Pa ge 1 Pa ge 99 American Journal of Food Science and Technology (AJFST) Antibiotic Susceptibility Pattern of Microorganisms Found in Fura De Nono Sold in Different Markets in Makurdi Benue State Ato Joy Iember1*, Ogbonna Innocent Okonkwo2 Volume 3 Issue 2, Year 2024 ISSN: 2834-0086 (Online) DOI: https://doi.org/10.54536/ajfst.v3i2.3738 https://journals.e-palli.com/home/index.php/ajfst Article Information ABSTRACT Received: September 02, 2024 Accepted: October 10, 2024 Published: November 30, 2024 Fura de Nono, a traditional fermented dairy and millet product popular in Nigeria, is widely consumed for its nutritional value. However, its production, often under unsanitary conditions, raises concerns about microbial contamination and public health risks. This study investigates the microbial load in Fura de Nono samples collected from local markets in Makurdi, Nigeria, assessing the presence of pathogenic bacteria and fungi. Using standard microbiological techniques, including colony counting and biochemical tests, the study identified harmful microorganisms such as Escherichia coli, Staphylococcus aureus, Proteus vulgaris, and Salmonella species. Antibiotic susceptibility tests revealed resistance in certain isolates, posing additional risks to consumers. The findings underscore the urgent need for improved sanitation practices among vendors, routine microbiological testing, and better regulatory enforcement to ensure food safety. The study concludes with policy recommendations to enhance hygiene practices and establish a certification system for vendors to protect public health. Keywords Antibiotic Resistance, Fura De Nono, Hygienic Practices, Microbial Analysis, Microbial Contamination 1 Department of Epidemiology and Evidence-based Medicine, F. Erismann Institute of Public Health, I. M. Sechenov First Moscow State Medical University, Moscow, Russia 2 Department of Microbiology, Joseph Sarwuan Tarka University, Makurdi, Benue State, Nigeria * Corresponding author’s e-mail: atojoyiember@gmail.com INTRODUCTION Milk is a liquid secreted by mammary glands of the female mammals. It serves as an important source of protein than most other foods. Milk is not 100 percent source of protein, since it is relatively poor in sulphureted amino acids such as methionine (essential) and cysteine (non- essential) as well as tryptophan and histidine. Curiously, children need less methionine and cysteine than adult. Because of this, cow milk meets the protein requirements of infancy quite well. According to Wodnerkos and Yitayal (2003), milk is primarily composed of water, protein in colloidal suspension, lactose and lipids in emulsion, inorganic salts in solution, vitamins, enzymes, gases, and other things. Along with being a great provider of calcium and phosphorus for healthy bones and teeth, milk also has notable levels of riboflavin, vitamin B1, vitamin B6, vitamin B12, and vitamin A (Yirsaw, 2004). Milk provides a range of essential nutrients, including carbohydrates, proteins, fats, vitamins, and minerals, all of which contribute to its high nutritional value. The specific composition of milk varies depending on the species, but cow’s milk typically contains around 3.4% protein, 3.6% fat, and 4.6% lactose, providing about 68 kcal per 100 g (Goff & Douglas, 2010). The carbohydrate lactose is the primary sugar found in milk, contributing to its sweet flavor and providing a readily digestible energy source. Additionally, lactose aids in calcium absorption, promoting bone health (Muir et al., 1992). The protein content in milk is composed of two main types: casein and whey. Casein accounts for about 80% of the protein in milk and is responsible for its white color and ability to form curds, which is essential in cheese production (Goff, 2010). Whey proteins, although less abundant, are water-soluble and offer a high biological value due to their complete amino acid profile, making them highly nutritious and easily digestible. Milk fat, meanwhile, varies depending on factors such as feed and environmental conditions but is rich in essential fatty acids and provides the characteristic texture and richness to dairy products (Goff, 2010). In many African countries, including Nigeria, Nono—a fermented milk product made from locally sourced cow milk—plays a significant role in local diets. Nono is made from non-pasteurized cow milk collected in a container called calabash, and allowed to ferment naturally for 24 hours (Uzeh, 2006). It is categorized into two types, with the term “nono” commonly referring to the type known as kindirimu. Another notable product, Fura da Nono, combines this fermented milk with a cereal mix made from millet, creating a nutritious beverage enjoyed for its taste and health benefits. Milk if not given the right conditions because of its high-water activity and nutritional content, it serves as a wonderful medium for many different types of microorganisms to grow. (Mesfin et al., 2017). If milk is not handled properly, it can become contaminated easily and deteriorate quickly. Fura da Nono is fermented milk-cereal mix that is highly nutritious on a two in one beverage product. It consists of a cereal Fura and Nono which is a locally fermented milk with a consistency that is thick but not quite as thick as yoghurt. Traditionally Fura da Nono is usually prepared by mashing millet into powdery form and mixing the powdered millet with hot water in a bowl to make dough, allowing the dough to cool and solidify and finally mixing the mashed millet (fura) with fermented milk (nono) and served to customer (Abbas et al., 2020). Pa ge 10 0 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 Fura is a porridge typically made from cereal grain millet (Pennisetum glaucum). There are a variety of millets that are specially used for Fura production. Millets ‘gero’ is one of the commonly used varieties. The preparation process involves powdering millet together with other spices like pepper and ginger, and then shaping the mixture into tiny balls by compressing between the two palms after adding a little amount of hot water. Next the dough is then boiled for about 30 minutes in boiling water and and coated with the remaining dried flour (Jideani et. al., 2010). The dough ball can be served by mashing them into fermented milk (nono) or water, sugar may be added to taste. This local milk product in Nigeria is mainly carried out by Nomadic Fulani’s from the Northern part of the country in villages where the producers are ignorant of the life and safety standard of the product but distribute it at will to consumers (Okonkwo, 2011). Fulani men milk the cows and thereafter, distribute the milk to the women in the encampment (Fulani farm stead) who later process the milk into various product like Fura da Nono, cheese and fermented milk (nono) etc. (Uzeh et al., 2006). The consumer’s expectation of processed milk is that it should have the typical appearance of milk, be free from extraneous matter and should have a clean and slightly sweet taste, with no abnormal odors or taints. Apart from the microbial flora of raw milk being of great importance as regards to hygiene and food safety, raw milk should be unadulterated and free of taints contaminants. Since nono is produced by illiterate Fulani’s in villages with poor knowledge of shelf-life and safety standards, handlers of these products may unknowingly introduce pathogenic microorganisms into the products during milking and also by using unsanitized milking equipment. since the products does not undergo further processing before being sold for consumption, it makes the food a potential source of health risk for consumers. Many dairy products, even those made from pasteurized milk have been associated with food borne diseases. In developing countries like Nigeria, it has been unusual to find microorganisms in Fura da Nono drinks. This is because milk is an excellent medium for microbial growth and also because of improper handling in Fura da Nono production. Raw milk and raw milk products can be contaminated with bacteria that can cause serious illness, hospitalization or death. These harmful bacteria include Campylobacter jejuni, Bacillus cereus, Shiga toxin producing E. coli (E.coli O157:H7), Coxiella burnetii, Listeria monocytogenes, Mycobacterium tuberculosis, Mycobacterium bovis, Mycobacterium avium subspecies paratuberculosis, Salmonella spp, Yersinia enterocolitica, and certain strains of Staphylococcus aureus which are capable of producing highly heat-stable toxins Brucella, streptococcus pyogenes (Anyanwu, 2019; Fagbemigun, 2021) Once the microbes get into the milk, they multiply rapidly because milk is an excellent medium for their growth (Okonkwo, 2011). The presence of coliform bacteria in Fura da Nono drinks could be due to the presence of dough. This is a source of concern in Nigeria because consumers have a strong preference for this traditionally produced and processed milk and millet products. It is therefore suggested that the microbial analysis of Fura da Nono should be carried out periodically as quality assurance measures. The addition of growth-inhibiting ingredients, adjusting the storage conditions to hinder the growth of microorganisms, and proper sanitation at every stage of handling and processing will help reduce the microbial level of Fura da Nono contamination (Shehu et al., 1990; Shehu et al., 2000). Relevant agencies like NAFDAC should put in place an irreducible minimum standard to producers and need to ensure and enforce strict compliance to this minimum standard all for production sectors in Nigeria. This research was aimed at evaluating the resistance or susceptibility pattern of microorganisms isolated from fura de nono using standard antibiotics kit. MATERIALS AND METHODS Sample Collection A total of 20 samples of nono and fura were purchased from Fulani hawkers using random sampling method from two different locations: North Bank and Wadata markets within Makurdi metropolis. purchase samples were transported to the university microbiology laboratory in sterile corked plastic tubes packed in iced container where analysis was carried out within two hours of collection. Microbiological Analysis Tenfold serial dilution of Fura da Nono samples were made before being place on the media to count for colonies. Inoculation of Fura samples was done using spread plate techniques while that of nono was done using the pouring plate techniques. Aliquot of 0.1ml of dilutions of 102 and 1010 of each Fura samples were pipette aseptically and dropped on a freshly prepared media and surface plated. The following media were used to isolate and count the organisms: Nutrient Agar (NA) (Biotech Lab. Ipswich, UK) for total aerobic and anaerobic microorganism counts, Eosine Methylene Blue Agar (EMBA) (Himedia Laboratories Pot Ltd, India) for E. coli, Desoxycholate Citrate Agar (DCA) for Salmonella and Shigella spp, Potato Dextrose Agar for Yeasts and mould counts, McConkey agar for total coliform counts, Man-Rogosa-Sharpe agar (MRS) for Lactic Acid Bacteria count, Mannitol Salt agar for staphylococcus aureus spp. All culture media were prepared according to manufacturer’s instruction. Media were sterilized by autoclaving at 121°C for 15 min except DCA which involved only boiling over gauze. Using a digital illuminated colony counter (Gallen Kamp, Loughborough, Leicestershire, UK), plates containing 30-300 colonies were counted after incubation. The computed results were expressed as colony forming unit (cfu) per mL by multiplying the average number of colonies from the observed triplicates by the reciprocal of the dilution factor. Number of organisms = number of colonies * dilution factor/Volume of Inoculum. Pa ge 10 1 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 Identification of Isolates Gram staining and microscopic identification: sub- cultured colonies on nutrient agar were gram stained. A small portion of 24 hours old colony was taken using a sterile wire loop and emulsified in a drop of distilled water on a clean glass slide and fixed by passing the smeared slide rapidly over a flame. The smear was covered with a crystal violet for about 30-60 seconds, rinsed with water and flooded with lugol’s iodine for 30-60 seconds and then rinsed under a running tap, rapidly decolorized with acetone (75% alcohol reagent) for 30 seconds and then rinsed with water and then counter-stained with safranine(red) for 1 minute and rinse under a slow running tap then the slides were air dried. A drop of oil immersion was added to the sliding containing the specimen, it was observed under the microscope using x100 objective lens (Cheesbrough, 2003). Gram positive appeared purple while gram negative organisms appeared pink. Biochemical Identification of Isolates The biochemical tests for the identification of the isolates were: Motility Test To determine whether an organism is motile or not. Catalase Test To determine the ability of an organism to produce catalase enzymes. Oxidase Test Use to identify bacteria that can produce cytochrome c oxidase, an enzyme of the bacteria electron chain. Coagulase Test To determine the ability of an organism to convert fibrinogen to fibrin in blood plasma. Citrate Test To determine if an organism is capable of utilizing citrates as its sole source of metabolism with resulting alkalinization of the medium. Indole Test To detect the ability of an organism to break down tryptophan to pyruvate and indole. Urease Test To determine the ability of an organism to split urea in the presence of water to release ammonia and carbon(iv) oxide. The ammonia combines with CO2 and H2O to form ammonium carbonate which turns the medium alkaline, turning the phenol red indicator from its original orange-yellow color to bright pink. The biochemical test were carried out on isolates according to Cowan and Steel (1965) and Cheesbrough (2004) procedures. Identification of fungi isolates Fungi isolates were done using the method described by Benson, (1990). The fungal isolates underwent additional microscopy using acetone and lactophenol on cotton blue stain placed on a clean grease slide. Each isolate was confirmed using Microgen Identification. The cultural and morphological traits of the isolates, such as hyphae (septation), reproductive structure (sporangia/conidia) in chain or single, type of spore, etc., were observed and served as criteria used for identification. Antibiotic Susceptibility Test The isolated organisms were tested against routinely used commercially available antibiotics. The multi-susceptibility Optudisc (manufactured by Optun Laboratories, Nigeria Ltd, Aba) for gram positive and gram-negative organisms were used. RESULTS AND DISCUSSION Table 1 describes the cultural and morphological characteristics of various bacterial isolates. Each bacterial species displays distinct features on nutrient agar (NA) and MacConkey agar (MA). For example, Escherichia coli appears flat and creamy with an even edge on NA, while Table 1: Cultural and Morphological Characteristics of the Bacterial Isolates NA MA Suspected Organism Flat and creamy with an even edge. Reddish pink, smooth circular LF colonies Escherichia coli Creamy smooth raised and very mucoid colonies Very mucoid, pink and smooth, LF Klebsiella sp Creamy flat smooth colonies Deep pink dull colonies with rough edges Lactobacillus sp Light green with fruits smell Pale NLF colonies Pseudomonas sp Golden yellow, smooth mucoid and raised Pink smooth LF colonies Staphylococcus aureus Creamy, shiny with putrefactive odour Pale smooth and discrete NLF colonies with irregular edges Proteus sp Creamy flat smooth Pale rough, shiny NLF colonies Salmonella sp Creamy brown flat and glittering colonies Pink flat and dull face LF colonies Streptococcus sp Flat, creamy small and slightly rough shiny Deep pink dull and small colonies Staphylococcus epidermidis Creamy and swarmy colonies Pale, mucoid and smooth NLF colonies Proteus vulgaris key: LF= lactose fermenting, NLF= non lactose fermenting, NGO= No growth observed, SP= Species Pa ge 10 2 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 on MA, it forms reddish-pink smooth circular colonies, indicating lactose fermentation (LF). Other organisms like Klebsiella sp. and Staphylococcus aureus are also lactose fermenters, showing pink smooth colonies. In contrast, Pseudomonas sp. and Proteus sp. are non- lactose fermenters (NLF), presenting pale, smooth, and discrete colonies on MA. The table effectively highlights the variability in colony morphology across different species, which aids in their identification. Microscopic and Biochemical Test Table 2: Microscopic and Biochemical Characteristics of the Bacterial Isolates Gram Reaction MOT CAT COA G CIT IND URE OXI Suspected Organism -ve rods in chains +ve +ve -ve +ve +ve -ve -ve Escherichia coli +ve cocci in clusters +ve +ve -ve - +ve - +ve Staphylococcus aureus +ve cocci in clusters +ve +ve -ve - +ve - -ve Staphylococcus epidermidis +ve rods in chains +ve +ve -ve +ve +ve +ve -ve Lactobacillus sp -ve rods in clusters and chains -ve +ve +ve +ve +ve +ve -ve Proteus sp -ve rods in pairs and short chains -ve +ve -ve +ve -ve +ve -ve Klepsiella sp -ve rods appearing singly and scattered -ve +ve +ve +ve +ve -ve -ve Salmonella sp -ve rods in clusters and chains -ve +ve +ve +ve +ve +ve +ve Proteus sp -ve rods slightly curved -ve +ve +ve +ve +ve -ve +ve Pseudomonas sp +ve cocci in chains -ve -ve +ve - +ve +ve +ve Streptococcus sp KEY: CIT = Citrate, IND = Indole, UREA = Urease, OXI = Oxidase, CAT = Catalase, MOT = Motility, SP Species COAG = Coagulase, -ve = Negative, - = Not done, +ve = Positive Table 2 details the microscopic and biochemical characteristics of various bacterial isolates, focusing on their Gram reactions, motility, and enzyme activities. The Gram stain distinguishes bacteria as either Gram- positive or Gram-negative, with the latter staining pink. The motility test identifies bacteria that can move, with different patterns observed. Enzymatic tests like the catalase test, which detects the production of the catalase enzyme, show bubbling in positive results. Similarly, the coagulase test indicates enzyme production by clumping of the organism. These initial tests provide foundational data for bacterial classification. Further biochemical tests like the indole, urease, and oxidase tests offer more precise identification. The indole test detects tryptophan breakdown, with a bright pink layer indicating a positive result. The urease test identifies bacteria that produce urease, turning the medium pink when positive. Finally, the oxidase test reveals cytochrome c oxidase activity, with a blue-purple color change within 10 minutes signaling a positive result. These biochemical characteristics are key to distinguishing species and are critical in clinical diagnostics Percentage of Occurrences of Bacteria Isolates Table 3: Percentage of Occurrences of Bacteria Isolates in Fura de Nono from Hawkers in North Bank Market Isolates Frequency of Bacterial Isolates (CFU/ml) Percentage Occurrence (%) Klebsiella sp 10 25.64 Proteus Vulgaris 12 30.76 Staphylococcus Epidermidis 2 5.12 E. Coli 4 10.25 Staphylococcus Aureus 2 5.12 Lactobacillus sp 7 17.94 Pseudomonas sp 2 5.12 Total 39 100 X2 = 28.00, DF = 16 (p<0.05) The Table 3 above from the study presents the percentage of bacterial isolates found in “Fura de Nono” from hawkers at North Bank Market. It lists seven types of bacteria, with Proteus vulgaris being the most prevalent at 30.76%, followed by Klebsiella sp at 25.64% and Lactobacillus sp at 17.94%. Other bacteria, such as E. coli, Staphylococcus Pa ge 10 3 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 epidermidis, Staphylococcus aureus, and Pseudomonas sp, show much lower occurrences, all hovering around 5.12%. The total frequency of bacteria isolates was 39 CFU/ml, and the statistical analysis yielded a Chi-square (X²) value of 28.00 with 16 degrees of freedom (p<0.05), indicating a significant distribution of these isolates. Table 4: Percentage of Occurrences of Bacteria Isolates in Fura de Nono from Hawkers in Wadata Market Isolates Frequency of Bacterial Isolates (CFU/ml) Percentage Occurrence (%) Klebsiella sp 3 7.31 Proteus Vulgaris 3 7.31 Proteus sp 5 12.19 Streptococcus 3 7.31 E. Coli 8 19.51 Salmonella sp 3 7.31 Pseudomonas 2 4.87 Staphylococcus Aureus 4 9.75 Staphylococcus Epidermidis 3 7.31 Lactobacillus sp 4 9.75 Total 41 100 X2 = 40.00, DF = 16 (p<0.05) The results from Table 4 provides data on the percentage occurrences of bacterial isolates in “Fura de Nono” sold by hawkers at Wadata Market. A total of 41 CFU/ml of bacterial isolates were recorded, with E. coli being the most prevalent at 19.51%. Other bacteria like Proteus sp accounted for 12.19%, while Staphylococcus aureus and Lactobacillus sp each made up 9.75% of the isolates. The remaining bacteria, including Klebsiella sp, Proteus vulgaris, Streptococcus sp, and others, each had lower percentages around 7.31% or less. The Chi-square (X²) value of 40.00 with 16 degrees of freedom (p<0.05) suggests significant variation in the distribution of these isolates. Result of Susceptibility of the Bacterial Isolates Table 5: Antibiotic Resistance Pattern of the Bacteria Isolated from the Fura de Nono Sample A nt ib io tic s C on c (m cg ) K le bs ie lla s p Pr ot eu s vu lg ar is Pr ot eu s sp St re pt oc oc us s p E .c ol i s p Sa lm on el la s p Ps eu do m on as St ap hy lo co cc us au re us St ap h. e pi de rm id is La ct ob ac ill us sp Tarivid 10 20 21 24 8 16 25 10 23 25 35 Reflacine 10 25 20 16 26 30 5 12 30 28 18 Ciproflox 10 17 27 32 35 18 6 25 17 11 10 Augmentin 30 17 14 15 19 23 25 16 10 15 14 Gentamycin 10 14 21 17 12 13 14 20 15 23 21 Streptomycin 30 15 20 14 22 16 10 24 21 30 25 Ceрorex 10 13 20 23 10 18 24 23 17 20 12 Nalidixic acid 30 15 24 14 20 10 12 11 29 22 15 Septrin 30 20 29 21 10 16 13 12 26 15 16 Amplicin 30 12 15 11 10 25 16 15 11 17 15 Pefloxacin 10 39 29 32 20 35 22 27 28 12 13 Gentamycin 10 20 15 18 10 17 30 35 14 12 19 Ampliclox 30 15 10 13 16 20 30 25 26 28 11 Zinnace 20 21 22 27 10 15 12 35 38 25 27 Amoxacilin 30 8 15 12 11 39 21 25 17 35 20 Ciproflaxacin 25 37 31 30 25 15 13 39 16 23 24 Pa ge 10 4 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 Microscopic and Biochemical Test Antibiotic resistance profiles of isolates show that Klebisiella sp was highly sensitive to Pefloxacin (39 mm), streptomycin (38 mm), Ciproflaxacin (37mm) and Rocephin (37 mm). It was highly resistant to Amoxacilin (8mm). Proteus sp were sensitive to streptomycin and resistant to most other antibiotic tested. S. aureus was more sensitive to Reflacine (30mm). Nalixidic acid (29mm). Zinnacef (38mm). Rocephin (36mm). The organism was resistant to Augmentin (10mm), Ampicilin (11mm) Streptococcus sp was more sensitive to ciproflox (35mm). Reflacin (26mm), Ciproflaxacin. (25mm). The organism was resistant to zinnacef (10mm), Gentamicin (10mm), Ampicillin (10mm), Sceptrin (10mm), Tarivid (8mm). E. coli was more sensitive to Reflacine (30mm), pefloxacin (35mm). Amoxicillin (39mm), Streptomycin (38mm), Sceptrin (30mm). The organism was resistant to Nalixidic acid (10mm). Salmonella sp was sensitive to Gentamicin (30mm), Ampiclox (36mm) and was resistant to Reflacine (5mm). Ciproflox (6mm). Pseudomonas was sensitive to Gentamycin (35mm), Zennacel (35mm). Ciprofloxacin (39mm), Erythromycin (30mm). The organism was resistant to Tarivid (10mm). The result from Table 6 outlines the macro and micro Streptomycin 10 38 33 15 18 38 25 20 18 15 34 Septrin 30 20 12 10 17 30 21 28 18 16 12 Erthromycin 10 20 14 12 11 15 17 30 19 17 35 Rocephin 25 35 16 20 23 14 27 25 36 13 14 KEY: CIT = Citrate, IND = Indole, UREA = Urease, OXI = Oxidase, CAT = Catalase, MOT = Motility, SP Species COAG = Coagulase, -ve = Negative, - = Not done, +ve = Positive Table 6: Macro and Micro Characteristics of Fungal isolates on Potatoes Dextrose Agar Colony Characteristics Microscopic Appearance Suspected Fungal Suspected Fungal Gray, blackish-brown or black surface with gray periphery, black on reverse Colorless, septet with chains of micro conidia Alternaria Bluish-green with Sulphur yellow areas on the surface Septate hyphae with chains of radiate conidia on conidiophores Aspergillus sp Creamy and yellowish colonies which are pasty and glittering Round conidia occurring singly and in small clusters with pseudo-hypae Candida sp White to dark gray mycelium with profuse cottony growth Elliptical spores, colorless and non-septet hyphae with sporangiospores. Mucor sp Blue to green moldy and velvety appearance with milky- white on reverse. Conidiophores bears sterigma with chains of conidia,parallel in arrangement. Penicillium sp White colony growth with black spots on the tip, some gray. Non-septet colorless hyphae and mycelium with rhizoid and solon sporangiospores present Rhizopus sp Rapidly growing, flat, smooth, moist, dull and creamy colonies which are large and circular. Unicellular multilateral and ellipsoid blastoconidia Saccharomyces’s characteristics of fungal isolates on Potato Dextrose Agar, detailing both colony characteristics and microscopic appearances of various fungi. For instance, Alternaria presents with a grayish-black surface, while microscopically showing colorless septate with chains of micro conidia. Aspergillus sp is bluish-green with sulfur- yellow areas and has septate hyphae with radiate conidia. Candida sp displays creamy yellow colonies, while Penicillium sp is blue-green with a velvety appearance. The table provides a clear comparison of colony morphology and microscopic traits, aiding in fungal identification based on both visual and microscopic features. Table 7: Mean Bacteria Count for Fura De Nono Samples Collected from Different Locations in Makurdi CFU/ml Sample S aureus Lactic acid bacteria Salmonella shigella Total viable counts Coliform counts Escherichia coli Total Fungal count Nono 1.00 ± 0.63° 42.40± 9.77° 6.00± 3.84° 7.00± 3.53° 1.40 ± 0.67° 1.80± 0.96° 6.00±1.14° Fura 4.20 ± 1.74° 44.20± 29.89° 179.40± 150.42° 224.601± 66.08° 158.80± 72.50° 98.80± 14.77° 6.60±1.88° Pa ge 10 5 https://journals.e-palli.com/home/index.php/ajfst Am. J. Food. Sci. Technol. 3(2) 99-106, 2024 Values for a given group in a column followed by same letter as superscript are not significantly different according to Duncan’s multiple comparison procedure (at p 0.05). The result from Table 7 compares the mean bacterial count of Fura De Nono samples collected from different locations in Makurdi, showing the presence of various bacteria such as S. aureus, lactic acid bacteria, Salmonella shigella, coliforms, E. coli, and total fungal counts. The bacterial counts vary significantly across samples. For instance, Nono 2 has the highest lactic acid bacteria count (233.80 CFU/ml), while Fura 2 exhibits the highest total viable counts (500.00 CFU/ml) and E. coli (455.20 CFU/ ml). These variations highlight the microbial differences based on sample location and type, with some counts exceeding typical safety thresholds, suggesting potential contamination risks. CONCLUSION The microbial analysis of Fura de Nono samples collected from various locations in Makurdi, Benue State, Nigeria, indicates significant bacterial contamination, posing a public health risk. High levels of bacteria such as Escherichia coli, Staphylococcus aureus, and Proteus vulgaris were found, with certain bacterial counts exceeding safe limits. These findings suggest inadequate hygiene practices during the preparation, storage, and sale of Fura de Nono by local vendors. The variability in bacterial counts across different samples highlights inconsistent sanitary measures and a lack of proper food safety protocols among vendors. This situation creates the potential for foodborne illnesses and underscores the need for immediate action to safeguard public health. Recommendations To address the public health risks posed by contaminated Fura de Nono in Makurdi, several policy measures should be introduced. First, it is essential to improve sanitation practices among vendors. The Benue State government, in collaboration with health authorities, should develop and implement food safety training programs targeting Fura de Nono producers and sellers. These programs should cover proper handling, personal hygiene, and the use of clean, potable water during preparation. Additionally, the enforcement of hygiene standards must be strengthened through regular inspections and the application of penalties for non-compliance. Second, the introduction of routine microbiological testing is necessary to ensure the safety of dairy products sold in local markets. Health authorities should conduct regular, random checks on Fura de Nono sold by vendors, testing for harmful bacteria such as E. coli and Salmonella. These tests will help in identifying contamination early and preventing unsafe products from reaching consumers. Public health awareness campaigns should also be launched to inform both vendors and consumers about the risks associated with poor hygiene in dairy production. Consumers need to be educated on recognizing safe products and purchasing from reliable vendors, while vendors should be encouraged to adopt best practices to maintain the safety of their products. Furthermore, a certification system should be established to regulate Fura de Nono vendors. Only vendors who meet the required food safety and hygiene standards should be granted a license to sell, ensuring that consumers can trust the quality of their purchases. This regulatory framework would not only improve food safety but also promote the economic well-being of certified vendors. Finally, cold chain infrastructure must be implemented to maintain the quality and safety of Fura de Nono throughout its transportation and sale. Keeping the product at lower temperatures can significantly reduce bacterial growth, thereby minimizing the risk of contamination. 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