lee, 2003. Purification and characterization of proteases from Bacillus amyloliquefaciens isolated from traditional soybean fermentation starter. J Agric Food Chem, 51:7664-7670.

[10.] Colins, C.H. and P.M. Lyne, 1984. Microbiological Methods 5th Ed. Butterworth and Co. Publ. Ltd., London, pp: 331-345.

[11.] Dakwa, S., E. Sakyi-Dawson, C. Diako, N.T. Annan and W.K. Amoa-Awua, 2005. Effect on the fermentation of soybeans into dawadawa (soydawadawa). International Journal of Food Microbiology, 104: 69-82.

[12.] Diawara, B., L. Sawadogo and I.Z. Kabore, 1992. Contribution a l'etude des procedes traditionnels de fabrication du soumbala au Burkina Faso. Aspects biochimiques, microbiologiques et technologiques. (CNRST: Ouagadougou, Burkina Faso).Sciences et Techniques, 20 : 5-14.

[13.] Harrigan, W.F. and M. Mc Cance, 1976. Laboratory Methods in Food and Diary Microbiology. Academic Press, London, pp: 225-231.

[14.] Hugh, R. and Leifson, 1953. The taxonomic significance of fermentative versus oxidative metabolism of carbohydrates by various Gram-negative bacteria. Journal of Bacteriology, 66: 24-26.

[15.] ICSMF, 1988. Micro-organism in Foods I. Their significance and Methods of Enumeration (Elliott, R.P., D.S. Clark, K.H. Lewis, H. Lundbeck, Olson, Je.J.C. and B.J. Simonsen). 15 University of Toronto Press, Toronto, pp: 105-155.

[16.] Ikenebomeh, M.J., R. Kok and J.M. Ingram, 1986. Processing and fermentation of the African locust bean (Parkia filicoidea Welw.) to produce dawadawa. J. Sci. Food Agric, 37: 273-282.

[17.] Jeyaram, K., W. Mohendro Singh, T. Premarani, A. Ranjita Devi, K. Selina Chanu, N.C. Talukdar and M. Rohinikumar Sing, 2008. Molecular identification of dominant microflora associated with 'Hawaijar'--A tradional fermented soybean (Glycine max (L.)) food of Manipur, India. International Journal of Food Microbiology, 122:259-268.

[18.] Jung, J.H. and H.C. Chang, 2009. Antifungal activity of Bacillus polyfermenticus CJ6 isolated from Meju. J. Korean Soc. Food Sci. Nutr, 38:509-516.

[19.] Kim, D., D. Lim, S. Bai and S. Chun, 1997. Fermentation characteristics of whole soybean meju model system inoculated with for Bacillus strains. Korean Journal of Food Science and Technology, 29: 1006-1015.

[20.] Kwon, G.H., H.A. Lee, J.Y. Park, J.S. Kim, J. Lim, C.S. Park, D.Y. Kwon, Y.S. Kim and J.H. Kim, 2009. Development of a RAPD-PCR method for identification of Bacillus species isolated from Cheonggukjang. Int J. Food Microbiol, 129: 282-287.

[21.] Mo, A.Y., B. Kwon, S. Kamala-Kannan, K.J. Lee, B.T. Oh, D.H. Kim, M.S. Yang, J.H. Kim and S.M. Park, 2010. Isolation and characterization of Bacillus polyfermenticus isolated from Meju, Korean soybean fermentation starter. World Journal of Microbiology and Biotechnology, 26: 1099-1105.

[22.] Mohamadou, B.A., C.M.F. Mbofung and D. Thouvenot, 2007. Functional potential of a product from traditional biotechnology: antioxidant and probiotic potential of Mbuja, produced by fermentation of Hibiscus sabdariffa seeds in Cameroon. J. Food Technol, 5 (2): 164-168.

[23.] N'Dir, B., R.D. Gningue, N.D.G. Keita, M. Souane, L. Laurent, C. Cornelius, and P. Thonard, 1997. Caracteristiques microbiologiques et organoleptiques du netetu du commerce. Cahiers Agricultures, 6: 299-304.

[24.] N'Dir, B., C. Hbid, C. Cornelius, D. Roblain, Ph. Jacques, F. Vanhentenryck, M. Diop et Ph. Thonart, 1994. Proprietes antifongiques de la microflore sporulee du netetu. Cahiers Agricultures, 3: 23-30.

[25.] Odunfa, S.A. and O.B. Oyewole, 1986. Identification of Bacillus species from iru, a fermented African locust bean product. Journal of Basic Microbiology, 26: 101-108.

[26.] Odunfa, S.A., 1981. Microorganisms associated with fermentation of African locust bean (Parkia filicoidea) during iru preparation. Journal of Plant Foods, 3: 245-250.

[27.] Ogbadu, L. and R.N. Okagbue, 1988. Bacterial fermentation of soya bean for daddawa production. J. Appl. Bacteriol., 65: 353-356.

[28.] Oguntoyinbo, F.A., M. Huch, G.S. Cho, U. Schillinger, W.H. Holzapfel, A.I. Sanni and C.M. Franz, 2010. Diversity of bacillus species isolated from okpehe, a traditional fermented soup condiment from Nigeria. Journal of Food Protection, 73: 870-878.

[29.] Omafuvbe, B.O., O.O. Shonukan and S.H. Abiose, 2000. Microbiological and biochemical changes in the traditional fermentation of soybeans for Fsoydaddawa--Nigerian food condiment. Food Microbiol, 17: 469-474.

[30.] Opai-Tetteh, S., 1999. The microbial activities involved in the alkaline fermentation of soybeans into dawadawa. MPhil. thesis. University of Ghana.

[31.] Ouoba, L.I.I., B. Diawara, W.K. Amoa-Awua, A.S. Traore and P. Lange Moller, 2004. Genotyping of starter cultures of Bacillus subtilis and Bacillus pumilus for fermentation of African locust bean (Parkia biglobosa) to produce Soumbala. International Journal of Food Microbiology, 90: 197-205.

[32.] Ouoba, L.I.I., C. Parkouda, B. Diawara, C. Scotti and A.H. Varnam, 2007. Identification of Bacillus spp from Bikalga, fermented seeds of Hibiscus sabdariffa: phenotypic and genotypic characterization. Journal of Applied Microbiology, 104: 122-131.

[33.] Ouoba, L.I.I., K.B. Rechinger, B. Diawara, A.S. Traore and M. Jakobsen, 2003. Degradation of proteins during the fermentation of African locust bean (Parkia biglobosa) by strains of Bacillus subtilis and Bacillus pumilus for production of Soumbala. Journal of Applied Microbiology, 94: 396-402.

[34.] Parkouda, C., B. Diawara and L.I.I. Ouoba, 2008. Technology and physico-chemical characteristics of Bikalga, alkaline fermented seeds of Hibiscus sabdariffa. African Journal of Biotechnology, 7: 916-922.

[35.] Parkouda, C., D.S. Nielsen, P. Azokpota, L.I.I. Ouoba, W.K. Amoa-Awua, L. Thorsen, J.D. Hounhouigan, J.S. Jensen, K. Tano-Debrah, B. Diawara and M. Jakobsen, 2009. The microbiology of alkalinefermentation of indigenous seeds used as food condiments in Africa and Asia. Critical Reviews in Microbiology, 35: 139-156.

[36.] Sarkar, P.K., B. Hasenack and M.J.R. Nout, 2002. Diversity and functionality of Bacillus and related genera isolated from spontaneously fermented soybeans (Indian Kinema) and locust bean (African Soumbala). International Journal of Food Microbiology, 77: 175-186.

[37.] Sopade, P. A., S. Ajisegiri Emanuel and B. Abass Adebayo, 1996. Moisture sorption isotherms of dawadawa, a fermented African locust bean (Parkia biglobosa Jacq. Benth). Food control, 7: 153-156.

[38.] Wang, J. and D.Y.C. Fung, 1996. Alkaline fermented foods: a review with emphasis on pidam fermentation. Critical Reviews in Microbiol, 22: 101-138.

[39.] Wu, X.Y., Walker, M.J., Hornitzky, M., Chin, J., 2006. Development of a group-specific PCR combined with ARDRA for the identification of Bacillus species of environmental significance. Journal of Microbiological Methods, 64: 107-119.

[40.] Yagoub, A.E.G.A., B.E. Mohamed, A.H. Ahmed and A.H.E. Tinay, 2004. Study on Furundu, a traditional Sudanese fermented roselle (Hibiscus sabdariffa) seed: effect on in vitro protein digestibility, chemical composition and functional properties of the total proteins. Journal of Agriculture and Food Chemistry, 52: 6143-6150.

(1) Savadogo Aly, (1) Ilboudo A. Jules, (2) Gnankine Olivier, (1) Traore Alfred S

(1) Laboratoire de Microbiologie et de Biotechnologie Centre de Recherche en Sciences Biologiques, Alimentaires et Nutritionnelles(CRSBAN). Departement de Biochimie-Microbiologie (DBM) ; Unite de Formation et de Recherche en Sciences de la Vie et de la Terre (UFR/SVT) ; UNIVERSITE DE OUAGADOUGOU, Tel/Fax (226) 50 33 73 73 03 BP 7'3', Ouagadougou, BURKINA FASO

(2) Laboratoire d'Entomologie Fondamentale et Applique/UFR-SVT

Corresponding Author

Savadogo Aly, Departement de Biochimie-Microbiologie /CRSBAN /03 BP 7131 Ouagadougou, Burkina Faso

Tel/Fax (226)50 33 73 73

E-mail: or
Table 1: Sampling of Bikalga and Soumbala

Samples Codes             samples    Number of sample

S1, S3, S4, S6, S9, S10   Soumbala   6
B1,B2, B5, B7, B10        Bikalga    5
S2                        Soumbala   1
S5                        Soumbala   1
S7                        Soumbala   1
B9                        Bikalga    1
S8                        Soumbala   1
B6                        Bikalga    1
B3, B4                    Bikalga    1
B8                        Bikalga    1

Samples Codes             Place of sampling

S1, S3, S4, S6, S9, S10   Zogona market
B1,B2, B5, B7, B10        (Ouagadougou)
S2                        Somgande market (Ouagadougou)
S5                        Goua market (Gaoua)
S7                        Nabigyaar market
B9                        (Ouagadougou)
S8                        Zone I market
B6                        (Ouagadougou)
B3, B4                    Karpala Market(Ouagadougou)
B8                        Goughin (Ouagadougou)
                          S=Soumbala, B= Bikalga

Table 2: Microbial analysis of Soumbala
after treatment at different temperature.

Soumbala samples   90[degrees]C        95[degrees]C

CFU [g.sup.-1]

S1                 2.76 x [10.sup.9]   2 x [10.sup.9]
S2                 2.81 x [10.sup.9]   1.8 x [10.sup.9]
S3                 2.96 x [10.sup.9]   2.56 x [10.sup.9]
S4                 2.89 x [10.sup.9]   2.69 x [10.sup.9]
S5                 3 x [10.sup.9]      2.78 x [10.sup.9]
S6                 2.45 x [10.sup.9]   2.37 x [10.sup.9]
S7                 1.84 x [10.sup.9]   1.24 x [10.sup.9]
S8                 2.84 x [10.sup.9]   1.65 x [10.sup.9]
S9                 1.4 x [10.sup.9]    1.28 x [10.sup.9]
S10                1.35 x [10.sup.9]   1.3 x [10.sup.9]

Soumbala samples   100[degrees]C       105[degrees]C

CFU [g.sup.-1]

S1                 1.51 x [10.sup.9]   3.5 x [10.sup.7]
S2                 1.56 x [10.sup.9]   1.10 x [10.sup.9]
S3                 2.02 x [10.sup.9]   1.54 x [10.sup.9]
S4                 2.27 x [10.sup.9]   5.8 x [10.sup.8]
S5                 2.19 x [10.sup.9]   1.21 x [10.sup.9]
S6                 1.98 x [10.sup.9]   6.7 x [10.sup.8]
S7                 6.2 x [10.sup.8]    3.8 x [10.sup.7]
S8                 7.1 x [10.sup.8]    5 x [10.sup.8]
S9                 8.9 x [10.sup.8]    6.2 x [10.sup.8]
S10                5.4 x [10.sup.8]    1.12 x [10.sup.8]

Values are weighted means of three replicates

Table 3: Microbial analysis of Bikalga
after treatment at different temperature.

Bikalga samples   90[degrees]C        95[degrees]C

CFU [g.sup.-1]
B1                2.3 x [10.sup.7]    2.2 x [10.sup.7]
B2                3.6 x [10.sup.7]    4.5 x [10.sup.7]
B3                1.9 x [10.sup.7]    2. [10.sup.6]
B4                4.5 x [10.sup.7]    7.1 x [10.sup.7]
B5                5 x [10.sup.7]      6 x [10.sup.7]
B6                3.5 x [10.sup.7]    2.2 x [10.sup.7]
B7                6 x [10.sup.7]      5.3 x [10.sup.7]
B8                10 x [10.sup.7]     6.9 x [10.sup.7]
B9                12.5 x [10.sup.7]   11.2 x [10.sup.7]
B10               12.8 x [10.sup.7]   10 x [10.sup.7]

Bikalga samples   100[degrees]C      105[degrees]C

CFU [g.sup.-1]
B1                2.1 x [10.sup.7]   1 x [10.sup.7]
B2                3.2 x [10.sup.7]   0
B3                0                  0
B4                5.6 x [10.sup.7]   4.4 x [10.sup.7]
B5                5.5 x [10.sup.7]   7 x [10.sup.6]
B6                1.3 x [10.sup.7]   1.2 x [10.sup.7]
B7                4.2 x [10.sup.7]   2.7 x [10.sup.7]
B8                6.8 x [10.sup.7]   2 x [10.sup.7]
B9                3.2 x [10.sup.7]   2.9 x [10.sup.7]
B10               9.6 x [10.sup.7]   7.6 x [10.sup.7]

Values are weighted means of three replicates

Table 4: Biochemical and morphology characters of
isolates of Soumbala and Bikalga.

Isolated         Isolated at 90[degrees]C

Characters       S1        B1          B2

Gram             +         +           +
Catalase         +         +           +
Sporulation      +         +           +
Proskauer        +         -           +
Lecihinase       -         -           +
hydrolysis       -         -           -
hydrolysis       +14mm     +10mm       +18mm
Urease           -         -           -
NaCl (5%)        +         -           -
NaCl (6%)        +         -           -
NaCl (7%)        +         -           -
NaCl (8%)        -         -           -
Presumptive      B.        B.          B.
Identification   pumilus   coagulans   cereus

Isolated         Isolated at 95[degrees]C

Characters       B3        S2         S3       B4

Gram             +         +          +        +
Catalase         +         +          +        +
Sporulation      +         +          +        +
Proskauer        +         -          -        +
Lecihinase       -         -          +        -
hydrolysis       -         +13mm      +13mm    +16mm
hydrolysis       +14mm     +10mm      +8mm     +10mm
Urease           +         -          +        -
NaCl (5%)        +         +          +        +
NaCl (6%)        +         +          -        +
NaCl (7%)        +         +          -        +
NaCl (8%)        +         -          -        +
Presumptive      B.        B.         B.       B.
Identification   pumilus   subtilis   cereus   licheniformis

Isolated         Isolated at 100[degrees]C

Characters       B5          S4        B6       B7

Gram             +           +         +        +
Catalase         +           +         +        +
Sporulation      +           +         +        +
Proskauer        +           +         -        +
Lecihinase       -           -         -        -
hydrolysis       +16mm       -         +17mm    +15mm
hydrolysis       +10mm       +9mm      -0 mm    +12mm
Urease           -           -         +        -
NaCl (5%)        -           +         +        -
NaCl (6%)        -           -         +        -
NaCl (7%)        -           -         -        -
NaCl (8%)        -           -         -        -
Presumptive      B.          B.        B.       B.
Identification   coagulans   pumilus   firmus   coagulans

Isolated         Isolated at 105[degrees]C

Characters       B8       S5       B9          B10      B11

Gram             +        +        +           +        11
Catalase         +        +        +           +        0
Sporulation      +        +        +           +        0
Proskauer        -        +        +           +        -
Lecihinase       -        +        -           +        -
hydrolysis       +16mm    -        -           -        -
hydrolysis       +4mm     +11mm    +14mm       +6mm     +6mm
Urease           -        +        -           -        -
NaCl (5%)        +        -        -           +        6
NaCl (6%)        -        -        -           +        -
NaCl (7%)        -        -        -           +        -
NaCl (8%)        -        -        -           +        -
Presumptive      B.       B.       B.          B.       B.
Identification   firmus   cereus   coagulans   cereus   coagulans

+ = positive reaction; -= negative reaction; B = strains from
Bikalga (B1, B2,B3, B4, B5, B6, B7, B8, B9, B10); S = strains
from Soumbala (S1, S2, S3, S4, S5)
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Title Annotation:Original Article
Author:Aly, Savadogo; Jules, Ilboudo A.; Olivier, Gnankine; Traore, Alfred S.
Publication:Advances in Environmental Biology
Article Type:Report
Geographic Code:1USA
Date:Aug 1, 2011
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