This study assessed vitamin content in Saccharomyces cerevisiae biomass cultured in cassava wastewater. The Saccharomyces cerevisiae was isolated from palm wine following microbiological techniques. The Saccharomyces cerevisiae was inoculated into the cassava wastewater and incubated at room temperature for 15 days. During the processes the medium was shaken intermittently between 7.00 to 19.00 hours’ daily. At the end of the experiment, the medium was decanted and then filtered using Whatman filter paper. The trapped biomass was washed with distilled water and then re-filtered. The resultant biomass in the filter paper was oven dried and analyzed for vitamins following standard protocol. The mean value of the vitamin were 40.73±7.94 mg/100g (vitamin D), 2280.37±105.85µg/100g (Vitamin A), 132.54±14.00µg/100g (Vitamin E), 6.88±1.62mg/100g (Vitamin C), 0.58±0.10mg/100g (Vitamin B12), 2.62±0.92mg/100g (Vitamin B3), 1.05±0.21 mg/100g (Vitamin B1). Based on the value obtained from the finding of this study, the biomass is a good source of vitamins that could be utilized in animal feed. Through utilization of the wastewater for Saccharomyces cerevisiae cultivation the attendant environmental impacts associated with the cassava wastewater could be minimized.

Keywords: animal feed, biotechnology, cassava wastewater, saccharomyces cerevisiae

Nigeria is the world leading producer of cassava accounting for over 20% of total global production.^1–16 The production of cassava increased in Nigeria in the early 2000 due to several developmental projects that requires cassava as major raw material. Cassava cultivation and processing is a major source of livelihood to several families especially in the rural areas in southern Nigeria.¹⁵ Cassava is processed into garri, fufu, lafun Kigigha et al.,¹⁷ and used to feed livestock. It has several industrial applications as well.

In Nigeria over 60% of the total cassava tuber produced is used for the production of garri. Over 80% of garri production is carried out by smallholder that use rudimentary equipment for processing. During cassava tuber processing into garri large volume of wastewater is produced in the dewatering stage.¹² This wastewater is often discharged into the ecosystem with little or no treatment.^3–5 The wastewater often percolates in the soil, and may drain to a nearby pit or surface water. In aquatic ecosystem, the wastewater is injurious to fishes especially fingerlings.^3,18 The wastewater also causes air pollution through the emission of offensive decomposing effluents.

Studies have shown that cassava wastewater has the tendency to alter soil microorganisms,¹⁵ soil heavy metals^8–10,12 and general physicochemical characteristics. Previous studies have also suggested the need to manage the environmental impacts of cassava wastewater through bioethanol production¹⁶ and Saccharomyces cerevisiae biomass production for potential utilization in animal feed.^5–7,13 To this effect, Saccharomyces cerevisiae biomass cultured in cassava wastewater have been studied with regard to amino acid,⁵ heavy metals,⁷ cations and cyanide.⁶ But information about the vitamin content of Saccharomyces cerevisiae is scanty in literature.

Typically, in living organisms, vitamins play essential role during metabolic processes. As such, the absence of certain vitamins could hinder metabolic processes leading to collapse of the actions. Vitamins are typically classified into two viz: water (vitamin C i.e. ascorbic acid, vitamin B1 i.e. thiamin, vitamin B2 i.e. riboflavin, Vitamin B3 i.e niacin, vitamin B12 i.e Cobalamin, vitamin B6, folic acid, pantothenic acid, and biotin) and lipid/fat (Vitamin A, D, E and K) soluble.¹⁹ The authors further reported that all water-soluble vitamins apart from Vitamin C have a catalytic function (that act as coenzymes of enzymes that is involved during the metabolism of carbohydrate, proteins and fats), and have specific functions in some specialized and differentiated tissues. On the other hand, lipid soluble vitamins aid in the maintenance of the integrity and structure of membranes, and they control the synthesis of some enzymes at the genetic level.¹⁹

Vitamin requirements tend to vary according to biological species and its genetic/biochemical variations as well as source of diets. This is because most of these vitamins are taken through vegetables including fruit of Synespalum dulcifcum (miracle fruit),²⁰ Seed of Aframomum melegueta and Garcinia kola.²¹ Like human, livestock need minerals at certain amount for optimal productivity. This study aimed at investigating the vitamin content of Saccharomyces cerevisiae biomass cultured in cassava wastewater.

Sample collection

Untreated Cassava wastewater containing palm oil was obtained in replicate from a smallholder cassava processor in Ndemili, Ndokwa west Local Government Area of Delta state, Nigeria. The raw wastewater that contain palm oil was transported to the laboratory under ice pack using plastic container. The samples were used immediately at the laboratory.

Isolation and identification of saccharomyces cerevisiae

The Saccharomyces cerevisiae used in this study was isolated from palm wine bought from Rumoumasi, Port Harcourt, Nigeria. The pure isolate was made following the pour plate method by Pepper and Gerba,²² Benson²³ using potato dextrose agar supplemented with chloramphenicol. The resultant pure isolate was identified following cultural, morphological, and physiological/biochemical characteristics (carbon fermentation and assimilation, glucose-peptone-yeast extract broth, lacto-phenol cotton blue stain and growth based on temperature) as previously described by Kurtzman and Fell,²⁴ Benson²³ and applied by Iwuagwu and Ugwuanyi,²⁵ Abioye et al.,²⁶ Okoduwa et al.²⁷ The features of the isolates were compared with the guide provided by Ellis et al.,²⁸ Kurtzman and Fell.²⁴

Growth of the biomass and vitamin determination

The Saccharomyces cerevisiae biomass was grown following the method previously described by Izah et al.,^3–7 Izah¹ (Figure 1). The water soluble vitamins such as Vitamin C were analyzed with the spectrophotometric method previously described by Drevanasiddappa and Veena,²⁹ while the other water soluble vitamins including thiamine (B1), Nicotinamide (B3) and Cyanocobalamin/cobalamin (B12) were analyzed based on the spectrophotometric method. Lipid and fat soluble vitamins such as Vitamin A, D and E was also analyzed following standard procedure.

Figure 1 Schematics of Saccharomyces cerevisiae biomass production from cassava mill effluents (Modified from Izah et al., 2017a-e; Izah, 2018a, e).

Statistical analysis

SPSS software version 20 was used to carry out the statistical analysis. The results were expressed as mean ± standard deviation.

The vitamin content in Saccharomyces cerevisiae biomass cultured in cassava wastewater is presented in Table 1. The concentration of vitamin D was 40.73±7.94 mg/100g. Typically, vitamin D is essential for bone health probably due to the fact that it aids in the maintenance of serum calcium levels,³⁰ proper mineralization of bone ¹⁹ as well as calcium and phosphorus ratio.³¹ Deficiency of vitamin D could lead to soft bone which could predispose the body to hunched back and rickets.

The level of vitamin A was 2280.37±105.85 µg/100g. Vitamin A is a fat soluble vitamin which play essential role in many animals including bone formation,^30,31 maintenance of epithelial cells and good vision.^19,31 Vitamin A is also essential for reproduction, embryonic development, growth and immune response.¹⁹ Vitamin A deficiency in animals leads to morphological changes in bone,³⁰ ocular disturbances that could lead to blindness and growth retardation.^19,31 Typically vitamin A is found in cassava and have been reported in the range of 5.0- 35µg in cassava root^32–36 and 8300 to 11800 µg in cassava leaves.^35–37

In this study, the concentration of Vitamin E was 132.54±14.00 µg/100g. Like other vitamins, Vitamin E is important for fatty acid metabolism and maintenance of cell membrane.³¹ Vitamin E is among the lipid soluble vitamins that have antioxidant potentials,³⁸ and has the tendency to interrupt free radical chain reactions and protect polyunsaturated fatty acids and cell membranes.¹⁹ Deficiency of Vitamin E could lead to muscle weakness,³¹ peripheral neuropathy and breakdown of red blood cells.¹⁹ The vitamin C concentration was 6.88±1.62 mg/100g. Vitamin C is a water soluble vitamin that is an essential cofactor for at least 8 enzymes.³⁹ Some of the cofactors are essential for the hydroxylation of lysine and proline, thus it is important for the synthesis of collagen³⁰ and dopamine.³⁹ Vitamin C also has anti-oxidant and wound healing potentials,³⁹ and could boost immune system and facilitate the absorption of non-heme iron (from plant foods).¹⁹ Deficiency of vitamin C could lead to slow wound healing processes, soreness and stiffness of joints, swollen and bleeding gums. Basically, vitamin C has been reported in the range of 14.9–50.0mg in cassava root^32–36and 60 –370mg in cassava leaves.^35–37

The level of Vitamin B12 in the Saccharomyces cerevisiae biomass was 0.58±0.10mg/100g. Typically, Vitamin B12 is essential for many physiological and metabolic responses including osteoblast function, iron metabolism,³⁰ glucose production, metabolisation of methionine and cell growth.³¹ Baigent and Carpenter¹⁹ also reported that Vitamin B12 is an essential cofactor for enzymes for the metabolism of amino acids and fatty acids. The authors further reported that it is needed for the synthesis of new cells and neurological functions. Deficiency of Vitamin B12 could lead to gastrointestinal disturbances.¹⁹ The vitamin B3 concentration in the Saccharomyces cerevisiae biomass was 2.62±0.92 mg/100g. Vitamin B3 is a water soluble vitamin that is an essential component of coenzymes required for cellular metabolism, oxidation of fuel molecules, and synthesis of fatty acid and steroid disturbances.¹⁹ The authors further reported that deficiency of Vitamin B3 could lead to skin lesions and gastrointestinal disturbances. In addition, vitamin B3 is found in cassava and have been reported in the range of 0.6 –1.09 mg in cassava root^32–36 and 1.3–2.8mg in cassava leaves.^35–37

The level of vitamin B1 in the yeast biomass was 1.05±0.21 mg/100g. Like vitamin B3, vitamin B12 and Vitamin C, vitamin B1 is water soluble and is an essential component of a coenzyme involved in the metabolism of carbohydrate metabolism.¹⁹ Deficiency of Vitamin B1 could lead to nerves impairment.¹⁹ Typically vitamin B1 have been reported in the range of 0.03 – 0.88mg in cassava root.^32–36 and 0.06 -0.31 in cassava leaves.^35–37

Typically, Saccharomyces cerevisiae have been reported to play essential role preventing diarrhea and mortality, enhancing immune system, boosting of performance, milk production, fiber degradation and nutrient digestability, adsorption of toxic metal, stabilization of rumen pH and microorganisms in livestock’s.⁴⁰ Based on the value of water soluble vitamins (B1, B3, B12 and C) and lipid soluble vitamins (A, D and E), introduction of Saccharomyces cerevisiae could also aid to improve the health status of livestock’s. In addition, most species of cassava are known to contain cyanide, but studies have indicated that heat treatment, additives such as palm oil and fermentation using Saccharomyces cerevisiae could significantly reduce to cyanide content.^41,42

Vitamins are an essential ingredient in diet due to their roles in growth and metabolic processes in living organisms. Diet is the major source through which humans and livestock’s obtain necessary vitamins required for normal health. This study assessed the vitamin content in Saccharomyces cerevisiae biomass cultured in cassava wastewater and the result revealed the presence of water soluble (Vitamin C, Vitamin B2, Vitamin B3 and Vitamin B12) and lipid soluble vitamins (Vitamin A, D and E). As such, the cassava wastewater is a promising raw material for the cultivation of Saccharomyces cerevisiae biomass for possible utilization in animal feed industry.

None.

The author(s) declares that there is no conflict of interest.

1. Izah SC. Estimation of Saccharomyces cerevisiae Biomass Cultured in Cassava Mill Effluents. Environmental Analysis and Ecology studies. 2018a;2(5):EAES.000547.
2. Izah SC. Estimation of Potential Cassava Mill Effluents Discharged into Nigerian Environment. Environmental Analysis and Ecology studies. 2018b;2(5):EAES.000550.
3. Izah SC, Bassey SE, Ohimain EI. Changes in the treatment of some physico-chemical properties of cassava mill effluents using Saccharomyces cerevisiae. Toxic. 2017a;5(4):28.
4. Izah SC, Bassey SE, Ohimain EI. Removal of Heavy Metals in Cassava Mill Effluents with Saccharomyces cerevisiae isolated from Palm Wine. MOJ Toxicology. 2017b;3(4):00057.
5. Izah SC, Bassey SE, Ohimain EI. Amino acid and proximate composition of Saccharomyces cerevisiae biomass cultivated in Cassava mill effluents. Molecular Microbiology Research. 2017c;7(3):20–29.
6. Izah SC, Bassey SE, Ohimain EI. Cyanide and Macro-Nutrients Content of Saccharomyces cerevisiae Biomass Cultured in Cassava Mill Effluents. International Journal of Microbiology and Biotechnology. 2017d ;2(4):176–180.
7. Izah SC, Bassey SE, Ohimain EI. Assessment of Some Selected Heavy Metals in Saccharomyces cerevisiae Biomass Produced from Cassava Mill Effluents. EC Microbiology. 2017e;12(5):213–223.
8. Izah SC, Bassey SE, Ohimain EI. Assessment of heavy metal in cassava mill effluent contaminated soil in a rural community in the Niger Delta region of Nigeria. EC Pharmacology and Toxicology. 2017f;4(5):186–201.
9. Izah SC, Bassey SE, Ohimain EI. Geo-accumulation index, enrichment factor and quantification of contamination of heavy metals in soil receiving cassava mill effluents in a rural community in the Niger Delta region of Nigeria. Molecular Soil Biology. 2017g;8(2):7–20.
10. Izah SC, Bassey SE, Ohimain EI. Assessment of pollution load indices of heavy metals in cassava mill effluents contaminated soil: a case study of small-scale cassava processing mills in a rural community of the Niger Delta region of Nigeria. Bioscience Methods. 2017h;8(1):1–17.
11. Izah SC, Bassey SE, Ohimain EI. Impacts of Cassava mill effleunts in Nigeria. Journal of Plant and Animal Ecology. 2018a;1(1):14–42.
12. Izah SC, Bassey SE, Ohimain EI. Ecological risk assessment of heavy metals in cassava mill effluents contaminated soil in a rural community in the Niger Delta Region of Nigeria, Molecular Soil Biology. 2018b;9(1):1–11.
13. Izah SC. Feed potentials of Saccharomyces cerevisiae biomass cultivated in palm oil and cassava mill effluents. Journal of Bacteriology and Mycology Open Access. 2018c;6(5):287‒293.
14. Izah SC. Variations in microbial density and in-situ water quality characteristics of cassava fermentation medium for fufu production. MOJ Toxicology. 2018d;4(6):386‒389.
15. Izah SC, Aigberua AO. Assessment of Microbial Quality of Cassava Mill Effluents Contaminated Soil in a Rural Community in the Niger Delta, Nigeria. EC Microbiology. 2017;13(4):132–140.
16. Izah SC, Ohimain EI. Bioethanol production from cassava mill effluents supplemented with solid agricultural residues using bakers’ yeast [Saccharomyces cerevisiae]. Journal of Environmental Treatment Techniques. 2015;3(1):47–54.
17. Kigigha LT, Nyenke P, Izah SC. Health risk assessment of selected heavy metals in gari (cassava flake) sold in some major markets in Yenagoa metropolis, Nigeria. MOJ Toxicol. 2018;4(2):47–52.
18. Seiyaboh EI, Izah SC. Mortality Rate of Juvenile Heterobranchus bidorsalis Exposed to Cassava Mill Effluents. Annals of Review and Research. 2018;4(1):555628
19. Baigent MJ, Carpenter K. Vitamin. Accessed 20th December, 2018.
20. Nkwocha C. Proximate and micronutrient analyses of synsepalum dulcificum pulp. Scientific Research Journal. 2014;2(1):71–74.
21. Okwu DE. Phytochemicals, vitamins and mineral contents of two Nigerian medicinal plants. International Journal of Molecular Medicine and Advance Sciences. 2005;1(4):375–381.
22. Pepper IL, Gerba CP. Environmental microbiology. A laboratory manual. 2nd edn. Elsevier academic press. 2004.
23. Benson HJ. Microbiological Applications: Laboratory Manual in General Microbiology/complete version, 5th edn. New York: McGaraw-Hill; 2002.
24. Kurtzman CP, Fell JW. The Yeasts: A Taxonomic Study. 4th edn, Elsevier Science, Amsterdam, The Netherlands. 1998.
25. Iwuagwu JO, Ugwuanyi JO. Treatment and Valorization of Palm Oil Mill Effluent through Production of Food Grade Yeast Biomass. Journal of Waste Management. 2014.
26. Abioye OP, Afolayan EO, Aransiola SA. Treatment of Pharmaceutical Effluent by Saccharomyces cerevisiae and Torulaspora delbrueckii Isolated from Spoilt Water Melon. Research Journal of Environmental Toxicology. 2015;9(6):188–195.
27. Okoduwa SIR, Igiri B, Udeh CB, et al. Tannery Effluent Treatment by Yeast Species Isolates from Watermelon. Toxics. 2017;5(1).
28. Ellis D, Davis S, Alexiou H, et al. Descriptions of Medical Fungi. 2nd edn. Printed in Adelaide by Nexus Print Solutions, Underdale, South Australia, 2007.
29. Drevanasiddappa HD, Veena MA. Sensitive spectrophotometric determination of ascorbic acid. E journal of Chemistry. 2008;5(1):10–15.
30. Palacios, C. The Role of Nutrients in Bone Health, from A to Z. Crit Rev Food Sci Nutr. 2006;46(8):621–628.
31. Black wood I, Duddy G. Assessing stock feed additives and mineral supplements. Prime facts. 2017.
32. Bradbury JH, Holloway WD. Cassava, M. esculenta. Chemistry of tropical root crops: significance for nutrition and agriculture in the Pacific. Australian Centre for International Agricultural Research, monograph nr 6, Canberra, Australia, 1988. p. 76–104.
33. Woot Tsuen WL, Busson F, Jardin C. Food composition table for use in Africa. FAO corporate document repository. Rome, Italy, 1968.
34. Favier JC. Valeur alimentaire de deux aliments de base Africains: le manioc et le sorgho. Paris, France: ORSTOM (editions de l’Office de la Recherche Scientifique et Technique Outre-mer). Travaux et documents nr 67, 1977.
35. Montagnac JA, Davis CR, Tanumihardjo SA. Nutritional Value of Cassava for Use as a Staple Food and Recent Advances for Improvement. Comprehensive Reviews in Food Science and Food Safety. 2009;8:181–194.
36. Salvador EM, Steenkamp V, McCrindle CME. Production, consumption and nutritional value of cassava (Manihot esculenta, Crantz) in Mozambique: an overview. Journal of Agricultural Biotechnology and Sustainable Development. 2014;6(3):29–38.
37. Lancaster PA, Ingram JS, Lim MY, et al. Traditional cassava-based foods: survey of processing techniques. Econ Bot. 1982;36:12–45.
38. Tufarelli V. Enhancing Egg Quality by Dietary Vitamin E and Selenium Supplementation. Vitam Miner. 2014.3:e131.
39. Chen Q. More Than a Vitamin. EC Nutrition ECO.01 2016. p. 04-05.
40. Izah SC. Growth Pattern of Saccharomyces cerevisiae in Cassava Mill Effluents. Journal of plant and Animal Ecology. 2018e;1(2):10–15.
41. Babalola OO. Cyanide content of commercial gari from different areas of Ekiti state, Nigeria. World Journal of Nutrition and Health. 2014;2(4):58–60.
42. Uhegbu FO, Akubugwo EI, Iweala EEJ. Effect of Garri processing effluents [waste water] on the cyanide level of some root tubers commonly consumed in the South East of Nigeria. African Journal of Food, Agriculture, Nutrition and Development. 2012;12(5):6748–6758.