An Assessment of Potassium Bromide and Aspergillus Flavus Contamination in Bakery Products Sold in Bamenda

IRASS Journal of Applied Medical and Pharmaceutical Sciences Abbriviate TitleIRASS J App Med Pharm Sci ISSN (Online) 3049-0901 https://irasspublisher.com/journal-details/IRASSJAMPS Vol-1, Iss-1(November-2024) © Copyright IRASS Publisher. All Rights Reserved 1 JOURNAL COVER PAGE An Assessment of Potassium Bromide and Aspergillus Flavus Contamination in Bakery Products Sold in Bamenda Esoh Rene Tanwieh1*, Fru Priscillia Masakoh2 Solomom Gyampoh3 Kiranjot kaur Rai4 Ngongpan Scott Nchatkang5 Ngongpan Bih Edwige6 1 Department of Medical and biomedical sciences, University of Bamenda-Cameroon. 2 Training School for Laboratory Technician-Nkwen Bamenda. 3 School of Pharmaceutical Sciences, Lovely Professional University. 4 Punjabi University patiala, Department of Biotechnology and food technology. 5 Ecole supérieur de management et de technologie applique (ESMATA)-Yaoundé Cameroon. 6 Ecole supérieur de management et de technologie applique (ESMATA)--Yaoundé Cameroon. Corresponding Author Esoh Rene Tanwieh Department of Medical and biomedical sciences, University of BamendaCameroon. Article History Received: 20 / 10 / 2024 Accepted: 02 / 11 / 2024 Published: 06 / 11 / 2024 Abstract: Bakery products especially bread are staple foods in the world and are recognized as semiperishable foods. Usually, spoilage is due to improper storage. This work was aimed at isolating and identifying Potassium bromide and Aspergillus flavus contaminants in bakery products sold in Bamenda. The study was a crosssectional study in which 25 samples of bakery products were conveniently purchased at random from different vendors and bakeries in Bamenda. The hypothesis of the study was prevalence of toxigenic fungi and potassium bromide in bakery products sold in Bamenda is high. The isolation and identification were done by their cultural and morphological characteristics by doing Lactophenol Cotton Blue staining of the various mould forms that grew on Saboraud Dextrose Agar and then examining under the microscope. The organisms found to be associated with spoilage of bakery products were strictly fungal organisms which included; Penicillium species (40%), Mucor species (26.67%), Aspergillus species (20%) and Fusarium species (13.33%). After analysing the samples, Penicillium species was found to be the most occurring in bakery products consistent with the study carried out by Legan J.D (1993), a clear prove that good hygienic handling of bakery products is essential, therefore, further investigation on isolation, identification and characterization with a larger sample size is very important. Qualitative determination of Potassium bromide showed that 40% of the bread samples were positive for potassium bromide consistent with the study carried out by H.B Alhanashi et al (2020), a clear prove that Potasssium bromate is used as a dough improver in backery products in Bamenda, therefore further investigation with a quantitative analysis and on a larger sample size is important. Keywords: Potassium bromide, bakery products, fungi contamination. Introduction 1.1 BACKGROUND OF THE STUDY In the majority of nations and civilizations, bakery goods are essential staple meals. Bread, buns, cupcakes, cookies, pizza bases, toasts, and many more are the most popular of these. The majority of our food calories and over half of our protein needs come from the grains used in baked goods, making them an important source of nutrients for our diet. Carbohydrates, proteins, fats, vitamins, calcium, iron, minerals, starch, and energy are the nutrients found in baked goods (V.S. Petil et al., 2020). One of the most significant products of the food business is said to be baked foods. Bread is consumed every day all around the world and is a staple cuisine in many nations. Bread production is a relative mainstay in the European Union (E.U.) and has grown slowly in the majority of western nations. The UK and Ireland consume less than 50 kilograms of bread annually on average, whereas the Germans and Austrians consume the most, averaging 80 kg per person annually (The Federation of Bakers, 2012). IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 2 In contrast to Europe, the consumption of bakery products shows an increasing trend in most developing countries (Elsanhoty et al, 2013). The majority of people in Cameroon rely heavily on baked goods to meet their nutritional needs. Because bakery products are ready to eat, whether they are hot or cold, they can be consumed right away at the point of sale without any additional processing. For this reason, products that are sent into the community to be sold should be carefully considered, and Hazard Analysis Critical Control Points (HACCP) handle this.Nowadays, this approach is regarded as a viable option for guaranteeing food safety. In order to prevent potential hazards, HACCP identifies Critical Control Points (CCPs) in the production process and puts monitoring procedures in place to stop them (Amaranta Carvajal Campos, 2019). Consumer behavior in the baked goods industry has evolved during the last few decades. Much attention has been paid to consumers' concerns regarding food additives and safety in particular. Furthermore, there are now more consumers who are concerned about their health. As a result, there is a strong market demand for "natural" and "wholesome" foods free of chemical additives and preservatives. As a result, bread producers have been making more and more "clean label" goods to cater to consumers' better lifestyles. These goods' food labels make statements like "natural" and "no preservatives." Health-focused new product developments are actually being driven by marketing the lack of additives and preservatives (21% of new products introduced in Europe 2013/2014) and the presence of wholegrain. In turn, such minimally processed foods without chemical preservatives or other artificial additives should still be of high quality and have an extended shelf life (Mintel, 2014). When Novozymes polled more than 4,000 bread consumers across Europe in 2011, they discovered that the primary reason bread was thrown out was because it had grown mold.Fungi are also in charge of producing mycotoxins and off-flavors, which may be created even before fungal proliferation is apparent, in addition to the unpleasant look of visible mold growth. Therefore, rotting bread poses a risk to the health of consumers (Magan et al, 2003b). Spoiled bakery products may be defined as bakery products that have been damaged or injured so as to make it undesirable for human consumption (V. S Patil et al, 2020). At several phases of bread preparation, including slicing and wrapping, the ingredients in bread promote the growth and multiplication of microbes. Moldiness and ropiness are the primary microbial deterioration kinds in bread that cause problems for bakers. A loaf of sliced bread is frequently the starting point for mold formation since the moisture content of the bread is higher than the surface, particularly in the creases (Salim-ur-Rehman et al, 2007). When contaminated, other bread components, tools, and packaging materials may also serve as a specific entry point for unwanted and spoilage-related microbes into the baking environment. Following regulations such as Codex Alimentarius, which include putting in place systems like Hazard Analysis Critical Control Point (HACCP), Good Manufacturing Practices (GMPs), Good Hygienic Practices (GHPs), and Good Agricultural Practices (GAP), is crucial to ensuring food safety and quality throughout the food production chain (Reale et al., 2013). Molds do not survive the bread-baking process in a typical bakery. On the other hand, because they are more heat stable, mycotoxins made by food-spoilage fungi and crop diseases pose a greater threat. According to Claudia Axel (2015), mycotxins have the potential to be immunosuppressive, nephrotoxic, neurotoxic, and/or carcinogenic, resulting in serious health issues. While monitoring takes place in most countries, mycotoxin levels from fungal contamination in cereals and cereal products continue to be a major global concern. They are often found in processed cereal products (Aldana et al, 2014; Claudia Axel, 2015). Physical techniques to eliminate post-baking pollutants in breads include ultraviolet light, infrared radiation, microwave heating, and ultra-high pressure treatment. Nevertheless, there haven't been many previous investigations using these techniques. Studies have increasingly established goals to replace conventional chemical preservatives with environmentally acceptable, "clean-label" alternatives in light of the ongoing consumer movement towards a healthier lifestyle. As a result, "bio preservation," which is the process of extending shelf life through the use of natural or regulated microbiota and/or their antimicrobial chemicals, has grown in importance as a research area (Stiles M.E, 1996). Filamentous fungi including Alternaria, Aureobasidium, Cladasporium, Clavicceps, Epicoccum, Fusarium, and Helminthosporium are examples of common phytogenic diseases; infections with Fusarium are thought to pose a serious risk. According to Claudia Axel (2015), the most prevalent fungi linked to losses in baked goods are Aspergillus, Cladosporium, Endomyces, Fusarium, Monilia, Mucor, Pencillium, and Rhizopus. The Use of Potassium Bromate in Baking In baking, potassium bromated is commonly used as a flour enhancer in the United States. It helps the dough rise higher and strengthens it. As an oxidizing agent, it will entirely react to a form with a lower oxidation state when the bread is baked under the correct conditions. A flour ingredient called potassium bromated gives the dough strength and raises its altitude. Additionally, it adds a pleasing white hue to the final bread. However, this additive is associated with cancer, which is an issue (Viswanath Pilla, 2016). In conclusion, it is important that all major players in the bakery industry should be aware of the risks posed by bakery products contamination as it relates to consumption of fungi toxins produced on contaminated products. Reports of loss of lives due to the activity of toxigenic fungus abound in the literatures and efforts should be made to reduce the risks. Therefore, there is need for systematic and universally applicable approach to food safety (Jay, 1996). Also, it is important that the use of potassium bromate which is now regarded as an adulterant, which is an oxidising agent and a lowcost dough improver should be strictly monitored in the bakery industry since higher levels of the substance in humans can cause detrimental issues of the respiratory tract ( Shanmugavel et al, 2020). Statement of the Problem The most common food pollutants worldwide are fungi. They are ubiquitous plant pathogens and major spoilage agents of food. Microbial contamination of bread may be as a result of unhygienic handling during production or the integrity of the raw material; unknown to the consumers, people that consume contaminated snacks may stand a risk of food borne illnesses and this may put a terrible economic burden on them, the society or country (WHO, 2000). IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 3 Snacks that are being sold may as well become compromised hygienically if safety regulations are not taken seriously during marketing, this exposes them to microbial contamination by fungi, bacteria, nematodes and many others (Johnathan et al, 2015, 2016). Invasion of food by various fungi may result in many results in remarkable rapid quality deterioration and consequently leading to food spoilage and sometimes production of secondary metabolites by the contaminants. Some moulds such as Aspergillus flavus,Aspergillus parasiticus and Penicilliium species have been associated with production of aflatoxins which have serious health effects (Johnathan et al,2016). Aflatoxins are capable of causing acute and chronic effects in man and animals ranging from disorders of central nervous system, cardiovascular systems, pulmonary systems, intestinal systems and even death (Johnathan et al, 2016). Also, not much attention has been given to fungi contamination in bakery foods like bread, cookies, cakes, etc as it has been done on other foods. Therefore, this study is to detect, isolate and identify the possible fungi that cause spoilage and contamination in bakery products. Due to nature of these food processing and also the ingredients needed in preparing the bakery products, microorganisms like fungi are paramount to breeding microorganisms in baked foods (Johnathan et al, 2015). The Federal Ministry of Health prohibited potassium bromate in 1993 after a research into the health risks of the ingredient in bread revealed the following negative impacts on consumers. Potassium bromate has been shown to cause thyroid follicular cell tumors, renal cell tumors, and peritoneal mesotheliomas. Furthermore, studies designed to clarify the mechanism of carcinogenesis have demonstrated that potassium bromated is a full carcinogen, having the ability to both initiate and promote kidney tumorigenesis in rats. This implies that potassium bromide poisoning is a risk that consumers face. There is therefore the need for continuous surveillance and enforcement of the ban on the use of potassium bromated in the bakery industry (A. I Abubakar et al, 2017). Research Questions 1. What are the possible fungal species in bakery products sold in Bamenda? 2. What is the prevalence of fungal species in bakery products sold in Bamenda? 3. What is the proportion of bakery products contain potassium bromide? 1.4.1 Main Objectives To isolate and identify the different fungal contaminants and potassium bromide in bakery products sold in Bamenda. 1.4.2 Specific Objectives 1. To isolate and identify the prevalence of Aspergillus flavus contamination in bakery products sold in Bamenda. 2. To determine the prevalence of potassium bromide toxicity in bakery products sold in Bamenda. 1.5 Hypothesis/assumption The bakery products in Bamenda contain potassium bromide and pathogenic fungi. 1.6 Significance of the Study  This study is to ensure that bakery products free of fungal and potassium bromide contaminants should be sold to the population of Bamenda to reduce the prevalence of fungal food poisoning.  Also, it is aimed at creating awareness of proper hygiene for bakers during preparation, packaging and storage and proper handling by the consumers after buying the products. Scope of the Study (Delimitation) The study was limited only to bakery products in Bamenda city which in Bamenda 1, 2 and 3 municilities. Limitation of the Study  The limitation to examination of end-product which is the bakery products and not the raw materials.  Also limited was the time period which was the same time for preparation for exams.  Also, as a limitation was the study area which was limited only to Bamenda 1, 2 1nd 3, this is not the whole region of the North West. Literature Review 2.1 Composition of Bakery Products and its Requirements Water, sugar, salt, eggs, fats, yeasts, baking soda, baking powder, yogurt, essence, cocoa powder, chocolate slabs, fruits, jams, and sweeteners are all necessary materials for bakeries. Starch, sugar chains, and proteins—that is, amino acids—make up the majority of wheat flour. (V. S. Patil and others, 2020). 2.2 Requirements Fresh whole grain four, sourdough, mixing bowl, bench knife, razor blade, bread knife, thermometer, scale, proofing basket, loaf pan (Diakonov,1999). 2.3 The Importance of Bakery Products Since baked foods, particularly breads, are a widespread and significant food from an economic and cultural standpoint, baking is still a fundamental skill and has nutritional value.(Figoni Paula, 2011) There are a vast array of varieties, shapes, sizes, and textures available worldwide as a result of the nearly endless combinations of various flours and varying ingredient proportions. Numerous techniques, ranging from the use of naturally occurring microorganisms to high pressure artificial aeration during preparation and baking, can be used to leaven (aerate) or leave it unleavened. Among the many additives that can be used are fruits, nuts, various lipids, and chemical additives that improve flavor, texture, color, and shelf life. Bread can be consumed as a snack, served in a variety of ways during meals throughout the day, or even utilized as a component in other recipes. Bread, a staple item around the world, has gained importance beyond its nutritional value and is now a part of religious ceremonies, secular culture, and language (Chevan et al., 1993). 2.4 Nutritional Values of Bakery Products  ENERGY Rida Safdar in 2019 indicated that Bakery products provide energy for daily leaving. One slice of Pat`s pan provides around 100kcal which is 5%of your recommended intake or RI (Reference intake) for a day which is 2000kcal). IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 4  Protein Bread is the third world largest contributor of proteins in our daily diet. Protein is essential for growth, development and repair of the body.  Fats Bread is naturally low in fats and form part of a healthy, balance diet.  Carbohydrates Bakery products produce complex carbohydrates which provide energy.  Vitamins Bakery products various B vitamins including Thiamine(B1) and Niacin(B3) which are essential for releasing energy from food.  Iron Iron is the key nutrient in wheat flour and essential for red blood cell formation which aids in oxygen transport around the body and is important for brain formation.  Calcium Flour is fortified with calcium in the form of calcium carbonate and calcium is known for its importance in formation of good teeth and strong bones, proper functioning of nerves, muscles, kidney and heart. 2.5 Nutritional Facts The following nutrition information is provided by the United States Department of Agriculture (USDA) for one slice (32g) of whole wheat bread: Calories________82g Fat___________1.1g Sodium________144g Carbohydrates__13.8g Fiber__________1.9g Sugars_________1.4g Protein________4g Bread nutrition varies by type. Nutrition experts recommend increasing your intake of whole grains. For example, the nutritional value of bread doubles that of sandwich or toast. Commercially prepared white bread provides about 75 calories and 1g of fats per slice, 15g of carbohydrates or more, but with less than 2g of fibre. Rye bread may or may not be made from whole grains depending on the brand. A typical slice of rye bread provides 83 calories, about 1g of fats, 16g of carbohydrates, and 1.9g of fibre and 2.7g of proteins (J. N Morris, 1981). 2.6 Role of Essential Ingredients in Baking  Yeast The key ingredient in the bread-making process is yeast. It is the key component that causes the dough to rise and lends the scent and delicious flavor of homemade bread.Each packet of yeast contains thousands of active microorganisms that resemble plants. Tiny carbon dioxide gas bubbles are released by the yeast when it is fed sugar or starch and activated by a warm liquid. After baking, this gas gives the dough its light texture and causes it to rise.  Flour The most popular kind of flour for baking bread is wheat. It contains whole wheat flour, bread flour, and all-purpose flour. Gluten, a protein found in wheat, provides dough its strength and elasticity. The gluten stretches and develops into a network that captures the carbon dioxide bubbles that the yeast produces when flour and yeast are combined with liquid and then kneaded or pounded.  Liquids Water It performs two vital functions, making it the most significant liquid.  The yeast is dissolved and activated by it.  It combines with the flour to form a dough that is elastic and sticky. Juice, cream, buttermilk, or milk may be added to improve texture or flavor. In a recipe, only warm liquids should be put to dry ingredients because  Yeast activity will be slowed down or stopped by an excessively cold liquid.  A beverage that is too hot can kill the yeast and stop it from rising. Sweetener Sugar gives a bread's crust flavor and a deep brown hue. You can also use jams, molasses, honey, and brown sugar.  Salt When baking bread, salt is a crucial component since it slows the rising period, which develops the dough's flavor and enhances the flavor of the finished product.  Eggs Eggs give breads their color, flavor, and nutritional value. They also aid in making the crust soft and the crumb fine. Eggs provide protein and richness.  Fats Bread becomes soft and moist with the addition of butter, margarine, shortening, or oil. Bread stays fresher for longer because fat prevents moisture loss. Potassium Bromate (KBrO3) Potassium bromate has a major impact on food biomolecules like protein and starch because it changes the degree of gelatinization, viscosity, and swelling properties of both gluten and protein. It also eliminates the sulfhydryl group and causes disulfide linkages to form, which enhances the qualities of bread. Nonetheless, numerous investigations clarify its detrimental effects on human health. Because of its strong oxidizing qualities and mutageneity in vivo, it is categorized under the class 2B category and considered a possible human carcinogen. In vivo, it produces mild effects on the kidney, liver, and brain. In 2020, Shanmugaval et al. In fact, the Food and Drug Administration states that there is insufficient proof of potassium bromate's harmful effects in humans, enabling additives to be used in bread baking at levels no higher than 75 parts per million (117). Therefore, regardless of whether potassium bromate has been used as a wheat processing additive, bromated levels in bread should be consistently and accurately checked (112). IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 5 Ideally therefore, the end product should contain no potassium bromate which has been broken down during the baking process into potassium bromide (KBr), a harmless byproduct (3). 2.7 Chemical Compisition of Bread According to P. Saranraj (2011), bakery goods have a high starch and simple saccharide content, which makes them high in carbs. Later carbohydrate levels are influenced by the amount of sugar added during the production process as well as the chemical makeup of the flour. As a result, rye bread, which isn't enhanced with additional carbohydrates, typically has fewer sugars than wheat bakery goods. In addition to fruits and vegetables, bakery goods are a significant source of dietary fiber. Its amount ranges from 0.3 to 1.5g/100g of fresh bread, depending on the type of flour. As a result, whole meal bread has higher dietary fiber content. Although the latter is not broken down in the human digestive system, it has a beneficial effect on intestinal peristalsis and makes it possible to remove toxic metabolites. Cellulose and hemicelluloses are main fractions of dietary fibre in wheat bakery products while in rye bread, it embraces pectin, gums, and mucus substances. Other nutrient concentration is also influenced by the types of flour and the amount of dietary fiber. Compared to white bread, whole grain and rye breads have two to five times the amount of iron, magnesium, manganese, copper, and zinc salts. Fresh bread's protein content ranges from 4.5 to 8.0g/100g. Rye breads provide less protein than wheat bakery goods, however rye proteins are more useful due to their higher concentrations of key amino acids. The amount of fat in baked goods is minimal, ranging from 0.7% to 2.5%. Bread can consequently be stored for a considerable amount of time. B-complex vitamins, including thiamine (vitamin B1), niacin, riboflavin, folic acid, and vitamin E, are abundant in bakery products. Wholemeal breads, especially yeast-fermented wheat bread, have higher concentrations of essential nutrients. Rye bread has a 50% humidity percentage, whereas wheat breads have a 45% humidity content (P. Saranraj, 2011). Bakery goods range in energy value from 874 to 1924 KJ, or 208– 459 kcal/100g of fresh goods, with the exception of pumpernickel and crisp breads, which have higher energy values and higher nutrient contents. It is estimated that bakery products satisfy about 50% of nutritional needs of humans and around 30% of energy needs (P. Saranraj,2011). 2.8 Sources of Contamination Ponte and Tsen, in 1978 realized that Although freshly baked products are free of viable vegetative moulds and mould spores, products soon become contaminated as a result of post baking contamination by mould spores from the air, bakery surfaces and equipment, food handlers and raw ingredients such as glazes, nuts, spices, and sugars. After baking, mold spores that are present in the air around loaves throughout cooling, slicing, packaging, and storage contaminate the bread. Because dry components, particularly flour, contain mold spores and flour dust travels rapidly through the air, the environment inside a bakery is not sterile. The naturally high spore content of flour, old bread deposits, polluted air conditioning systems, etc., are the causes of extremely high levels of air contamination. Spores are dispersed throughout all manufacturing and storage areas, especially by air conditioning systems. As a result, the baked goods get significantly contaminated with fungus spores. Reiss (1981). Ponte et al, in 1978 discovered that Industrially produced bakery products emerge from baking process with surface that is essentially sterile but post bake handling can quickly lead to fungal contamination as a result of exposure to airborne contaminants as well as equipment contact as follows;  Slicing machine  Bread coolers  Conveyor belts and racks have been identified as potential sources. 2.9 Microbial Spoilage of Bakery Products Within the European union (EU) the production of bread is relatively stable showing low growth in most western countries. (Elsanhoty et al, 2013). However, bakery goods are susceptible to microbiological, chemical, and physical deterioration, just like a lot of processed meals. At several phases of bread preparation, including slicing and wrapping, the ingredients in bread promote the growth and multiplication of microbes. Moldiness and ropiness are the two primary microbial bread deterioration kinds that cause problems for bakers. Since moisture is more readily available within a loaf of sliced bread than on the outside, particularly in the crease, mold development frequently starts there. (Axel, Claudia, 2019). The main cause of bakery items' short shelf lives is frequently microbial spoiling. Both manufacturers and customers suffer financial losses as a result of microbial growth-induced spoilage. These losses may result from a variety of specific factors, including product turnover, storage conditions, hygienic manufacturing practices, and packaging. (Needham Rachel et al., 2004). Products from bakeries, particularly bread, have an intermediate moisture level and are highly perishable. Microbial spoiling and moisture loss are the two most prevalent types of bread deterioration. More than 90% of all microbial contamination occurs when fungus colonize and thrive on wheat bread and other bakery products (Arroyo et al., 2008). Moreover, during bread preparation such as baking, only vegetative forms of micro-organisms are removed, but bacteria and fungi spores are to survive. Due to its widespread consumption, bread quality is important. Due to unfavorable changes that start shortly after baking and degrade the crumb's texture and sensory qualities, bakery goods have a short shelf life. Reduced humidity, the development of filamentous fungus and yeast, and stalling are signs that bread is getting older (Claudia Axel, 2009). Molds, yeasts, and bacteria can all lead to bread spoilage. However, fungal spores that are deposited from the baking environment are the main source of contamination after baking (Claudia Axel, 2015). The most important variables influencing the growth of unwanted fungi on food items are water activity (aw), pH, temperature, and oxygen.At a pH of roughly 6, bread typically has a reasonably high moisture content and water activity between 0.94 and 0.97. Sliced, packaged, and wrapped bread are among the bakery goods most vulnerable to mold deterioration (Magan et al, 2003a, Claudia Axel, 2015). 2.9.1 Mould Spoilage For bakers, mold deterioration is a major and expensive issue. Rhizopus species, Mucor species, Penicillium species, Eurotium species, Aspergillus species, and Monilia sitophilia are filamentous IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 6 fungus that cause bread to deteriorate. Bread spoilage in low humidity conditions inhibits the growth of mold. However, the most frequent cause of spoiling in dried grains that may grow at 0.75aW is fungi such Eurotium species (Cisarova et al, 2018). In fresh bread and other baked goods, the baking process usually eliminates mold spores. Bakery items must therefore be contaminated—either by the air, bakery surfaces, equipment, food handlers, or raw ingredients—or after baking, during the chilling, slicing, or packaging processes, in order to develop mold. This indicates that all mold-related spoiling issues arise after baking. Because of airborne contamination brought on by warmer temperatures and more humid storage conditions, fugal spore counts are higher in the summer than in the winter. The product's surface is frequently affected by fungal spoiling, which produces an unwanted odor (V. S. Patil et al, 2020). Various mycotoxins, or poisons, are also produced by molds and fungus and have been scientifically connected to a wide range of illnesses, such as diabetes, cancer, damage to internal organs, etc. Large amounts of mycotoxins that cause illness and death can be produced by even tiny amounts of these fungus, which can grow rapidly inside the body. Water activity is therefore the most important element influencing the kind and rate of spoiling in various bread items (Ambreen Akhtar Saddozai and Samina Khalil, 2009). Microorganisms (bacteria, yeast and moulds) can grow well when water activity and moisture content is high. Some strains cause a defect called ropiness, a soft sticky texture caused by starch degradation and slimy exopolysaccharide often accompanied by a fruity odour. Furthermore, depending on the product type, season, and processing method, product losses from mold deterioration range from 1 to 5%. Hickey (1998) states that the bakery business loses an average of 200 million pounds of goods annually as a result of mold deterioration (P. Saranraj M. Geetha, 2012). 2.9.2 Mycotoxins Because mycotoxin-contaminated staples have harmful effects on both humans and vertebrates, they pose a serious public health issue. Since their discovery, the main mycotoxins have been identified based on geographic locations, the lowest toxicity levels, the fungi that produce them, and the need to develop control measures to prevent negative effects on both human and animal health as well as to minimize financial losses. It is rarely advised to remove staples from the food chain after the mycotoxin content surpasses the threshold allowed by the laws (Amaranta Carnaval Campos, 2019). One of the most commercially significant groupings of molds is Aspergillus section flavi; its harmful effects are a major public health concern, and the taxonomy's stability is a practical one.The species in this region are capable of producing a variety of mycotoxins, but aflatoxins are particularly harmful due to their detrimental effects on vertebrates (Amaranta Carnaval Campos, 2019). Figure 1: Structure of Aspergillum (Amaranta Carnaval Campos, 2019) These species primarily flourish in tropical and subtropical climates across the world due to their physiological requirements. They are an issue in these regions because the conditions for harvest and storage are not always the best for the growth of mold and the creation of mycotoxin, and environmental factors also typically play a role in their production. resulting in two major problems: first, the health risks to humans and animals; second, contaminated staples cannot be exported, which has a detrimental impact on the economics of some nations that rely on exporting (Perrone et al, 2014). Best known mycotoxins include aflatoxins, ergot alkaloids, fumonisins, ochratoxins, patulin, trichothecenes and zearalenone. IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 7 Figure 2: Reproductive Life Cycle of Aspergillus Species (Amaranta Carnaval Campos, 2019) Table 1: Typical Sources, Impacts, and Principal Mycotoxin and Generating Species (AFSSA, 2009). The main Manufacturers of Mycotoxins are Indicated in Red. Mycotoxin Type Main producer Contaminated products effects Chemical nature Aflatoxins B1, B1, G1, G2 Flavus Apergillus A. parasiticus A. namius A. section flavi contains several species. Cereals, rice, sorghum, wheat, corn, coconut, pistachios, almonds, spices, nuts, and dried fruit Acute toxicity, hepatotoxic, carnogenic, immunogenic, and teratogenic Polyketide Trichotecenes T-2 toxin, HT-2 F. langsethiae and F. triciniae F. F.sporotrichioides F. equiseti F. poae Cereals, maize, wheat, rice, sorghum Genetoxic, Imminotoxic, Reprotoxic, neurotoxic Terpene deoxyrivalenol Fusarilum tricinctum, F. langsethiae, F. culmarium, F.sporotrichoides, F. poae, F. equiseti, and F. solani Sorghum, rice, wheat, and cereals Haematopoietic, digestive, and imminotoxic terpene Fumonisis B1, B2, B3 F. verticillioides, F. proliferation Cereals, maize, rice, sorghum Carcinogenic, neurotoxic Poliketide Ochratoxin A A. ochraceus, P. nardicum, Coffee, wine, grape juice, cereals, Nephrotoxic, polyketide IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 8 A. carbonarius, and Penicillium verrocosum chocolate, and spices Immunotoxic, teratogenic Zearalenone F-2 toxin F. graminearum, F. culmarum, F. crookwellense Cereals, wheat, rice, oats, maize, soy, and sorghum Reprotoxic, immunotoxic Polyketide Patulin P. expansum, Byssochlamys nivea Juices from apples, pears, and derivatives Neurotoxic, Genotoxic, Cytotoxic. polyketide Ergot alkaloids Claviceps purpurea, C. paspali, C. africana, C. fusiformis Rye, wheat, triticale Vasoconstriction, digestive issues, and neurotoxicity Alkaloids Mycotoxins are categorized according to their chemical makeup, manner of action, and/or the fungus that produces them. Since the majority of them are heat stable and produce hazardous chemicals when degradation techniques are used, it is difficult to degrade them. With the exception of fumonis, these substances are often hydrophobic, which enables them to build up in plant and animal lipophylic tissues (Amaranta Caarnaval Campos, 2019). The chemical composition of the mycotoxin, the length and dosage of exposure, the organism that consumes the microbe (species, sex, age, health, and nutrition), and the interaction of the mycotoxin's effects with other xenobiotics are some of the interrelated factors that affect mycotoxicosis symptoms. Mutagenic, teratogenic, carcinogenic, nephrotoxic, hepattoxic, immunotoxic, estrogenic, and chronic (low doses, long periods of time) or acute toxic (high doses, short periods of time) are some of the possible effects on vertebrates. Mycotoxins and the creatures that ingest them are essential to the neurological, digestive, endocrine, immunological, liver, lungs, and kidneys. Antagonists, additives, or synergy are associated with the type of mycotoxin, the host species' decontamination process, exposure duration, and mycotoxin dosages and ratios (Amaranta Carnaval Campos, 2019). Humans have been using mycotoxins for food preservation for as long as agriculture has been, possibly even before memory began to be formed. Mycotoxicosis occurrences can be traced back to folklore, literature, and the arts. For example, they may be seen in the Dead Sea Scroll or in the Bible as part of the seven plagues of Egypt (Richard, 2007). Furthermore, fusariotoxins (toxin T2 and ZAE) may have contributed to the fall of the Etrusan Civilization in the fifth century B.C. Jouany and Yiannikouris (2002). However, the most well-known instances of mycotoxicosis in antiquity may be the hallucinations of "Saint Antony's fire" of ergotism (11th century) caused by the alkaloids of Claviceps purpurea on rye. Delirium, prostration, agonizing agony, limb abscess and gangrene, and even death are all signs of ergotism (Claudia Axel, 2015; Richard, 2007). Similarly, another well-known instance of mycotoxicosis epidemic is "Shoshin-Kakke," also known as "yellow rice disease," which was documented in Japan and primarily affected the colder regions. It causes acute cardiac beri-beri. Exposure to Citrovirirdin, a mycotoxin produced by Penicillium citreonigrum, is the cause of this sickness. Due to inadequate circumstances and procedures, this fungus infiltrated rice while it was being stored (Claudia Axel, 2015). Mycotoxins and their harmful consequences were first recognized in London, England, in 1962, despite the fact that fungal contamination did occur and that they were thought to be related to certain disorders. A unusual ailment known as "Turkey X syndrome" was found in poultry, killing at least 10,000 birds. Curiously, upon investigating the cause of the disease, it was found that peanuts used to feed chickens were tainted with aflatoxins, which are secondary metabolites of Aspergillus flavus. It was discovered years later that cyclopiazonic acid also contributed to this outbreak.The most well-known mycotoxins are Trichothecenes, Patulin, Zearalanone, Ergot alkaloids, Fumonisis, Aflatoxins, and Ochratoxins (Claudia Axel, 2015). 2.9.2.1 Aflatoxins Aspergillus species create a class of closely related and well studied mycotoxins known as aflatoxins. Before harvest, aflatoxins can be identified in the field, and contamination can rise during post-harvest processes such crop drying or storage. But even when there is no field contamination, aflatoxins can nevertheless contaminate products that have been stored. Aflatoxin production is limited to a small number of fungal species, all of which are members of the Aspergillus section flavi. Aspergillus flavus, Aspergillus paraciticus, Aspergillus nomius, Aspergillus psuedotamarii, Aspergillus bombycis, and Aspergillus ochraceoroseus are the species in question. While some of them are less common, Aspergillus flavus and Aspergillus paraciticus are economically significant and occupy overlapping habitats. Aspergillus flavus is one of the most prevalent and widespread soil-borne molds in the natural world. Nonetheless, within each aflatoxin species, the toxigenic capacities of distinct strains vary both qualitatively and quantitatively. Only half of the A. Flavus stains are reportedly capable of producing more than 106 μg/kg aflatoxins. While Aspergillus paraciticus produces both B and G aflatoxins (AFG1 and AFG2), Aspergillus flavus typically only produces B aflatoxins (AFB1 and AFB2). Conidia, asexual spores, and scleriotia, an asexual fruiting body with a resistant structure that allows the strains to endure in hostile environments, are all produced by Aspergillus flavus. In people and animals, Aspergillus flavus also produces mycoses, which are fungal infections as opposed to mycotoxicoses, which are illnesses brought on by ingesting fungal toxins. Aspergillus flavus is unique in that it is an opportunistic pathogen of both plants and animals (Yogendrarajah P., 2015). 2.9.2.2 Ochratoxins A dihydroisocoumarin bound to phenlalanine makes up this class of pentaketide metabolites. The primary producer of the most prevalent congener, ochratoxin A (OTA), is Aspergillus ochraceus. IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 9 Ochratoxins A, B, and C are known to be produced by Aspergillus species, including Penicillium verrucosu. Grain, coffee, chocolate, spices, wine, beer, and pigs are frequently contaminated with ochratoxins. It is a strong nephrotoxin that is implicated in the development of Balkan endemic nephropathy, a disease that affects humans. It has also been demonstrated to be immunotoxic, teratogenic, and hepathotoxic. 2.9.2.3 Trichothecenes Based on the differences in the functional hydroxyl and acetoxyl side groups, there are over 170 species that have been found and are divided into four kinds (A-D). Fusarium species create a number of naturally occurring trichothecene mycotoxins in food and feed. Although the most prevalent kind in grains is not considered carcinogenic to humans, it can have negative health effects such as immunological changes, endocrine dysfunction, weight loss, anorexia, and malnutrition (Pestka, 2010). 2.9.2.4 Fumonisins They are made up of a collection of compounds that have two tricarballytic acid groups esterified at positions C14 and C15 on the carbon chain backbone. Fusarium proliferatum and Fusarium verticilloides are the primary producers of them. It is categorized as a group 2B potential human carcinogen and is nephrotoxic and hepatotoxic. In high-risk groups, it has been linked to oesophageal and liver malignancies (Persson et al, 2012). 2.9.2.5 Zearalenone This estrogenic mycotoxin, which is mostly produced by Fusarium graminearum and Fusarium culmoru, is present in high moisture corn, wheat, barley, and sorghum. It has also been discovered in moldy hay and palletized feeds. Although it is not considered harmful to humans, its estrogenic activity makes it powerful. It binds to the estrogen receptors in mammalian target cells because it closely resembles 17-β-estradiol, the main hormone generated in human ovaries. Accordingly, it may cause girls to reach puberty too soon (Yogendrarajah P., 2015). 2.9.3 Fungi An estimated 3.5 to 5.1 million creatures, ranging from unicellular to tiny multicellular, make up the complex eukaryotic kingdom of fungi, which occupy a variety of ecological niches across the globe. These species, which include saprophages, symbionts, and pathogens, are important decomposers in the nutrient cycle. As heterotrophs, fungus produce enzymes that help them break down and absorb nutrients from the extracellular digestion of other species. In order to develop, fungi need specific elements that are utilized in their primary and secondary metabolisms. These elements are primarily sources of carbon and nitrogen, with trace amounts of potassium, phosphorus, and magnesium also being needed. Furthermore, their development depends on environmental elements including pH, light, temperature, and water availability (Amaranta Carnajal Campos, 2019). Figure 3: Fungi Classification (Pitt and Hocking, 2009) IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 16 18 Medium Raised Yellow/ White Mucoid Yellow Negative Negative Positive 19 Medium Raised White Non Mucoid Green Positive Negative Positive 20 Medium Raised White/ Yellow Mucoid Green/ White Penicillium Negative Negative 21 Medium Raised White Mucoid Absent Negative Negaive Negative 22 Medium Raised Yellow/ White Mucoid White Negative Mucor Positive 23 Large Flat White Mucoid Absent Negative Negative Negative 24 Small/ Medium Raised Yellow/ White Non Mucoid White/ Blue Negative Mucor Positive 25 Small Raised White Non Mucoid Absent Negative Negative Positive Table 5: Distribution Species in Bakery Products Sold in Bamenda of Fungal Fungal species Frequency Percentage Aspergilus 3 20% Mucor 4 26.67% Penicillium 6 40% Fusarium 2 13.33% Total 15 60% From the above table, 60% of the samples was positive for different fungal species from which 20% 0f fungal species was Aspergillus, 26.67% was Mucor species, 40% was Penicillium species and 13.33% was Fusarium species. 5.1 Discussion The prevalence of fungi in a majority of the samples may be due to the fact that these microorganisms are widely spread in the environment. This work gives a description of the isolation and identification of four different fungal species which were able to grow on SDA. They belong to the genera: Penicillium, Aspergillus, Fusarium and Mucor speies. The main contaminating fungus was from the genus Penicillium with a percentage of 40%, this finding as consistent with (Legan, 1993) who found out that Penicillium is the most common species. Potassium bromide was also present in most of the samples. This confirms that potassium bromate is still used as a bread improver in some bakeries in Bamenda. Germ tube technique showed four positive samples giving a percentage of 16% indicating the presence of Candida species in some bread samples, this finding consistent with Reynolds and Braude (1956). 5.2 Conclusion In conclusion, the study's findings of a 40% prevalence of Penicillium, 26.67% mucor, 20% aspergillus, and 13.33% Fusarium clearly demonstrate the importance of hygienic handling of bakery products. As such, bioaerosols in the bakery production environment should be managed with the use of suitable filters in conjunction with production procedures that reduce aerosols, as is typically done by HACCP. The health of employees depends on the control of airborne illnesses. The overall quantity of molds has increased as a result of poor worker, equipment, and air hygiene (working environment). Since most fungal species are common and found in the air, they readily settle on products when exposed, which is why the high prevalence of fungal species found on bakery products at roadside locations is a reflection of the lack of health policies to ensure adequate coverage of these products to prevent contamination. The high prevalence of potassium bromide identified in bakery products sold in Bamenda is an indication that high quantities of potassium bromated is used in most bakeries in Bamenda during bread baking process. These findings agree with the hypothesis that the prevalence of toxigenic fungal and potassium bromide in bakery products sold in Bamenda is high. 5.3 Recommendation From the results obtained, the following recommendations were made: To the Government  Strategies for routine sanitary inspection in bakeries, roadside vendors and bakery product retailing stations should be put in place to reduce fungal contamination.  Government should implement the control in the use of potassium bromate as a rising agent in IRASS Journal of Applied Medical and Pharmaceutical Sciences Vol-1, Iss-1 (November2024): 1-22 © Copyright IRASS Publisher. All Rights Reserved 17 bakery products and ensure that bakeries always comply with the safety guidelines.  Government should also adopt sensitisation strategies, for example, radio and television talks, health education on the prevention of fungal contamination of bakery products. To the Population  The population should adopt healthy attitudes in handling bakery products by properly covering them and avoid eating bakery products with visible mould.  Bread makers should use alternative flour improvers that are much more safe to humans. To Health Sector/Students  Students can research on the quantitative aspect of potassium bromide contamination in bakery products.  Students can equally research on isolation and determination of the total number of mould types in each bakery products. References 1. Abdulrazzaq et al (2004). Morbidity in neonates of mothers who have ingested aflatoxins. Annals of tropical paediatrics, 24: 145151. 2. Ade Kakure and J. M Serret (2020). The analysis of potassium bromated in bakery products. 3. AFSSA (2009). Evaluation des risqué lies a la presence de mycotoxines dans les chaines alimentaires humaine et animale. 4. Acharya Tankeshwar (2014). 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