Ablo is wet bread, slightly salty and sweet, steamed and sold in the form of pellets. The objective of the study was to assess the microbiological quality of millet-based Ablo and sorghum-based Ablo. The methodology adopted was to perform production tests followed by analyses in the laboratory. The results showed that the lactic acid bacteria, yeasts and moulds were the dominant micro flora of the millet- based Ablo and sorghum-based Ablo. Furthermore, analysis showed that the millet-based Ablo and the sorghum-based Ablo doesn’t contain either total coli forms, coli forms thermo tolerant or consequently Escherichia coli.

Published by the  International journal of Microbiology and Mycology (IJMM

Adoukonou Awo Sagbadja Hubert

Assogba Karl

Banon Sèmèvo B. Jultesse

Bleoussi Roseline

Bokossa Yaou Innocent Padonou

Djogbe Anayce

Sachi Pivot

Tchekessi Comlan K. Célestin

English

ZENODO

 7 Jultesse et al.   International Journal of Microbiology and Mycology | IJMM | pISSN: 2309-4796 http://www.innspub.net Vol. 5, No. 6, p. 7-13, 2017   Microbiological characterization of the millet-based (Pennisetum glaucum) Ablo and sorghum-based (Sorghum bicolor) Ablo produced in Benin  Banon Sèmèvo B. Jultesse*1, Bokossa Yaou Innocent Padonou1, Adoukonou Awo Sagbadja Hubert2, Tchekessi Comlan K. Célestin1, Bleoussi Roseline1, Sachi Pivot1, Djogbe Anayce1, Assogba Karl1  1Unité de Recherche en Sécurité Sanitaire des Aliments (URSSA), Département de Biologie Végétale, Faculté des Sciences et Techniques (FAST), Université d'Abomey-Calavi (UAC), Bénin 2Laboratoire des Ressources Génétiques et d’Amélioration des Espèces (LaREGAME), Département de Génétique et des Biotechnologies, Faculté des Sciences et Techniques (FAST), Université d'Abomey-Calavi (UAC), Bénin  Keywords: Ablo, Microflora, Millet, Sorghum, Benin Publication date: June 30, 2017 Abstract Ablo is wet bread, slightly salty and sweet, steamed and sold in the form of pellets. The objective of the study was to assess the microbiological quality of millet-based Ablo and sorghum-based Ablo. The methodology adopted was to perform production tests followed by analyses in the laboratory. The results showed that the lactic acid bacteria, yeasts and moulds were the dominant micro flora of the millet- based Ablo and sorghum-based Ablo. Furthermore, analysis showed that the millet-based Ablo and the sorghum-based Ablo doesn’t contain either total coli forms, coli forms thermo tolerant or consequently Escherichia coli. * Corresponding  Author:  Banon Sèmèvo B. Jultesse  bbanonjultesse@yahoo.fr                    Open Access                                                                                          RESEARCH  PAPER                8 Jultesse et al.  Introduction In Benin, corn, main cereal in the food system, investigated a multitude of transformations leading to more than 40 different products (Nago, 1989). Among these many products, Fig. the Ablo, a dough drying corn, slightly salty and sweet of origin Mina prepared from the mawe, sometimes added to wheat flour and of various ingredients (yeast, salt, sugar). Traditionally consumed in Benin, Togo and Ghana, he is poised to win the West-African (Akakpo, 2013). However, the product of our days, is subject to significant changes in its preparation, mainly regarding the raw (maize, rice and wheat). The partial introduction or the substitution of rice in the preparation of the Ablo is a fairly recent but increasingly observed practice in urban areas. These types of Ablo meet consumer’s acceptability and allow satisfying the desire of the customer in relation to its requirement related to its new styles and eating behaviours (Akakpo, 2013).  The expansion of cities and the distance between the home and the workplace promote the increase of consumption outside the home, in the small restaurants and more particularly with the street vendors (Banon, 2012). Today, street food receives a more numerous and diverse clientele including all socio-professional categories (Bokossa et al., 2013). It is an important area of activities involving large sums of money and source of jobs in a large proportion of the population, including women and families (Aholou-yeyi, 2007). In urban areas, low productivity of these traditional crafts enterprises and the tedium of some unit operations induce profound changes at the level of processes. A thorough knowledge of the endogenous technologies for food processing and their variability would contribute to their optimization.  This study focused on two new types of Ablo namely: the sorghum-based Ablo and millet-based Ablo. The presented information is gotten following experiences of consistent production of analyses in the laboratory. The study permitted us to master the microbiological quality of the two new types of Ablo. The study aimsto assess the microbiological quality of the sorghum-based Abloand millet-based Ablo.   Materials and methods The productions were made in the research unit in safety health food (URSSA) of the laboratory of Microbiology and of the food Technologies (LAMITA) in the Faculty of science and technology (FAST) of the University of Abomey-Calavi (UAC).  Materials Plant material  Sorghum (Sorghum bicolor) of red color designated in local language fon by "abokun" and the small millet (Pennisetum glaucum) greenish color called "likun" in fon were used. Wheat flour also served as plant material. These cereals were purchased at the Dantokpa Cotonou's international market.   Biological material  The instant yeast (Saccharomyces cerevisiae) of trademark PASHA made in Turkey by DOSU MAYA MAYACILIK A.S. company certified ISO 9001: 2008 has been used. It was purchased at the Dantokpa Cotonou's international market. It is used as a leaven in the manufacturing technology of the Ablo (Ahokpe, 2005; Aholou-yeyi, 2007; Bokossa et al., 2013).  Laboratory equipment The material of analysis consists of classical material used for microbiological handling.  Other materials  The material used for the different manufacturing was constituted of ingredients (sugar, salt) and usual equipment of for the production of Ablo such as basins, plastic buckets, pots, a spatula, a whip, grinding wheels, a sieve and a fireplace. The water of the national society of waters in Benin (SONEB) was also used.   9 Jultesse et al.  Methods Experimental method The production tests were conducted according to the original method described by Aholou-yeyi (2007) modified. The difference in this technology was the use of other types of cereals such as millet and sorghum and the reduction of the fermentation time. Each test was repeated three times in the laboratory. We realized samples at the end of the fermentation and after cooking. The various microbiological analyses on these samples.  Methods of analysis Microbiological analyses consisted of counting the total flora, of yeast and moulds, lactic acid bacteria, total and thermo tolerant coli forms by crops on synthetic media. The counts were taken by counting the colonies (Guiraud and Galzy, 1980). Microbiological analyses were conducted in three repetitions on each sample of the products.  Preparation of suspensions mothers and decimal dilutions Before seeding, it was imperative to sterilize glassware and prepare the suspension-mother and the successive decimal dilutions thereof. Here, all operations were conducted in the field of the flame of the Bunsen burner or under the hood. According to the AFNOR (1999), test socket for the preparation of suspensions-mothers was 10g for each sample. To get the suspension-mother, these 10g of sample were collected aseptically in a previously sterilized erlenmeyer. Then, 90ml of water solution sterile peptone (EPS) prepared following the indications of the manufacturer have been added. The mixture was homogenized with the brand vortex Homogenizer. One milliliter of this suspension-mother has been taken and added to the first tube of a series of test tubes to screw containing 9ml of the solution of dilution and homogenization has been carried out. The same procedure was repeated by taking 1ml of the resulting suspension this time, and by adding it to the tube in the series and so on until the last decimal dilution desired.  Count of total flora (ISO 4833, 2003) From each prepared decimal dilution, 1ml was taken and introduced into a sterile Petri dish box. 10 to 15ml of the previously melted plate Count Agar (PCA Oxoid CM 0325) solid medium were added, and then all was perfectly homogenized. After complete solidification, plates were incubated at 30°C for 48 to 72 h.  Enumeration of yeasts and moulds (NF ISO 21527-2, 2008) The medium that was used for their research was the word Dextrose Agar (Oxoid CM 0041) to chloramphenicol (0.05 g/L). Previously prepared and sterilized, 10 to 15 ml of Word have been sunk in sterile boxes of Petri dish and left to solidify. From each prepared dilution of decimal, 0.1ml was collected and spread on the surface of the agar using a spreader rake. The counting of colonies was made after incubation plates at 25°C for 3 to 5 days.  Enumeration of lactic acid bacteria (NF ISO 15214 (V 08-030), 1998) Lactic bacteria count was based on a culture in depth of 1ml of each prepared dilution of decimal. So, 1ml of each prepared dilution was placed in sterile Petri dish boxes. Then, 10 to 15 ml of medium of Man Rogosa Sharpe Agar (MRS Agar CM 0361) have been added. After solidification, a second layer was conducted. Petri boxes were incubated at 30°C for 48h.  Enumeration of total coliforms (NF ISO 4832 (V 08-015), 2006)  One (1) ml of each prepared dilution was placed in sterile Petri dish boxes. There was paid then the violet Red Bile Agar (VRBA Oxoid CM 0107). After solidification, a second layer was conducted. Boxes of Petri dishes were incubated at 37°C for 24 h.  Enumeration of thermo tolerant coli forms (NF V08-060, 2009) One (1) ml of each prepared dilution was placed in sterile Petri dish boxes. There was paid then the violet Red Bile Agar (VRBA Oxoid CM 0107).   10 Jultesse et al.  After solidification, a second layer was conducted. Boxes of Petri dishes were incubated at 44°C for 24-48h.   Expression of the results The results were obtained from the count of net settlements. The counts were taken by dials. The results were expressed in cfu per gram of product according to the method described by Bokossa in 2007.  Statistical analyses of the data The collected data were analyzed using SPSS 16 and MINITAB 14 software. MINITAB 14 software was used to verify the conditions of application of the statistical tests.   These were made with the software SPSS 16 which has to do the analyses of variance (ANOVA) and Tukey test for the comparison of averages. The chosen significance level was 5% (p < 0.05).  Results Results of microbiological analyses of the fermented dough of the two types Ablo have been indicated in table I and table II. Microbiological analysis has revealed the absence of germs such as total coli forms and thermo tolerant coli forms. Load the total flora of lactic acid bacteria, yeasts and moulds was higher in the fermented pulp (millet and sorghum) uncooked in the two types of Ablo after cooking.   Thus, the total flora spent of 7.44 ± 0.12 to 2.95± 0.17 Log10 cfu/g, lactic acid bacteria of 6.073±0.28 to 4.80±0.64Log10 cfu/g and the yeast and moulds of 7.50±0.14 to 4.22±0.45Log10 cfu/g (Table I).  In the fermented pulp of uncooked sorghum and the sorghum-based Ablo, the load of the total flora, of lactic acid bacteria and yeasts and moulds ranged respectively from 6. 42±0.23 at 2.03±0.13Log10 cfu/g, of 8.04±0.66 to4.99±0.73Log10 cfu/g and of 8.53±0.32to5.42± 0.78 Log10 cfu/g (Table II). Table I. Microbiological characteristics of millet fermented dough and that of millet-based Ablo. Product average values of the desired microorganisms (Log10 cfu/g)   Total flora Yeasts and moulds Lacticacid bacteria Total coliforms Thermotolerant coliforms Millet fermented dough 7.44±0.12a  7.5±0.14a 6.73±0.28a Absenta Absenta Millet-based Ablo 2.95±0.17b 4.22±0,45b 4.80±0,64b Absenta Absenta Mean values with the same letter in the same column are not significantly different at the 5% level. Data represents in table is mean of three replications. ± Standard deviation.  Table II. Microbiological characteristics of the fermented dough of sorghum and the sorghum-based Ablo. Product average values of the desired microorganisms (Log10 cfu/g)  Total flora Yeasts and moulds Lacticacid bacteria Total coliforms Thermotolerant coliform  Sorghum fermented dough 6.42±0.23a  8.53±0.32 a  8.04±0.66a  Absent a  Absent a  Sorghum-based Ablo 2.03 ±0.13 b 5.42± 0.78b  4.99±0.73 b  Absent a  Absent a  Mean values with the same letter in the same column are not significantly different at the 5% level. Data represents in table is mean of three replications. ± Standard deviation.  Discussion  The most common microorganisms in fermented doughs, on the millet-based Ablo and sorghum-based Ablo are yeasts, mould and lactic acid bacteria. The same comments were made by Ahokpe (2005), by Aholou-yeyi (2007), by Banon (2012) and by Bokossa et al. (2013) who said that the dominant microflora of the Ablo consists of lactic acid bacteria and yeasts and moulds.  11 Jultesse et al.  These microorganisms come from the commercial yeast added but also from the contamination of flour during milling. The results are consistent with those obtained by Ahokpe (2005) and Aholou-yeyi (2007) on the Ablo corn who pointed out that the micro flora present in corn grain, the equipment used and in the mawe or the flour contaminated during grind play a role in the fermentation.  The Two types of Ablo developed contain neither total coliform, fecal coliform nor Escherichia coli. These results mean that good hygiene practices are met during production and indicate that the manufactured Ablo present no danger to human health. The same comments were made by Tchekessi et al., 2013, Banon (2012), Tchekessi (2012), Clabots (2007), Leclerc et al. (1977) reported that the absence of enterobacteria is a good indicator of the level of acceptable hygiene in the production of food. They said that the enterobacteria are witnesses of contamination after heat treatment and that their presence in large numbers indicates a failing hygiene during production but also an ability of the product to promote their development.  The total flora found in the two types of Ablo is due to contamination after cooking. These results are consistent with those of Turpin (2010) and Tchekessi (2015) which attested that the total flora encountered in the food after cooking comes from the environment. The reduction of the microbial load on products after cooking is explained by the fact that microorganisms are partly destroyed by heat. Cooking at the same time plays the role of pasteurization. Alais (1984) and Vignola (2002) show that the destruction of the microorganisms is function of two parameters such as the temperature and the duration of the heat treatment. Similarly, Leclerc et al. (1977), Akapko (2013), Agro (2013) and Bokossa et al. (2016) have mounted that a heat treatment for a few seconds at 72°C is sufficient to destroy the vegetative forms of microorganisms in food products.  Conclusion Yeasts, moulds and lactic acid bacteria are predominant germs of millet-based Ablo and sorghum-based Ablo. The absence of total coli forms and thermo tolerant coli forms is a good indicator of the level of acceptable hygiene during the production of the two types of Ablo. We can say that the millet-based Ablo and sorghum-based Ablo are healthy foods in microbiological terms and therefore present no danger to human health.  References AFNOR NF V 08-051 (Association Française de Normalisation, Norme Française Version 08-051). 1999. Microbiologie des aliments: dénombrementdes microorganismes par comptage des colonies obtenues à 30 degrés Celsius Méthode de routine. Firm Performances: Evidence From. Journal of Accountingand Economics 26, 149-178.  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Microbiological characterization of the millet-based (Pennisetum glaucum) Ablo and sorghum-based (Sorghum bicolor) Ablo produced in Benin

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Microbiological characterization of the millet-based (Pennisetum glaucum) Ablo and sorghum-based (Sorghum bicolor) Ablo produced in Benin

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