Background

Nigeria, like most developing countries of the world is facing food crisis. The diet of most of her populace is often protein deficient (Igene et al., 2016) and according to FAO (2008), its average animal protein consumption per person per day is 7.9 g which is far below Food and Agriculture Organisation recommendation of 35 g as the average animal protein consumption per person per day (FAO, 2000). This low level of animal protein intake by Nigerians has generated concern as it affects physical, physiological and mental development of Nigerian youth and labour force in the country (Akinmutimi, 2004). There is, therefore the need for conscious efforts toward adequate supply of animal protein from animal products that are rich in protein, energy, vitamins and minerals required for adequate nutrition. Fish are source of protein rich in essential amino acids, micro and macro elements (calcium, phosphorus, fluorine, iodine), fats that are valuable sources of energy, fat-soluble vitamins, and unsaturated fatty acids that, among other benefits, have a hypocholesterolic effect (anti-arteriosclerosis) (Usydus et al., 2008).

In the last decade, there has been an increase of protein-based fish products such as: fish sausage, fish nuggets, fish cake, fish balls, etc. The quality of these products depends on the raw materials used in their manufacture. Thus, various protein-based fish products will depend on the amount of myofibril proteins that are present in the fish species (Darmanto et al., 2014).

Consumption of snack food is a worldwide practice (Osibona et al., 2009). Snack foods are important vehicles for delivery of essential nutrients because of the growing change in eating habits (Henshaw and Agunbiade, 2004). In Nigeria, there have been limited studies on the utilization of fish as fish based snack (suya). Plants have been reported to have beneficial effects on human health especially in managing and preventing non communicable diseases such as diabetes, obesity, coronary heart diseases etc. Plant crude extracts have the advantage to have been consumed by humans for thousands of years with little or no adverse effect, and besides antimicrobial importance, several plants are being used in different areas of human health such as traditional medicine, functional foods, dietary supplements and recombinant protein manufacturing (Carmen et al., 2014) Their role as antimicrobials derives from their capacity to control natural spoilage processes (food preservation) and to prevent/control growth of microorganisms, including pathogenic micro-organisms (Tajkarimi et al., 2012).

Apart from the use of herbs for antimicrobial purposes, herbs and spices are also of medicinal importance to human health. Bitter leaf (Vernonia amygdalina) and scent leaf (Ocimum gratissimum) as herbs have been reported by many researchers to be highly medicinal. Vernonia amygdalina has been found useful in the ethno therapy of asthma, schistosomiasis, malaria, measles, diarrhoea, tuberculosis, abdominal pain and fever. Arhoghro et al., (2009), reported that O. gratissimum crude extracts significantly reduced and even reversed carbon tetrachloride (CCl₄)-induced liver damage in Wister rats after establishment. Hence O. gratissimum might be an effective hepatoprotector in the diet of patients with hepatopathies (Arhoghro et al., 2009). Spices (Ginger, Garlic, African negro pepper, Peanuts) have been reported as a remedy for various ailments such as arthritis, muscle soreness, chest pain, low back pain, stomach pain, and menstrual pain, dyspepsia, acute or chronic gastritis, diarrhea, cardiovascular disease, arterial sclerosis and other disease related to oxidative stress (Manach et al., 2005). It can be used for treating upper respiratory tract infections, cough, and bronchitis. As an anti-inflammatory agent, it is recommended for joint problems (Shukla and Singh, 2007).

To gain consumer’s acceptance of a fish based product rather than the carbohydrate snacks, the consumer would have to be convinced of the quality and possible nutritional advantages with respect to the standard product it is replacing. This study examines the effect of plants (Ocimum gratissimum and Vernonia amygdalina) crude extract on the nutritional quality of Sarotherodon galilaeus snack.

1 Materials and Methods

1.1 Source of raw materials

The fish sample used for this study (Sarotherodon galilaeus) were procured from fish mongers at the landing site of the fishermen at Egbe reservoir, Ode-Ekiti, Ekiti State, South west Nigeria. Nine hundred and forty grams of Vernonia amygdalina and 827 gms of Ocimum gratissimum leaves and ingredients for the composite spice (suya spice) and other materials used were procured from Akungba Akoko, Ondo state, Nigeria.

1.2 Production of spices

Table 1 shows the the proportions and grams of different ingredients used in the spice. The following ingredients are; Human edible Groundnut cake locally called “kulikuli”, Ginger, Garlic, Cayenne pepper flakes, Black Pepper, Cooking salt and African Negro pepper. These ingredients were ground into powdery form to increase surface area and improve attachment. The spice was similar to “Yaji” that is “suya” spice.

1.3 Preparation of brine and extracts

Nine hundred and forty grams of Vernonia amygdalina and 827 g of Ocimum gratissimum leaves were quickly washed in jet of tap water and rid of excess water before squeezing into separate bowls to obtain crude extract. The extracts were diluted to acceptable taste based on taste range finding test. Ocimum gratissimum was diluted with potable water to give 54.5 v/v concentration; also, Vernonia amygdalina was diluted with potable water to give 23 v/v concentration while equal volume of Vernonia amygdalina and Ocimum gratissimum extract mixed together was diluted to give 45.5 v/v concentration. Eight grams of salt was added to Vernonia amygdalina extract, 6 gms of salt to Ocimum gratissimum extract and 8 gms of salt into mixture of Vernonia amygdalina and Ocimum gratissimum extract for brining. A five man taste panelists was used to determine the acceptable level of extract and salt concentration.

1.4 Preparation of fish snack

The fish samples were washed in jet of tap water to remove dirt and thereafter, allowed to drain. The fish were scaled, gutted and washed clean again after gutting to prevent contamination of the muscle. Thereafter, they were filleted, and the weight of the fillets taken. The fillets were cut into smaller pieces (2.5 cm x 6 cm), the pieces were staked on sterilized palm frond sticks. Staked fillets pieces were weighed and soaked in brine and mixture of plant extracts and brine for two minutes, the brined/brine plus extracts stakes were hung to drain excess solution after which oil and spice was applied. Smouldering fire from wood and charcoal combustion was used to cook dry the snacks. The stakes were arranged in a circular form around the smoldering fire at an average distance of 30 cm away from direct heat.

1.5 Proximate composition of the snack

Proximate composition of the snack was determined by conventional method of Association of Official Analytical Chemists.

1.6 Moisture content

Moisture content of the snack was determined according to the method of Association of Official Analytical Chemists (AOAC, 2005). One gram of sample was dried in moisture dish in an oven at 105°C until constant weight was obtained.

Where: W₀=weight of crucible, W₁=weight of crucible+sample before drying, W₂=weight of crucible+oven dried sample W₁

1.7 Ash content

Ash content of the snack was determined according to the method of Association of Official Analytical Chemists (AOAC, 2005). Pre-dried samples obtained from moisture content analysis were ashed in furnace at 550°C overnight.

Where; W₀=weight of empty crucible, W₁=weight of sample+weight of the crucible (initial)

W₂=final weight of the ash residue+weight of crucible

1.8 Crude protein

Crude protein content of the snack was determined according to the method of Association of Official Analytical Chemists (AOAC, 2005). Briefly, one gram of the snack was weighed into digestion tubes. Two Kjeltabs Cu 3.5 (catalyst salts) were added into each tube. About 20 mls of concentrated tetraoxosulphate (VI) acid (H₂SO₄) was carefully added into the tube and then shaken gently. Digestion procedure was carried out. Digested samples were cooled for 10-20 minutes. Distillation procedure was then performed using distillation unit and the distillate was titrated with 0.025 N tetraoxosulphate (VI) acid (H₂SO₄) until the end point changes from green to pink. Volume of acid used for the titration was recorded. Blank was prepared with the exclusion of sample. The percentage of protein content was calculated according to the equation below.

% Nitrogen= 0.014×VD×N×100×TV

Weight of sample×AD

% Protein=% N×F

Where,

VD=Volume of digest, N=Normality of acid, TV=Titre value,

AD=Aliquot of digest

F=Conversion factor for nitrogen to protein (6.25)

1.9 Lipid

Lipid value was obtained by exhaustively extracting 2.0 g of each sample in a Soxhlet apparatus using petroleum ether (b.p. 40-60°C) as the extractant.

Where; W1=weight of samples, W2=weight of extraction cup (empty (g)), W3=weight of extraction cup+residue (g)

1.10 Fibre content

One gram of dried sample was digested with 0.128 ml of tetraoxosulphate (VI) acid (H₂SO₄) with two drops of octanol to prevent foaming. The content was boiled for 30 min, and then filtered and washed with hot water to remove acid. The residue was boiled with 0.22 ml of potassium hydroxide (KOH) for 30 min, and then washed in boiling water and acetone. The residue was dried and ignited in furnace. The loss of weight represents the crude fibre (Fawole et al., 2013).

1.11 Carbohydrate

Carbohydrate content was calculated based on difference calculation [Carbohydrate=100%-(% moisture+% ash+% crude protein+% fat)] (Olagunju et al., 2012).

1.12 Mineral concentration

The percentage mineral elemental concentration (macro and micro) including trace elements concentrations were determined using Atomic Emission Spectrophotometer (AES) and Atomic Absorption Spectrophotometer (AAS) (Fawole et. al., 2007).

1.13 Energy value

The total energy value of the snack was calculated using the method of (Palani et al., 2014). The crude values of protein, fat and carbohydrate as obtained from the proximate analysis were used as hereunder：

Energy value Kcal/100 g=Px4.00 Protein+Cx4.00 NFE Kcal/100+Fx9.00 Fat Kcal/100 g

1.14 Statistical analysis and data presentation

Data were subjected to one-way analysis of variance (ANOVA) and the means separated using Duncan’s Multiple Range. Statistical Analysis System (SAS^®), version 9.1 package was employed for statistical analysis and at 5% confidence level. Data were presented in mean±, percentile, as well as frequency, pie and bar chats as appropriate.

2 Results

2.1 Nutrient quality

2.1.1 Proximate composition

The proximate composition of snacks produced from S. galilaeus and S. galilaeus fortified with plant crude extracts is shown in Table 2. The percentage protein content obtained for the snacks ranged from 48.35±0.01% to 60.87±0.01%. The highest protein value was obtained from unfortified S. galilaeus snacks and it is significantly (p>0.05) higher than values obtained for S. galilaeus snacks fortified with crude extracts.

For moisture, the value obtained ranged from 7.34±0.01% to 8.45±0.01%, the highest value was recorded for snacks fortified with Vernonia amygdalina and mixture of Vernonia amygdalina plus Ocimum gratissimum extracts (8.45±0.01%).

Lipid content ranged from 12.71±0.23% to 13.15±0.05%. The unfortified snack had significantly higher value (13.15±0.05%).

Value for Ash ranged from 3.57±0.01% to 4.34±0.04%, the unfortified snacks recorded significantly lower value (3.57±0.01%) than the fortified snacks.

The total value of crude fibre ranged from 1.55±0.01 to 2.37±0.01 for all the snacks (fortified and unfortified). Snack fortified with Vernonia amygdalina plus Ocimum gratissimum extract recorded the highest value for crude fibre 2.37±0.01 which was significantly higher than unfortified snacks and snacks fortified with Ocimum gratissimum but not different from value obtained for Vernonia amygdalina fortified snack. 

Carbohydrate content of the snacks ranged from 13.64±0.03% to 23.70±0.04%, and significantly varied between the snacks, snacks fortified with Ocimum gratissimum extract had the least value (13.64±0.03%), the highest value (23.70±0.04%) was recorded for snacks fortified with Vernonia amygdalina extract.

2.2 Mineral composition

The concentrations of the selected elements obtained for the different snack samples are shown in Table 3. The values for Iron, Sodium, and Selenium 13.23±0.015 mg/100 g, 702.2±0.004 mg/100 g, and 2.44±0.072,009 mg/100 g respectively were significantly higher in snacks fortified with Vernonia amygdalina extract.

Snacks treated with Ocimum gratissimum recorded the highest value (3.07±0.009 mg/100 g) for Zinc; the least value (0.64±0.002 mg/100 g) was obtained for snacks fortified with Vernonia amyygdalina plus Ocimum gratissimum mixture extract.

Concentration value of Calcium showed variation among the crude plant extracts with values ranging from 80.82±0.029 mg/100 g to 360.9±0.121 mg/100 g, the highest concentration 360.9±0.121 was obtained in snacks fortified with Ocimum gratissimum while the unfortified snacks had the least concentration 80.82±0.029 mg/100 g with significant difference.

The result ranged from 75±0.006 mg/100 g to 130±0.002 mg/100 g. The concentrations of Magnesium obtained from both fortified and unfortified snacks were in the order of snack fortified with the mixture of Vernonia amyygdalina plus Ocimum gratissimum<Ocimum gratissimum<Vernonia amygdalina. All the samples were significantly different from each other.

Manganese, Copper and Potassium values 1.17±0.026 mg/100 g, 0.48±0.001 mg/100 g and 102.9±1.093 mg/100 g were significantly lowest in snacks fortified with the mixture of Vernonia amyygdalina plus Ocimum gratissimum extract. 

The nutritional profile of commercially sold snacks and Sarotherodon galilaeus snack is showned in Table 4.

3 Discussion

Fish is a highly proteinous food consumed by the populace; a larger percentage of consumers do eat fish because of its availability, flavors, palatability while fewer percentages do so because of its nutritional value. The nutritional value of fish meat comprises the contents of moisture, protein, lipids, vitamins and minerals plus the caloric values of the fish (Olopade et al., 2013). The crude protein content of samples fortified with Ocimum gratissimum (60.38%) crude extract and unfortified (60.87%) were higher than the crude protein content of samples fortified with the mixture of Ocimum gratissimum plus Vernonia amygdalina crude extract and Vernonia amygdalina crude extract. Generally, the protein content of the snacks were higher than the crude protein of the species in dried form reported by Olopade et al., (2013), protein content for O. niloticus reported by (Zenebe et al., 1998), protein content of minced fish reported by Hosseini-Shekarabi et al., (2014), and protein content of deep fried spiced minced fish cake reported by Osibona et al., (2009). Protein content of snack samples fortified with the mixture of Vernonia amygdalina plus Ocimum gratissimum and Vernonia amygdalina crude extract were 48.48% and 48.35% respectively, these values were higher than value reported by Fawole et al., (2007), for dried S. galilaeus reported by Fawole et al., (2013), and similar to those reported by Fashina-Bomobata and Megbewon (2012) for Tilapia species. The relatively high to moderate percentage crude protein could be attributed to the fact that; fishes are good source of pure protein; the differences observed in snack fortified with V. amygdalina could be due to the effect of alkaloids in Vernonia amygdalina crude extract on the protein. The amino group in the alkaloids could have reacted with the amino acid group of the fish protein thus leading to reduction of available protein in samples fortified with Vernonia amygdalina and mixture of Vernonia amygdalina plus Ocimum gratissimum crude extract. The relatively higher protein content of the snacks might be due to the effect of groundnut cake in the spice which is a very good source of protein.

The moisture content in this study ranged from 7.34±0.02% to 8.45±0.01%, which was higher than published work by Fawole et al (2007) on dried S. galilaeus, (5.80), the relatively higher moisture content in the present study could be due to the fact that the snack was processed to be fairly juicy, also the spice could also reduce the rate at which water is removed during processing.

The lipid content of 13.5±0.05 in the unfortified snack was significantly highest (p>0.05) while snack fortified with mixture of Vernonia amygdalina plus Ocimum gratissimum crude extract was lowest (12.71±0.23), (p<0.05) different from the lipid level of other snacks. The lipid content of the fish snack products were higher than values recorded in some literatures for S. galilaeus; (Fawole et al., 2013; Olopade et al., 2013; and Adewumi et al., 2014). The high lipid content obtained in this study could be due to the relatively high lipid content of groundnut cake contained in the composite spice. ‘Kilishi’ from beef and pork in other studies Prabhakaran and Mendiratta, 2013; Ogunsola and Omojola, (2008) recorded a slightly higher lipid content than that of S. galilaeus snack in this study, this could be due to the fish belonging to low fat group of fishes. However, fish lipid is largely composed of unsaturated fatty acids, which supplies Omega-3 PUFAS for lowering blood cholesterol level and high blood pressure (Oladipo and Bankole, 2013). 

Ash content of the snack ranged from 3.57±0.01% to 4.34±0.04%, snacks fortified with crude extract of Vernonia amygdalina and mixture of Vernonia amygdalina plus Ocimum gratissimum had the highest ash content 4.33±0.01% and 4.34±0.04% respectively. Values obtained in this study is lower than ash content reported by Olopade et al., (2013) for dried S. galilaeus but similar to value obtained from fish cake made from shrimp by-catch (Osibona et al., 2009).

The values obtained for fibre was higher than values (1.06±0.08; 0.12) reported by Fawole et al., (2007; 2013) and 0.71 reported by Idah et al., (2013) for 0.71 tilapia dried at 60°C for ten hours. The fibre content ranged from 1.55±0.01 to 2.37±0.01 with the unfortified snacks having significantly lower value (figure) than other snacks. The higher fibre value of the treated snack might be due to the fibre content of the plant extracts. However, this is not likely to result in adverse effect to consumers but rather might aid bowel evacuation and thus prevents constipation.

Carbohydrate content in this study ranged from 13.64±0.03% to 23.70±0.04%, the seemingly higher carbohydrate content may not be unconnected with the carbohydrate content of the spice, generally, as fish is low in carbohydrate. The carbohydrate level is advantageous because it will supply energy and allow the use of its protein for body maintenance and growth rather the protein being speared for energy through deamination which could over work the liver with consequence adverse effect on the health of the consumers.

The mean energy value (409.22±5.83 Kcal/100 g) of the snacks in this study is lower than energy value of some commercial snacks like ‘Top’ biscuits (523 Kcal/100 g), ‘Bigi’ sausage roll (477 Kcal/100 g), ‘Mimi chinchin’ (510 Kcal/100 g), ‘Oxford’ biscuit (420 Kcal/100 g) etc. Snacks treated with mixture of Vernonia amygdalina plus Ocimum gratissimum crude extract had the least energy value of 403.57 kcal/100 g and this can be recommended for hypertensive patients, more especially that it is also low in Sodium.

Studies have shown that Iron (Fe) is one of the most abundant metals in the earth crust (Ibrahim and Tayel, 2005). Iron is important in the formation of heamoglobin and its deficiency leads to anaemia (Sodamade, 2013). The high value of Fe observed in this study compared to other metals might not be unconnected with its availability and consequent uptake by the fish as well as availability in the spice ingredients. However, the values obtained for Fe in this study were significantly different between the treatments. The value ranged from 6.07±0.038 mg/100 g to 13.23±0.015 mg/100 g, the iron in the snack per stake is capable of supplying 20.6% of the daily recommended intake of 15 mg (NRC, 1989). Consumption of the snack could thus prevent anaemia.

Calcium is a macro element which is needed in large quantity in the body; it is involved in enzymatic systems and plays a role in regulating muscle contraction, transmitting nervous impulse, acts as an agent of blood coagulation, and also helps in the formation of strong bones (NRC, 1989). Calcium content of the snack ranged from 80.82±0.029 mg/100 g to 360.9±0.121 mg/100 g. A stake averagely weighing (33.7±2.18 g) of the snack is capable of supplying minimum of 8.14% of recommended daily intake of calcium (1,000) mg (NRC, 1989). Consumption of the snack will aid the formation and development of strong bones hence could be recommended for both adults and children (NRC, 1989).

Copper is an essential element in the body and is important in the production of red and white blood cells, triggers the release of iron to form heamoglobin. It is also involved in infant growth, brain development, boost the immune. The copper content (0.38±1.18 mg/100 g) obtained in this study is higher than the value (30 µg/g) obtained by Fawole et al., (2013), this could be due to the groundnut cake in the spice and the fact that nuts are good source of copper thus; consumption of this snack will enhance good health (NRC, 1989). Copper content in one stake of S. galilaeus snack is capable of supplying 6.4% to 2 mg of the recommended daily intake of this trace element.

Manganese content of the snacks ranged from 0.46±0.025 mg/100 g to 1.17±0.026 mg/100 g, the recommended daily intake of this trace element is 5 mg and S. galilaeus snack in this study is capable of supplying 6% of recommended daily intake. Manganese is important in making and activating some enzymes in the body but taking high dose of manganese for too long can lead to muscle pain, nerve damage and neurological symptoms.

Potassium value ranged from 479.6±0.124 mg/100 g to 884.6±0.061 mg/100 g, these values implies that consumption of one stake of S. galilaeus snack will supply 9.72% to the recommended daily intake of 2,000 mg (NRC, 1989). Potassium plays important functions such as controlling the balance of fluids in the body and also corrects the functioning of the heart muscles. Excessive intake of potassium could cause stomach pain, nausea and diarrhea, results of potassium content in this snack is not likely to cause any of these ailments.

The values obtained for Selenium ranged from 0.65±0.008 mg/100 g to 2.44±0.072 mg/100 g. Fish is a very rich source of Selenium, S. galilaeus snack in this study is capable of supplying the required amount of 0.7 mg (NRC, 1989) needed by the body in a day. Selenium is known to play an important role in immune system function and in reproduction; it also helps to prevent damage to cells and tissues. A stake of the snack is capable of supplying 72% of the recommended daily intake of 0.7 mg.

Sodium is important in neuron functions and osmoregulation between cells and the extracellular fluid. Salt is the main source of sodium, value obtained ranged from 420±0.008 mg/100 g to 702±0.004 mg/100 g, the recommended daily intake is 2,400 mg. High sodium content of food is of great concern for health because too much of sodium could lead to high blood pressure. This result indicated that the sodium content of the fish snack could not lead to high blood pressure, thus, hypertensive patient could take the snack without fear of aggravating the condition. A stake of this snack is capable of supplying 8.3% of the recommended daily intake of 0.7 mg.

Magnesium is a mineral that among other functions helps turn the food we eat into energy, and also helps to make sure that parathyroid gland which produce parathyroid hormone (PTH) or parathormone or parathyrin that are important for bone health, work normally. Values obtained in this study ranged from 75±0.006 to 130±0.002 and is capable of supplying 12.22% of recommended daily intake (300 mg).

Zinc is known to be involved in most metabolic pathways in humans which include among others; healing of wounds, making new cells and enzymes, zinc deficiency can lead to loss of appetite, growth retardation, skin changes and immunological abnormalities (Malakootian et al., 2011). The value of zinc in this study ranged from 0.64±0.002 mg/100 g to 3.07±0.009 mg/100 g and is capable of supplying 7.8% to 9.5 mg recommended daily intake. Excess zinc have been implicated in copper absorption, zinc content of the snacks could not interfere with body copper absorption.

Most of the elements were found to be higher in snacks treated with Vernonia amygdalina crude extract; these values could have been influenced by its (Vernonia amygdalina) high elemental concentrates as reported by Sodamade, (2013). It was observed that the snacks treated with the mixture of Vernonia amygdalina and Ocimum gratissimum crude extract had the least elemental value. This could probably be due to the effect of counter action of compounds present in the two crude extracts.

The values obtained in this study although vary between the treatments, are within the recommended daily intake of minerals for maintenance and promotion of good human health thus indicating that S. galilaeus fish snack could enhance healthy diet.

Fawole et al., (2007) opined that microelements tend to become harmful when their concentrations in body tissue exceed metabolic demands. Their concentration in the fish snack is within the recommended daily limit. This study therefore, showed that the snacks are good sources of nutrient and minerals which could be attributed to the fact that fresh water fishes are rich in nutrients and mineral (Fawole et al., 2013), and Vernonia amydalina plant crude extract has the potential to balancing the recommended daily intake of nutrients and minerals to the benefit of human health.

4 Conclusion/Recommendation

Processing fish into ‘suya like’ snack improves the nutrient composition, especially, the protein, fibre and ash thus making it nutrient dense. Addition of plant extracts and composite spice to the fish fillet have been shown to have an impact on food quality in increasing the nutritional quality of the snack.

The processing of fish species into fish snacks is recommended because it could help in promoting cottage industry to enhance food and employment security more especially that the takeoff capital and technology is not much, coupled with availability of raw material and market. Women entrepreneurs should be encouraged to venture into fish snacks production to provide them with a viable livelihood. The use of polythene material for packaging should be encouraged so as to make it attractive and enhance its shelf life.

The potential of the use of the snacks as instant energy food product during military/security operations as well as food for famished refuges should also be exploited.

To ensure economic viability and sustainability, the production of the snack should include condiments production from the remnants which accounts for about fifty percent of processing yield.

Authors' Contributions

Ileogben T.A. is the student who carried out the research and wrote the paper. Oginni O., T.A. is the supervisor who supervised the work all through and also made sure all necessary corrections were effected. All authors involved in draft manuscript preparation and approved the final manuscript.

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