Study of the Bacteriological Profile of the Drink Tchakpalo (Sugar Must) Sold In Cotonou Schools in the Republic of Benin

1National University of Sciences, Engineering and Mathematics/ Department of Human Biology/ Laboratory of Clinical Biochemistry and Medical Microbiology, BENIN 2Abomey Calavi University, Senior Lecturer / General Medical Microbiology and Hospital Hygiene Abomey Calavi Politechnical School / Human Biology Engineering, BENIN 3Regional Institute of Public Health Ouidah BENIN 4Medicine Faculty, University of Parakou, Benin, 5Department of Clinical Biochemistry / Assistant, BENIN 6Department of Clinical Biochemistry / Assistant, BENIN 7Department of Clinical Biochemistry / Assistant, BENIN 8Full professor, Department of Biochemestry/ UAC, BENIN Abstract The objective of this study is to identify the germs present in the drink (sweet must) sold in Cotonou schools in the Republic of Benin. One hundred (100) samples of the sweet must drink were used for this study. All samples of the sweet must drink were taken from sterile 40 ml bottles. After carrying out the various bacteriological examinations, it emerged from the tests that the germs isolated belong to the genera Staphylococcus and Klebsiella. We obtained the following percentages: 52% Staphylococcus aureus; 28 % Staphylococcus sp; 12 % Klebsiella pneumoniae and 8 % Klebsiella oxytoca. It is therefore essential to raise awareness among the saleswomen on the respect of hygiene rules and standards.


INTRODUCTION
M an has been using fermentation for thousands of years to obtain food with improved nutritional value (1). In Africa, certain cereals such as sorghum, maize and millet are often made into a drink that includes an essential alcoholic fermentation step (2, 3). Indeed, often linked to traditions of hospitality and conviviality, they are part of the way of life of most families and serve to seal relationships between individuals (4). The empirical technological process involves a double fermentation: an alcoholic fermentation and a natural lactic fermentation (5). In addition, the production process suffers from a crucial lack of precision measuring instruments, good practices and hygiene standards (6).
Louis Pasteur's work has revolutionized bacteriology (7). Thus he demonstrated in 1859 that fermentation processes are caused by microorganisms and that this growth was not due to spontaneous generation. Bacteria are ubiquitous and are present in all types of biotopes found on earth. They can be isolated from soil, fresh, marine or brackish water, food, air, ocean depths, radioactive waste, the earth's crust, plants, on the skin and in the intestines of animals. In humans it has been calculated that 1012 bacteria colonize the skin, 1010 colonize the mouth and 1014 live in the intestine, which means that there are ten times more bacterial cells than human cells in the human body. There are about 40 million bacterial cells in one gram of soil and one million in one milliliter of fresh water (8). Thus the reservoirs of microorganisms that cause infections are multiple. They can be classified in two categories: environmental and human. The environmental reservoir is the place where the sweet must is prepared for drinking, the field where cereals (maize, millet and sorghum) are grown. The human reservoir, which is the most important source, has a variety of origins : on the one hand, by the Tchakpalo (sweet must) sellers, which causes their uncleanliness, for example faecal contamination; on the other hand, by the farmers, which causes the uncleanliness of the granaries, using water from rivers and lagoons, which the farmers use to water their fields; it should

Supplementary information
The online version of this article (https://doi.org/10.15520/ijcrr.v12i01.87 8) contains supplementary material, which is available to authorized users.

Corresponding Author: Dr AÏKOU Nicolas
National University of Sciences, Engineering and Mathematics/ Department of Human Biology/ Laboratory of Clinical Biochemistry and Medical Microbiology, BENIN Email: aikounicolas@yahoo.fr be noted that it is in these same waterways that these farmers do their washing, wash themselves, and do all their needs (9, 10). In view of the place this drink occupies in the batch of drinks we consume daily; in view of the number of people who consume it; in view of the praise it receives from the mass media, an indispensable study has been carried out on the bacteriological profile of the Tchakpalo drink sold in schools in Cotonou (Republic of Benin). In order to study the bacterial flora that this drink may contain, various bacteriological tests were carried out to determine the germs present in the Tchakpalo drink (sweet must).

PATIENTS AND METHOD
The SPACE-LABM laboratory in Cadjèhoun was used as a study framework. The laboratory is multidisciplinary and carries out research and service work in the following fields: Biochemistry -Clinical; General and Medical Microbiology; Hospital Hygiene; Biomedical Waste Management; Hospital Quality and Patient Safety.
This is a prospective study that lasted from October 2018 to March 2019. One hundred (100) 40 mL samples were taken from different sites and analyzed during this study. Various examinations were carried out, including macroscopic examination and microscopic examination: fresh and coloured. Biochemical tests including catalase, staphylocoagulase, oxidase, Dnase and Leminor's rapid gallery were carried out to identify the germs present in this drink. The antibiogram was carried out to observe the resistance of the isolated germs.

DISCUSSION
The objective of this study is to carry out the various bacteriological examinations that contribute to the determination of the germs present in the drink of   Tchakpalo (sweet must) in order to evaluate the quantity of these germs in the drink and to determine the antibiotic resistance profile.
From the analysis of Table 1 on the distribution of samples according to the sampling sites, we note a predominance of sampling in the zones of Cadjèhoun 30% and Agla 25%. We recall that these zones are floodable during the rainy season.
The majority of the sweet must samples have a light brown color, i.e. 90.0%; all the sweet must samples have a fermentation odor; 76% have a very sweet taste; and note that all the samples have a cloudy   Gram staining shows the presence of Gram-positive cocci observed in a high proportion of 54%, Gramnegative bacilli 13% and samples with Grampositive cocci and/or Gram-negative bacilli at 33%. No Gram-negative cocci or Gram-positive bacilli were found Table 4. Biochemical identification tests revealed the presence of Klebsiella including Klebsiella pneumoniae and Klebsiella oxytoca which are Enterobacteriaceae. Likewise the presence of Staphylococcus aureus, Staphylococcus sp Figure 1 All strains isolated from Enterobacteriaceae show no resistance to antibiotics; however, strains isolated from Staphylococci show resistance to the same antibiotic as Nitrofurantoin but in small quantities Table 5. The presence of germs is a public health problem, and appropriate measures must be taken to prevent the population from being contaminated by bacteria with potentially endemic diseases such as Salmonella, Shigella or even vibrio's. These appropriate measures must be limited to the strict respect of cleanliness and hygienic conditions by the sellers of these drinks and even the growers of these cereals on the measures of conservation of these cereals (maize, millet and sorghum) so that each and every one is in good health. Otherwise the drink would be unfit for consumption and its sale should be prohibited.

CONCLUSION
At the end of this work, the germs isolated from samples of Tchakpalo (sweet must) drink are enterobacteria including Klebsiella pneumoniae, Klebsiella oxytoca and staphylococci including Staphylococcus aureus, Staphylococcus sp. These identified pathogenic germs pose a public health problem.
The presence of the germs may be due to the uncleanliness of the vendors, the serving bowls, the environment in which these drinks are sold, the substandard hygienic conditions put in place during the manufacture of the sweet must and even the origin of these maize, millet and sorghum cereals. Appropriate measures must be taken in order to prevent the population from being contaminated by bacteria and the occurrence of diseases with endemic potential such as Salmonella, Shigella and even vibrio's. However, it is recommended that women pasteurize the drink and strictly observe cleanliness and hygienic conditions.