Effect of Solid State Fungal Fermentation on the Chemical Composition of Adansonia Digitata Seed

Covenant Journal of Physical & Life Sciences (CJPL) Vol. 7 No. 1, March, 2019 (SE)

An Open Access Journal Available Online

Effect of Solid State Fungal Fermentation on the Chemical Composition of Adansonia digitata Seed

1*Majekodunmi Racheal Adedayo & 2Alhassan Sani

1Microbiology Unit, Biosciences and Biotechnology Department, College of Pure and Applied Sciences, Kwara State University, Malete, Nigeria. 2Department of Microbiology, Faculty of Life Sciences, University of Ilorin, Ilorin, Nigeria. *[email protected] [email protected] [email protected]

Received: 24th October, 2018; Accepted: 10th February, 2019; Online Published: May 7st, 2019. URL http//Journal.covenantuniversity.edu.ng/cjoe/

Abstract: The seed of Adansonia digitata was fermented with the aim of producing additional plant protein food and feed. The seed was subjected to natural fermentation for 120 hours under laboratory condition. Nine moulds and two yeasts were isolated and characterized macroscopically and microscopically as Aspergillus niger, Aspergillus flavus, Penicillium citrinum, Penicillium chrysogenum, Mucor racemosus, Mucor hiemalis, Rhizopus stolonifer, Alternaria tenuis, Scopuloriopsis brevicaulis, Saccharomyces cerevisiae and Schizosaccharomyces pombe. Spores of isolated fungi were used as starter cultures in the fermentation of the seed using solid-state fermentation method for 120 hours. The fermented products were analyzed for proximate and anti-nutrient content using standard methods. The result showed a significant increase (p<0.05) in crude protein, total ash and carbohydrate but decreased significantly (p>0.05) in crude fat and crude fibre. The anti-nutrients in form of total tannins, saponins, oxalate and phytate content were significantly decreased after fermentation. From the findings in this research, it was concluded that fermented A. digitata seed can serve as additional plant protein food source in food and feed formulation for livestock. Key words: Adansonia digitata, anti-nutrients, nutrients, fermentation, fungi.

Introduction fermentation. Fermentation is one of the The growing concern for the acute food oldest methods and widely used process shortages for the world’s expanding of producing and preserving food on local population has led to the exploitation of and industrial levels [8]. Fermentation for non-conventional food sources as food production may either be anaerobic potential alternatives [1]. The solution to or aerobic or both. Fermentation changes the food problem must be sought through the characteristics of the food by the a combination of all available sources. action of the enzymes produced by the Due to the increasing demand for protein fermenting microbes whether bacteria, and energy to support the ever-increasing mould or yeasts. The term solid-state world population, efforts are been directed fermentation (SSF) has been variously at exploring new and nonconventional defined as the cultivation of sources of food that grow in the arid and microorganisms on solid, moist substrates semiarid land regions of the world. Food in the absence of free aqueous phase [9], and agricultural scientists are now or cultivation of microorganisms in the beginning to screen wild and under- presence of a liquid phase at maximal exploited native plants for possible solid substrate concentrations or on inert potential sources of food in an attempt to carriers supporting moist substrate [10]. It widen the narrow food base [2, 3]. Several has several advantages over liquid state or reports have also indicated that lots of submerged fermentation. Adansonia lesser-known native crop species are high digitata, the baobab tree, is a member of in nutrients and could possibly relieve the Bombacaceae family, which consists critical food shortages if given adequate of around 20 genera and around 180 promotion and research attention [4, 5]. species [11]. Authors [12] reported that However, prior to utilization of such the acceptability and optimal utilization of unconventional resources, data indicating A. digitata seed as a protein source is the nutrient composition and toxic factors limited by the presence of inherent anti- should be available. Toxicological nutrients such as protease inhibitors, evaluation of possible epidemiological tannins, phytic acid and amylase response to the ingestion of novel food inhibitors. Phytochemicals (anti-nutrients) sources and the methods of processing have been reported to be toxic at 5 g per that will enhance their utility as food or serving [13]. The acceptability and feed ingredient are all necessary in order optimal utilization of A. digitata seed as a to achieve optimal utilization [6]. protein source has been reported to be Competition between man and his limited by the presence of anti-nutritional livestock for food sources has been factors such as trypsin inhibitors, protease recognized as a major cause of the inhibitors, tannins, phytic acid, oxalate, increase in the cost of ingredients used in alkaloids, phytate and amylase inhibitors compounding livestock feed. This has [14, 15, 16]. [17] suggested that though been recognized to account for more than processing techniques may rob a food 70% of the total cost of animal production item of some nutrients, processing thus seriously reducing the return and systems may also enhance food nutritional marginal profit [7] in the developing quality by reducing or destroying the anti- countries essentially in Africa and nutrients present. The aim of this work particularly in Nigeria. Several methods therefore, is to evaluate the proximate and have been employed to improve the antinutrient content of A. digitata seed nutritional quality of legumes, cereals and fermented with mono-culture fungi under other form of seeds, essentially

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE) solid state techniques with a view to fungal isolates were preserved on agar determining their nutritive potentials. slant at 4 oC for further use. Materials and Methods Fungal spore preparation and Collection and authentication of seed monoculture fermentation of seed Mature, dried A. digitata pods were Fungal spore suspension of actively collected from the premises of University growing mid log phase culture of the of Ilorin, Ilorin, Kwara State, Nigeria was fungal isolates were prepared according to authenticated at the Department of Plant the method described by [21]. An agar Biology with voucher number of slant of four days old pure culture of each Adansonia digitata (UIH 1048). of the organisms was used. Sterile Preparation of seed distilled water (10 ml) was added to the The pods were cracked manually to slant and shook well to wash the spores. release the seeds. The seeds ebbed in pulp The spore suspension was counted using were washed with plenty of clean water to the Neubauer counting chamber. A spore remove the pulp before drying to remove suspension of about 5x104 spore/ml was the wetness. The seeds were pulverized used in each case for inoculation. Twenty with an electrical grinder to rough grams (20 g) of the seed samples were particles sizes of about 2 mm in diameter, measured separately into 250 ml thereafter they were stored in airtight Erlenmeyer flasks, plugged with cotton container for further use. wool, wrapped with aluminium foil and Isolation of organisms from naturally sterilized in the autoclave at 121 oC for fermented A. digitata seed 15minutes. The sterile samples were The pulverized seeds were subjected to mixed with 20ml of sterile distilled water natural fermentation as follows. Precisely and stirred properly until uniform mashes 250 g of the pulverized seeds were mixed were obtained in each case. Two millilitre with 250 ml of sterile distilled water in (2 ml) from each of the monoculture plastic fermentors of two litre capacities. suspension was used as fermentation The mixtures were stirred properly until a starter to inoculate each of the samples in uniform mash was obtained, covered and the fermentors. The mixtures were allowed to ferment at room temperature allowed to ferment for 120 hours at room (28±2 oC) in the laboratory for seven days or ambient temperature (28±2oC) [18, 19]. [18, 19]. Fungi were isolated from the Fermented samples were taken daily, naturally fermented seeds through serial dried at 60 oC in the oven for 4hours to dilution and pour plate method using safe moisture content and used for Potato- Dextrose agar into which 10 % analysis of proximate (moisture content, Streptomycin has been added to inhibit crude fibre, crude protein, crude fat, ash bacteria growth. Culturing was done in content, carbohydrate and glucose), duplicates. phytochemical (saponins, tannins, Identification of the Isolates phytates and oxalates) analysis were done The fungi isolated from the fermenting on fermented products after 120 hours. mixture were sub cultured until pure Determination of proximate content of isolates were obtained. Morphological and the seed microscopical analyses for the The crude protein content of the samples identification of the isolates were carried was determined following the Kjeldahl out and result obtained compared with method. literature to identify the organisms as The moisture content, total ash, crude described by [20]. Pure cultures of the fibre, crude fat (Soxhlet extraction 13

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE) method) content of the seed were the proximate composition of the seed. determined following [22]. The total The results obtained are significantly carbohydrate or Nitrogen Free Extract (p<0.05) different from the unfermented (NFE) was determined by the difference sample. Crude protein, carbohydrate and method.The reducing sugar content of the total ash were significantly (p<0.05) seed wasdetermined quantitatively by the increased after fermentation while the 3,5- Dinitrosalicylic Acid (DNSA) as crude fat and crude fibre were described by [22]. The water holding significantly (p<0.05) decreased. The capacity of the seed was determined highest increase in protein (31.29%) was according to the method described by recorded in sample fermented with [23]. Penicillium citrinum while the lowest Determination of the some anti- (28.80%) was recorded in sample nutritional factors in the seed fermented with Mucor racemosus (Table The antinutrient content of the seed was 2). determined quantitatively. Total saponins Reducing sugar (glucose) content of was determined by the method of [24], fermented seed oxalates by the method of [25], total The effect of fermentation on the reducing soluble tannin by the method described by sugar content of monoculture-fermented [26] and the phytate content was seed is as presented in Figure 1. The value determined by the method of [27]. of reducing sugar increased significantly 3. Results (p<0.05) in all the fermented products Isolation of Fungi compared to the unfermented sample. The Nine moulds were isolated and identified highest increase (16.5 mg/g) was recorded during fermentation as Aspergillus niger, in sample fermented with Aspergillus flavus, Penicillium Shizosaccharomyces pombe while the chrysogenum, Penicillium citrinum, value was insignificantly reduced Rhizopus stolonifer, Mucor racemosus, (p<0.05) to 2.5mg/g recorded in sample Mucor hiemalis, Alternaria tenuis, fermented with Rhizopus stolonifer. Scopulariopsis brevicaulis, Effect of monoculture fermentation on Saccharomyces cervisiae and some anti-nutrients in the fermented Schizosaccharomyces pombe (Table 1). seed. Proximate content of fermented The effect of monoculture fermentation products on saponins, oxalates, tannins and The effect of monoculture fungal phytates content of A. digitata seed is fermentation on A. digitata seed at presented in Figures 2 to 5. The values of ambient temperature for 72 hours is the phytochemicals significantly (p<0.05) presented in Table 2. Fermentation with reduce after fermentation. each of the fungus has various effects on

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

Macroscopy and microscopy description of isolates Fungal isolates

Filaments were white fast spreading with brown- Aspergillus niger black spores at the tip Colonies were greenish yellow with rough edges, Aspergillus flavus flat and fast spreading The filaments were bluish green or pale with broad Penicillium chrysogenum white margin Filaments were whitish on top but leathery and Penicillium citrinum orange yellow underneath Colony appeared fluffy white but turned black with Rhizopus stolonifer age Filaments appeared gray, fluffy white but turn dirty Mucor racemosus white with age Mycelia were grayish or slightly yellowish turning Mucor hiemalis grayish brown with age Colony has blackish aerial mycelium; spores were Alternaria tenuis dark, brown borne in chains Colony has brownish loose hyphae carrying lemon shaped spores Scopulariopsis brevicaulis Colonies were creamy with rough edges; cells were Saccharomyces cervisiae ovoid or ellipsoidal Colony was round and smooth at the edge; cells Schizosaccharomyces were cylindrical pombe

Table 1: Fungal isolates from naturally fermented pulverized A. digitata seed

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

Organisms MC PRO FAT FIB CHO TA BTe 6.98±1.39ab 4.32±0.22ab 31.50±2.06c 11.92±0.64a 4.46±0.7ab 40.84±0.88ab

BSt 5.87±0.00a 3.96±0.00a 29.43±0.00ab 12.57±0.00ab 4.96±0.00ab 43.21±0.00bc

BRa 7.43±1.28bc 4.45±0.40ab 28.80±1.12ab 11.85±0.13a 4.88±0.21ab 42.6±0.24bc

BHe 6.29±0.05ab 4.44±0.05ab 28.92±0.33ab 11.91±0.60a 4.48±0.62ab 43.98±1.55bc

BCh 8.20±3.34c 6.75±2.02b 30.83±0.34bc 11.77±0.46a 3.74±1.90a 38.76±7.40a

BCt 8.27±3.29c 6.61±2.25ab 31.29±2.60c 12.25±0.41a 3.45±1.34a 38.14±7.21a

BNi 7.25±1.89bc 5.52±0.97ab 30.36±2.04bc 13.31±0.89ab 3.71±1.91a 39.87±1.95a

BFl 7.91±3,10bc 3.89±0.74a 30.60±1.68bc 12.59±0.21ab 3.65±1.91a 41.36±3.39bc

BCe 7.50±0.00bc 5.00±0.00ab 31.00±0.00c 13.50±0.00ab 3.50±0.00a 40.20±0.00ab

BPo 8.00±0.00c 5.00±0.00ab 30.00±0.00bc 13.00±0.00ab 3.00±0.00a 40.10±0.00ab

BAo 7.64±2.11bc 4.39±1.61ab 19.86±1.30 a 18.60±8.46b 8.87±5.31b 40.01±1.35ab

Table 2: Proximate composition of Adansonia digitata fermented with mono-culture of fungal isolates.

Data are mean of two replicate ± SEM. Mucor racemosus, BHe= baobab Mean within the same row carrying fermented with M. hiemalis, BCh= different superscripts are significantly baobab fermented with P. chrysogenum, different at (p<0.05). (MC= moisture BCt= baobab fermented with P. citrinum, content, TA= total ash, CPRO= crude BNi= baobab fermented with A. niger, protein, CFAT= crude fat, CFIB= crude BFl= baobab fermented with A. flavus, fibre and CHO= total carbohydrate.) BPo= baobab fermented with S. pombe, (BSt= baobab fermented with BCe= baobab fermented with S. R.stolonifer, BTe= baobab fermented with cerevisiae and BRo= unfermented baobab A. tenuis, BRa= baobab fermented with seed)

8020 7018 f Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

6016 e e 14 d 50 12 40 c 10

308 SaponinGlucose b

Saponin Saponin content (Mg/g) 206 Concentration (mg/g) b a b b b 4 10 a 2 0 0 B Ao B Te BSt B Ra B He BCh BCt B Ni BFl BBr B Po Bce B Ra B He BFl B Ni BCh BCt BSt B Te B Ce B Po BBr Fungi used for fermentation Monoculture fermented samples

Data are mean of two replicate ± SEM.

Figure 1: Glucose content of monoculture fungal fermented Adansonia digitata

(BAo: unfermented baobab; BTe:Moringa fermented with Aspergillus niger; BFl: fermented with Alternaria tenuis; BSt: baobab fermented with Aspergillus baobab fermented with Rhizopus flavus; BBr: baobab fermented with stolonifer; BRA: baobab fermented with Scopuloriopsis brevicaulis; BPo: baobab Mucor racemosus; BHi: baobab fermented with Schizosaccharomyces fermented with Mucor hiemalis; BCh: pombe; BCe: baobab fermented with baobab fermented with Penicillium Saccharomyces cerevisiae) Data are mean chrysogenum; BCt: baobab fermented of two replicate ± SEM. with Penicillium citrinum; BNi: baobab

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

Organism MC PRO FAT FIB CHO s TA BTe 6.98±1.39ab 4.32±0.22ab 31.50±2.06c 11.92±0.64a 4.46±0.7ab 40.84±0.88ab

BSt 5.87±0.00a 3.96±0.00a 29.43±0.00a 12.57±0.00a 4.96±0.00ab 43.21±0.00bc

b b BRa 7.43±1.28bc 4.45±0.40ab 28.80±1.12a 11.85±0.13a 4.88±0.21ab 42.6±0.24bc b BHe 6.29±0.05ab 4.44±0.05ab 28.92±0.33a 11.91±0.60a 4.48±0.62ab 43.98±1.55bc

b BCh 8.20±3.34c 6.75±2.02b 30.83±0.34b 11.77±0.46a 3.74±1.90a 38.76±7.40a c BCt 8.27±3.29c 6.61±2.25ab 31.29±2.60c 12.25±0.41a 3.45±1.34a 38.14±7.21a

BNi 7.25±1.89bc 5.52±0.97ab 30.36±2.04b 13.31±0.89a 3.71±1.91a 39.87±1.95a

c b BFl 7.91±3,10bc 3.89±0.74a 30.60±1.68b 12.59±0.21a 3.65±1.91a 41.36±3.39bc

c b BCe 7.50±0.00bc 5.00±0.00ab 31.00±0.00c 13.50±0.00a 3.50±0.00a 40.20±0.00ab b BPo 8.00±0.00c 5.00±0.00ab 30.00±0.00b 13.00±0.00a 3.00±0.00a 40.10±0.00ab c b BAo 7.64±2.11bc 4.39±1.61ab 19.86±1.30 a 18.60±8.46b 8.87±5.31b 40.01±1.35ab

Table 2: Proximate composition of Adansonia digitata fermented with mono-culture of fungal isolates.

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

20 18 f

16 e e 14 d

12 c 10

8 Glucose b 6 Concentration (mg/g) b Data are mean aof twob replicate ± SEM. b b Figure 1: 4Saponin content of mono-culture fungal fermenteda Adansonia digitata 2 (BAo: unfermented baobab; BTe:Moringa with Penicillium citrinum; BNi: baobab fermented0 with Alternaria tenuis; BSt: fermented with Aspergillus niger; BFl: baobab fermentedB Ra B He withBFl B NiRhizopusBCh BCt BStbaobabB Te fermentedB Ce B Po BBr with Aspergillus stolonifer; BRA: baobab fermentedMonoculture with fermentedflavus; samples BBr: baobab fermented with Mucor racemosus; BHi: baobab Scopuloriopsis brevicaulis; BPo: baobab fermented with Mucor hiemalis; BCh: fermented with Schizosaccharomyces baobab fermented with Penicillium pombe; BCe: baobab fermented with chrysogenum; BCt: baobab fermented Saccharomyces cerevisiae)

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

30 Saponin

Saponin Saponin content (Mg/g) 20

0 B Ao B Te BSt B Ra B He BCh BCt B Ni BFl BBr B Po Bce Data are mean of twoFungi replicate used ± for SEM. fermentation

Figure 2: Saponin content of monoculture fungal fermented Adansonia digitata

(BAo: unfermented baobab; with Penicillium citrinum; BNi: baobab BTe:Moringa fermented with Alternaria fermented with Aspergillus niger; BFl: tenuis; BSt: baobab fermented with baobab fermented with Aspergillus Rhizopus stolonifer; BRA: baobab flavus; BBr: baobab fermented with fermented with Mucor racemosus; BHi: Scopuloriopsis brevicaulis; BPo: baobab baobab fermented with Mucor hiemalis; fermented with Schizosaccharomyces BCh: baobab fermented with Penicillium pombe; BCe: baobab fermented with chrysogenum; BCt: baobab fermented Saccharomyces cerevisiae)

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

Oxalate 2

Oxalate content (Mg/g) 1

0 B Ao B Te BSt B Ra B He BCh BCt B Ni BFl BBr B Po BCe Fungi used for fermentation

Data are means of two replicates ± SEM. Figure 3: Tanin content of mono-culture fungal fermented Adansonia digitata

(BRO: unfermented baobab; with Penicillium citrinum; BNi: baobab BTe:Moringa fermented with Alternaria fermented with Aspergillus niger; BFl: tenuis; BSt: baobab fermented with baobab fermented with Aspergillus Rhizopus stolonifer; BRA: baobab flavus; BBr: baobab fermented with fermented with Mucor racemosus; BHi: Scopuloriopsis brevicaulis; BPo: baobab baobab fermented with Mucor hiemalis; fermented with Schizosaccharomyces BCh: baobab fermented with Penicillium pombe; BCe: baobab fermented with chrysogenum; BCt: baobab fermented Saccharomyces cerevisiae)

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

2.5

1.5

1 Tannins

Tannin content (Mg/g) 0.5

0 B Ao B Te BSt B Ra B He BCh BCt B Ni BFl BBr B Po Bce

Fungi used for fermentation

Data are means of two replicates ± SEM. Figure 4: Tanin content of mono-culture fungal fermented Adansonia digitata

(BAo: unfermented baobab; BTe:Moringa with Penicillium citrinum; BNi: baobab fermented with Alternaria tenuis; BSt: fermented with Aspergillus niger; BFl: baobab fermented with Rhizopus baobab fermented with Aspergillus stolonifer; BRA: baobab fermented with flavus; BBr: baobab fermented with Mucor racemosus; BHi: baobab Scopuloriopsis brevicaulis; BPo: baobab fermented with Mucor hiemalis; BCh: fermented with Schizosaccharomyces baobab fermented with Penicillium pombe; BCe: baobab fermented with chrysogenum; BCt: baobab fermented Saccharomyces cerevisiae)

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

10 Phytates

Phytate content (Mg/g) 5

0 B Ao B Te BSt B Ra B He BCh BCt B Ni BFl BBr B Po Bce Fungi used for fermentation

Data are means of two replicates ± SEM.

Figure 5: Phytate content of mono-culture fungal fermented Adansonia digitata

(BAo: unfermented baobab; BTe:Moringa be attributed to the amylolytic and fermented with Alternaria tenuis; BSt: proteolytic activities of the fungi involved baobab fermented with Rhizopus during growth in the process of stolonifer; BRA: baobab fermented with fermentation. These enzymes were Mucor racemosus; BHi: baobab involved in breaking down complex fermented with Mucor hiemalis; BCh: organic molecules into simpler ones such baobab fermented with Penicillium as glucose and amino acids. The product chrysogenum; BCt: baobab fermented of the enzymes is also utilized by the with Penicillium citrinum; BNi: baobab organisms for their own metabolic fermented with Aspergillus niger; BFl: activities [28]. Moreover the noticeable baobab fermented with Aspergillus increase in crude protein in the mono- flavus; BBr: baobab fermented with culture fermented seed could also be Scopuloriopsis brevicaulis; BPo: baobab attributed to the addition of mycoprotein fermented with Schizosaccharomyces (single cell protein), non-protein nitrogen pombe; BCe: baobab fermented with amide and nucleic acid synthesized by Saccharomyces cerevisiae) fungal cells during growth [19]. The

Discussion improvement observed could be due to the The valuable nutrients in the fermented breaking down of the complex organic seed such as crude protein, mineral ash carbon compound in the seed hence the and total carbohydrate were increased increase in glucose and decrease in crude after fermentation. This improvement can fiber and fat content [29]. However the

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE) available carbohydrate and lipid in the the availability of proteins, carbohydrates fermented seed was still sufficient to meet and minerals by forming indigestible the daily requirement for animal feed, also complexes with the nutrients. The they are now in a readily accessible form reduction in tannin level due to upon consumption by animals when fermentation could improve the incorporated into food and feeds items. availability of nutrients in the seed [36, The increase in carbohydrate content may 37, 31]. However, this result is contrary to also be attributed to the reduction in the the report of [30] on tannin content after crude fibre content. During fermentation fermentation of A. digitata seed. polysaccharides including cellulose, Fermentation has been reported as a pectin, lignocellulose and starch are means to improve the nutritional quality broken down by microorganisms thereby and drastically reduced anti-nutritional reducing the fibre content of such seed. factors to safe level because the process This result was in contrast to that obtained produces enzymes that break down by earlier author on carbohydrate content protein-tannin complexes to release free of fermented A. digitata seed [30, 31] but tannins [36]. Similarly the reduction in was similar to that obtained by [19, 32, phytic acid is attributable to increase in 33] on effect of fermentation on the activities of phytase during Mangifera indica and pigeon pea seed. fermentation. According to earlier report, The improvement in total ash recorded the series of enzymatic actions can be attributed to the breakdown of terminating with the formation of inositol organic complexes like protein tannin and phosphoric acid releases certain complex or calcium oxalate complex in metals to increase their availability and the seed to release the minerals into caused subsequent decrease in phytate and biologically available form [34]. The increase in total ash [31, 38]. Moreover, inherent antinutrients content was lowered reduction of phytic acid content of some significantly (p<0.05) after fermentation. plant products after processing has been The decrease in tannin, saponins, oxalates reported [39, 40]. Phytic acid has been and phytate content of A. digitata seed reported to form complexes with proteins could be due to degradation by microbial (protein-phytate complex) [41] and enzymes that are secreted by the chelates essential dietary minerals such as fermenting fungi during growth and iron, zinc, calcium and magnesium, thus metabolism in the process of [35, 31]. decreasing their utilization. These antinutrients are known to occur in Furthermore, saponins has been shown to complex compounds (protein-tanin affect animal nutrition, performance and complexes, calcium- oxalate complex, metabolism through erythrocyte phytate and saponin protein complexes haemolysis, depression of growth rate, and other forms of complexes) in plants bloat (ruminants), inhibition of smooth where they occur. The formation of these muscle activity, alter cell wall complexes binds protein and minerals in permeability and therefore produce some the seed and renders them biologically not toxic effects when ingested, cause enzyme available to plants after consumption. inhibition and reduction in nutrient The breaking down of these complexes by absorption if it occurs beyond certain various amylolytic, proteolytic and level in feed. [42]. Saponin content of A. lipolytic enzymes produced by these fungi digitata seed was significantly (p<0.05) probably led to the reduction in anti- decreased after fermentation. The finding nutrients. Tannins are known to reduce was similar the report of [43, 38]. 24

Majekodunmi Racheal Adedayo & Alhassan Sani CJPL (2019) 7(1) 11-28 (SE)

High occurrence of oxalates in feeds, has oxalate content of mango kernel cake been previously shown to bind minerals fermented by A. niger. like calcium and magnesium and interfere Conclusion with their metabolism, cause muscular Monoculture fungal fermentation has weakness and paralysis, gastrointestinal desirable effects on the seed of A. tract irritation, blockage of the renal digitata. Fermented seed could therefore tubules by calcium oxalate crystals, be considered as an alternative, development of urinary calculi and supplement or additional protein source hypocalcaemia [44]. Oxalates content 22 of for propounding animal feed. This will fermented A. digitata seeds reduced tremendously reduce the cost of feeding in significantly after fermentation with the animal production and also reduce the isolated fungi. Fermentation and other competition between man and animal for methods of processing have been reported the available staple food. However only to decrease the oxalate content of food fungi without history of poisoning or [45]. [19] however reported an increase in production of harmful metabolites should be employed for the fermentation.

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