Moringa oleifera leaf meal as partial replacement of soybean meal in diets of Clarias gariepinus juveniles

Irabor Arnold Ebuka, Ekokotu Paterson Adogbeji, Obakanurhe Oghenebrorhie¹, Adeleke Mosunmola Lydia², Obugara Joan Ejovwokoghene and Ayamre Efevogho Ufuoma

Department of Fisheries Delta State University, Asaba Campus. Nigeria
obakaoghenebrorhie@gmail.com
¹ Department of Animal Science Delta State University, Asaba Campus. Nigeria
² Federal University of Technology, Akure. Nigeria

Abstract

Performance of Clarias gariepinus was evaluated as soybean meal was supplemented in part by moringa leaf (ML) at 0%, 10%, 20%, 30% and 40%. Fish samples with initial average weight 9.14g were stocked randomly into fifteen tanks (A_1-3, B_1-3, C_1-3, D _1-3, and E_1-3) at a density of 20fish/tank and were fed twice daily. At day 56, parameters revealed increase in the values as inclusion level increased, although p< 0.05 was observed at 30% and above. M20% had best mean values for body weight (34.26g) and feed conversion ratio (2.11). Haematological parameters revealed high mean values of PCV (28.12) and Hb (8.20) for M10%, while WBC (7.41) and LYMPH (71.02) were higher in M40%. MCH (54.71) and MCV (159.62) were higher in M30%, while MCHC was highest in M20%. Results on serum enzymes activities revealed proportionate increase between mean values of alkaline phosphatase, alanine aminotransferase and aspartate aminotransferase and inclusion levels. Best performance of C. gariepinus after 56days was observed in 20% inclusion level of MOLM.

Keywords: growth, haematology, serum, tanks

Introduction

The fish species Clarias gariepinus is a freshwater catfish species known for its capability of using atmospheric oxygen, which gives it the potential to walk on land for several hundred meters aided by its pectoral spines (Sharma et al 2019). It is widely seen as an essential tropical catfish species for aquaculture and generally recognized in both commercial and artisanal fisheries and profitable species in Nigeria where it serves both socio-cultural and study purposes in most regions. It commands high market price and grow well under high stocking density. Fagbenro (2010) reported that high stocking density does not negatively affect growth performance in Clarias gariepinus under culture condition. It has excellent feed acceptability (both artificial and natural food) and can adapt to varieties of feeding mode in expanded niches. Other attributes include desiccation, ability to endure severe drought and food scarcity (high survivability), good meat quality and can be produced all year round (Sharma et al 2019). The success of aquaculture in a dynamic global food production environment has always been a major challenge due to inadequate nutritious feeding stuffs (Kong et al 2020). Thus, the substitution of protein feed ingredients in fish diets have been considered by fish nutritionists world-wide. The high cost and fluctuating quality as well as the uncertainty in the availability of soya bean meal have led to the need to look into alternative protein sources for fish feed formulation (Mmanda et al 2020).

Therefore, to attain a more frugal, justifiable, approachable and practicable production of fish, studies have been focused towards the assessment and use of non-conventional protein source derived from plant. Numerous studies have revealed that the required protein for optimum fish performance is also derived from plant sources (vegetable protein) such as moringa (Moringa oleifera) (Seyed et al 2018; Montoya-Camacho et al 2019), sweet potato and coffee bush leaves (Dorothy et al 2018; Low et al 2020).

M. oleifera belongs to the single genus monogeneric family Moringaceae and is well cultivated and distributed in Africa and Asia (predominantly the dry topical regions) (Nouman et al 2013; Marrufo et al 2013). Moringa has medicinal values, aside from being a healthy source of proteins, vitamins and amino acids (Montoya‐Camacho et al 2019) antioxidants, antimicrobial and therapeutic qualities (Obakanurhe and Okpara 2016; Seyed et al 2018; Batool et al 2020). This supports its potential to replace traditional feeds (Sarwart et al 2002). The wellbeing of fish is reflected significantly on its blood profile, this have made haematology a major monitoring indicator in fish culture (Jan and Ahmed 2021). Although, physiological factors such as stress incurred from environmental toxicity, handling/sampling and transportation are primary influencers of the blood profile of fish, in culture fisheries priority for haematological examination is due to feed input (Jimoh et al 2020).

This research established the performance and blood indices of C. gariepinus fed with M. oleifera leaf meal as an additive.

Materials and methods

Study Area

This research was carried out in the Department of Fisheries, Delta State University, Asaba Campus. The town Asaba is in Oshimili South Local Government Area of Delta State, Nigeria. It lies between longitudes 6 ⁰E and 8 ⁰E and between latitude 4 ⁰N and 10 ⁰N.

Source of ingredients/ preparation of experimental diets

Moringa leaves were collected from Delta State University Abraka Asaba Campus. While other ingredients such as soyabeans, groundnut cake, maize, vitamin premix, lysine, methionine, salt and binder were procured from Hausa market in Asaba. The ingredients were ground into powder and mixed together. The moringa leaves were harvested from the agronomy department farm of Delta State University and were washed to remove impurities before being dried under room temperature. This was to retain its green coloration as well as reduce the antinutritional factors contained in the leaves. The dried leaves were further processed using hammer mill to attain 3mm leaf meal. This was introduced into the feed at various inclusion levels (M10%, M20%, M30% and M40%) respectively and pelleted into 2 mm and 3 mm sized feeds using a locally fabricated flat-die pelletizer. The pelleted diets were dried at room temperature for five days and stored in air tight plastic container.

Experimental Fish and Experimental Set Up

Three hundred C. gariepinus juvenile were procured from a notable fish farm in Asaba and transported to the Fisheries Laboratory, Delta State University, Asaba Campus, where the initial length (total and standard) was measured with meter rule and body weight was measured with triple bean balance. The juveniles measured an average initial length and weight of 8.12 cm and 9.14 g respectively. At the laboratory, fish samples were disinfected with potassium per manganate (2 ppm) for 20 minutes. Thereafter, stocked in a tank of (2 m x 2 m x 0.4) and fed twice daily, in the morning at 08.00 hour and evening at 20.00 hour with 2 mm Coppens feed at 5% body weight and proper management routine was carried out. This process lasted for two weeks prior to the transfer of samples to experimental tanks.

The already acclimated fish samples were separated into fifteen tanks of 1.6 m² (2 x 2 x 0.4) water holding capacity and each tank was stocked with twenty (20) C. gariepinus juveniles. The tanks A_1-3 (control) had no moringa leaf meal inclusion diet, while B_1-3, C_1-3, D_1-3, and E_1-3 were fed with the experimental diets respectively.

Duration of study

For a total of 8 weeks this study was performed, across January to March, 2021.

Data collection

Total length, standard length and body weight measurements were done each week. Using a graduated measurement board, length to the closest 0.1 cm while, weight measurement was taken to the nearest 0.1 g.

Growth performance and feed utilization

The growth performance indices were calculated using Zhang et al (2019) methods

Mean body weight gain

This was calculated as the difference between the initial and final body weights of fish.

Wg = W₂ – W₁

Where W₂ = final body weight

W₁ = Initial body weight

Feed intake

The feed intake was calculated per day to ascertain the amount of feed taken per fish.

Feed given divided by the number of fish stocked per treatment.

Feed Conversion Ratio (FCR) = Food Intake (g) / Weight gain (days)

Fluid (blood and serum) collection and biochemical examination

Blood was extracted from sampled fish and examined to ascertain blood profile (Packed cell volume, White blood cell, Haemoglobin, mean corpuscular volume, mean corpuscular haemoglobin concentration, mean corpuscular hemoglobin, and red blood cell), using standard procedures of Kone et al (2017).

Serum (clear fluid) separated from the extracted blood sample was subjected to biochemical examination to evaluate serum enzymes indices (Alanine aminotransferase, Aspartate aminotransferase and Alkaline phosphatase) using Hekmatpour et al (2019) methods.

Water Quality Monitoring

Water quality parameters in each tank were recorded once during the experimental period. The pH was recorded by a pH meter (Schott Greate, Florida state, USA) and dissolved oxygen (DO mg/L) by Winkler titration (Kyle et al 2019). Temperature (ºC) was recorded daily with mercury in glass thermometer at 8.00 h and 14.00 h.

Data Analysis

Data collected were analyzed using Analysis of Variance (ANOVA) in an entirely randomized strategy. Comparisons among diets means was carried out by Duncan Multiple Range Test at p<0.05. All computation was performed using statistical package SPSS version 23.

Results

The proximate composition of ML was examined and presented as follow;

The proximate composition of diets recorded in Table 3 revealed highest vales of crude protein (40.67%) and NFE (36.53) in M10%, while the lowest was recorded in M30% (39.07%) and M40% (35.26) respectively. The values for crude fibre and total ash were higher in M40% (7.17% and 5.68%) respectively, however M0% had the least value for crude fibre (3.87%) and M10% (4.79%) for total ash.

The growth indices of C. gariepinus at M20% revealed that the mean weight gain (34.42g) and feed conversion ratio (2.11) showed better performance compared to the other treatments.

M: Moringa oleifera leaf meal, WG: Weight gain, FI: Feed Intake, LWG: live weight gain, FCR: Feed conversion ratio

Figure 1. Weight gain of C. gariepinus at 56 days	Figure 2. Feed conversion ratio of C. gariepinus at 56 days

A curvilinear relationship was revealed between weight gain and inclusion levels of moringa (Figure 1).

There was a curvilinear relationship between feed conversion and inclusion levels of moringa (Figure 2).

The proximate composition of C. gariepinus carcass at 56days old showed that crude fibre, total ash and NFE increased as moringa levels increased among other parameters (Table 5).

Among the blood profile parameters only WBC and LYMPH revealed increases as inclusion levels increased (Table 6).

The serum liver enzyme activities increased as moringa levels were increased (Table 7).

The pH and temperature increased as inclusion of moringa levels increased while dissolved oxygen decreased (Table 8).

Discussion

The acceptability and characteristics of feed materials such as leaf meals substitutes is ascertained using its proximate structure. The proximate analysis of MOML across treatments revealed (p < 0.05) difference in all parameters as inclusion level increased. This was due to the high nutrient potential of the additive and this confirms the findings of Hiam et al (2019) who reported increased primary nutrients with increased MOLM inclusion in O. niloticus formulated feed. Also, other leaves such as sweet potato and coffee bush leaves with feedstuff potentials have been reported to contain high nutritive values (Dorothy et al 2018; Low et al 2020).

From the result, increased growth performance was observed as inclusion level of moringa leaf meal increased, but a decline was also observed at 30% and above inclusion level. This result is in consistent with Xuhui et al (2020) who reported a reduction impact on the growth efficiency upon higher replacement level of fish meal with Moringa leaf meal. This could be as a result of the presence of anti-nutritional factors such as phenol, tannin and phylates during processing. The effect of these substances on fish can reduce palatability, altered nutrient balance of the diet, disturb digestive process, growth and decreased feed efficiency (Zhang et al 2019; Su and Chen 2020). This is also in agreement with Albert et al (2020) and Karimi et al (2019) who reported that variation in condition factors can be as a result of food, maturity, stages and environmental factors.

Fish fed with (M40%) had the worse feed conversation ratio. This finding is in agreement with Hassaan et al (2019), who established decreasing trend in feed conversation ratio corresponding with increased inclusion levels of seed meal in diet. The reason in this present study could be as a result of higher fiber content in the diet of M 40%.

The carcass of the sampled fish evaluated after the feed trail revealed the effect of the additive on the nutritive components of the fish. As inclusion level increased, there was progressive p < 0.05 difference observed across treatments especially in total ash, crude fibre, moisture content and NFE which indicated the dietary effect of MOLM on the fish which could be due to the rich moisture, NFE and fibre nature of plant feed sources. This confirmed the report of Dienye and Olumuji (2014), where increased fibre, moisture, ash and NFE in carcass quality of fish fed M. oleifera was revealed.

Evaluation of the blood parameters of fish exposes its health status in relation to ingest constitutes in feeds especially in aquaculture (Muttappa et al 2015; Indranath et al 2020). The observed differences exhibited in various treatments showed the antibiotic potential of the additive. The observed changes of PCV and RBC which revealed a decline as inclusion level increased could be due to absence of stress induced from the test ingredient (antioxidant). This confirms the report of Kakwi and Olusegun (2020), where decreased PCV and RBC was observed in Cyprinus carpio fed different level of inclusion of Mucuna pruriens. Also, the immune-stimulatory capability of the test ingredient was revealed with increased WBC count. Increased WBC could also reveal the reaction of the fish to the toxic nature of test ingredient. In the same vein, WBC increase in C. carpio was attributed to Sesbania leaf meal additive in the feed (Anand et al 2020).

Similar trend of increase was observed in other parameters (LYMPH, MCH and MCV) which could have resulted from physiological stress. This affirmed the finding of Rezaei et al, (2020), who reported increased blood parameters (LYMPH, MCH and MCV) in C. carpio exposed to carbamazepine. Dicentrarchus labrax fed varying inclusion levels of garlic and chitosan powders also revealed similar trend (Abdel-Tawwab et al 2020). Although, these changes are within acceptable limits therefore it posed no threat to the wellbeing of the cultured fish species.

Serum analysis revealed (p < 0.05) difference across treatments which indicated the influence of varying levels of the test ingredients on the activities of the enzymes. The increased values of serum enzymes activities as test ingredient appreciates may be associated to antioxidant and hepatoprotective potentials of moringa leaf thereby suppressing liver damage through prevention of fatty acid peroxidation. This confirms the results of Xuhui et al (2020), who reported increased serum enzyme activities in gibel carp juvenile fed varying inclusion of fermented ML.

Water quality parameters observed in this study falls within an acceptable range and with limits of national standard (Dinesh et al 2020). It was observed that at low range of dissolved oxygen (6.01 and 6.02), growth performance was low. This finding is in agreement with Dinesh et al (2020) they establish that growth performance decrease with decrease level in dissolved oxygen and also Bueutello et al (2000) established that as dissolved oxygen declined from 100% to 30% there was a progressive reduction in growth performance.

Result has shown that low level of pH (7.9) resulted to lower growth performance. This finding is in agreement with Daniel et al (2020) they established that at low level of pH there is a reduction ability of fish oxygen carrying capacity, of which result to low level of growth performance. Higher temperature was observed in tank D, of which resulted to reduction of dissolved oxygen level. Similar result was observed by Liyu (2019), who established that water temperature directly impacts the level of dissolved oxygen retention.

Conclusion

• Soybean meal is one of the most important fish feed ingredients. It is also an expensive ingredient due to its nutritional value and can be very scarce in some seasons. Moringa leaf is one of the cheapest close substitutes, being a plant feed stuff.
  
  
• It has a nutrient profile that can substitute adequately to what is obtained in soybean meal and it can also be made available all year round to support the aquaculture industry.
  
  
• This study has shown that 20% substitution level of moringa leaf meal in C. gariepinus diet produces better result than without substitution.

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