Masa is a fermented bread-like product, which is round in shape with brown colour and smooth surface made from millet, maize, or rice flour. The total bacterial aerobic plate count, coliforms, mould, yeast and staphylococcal counts of Masa were determined. Discrete colonies appearing on incubated plates were isolated and identified. The results showed that South-east of University area had the highest bacterial counts, ranging from 4.26x10² to 8.33x10²CFU/g and coliforms counts (1.331x10² to 3.14x10²CFU/g). North-east sample had the highest yeast count (3.2x10² to 1.0x10² CFU/g). Mould count was the highest in South-west ranging from 1.03x10² to 2.331x10²CFU/g, while staphylococcal count was high (2.041x10² to l.061x10²CFU/g). The result indicates that Masa contain different type of microorganism like Shigella spp, Salmonella spp, Pseudomonas sp, E coli and Staphylococcus. Hence Personal and environmental hygiene is required during production of Masa to avoid food borne illnesses.

Keywords: masa, microbial load, evaluation, process sold, pathogens

Masa is a traditional fermented snack usually prepared and consumed in Northern and some part of Western states in Nigeria.¹ It can be produced from maize (Zee mays), millet (Pennisetumtyphoideum), sorghum (sorghum vulgare) and rice (Orizasativa). According to the types of cereals used in preparing Masa, we have Masa of different types such as; Masa Shinkafa, Masa Masara, Masa Gero. Masa Dawa. Good quality Masa is round in shape with brown colour and smooth surfaces. Masa is a Spanish word mostly referred to all kind of dough.^2,3 It is also a popular food among Hausa and Fulani tribes of Nigeria.⁴

The consumption of Masa brings about some certain health benefit to consumers because it contained high calories and calcium minerals. It is also a good source of energy and fibre which helps to control appetite and prevent constipation.⁵

Cereal grains are fruit of cultivated grasses of the monocoteledoneus family.⁶ Its demand for food makes it abundant in East Asia and Sub-Sahara West Africa. Many researchers have carried out studies on fermentation of cereal product. It is noted that Masa is one of the major food that is mostly consumed by student because it is one of the common ready-to-eat food. It is also a fast food with low cost compared to other foods.

It was reported that unhygienic practices among the local food processor usually brings about the contamination of food by microorganisms. This may pose significant health effects to the consumers. In Maiduguri, the outbreak of food borne diseases due to the use of contaminated water has caused the death of many children.⁷ In this region of the country, cause of clean water has been advocated. Since it was known that the use of clean water for food preparation influences the outcome on the final products. This includes shelf-life stability, acceptability in terms of sensory qualities and products which pose no health hazards to the consumers. Similarly, the carelessness of the local food vendors, have also been reported.⁸ Carelessness in food practices such as the use of unclean utensils and equipment; raw materials selection; selling of foods in open places are potential treats and so increase risk of public health.

Therefore, there was a need to investigate the microbiological quality of Masa that is being sold within the University of Maiduguri. This also ensured and ascertained the type of foods produced and sold within the University community.

The method of preparing and selling of Masa in an open space in schools may cause contamination to foods. This contamination can also occur due to the use of unclean hands. Water, equipment and utensils during preparation. So there was need to know the types of microorganisms associated with the Masa that is being consumed by the students within the university premises to ascertain its safety.

Study area

Handling of foods by traditional food processors and hawkers who do not adhere to strict hygiene practices could pose a threat to public health. There is the risk of foods contaminated with pathogenic microorganisms which could lead to outbreak of food borne illnesses such as staphylococcal food poisoning, shigellosis, salmonellosis and other microbial foodborne illnesses and spoilage microorganisms. Therefore, it became necessary to sample foods from processors and hawkers for microbiological assessment. It was against this programme that this study enumerated, isolated and identified of microorganisms of significance from masa, a common convenient food processed and hawked within university of Maiduguri masa.

The study area where food samples (Masa) were collected is within different locations such as commercial area, hostel area complex area with much population because of the high demand by the student and staff quarters in the university of Maiduguri campus, Borno state.

Description of food

Masa (Figure 1) is a fermented bread-like product, which is round in shape with brown colour and smooth surface made from Millet, maize and rice floor. It is like the "india idle" in shape used in tortilla preparation it is also consumed by all age group in Northern Nigeria, with population of about 47million.⁹

Figure 1 Annular, erythematous lesions involving the skin of the abdomen and groin (tinea corporis and tinea cruris) in a case of dermatophytosis

Samples and sampling

A total number of 12 Masa samples were collected from vendors, 3 from each of the four cardinal point of the university of Maiduguri campus. The samples were collected in sterile containers and labelled. It was taken immediately to the laboratory for analysis.

All the glass wares and materials that were used for the analysis in the laboratory were washed and sterilized in a hot-air-oven at 1600C for one hour.¹⁰

Preparation of culture media

Culture media were prepared in accordance with the methods described by Collins and Lyne (1970), Harrigan and McCance (1976) Nkama, Badau, Chesbrough, et al.¹¹⁻¹⁵

The culture media prepared were Nutrient (NA), Eosin methylene blue (EMB), Mannital salt agar (MSA), Macconkey agar (MA). potato Dextrose agar(POA) and corn meal agar. The media were prepared according to the methods described by Chesbrough.¹⁵

 Preparation of nutrient agar (NA)

Nutrient agar (NA) was prepared by weighing fifty-two grams of the power and dispensing into 1liter of sterile distilled water in a conical flask and was allowed to dissolve completely by swirling. It was sterilized by autoclaving at 12I°C for 5 minutes at 15pounds pressure. It was allowed to cool to a temperature of 45°C and poured into sterile peptic dishes and allowed to gel (solidity). The surface was dried in hot-air-oven before inoculation.¹⁵

Preparation of eosine methylene blue agar (EMB)

Eosine methylene blue (EMB) was prepared by weighing thirty six grams of EMB powder (Himedia laboratory) into Ilitre of distilled water in a clean control flask and was heated to dissolve completely. It was then sterilized by autoclaving at 121°C at 15psi pressure for 15 minutes. The culture medium was allowed to cool to 45°C before pouring into petri-dishes.¹⁵

 Preparation of mannitol salt agar (MSA)

Mannitol salt agar (MSA) was prepared by dissolving 11grams in 1liter of distilled water, followed by gentle heating to dissolves the medium completely and sterilized by autoclaving at 12I°C, 15 minutes.¹⁵

 Preparation of macconkey agra (MA)

MacConkey agar (MA) was prepared by weighing fifty-two grams of the powder into sterilized flask containig I liter of sterile distilled water. It was sterilized by autoclaving at 121°C, 15psi pressure for 15 minutes. Thereafter mixed well before pouring into petri-dishes and allowed to gel (solidity). The surface was dried in hot-air-oven before inoculation.¹⁵

 Preparation of potato dextrose agar (PDA)

Potato Dextrose agar (PDA) was prepared by dissolving 39grams of media into sterilized conical flask containing 1litre of distilled water and heated to dissolve the medium completely. It was sterilized by autoclaving at 12I°C 15psi pressure for 15minutes.¹⁵Preparation of corn meal agar (CMA)

Corn meal agar was prepared by dissolving 17grams of the power into 1litre of sterile distilled water in conical flask and gently heated to dissolve the medium completely, then sterilized by autoclaving at 12I°C 15psi pressure for 15 minutes. This was allowed to cool 45°C before pouring into petri-dishes and allowed to solidity befor use.¹⁵

One gram of each sample of Masa was weighed and dispensed into 9ml of sterile distilled water as a diluent. Serial dilution was conducted by adding 1ml from tube one into the second test tube and mixed carefully by shaking it gently. The same procedure was made several times.⁴

Plating, culture incubation

A pour plate technique was used for plating the Masa culture. A triplicate of sterile petri- dishes were labelled corresponding to the number of tube containing the culture. Separate sterile pipettes was used to transfer 1ml of sample from the dilution tube to the corresponding petri-dishes before pouring molten nutrient agar that has been cooled to 45°C after sterilization. This was swirled gently to mix the agar and allowed to solidify before it was incubated at 37°C for 24hours.¹⁵

 Viable cell count

The total plate count for the growth of bacterial colonies on plates was observed by colony counter to count and record the number of colonies on each plate. The actual number of bacterial colonies was calculated as colony forming unit (CFU/ml) as described by Harriggan and McCance.¹⁶

 Identification of bacteria

 Isolation and identification of microorganisms: Microorganisms were isolated and identified based on cultural and morphological characteristics using method described by Harrigan et al.¹⁶ Biochemical tests such as catalase test, coagulase test, indole test, methyl red (MR) test, Voges-Proskauer test, citrate utilization test, urease test and oxidase test were carried out as described by Harrigan & McCance,¹⁶ and Cheesbrough.¹⁵ Results obtained from these tests were compared with literature standards using diagnostic tables showing the biochemical reactions identifying many genera and species of bacteria.¹⁷

Data analysis

The data obtained were analysed to find the mean using Microsoft word excel (2010).

The result of microbial load of Masa obtained from four cardinal points within university of Maiduguri campus is shown in Table 1. The total bacterial count ranged from 4.26x10² to 6.39x10², 6.80x10² to 7.46x10², 4.27x10² to 5.6x10² and 5.16x10² to 8.33x10² CFU/g of masa from North West, North East, South West and South east of University of Maiduguri campus, respectively. Coliform counts of masa from North West ranged from 2.40x10² to 2.77x10² CFU/g, North East masa had coliform count of 1.33x10² to 2.66x10² CFU/g. Coliform counts of masa collected from South West and South East ranged from 1.71x10² to 2.35x10² and 1.39x10² to 3.41x10²CFU/g, respectively.

The mould count of masa obtained from North West ranged from 1.10x10² to 2.18x10²CFU/g, North East masa ranged from 1.16x10² to 2.04x10²CFU/g, South West had mould count of 1.20x10² to 2.33x10²CFU/g and South East masa had 1.15x10² to 2.17x10² CFU/g of mould count. The yeast count of masa obtained from North West, North East, South West and South East ranged from 1.01x10² to 3.02x10², 1.15x10² to 3.21x10², 1.06x10² to 2.12x10² and 1.04x10² to 3.01x10², respectively. Whreeas, staphylococcal count of masa from North West of university of Maiduguri campus ranged from 1.10x10² to 2.01x10², North East, ranged from 1.08 x10² to 2.04x10², South West, ranged from 1.15x10² to 2.04x10² and South East, ranged from 1.06x10² to 1.18x10² CFU/g.

Table 1 above presents the results of the mean value of microorganisms of Masa sample from different locations in the University of Maiduguri. The result revealed the total bacterial count from south east of 8.33x10² had the highest bacterial load count while Northwest had the lowest bacterial count of 4.26x10². Colifonns count had the highest count of 3.14x10² from south-east and the lowest count at 1.33x10² from North-east. Yeast count which is 3.2 Ix10² had the highest count. While 1.01x10² from north- west had the lowest count. Mould count 2.33x10² had the highest count from south west. Staphylococcus count from north-east with 2.04x10² had the highest count while south-east had the lowest count of 1.06x10².

The result obtained from the analysis showed that bacterial count from south-east had the highest count compared to others, which could be as a result of poor processing method, poor hygiene practice, improper and unhygienic handling of the product, bad sanitation operations and use of unclean utensils. This agrees with the fact that immense microbial contamination of food is linked to poor post processing handling practices¹⁸ and the Masa from North-west had the lowest bacterial count and it could be as a result of the differences in processing practices.

The presence of coliforms from south-east is high compared to other locations and it can be as a result of unhygienic practices related to feacal contamination, the great increase in the count could be as the location of the retail outlet in which basically heavy contamination occurs. This finding suggests food was exposed to feacal or sewage contamination through the use of contaminated water or contamination from the unsanitary environment, equipment and carriage.⁴

Yeast count is indicated to be the highest from North-east due to direct exposure to air, product is opened as often as the customer’s demand and also the handling processes by the vendors,¹³ started that the dusty unhygienic environment coupled with the poor handling by vendors are factors contributing to the high microbial load. Masa from North-west is lower in yeast count and it could be due to environmental factors.

Mould count is high from south-west which could be as a result of the environmental factors due to dust and open air; therefore, enviromental contaminants have also been implicated as food borne pathogens.¹ North-west had the lowest count which indicates that the environment where the Masa is processed is safer.

Staphylococcus had the highest count from North-east which could be as a result of contaminants from personnel through dirty hands, cloths and utensils that comes in contact with the food. Staphvlococcus is common organisms found in all individuals and expelled from the respiratory tract, nose, mouth, clothing, hand and skin.¹

From the result obtain in Table 2 salmonlla is known as a pathogen that causes typhoid, fever and food poisoning.¹⁹ Pseudomonas sp is an opportunistic pathogen that cause bacteremia and gastrointestinal infections²⁰ shigella is a pathgen and its presence has been contributed to unhygienic practices related to faecal contamination.^21,22 E coli is an indicator of postprocessing contamination. Staphylococcus is common organisms found in all individuals and expelled from the respiratory track, nose, mouth, clothing, hand and skin. Aboloma have reported that the incidence of staphylococcus in food is an indication of environment and human contamination. It is a pathogen of public health importance.

The percentage occurenrance of microorganisms for "Masa" is shown in Table 3. Salmonella, Shigella and Rhizopus Oryza had 37.5% occurrence in "Masa" but the occurrence of S. aureus, E coli, Pseudomonas, Candida albicas, Saccharomyce cerevisie, is 25% in Masa. The occurrence of all this microorganism is a result of contamination from air could be as a result of unhygienic practices and poor sanitation in and around the environment.^23–25 The dusty, unhygienic environment coupled with the poor handling by vendors are factors contributing to the high microbial load.

The analysis of Masa for the isolation and identification of microorganisms has shown that there is higher amount of microbial contaminants in Masa due to the way of distribution, use of contaminated water, dirty utensils, the method of service and display in an open air which may be potentially pathogenic to human being and could be a health safety problem.

Technical assistance rendered by laboratory staff of the Department of Food Science and Technology is gratefully acknowledged.

Author declares that there is no conflict of interest.

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