Potential of Moringa oleifera Leaf Meal as Substitute of Fishmeal on Performance of Catla catla Fingerlings Reared in Earthen Ponds

Muhammad Faisal1, Syed Makhdoom Hussain1*, Majid Hussain2,

Muhammad Zubair ul Hassan Arsalan3, Zeeshan Yousaf1 and Muhammad Amjad1

1Fish Nutrition Lab, Department of Zoology, Government College University, Faisalabad, Pakistan

2Department of Fisheries and Aquaculture, University of Okara, Okara, Pakistan

3Department of Life Sciences, Khwaja Fareed University of Engineering and Information Technology, Rahim Yar Khan, Pakistan

ABSTRACT

This study was carried out in a semi-intensive rearing system in order to assess the impact of Moringa oleifera leaf meal (MOLM) based diets on the overall performance of Catla catla fingerlings cultured in 6 earthen ponds. In order to formulate experimental diets, MOLM was used as test ingredient. MOLM was used to replace fish meal (FM) in the diet at varying levels of 0%, 10%, 15%, 20%, 25%, and 30%, respectively. There were five experimental diets and one control diet used during the trial. Fingerlings were placed in a cemented pond for fifteen days under laboratory conditions for acclimatization. There were 15 fingerlings put into each of the six earthen ponds, for a total of 90 fingerlings. Analyses showed that fish fed with 10% MOLM-based diet had the highest growth performance when compared with other experimental and control groups. Best results for carcass composition, hematological analysis, and body mineralization were noted in fingerlings given a diet containing a 10% substitution of FM with MOLM. In the present investigation, it was determined that replacing 10% of FM in the diet with MOLM improved the growth, body composition, hematological parameters, and body mineralization of C. catla fingerlings.

Article Information

Received 02 December 2022

Revised 15 December 2022

Accepted 02 January 2023

Available online 18 March 2023

(early access)

Published 16 May 2024

Authors’ Contribution

MF conducted the study and wrote the manuscript. SMH administered and supervised the project, and acquired funds. MH, ZY, MA and MZHA curated the daata and helped in writing, editing and reviewing the manuscript.

Key words

Moringa leaf meal, Catla catla, Growth performance, Body mineralization

DOI: https://dx.doi.org/10.17582/journal.pjz/20221202061259

* Corresponding author: [email protected]

0030-9923/2024/0004-1509 $ 9.00/0

Copyright 2024 by the authors. Licensee Zoological Society of Pakistan.

This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

INTRODUCTION

All living organisms, including fish, need nutritious food to survive, reproduce, and maintain their bodies (Adebayo et al., 2020). The aquaculture industry is growing at a rate of around 5.8% each year as a result of the tremendous growth and development occurring throughout the sector (FAO, 2020). It is the fastest developing industry and provides 50% of entire worldwide food fish utilization (FAO, 2010). There has been a rise in the demand for food and the need of a healthy diet in developing nations as a consequence of their rapid population expansion (Abdulkadir et al., 2016). Fish and fish by products play an important part in meeting the needs of the human population in terms of food security as well as the nutritional requirements of both emerging and developed nations (FAO, 2014). It accounts for 17% of animal protein and 7% of entire protein consumed worldwide (FAO, 2020).

Fish meal (FM) is a significant contributor of protein to the diets of farmed fish (Dawood et al., 2015). The aquaculture industry utilizes 3.06 mt of it since it is a significant source of protein (FAO, 2008). In aquaculture, fish feed contributes around 40-60 % of production costs (Nasr et al., 2021). Furthermore, from last 30 years, FM cost have also increased in right terms. The rapid rise in the price of fish oil and FM may possess a negative impact on commercial aqua-operations (Naylor et al., 2000). Unpredictable availability, rising demand, and a greater value for fishmeal lead it necessary to look for new protein sources (Hussain et al., 2015a).

Plant-based protein sources have largely replaced fish meal as fish feed (Elumalai et al., 2021) because they are more readily accessible, less harmful to the environment, more sustainable, and cheaper (Hardy, 2010). One excellent protein source is the Moringa oleifera (miracle tree), which can be incorporated into aquaculture diets. This plant is often found in tropical and subtropical regions and is also known as drumstick tree. It is used in variety of applications, both in industry and medicine (Saini et al., 2016; Falowo et al., 2018). The most commonly used part of this plant is its leaves that are important source of polyphenols, flavonoids, alkaloids, saponins, tannins, glucosinolates, carotenoids and vitamins (Leone et al., 2015). For the purpose of improving growth and immunological function, M. oleifera leaf meal (MOLM) has been used in aquaculture diets as a research subject (Puycha et al., 2017). It has been found that moringa leaves have high ratio of vitamins A and C, iron, potassium, calcium and proteins (25 to 32%) than other products of food like banana, orange, yoghurt, carrot and milk (Soliva et al., 2005; Gopalakrishnan et al., 2016). In aquaculture technology, moringa has become possible substitute plant protein source, moreover, its leaves contain important amino acids, especially tryptophan, lysine, methionine and cysteine (Sherif et al., 2022).

Catla catla feeds on surface and called as surface feeder that is commonly used in Pakistan’s polyculture technique and is grown along with other varieties (Aslam et al., 2016). Its output increased in the first decade of the first-twenty century, reaching roughly 2.8 mt per year in 2012 (FAO, 2010). As a result, the current study aimed to examine the efficacy of MOLM to serve as an alternative source of protein in formulated diets of fingerlings as well as to evaluate its effects on growth performance, body composition, hematology and mineral status of C. catla fingerlings when cultured in earthen ponds.

Materials and Methods

Study area and pond preparation

Six-month feeding trial was done between February and August 2021 in six earthen ponds using a semi-intensive rearing method at the Fisheries Research Farms, Department of Zoology, University of Agriculture Faisalabad, Pakistan. Furthermore, analysis of feed ingredients and fish body were carried out at Fish Nutrition Laboratory, Government College University, Faisalabad, Pakistan. Each pond had a rectangular form. The ponds were irrigated by a nearby groundwater tube well. Every pond had an entrance and an outlet for water supply. Prior to the start of the experimental trial, ponds were rebuilt, aquatic flora, fauna and fish were removed. Before fertilization, agricultural lime (CaCO3) was provided to all ponds at a rate of 250 kg/ha to maintain pH of the ponds and filled with water for 7 days. After that, ponds were fertilized with both organic and inorganic fertilizers to keep planktonic biomass for fingerlings. Both organic and inorganic fertilizers were uniformly applied to each pond on fortnightly basis for a period of six months.

Fish and experimental conditions

Fingerlings were purchased from Government Fish Seed Hatchery, Satiana Road, Faisalabad and transported to Department of Zoology, Government College University, Faisalabad. Fingerlings were placed in a cemented pond for 15 days under laboratory conditions for acclimatization. At 10:00 A.M. the fingerlings were fed to satiation with the feed. An air pump provided continuous air to the fingerlings via capillary system. After acclimatization, the fish were divided into groups and then transported to the Fisheries Research Farms, Department of Zoology, Wildlife and Fisheries, University of Agriculture, Faisalabad. NaCl (5g/L) was applied to fingerlings before to the start of feeding experiment to kill any ectoparasites present and prevent any fungal infections (Rowland and Ingram, 1991). In order to maintain the level of dissolved oxygen (DO), air was provided to each pond by changing water. Physical and chemical parameters were monitored on regular basis, including temperature, pH, DO, and carbon dioxide. Prior to the start of the experiment, wet weight, fork length and total length of all groups of fingerlings were measured.

Experimental design

In order to formulate an experimental diet, MOLM was used as test ingredient. One group was kept as control for this experiment and received only organic and inorganic fertilizers. However, the other groups were treated with different levels of MOLM based diets along with fertilizers. During the trial, total 90 fingerlings were distributed among 6 earthen ponds. Each pond received fifteen fingerlings, which were nourished at a ratio of 3% of their biomass each day. The six-month feeding study was conducted.

Moringa oleifera leaf processing and other feed ingredients

The feed ingredients were transported from Multan in southern Punjab. The crushed leaves were first treated by soaking in tap water at room temperature for three days. In order to reduce anti-nutritional substances such as saponins, the mixture was stirred daily for one hour while soaking (Tacon and Jackson, 1985; Wee and Wang, 1987). The mixture was soaked and then the water was removed by squeezing it through a cloth. The grinded materials for the experimental diet were tested for their chemical make-up before they were blended (AOAC, 2005). Chrome oxide served as an inert marker in the experimental diets. Six test diets were prepared using MOLM to replace fishmeal at levels of 0%, 10%, 15%, 20%, 25%, and 30%.

 

Table I. Composition of ingredients (%) in test diets.

Ingredients	Test diet-I (control)	Test diet-II	Test diet-III	Test diet-IV	Test diet-V	Test diet-VI
Replacement level	0 %	10 %	15 %	20 %	25 %	30 %
MOLM*	0	10	15	20	25	30
Fish meal	50	40	35	30	25	20
Wheat flour**	17	17	17	17	17	17
Corn gluten	20	20	20	20	20	20
Fish oil	9	9	9	9	9	9
Vitamin premix	1	1	1	1	1	1
Mineral premix	1	1	1	1	1	1
Ascorbic acid	1	1	1	1	1	1
Chromic oxide	1	1	1	1	1	1

 

MOLM*, Moringa oleifera leaf meal.

 

Formulation and processing of experimental diets

A 0.5 mm sieve was used to finely grind the feed materials. After 5 min of mixing all of the contents in a mixer, fish oil was gradually added. After adding 10–15% water to the feed ingredients, the mixture was incorporated into the mixer to generate dough with an appropriate texture, which was then processed through an experimental extruder to produce pellets that floated (Lovell, 1989).

Proximate analysis

The whole body and experimental diet samples were individually homogenized using a motor and pestle prior to being analyzed utilizing standard procedure. Crude protein (N×6.25) was assessed by micro Kjeldahl apparatus; moisture by oven drying at 105°C for 12 h; crude fat by petroleum ether extraction procedure using Soxtec HT2 1045 system and ash was calculated by electric furnace ignition at 650°C for 12 h (Eyela-TMF 3100). Mineral analysis of fingerlings was determined by atomic absorption spectrophotometer (AOAC, 2005).

Growth study

After the completion of trial, all fish were weighed to assess growth.

Hematological parameters

Blood was obtained from the caudal vein of C. catla fingerlings using a syringe (heparinized) and delivered to the Department of Zoology, UAF, for hematological investigation. The micro-hematocrit approach was applied to assess hematocrit using capillary tubes (Brown, 1988). To count RBCs (red blood cells) and WBCs (white blood cells), a hemocytometer with an authorized Neubauer counting chamber was utilized (Blaxhall and Daisley, 1973). As indicated by Wedemeyer and Yastuk (1977), Hb (hemoglobin) concentration was determined. Standard formulae were used to compute MCHC (mean corpuscular hemoglobin concentration), MCH (mean corpuscular hemoglobin), and MCV (mean corpuscular volume) (mean cell volume).

MCHC = Hb/PCV x 100

MCV = PCV/RBC x 10

MCH = Hb/RBC x 10

Data analysis

The data on different parameters of growth, body composition, hematological parameters and body mineralization were statistically analyzed using a microcomputer, as described by Steel et al. (1996). One-way analysis of variance (ANOVA) was used to analyze the data (Steel et al., 1996). Tukey’s Honesty Significant Difference Test was used to compare the differences among means (Snedecor and Cochran, 1991). For statistical analysis, the Co-Stat computer software (Version 6.303, PMB 320, Monterey, CA, 93940 USA) was used.

RESULTS

Growth performance

Table II describes the growth parameters of C. catla fed with MOLM based diet, in terms of final weight and weight gain. Maximum value of final weight (677.42 g) was obtained when fingerlings given 10% MOLM based diet while minimum value of final weight (438.39 g) was recorded at group fed 30% inclusion of MOLM based diet. Highest weight gain (662.09 g) of fingerlings was observed when fingerlings fed at 10% of MOLM based diet while lowest value of weight gain (432.15 g) was recorded when fingerlings fed at 30% inclusion of MOLM based diet. It was indicated that, there was an increasing tendency in final weight and weight gain when fingerlings fed 10% MOLM based diets. Moreover, significant difference was observed in the final weight and weight gain when compared to control diet and all other test diets. Current study revealed that 10% inclusion of MOLM in the diets improved the growth performance of fingerlings.

Body composition

Table III provides an explanation of the body composition of C. catla fingerlings treated for 90 days on various MOLM based diets. The best value of CP (20.81%) and poor value of CF (2.04%) in the fingerlings body were observed at group II having 10% substitute level of MOLM based diet. These values were statistically significant (p<0.05) from fingerlings treated 0%, 20%, 25%, and 30% replacing level MOLM based diets as represented in table. Lowest value of CP (16.14%) and greatest value of CF (6.56±0.14%) were found in the group VI that fed on the 30% replacement of MOLM based diet. Highest value of ash (3.10%) was found in the group II having replacement level of 10% MOLM based diet. While lowest value of ash (2.41%) content was observed in the group VI having substitute level of 30% MOLM based diet. It was also noticed that protein and ash contents were highest in group II having 10% substitution level of MOLM based diet as compared to other diets such as 0%, 20%, 25% and 30% replacement levels of MOLM based diets. The greatest value of moisture (74.88%) was found in group VI fed 30% replacement level of MOLM based diet. While lowest value of moisture (74.03%) was found in group II fed 10% replacement level of MOLM based diet. It was observed that substitution of 10% MOLM based diet improved the body composition of fingerlings, while increased level was non-significant.

Hematological parameters

Table IV is representing the results of hematological parameters fed fingerlings with inclusion of MOLM based diet. Maximum value of RBCs (2.80×106 mm-3) was showed by fingerlings fed 10% MOLM based diet while lowest value of RBCs (2.32×106 mm-3) was recorded in the fingerlings when fed with 30% MOLM based diet. Maximum WBCs (7.82× 103 mm-3) were recorded in the group II having 10% replacement of MOLM based diet while lowest value of WBCs (7.17× 103 mm-3) was found in the group VI having 30% MOLM based diet. The best values of RBCs and WBCs fed 10% MOLM based diets were statistically significant from control diet and all other diets. Highest value of PLT (64.79) was observed in the

 

Table II. Analysis of final weight and weight gain of C. catla fingerlings fed with MOLM based diets.

Experimental diets	MOLM replacement level	Initial weight (g)	Final weight (g)	Weight gain (g)
Group I (Control)	0%	15.34±0.03	514.43±0.24d	499.08±0.25d
Group II	10%	15.33±0.06	677.42±0.19a	662.09±0.24a
Group III	15%	15.25±0.03	596.40±0.23b	581.15±0.20b
Group IV	20%	15.34±0.03	553.63±0.16c	538.29±0.14c
Group V	25%	15.22±0.03	476.42±0.03e	461.19±0.05e
Group VI	30%	15.24±0.05	438.39±0.14f	432.15±0.09f

 

Means within rows having different superscripts are significantly different at (p< 0.05).

 

Table III. Body composition of C. catla fingerlings fed on various replacement levels of MOLM based diets.

Experimental diets	MOLM replacement level	CP (%)	CF (%)	Ash (%)	Moisture (%)
Group I (Control)	0%	19.30±0.22c	3.38±0.15d	2.96±0.01b	74.35±0.08bcd
Group II	10%	20.81±0.13a	2.04±0.09f	3.10±0.01a	74.03±0.03d
Group III	15%	18.40±0.13d	4.34±0.13c	2.83±0.04c	74.41±0.05bc
Group IV	20%	20.01±0.10b	2.81±0.30e	2.98±0.06bd	74.18±0.18cd
Group V	25%	17.33±0.22e	5.38±0.18b	2.69±0.02d	74.59±0.08ab
Group VI	30%	16.14±0.09f	6.56±0.14a	2.41±0.04e	74.88±0.20a

 

Means within rows having different superscripts are significantly different at (p< 0.05). Data are means of three replicates.

CP, crude protein; CF, crude fat.

 

Table IV. Hematological parameters C. catla fingerlings fed on various replacement levels of MOLM based diets.

Experimental diets	MOLM (%)	RBCs (10-6 mm-3)	WBCs (10-6 mm-3)	PLT	Hb (g/100 ml)	MCHC (%)	PCV (%)	MCV (fl)	MCH (pg)
Group I (Control)	0	2.42± 0.01c	7.36± 0.07c	58.45± 1.00d	7.53± 0.05d	28.31± 0.09d	24.21± 0.09c	107.68± 4.07d	38.13± 0.57d
Group II	10	2.80± 0.08a	7.82± 0.08a	64.79± 0.17a	8.76± 0.05a	34.81± 0.08a	26.63± 0.02a	178.27± 2.09a	53.92± 0.95a
Group III	15	2.65± 0.02b	7.69± 0.02a	62.56± 0.11b	8.32± 0.03b	33.02± 0.68bb	25.35± 0.06b	153.79± 1.96b	48.76± 1.04b
Group IV	20	2.55± 0.02b	7.52± 0.02b	60.73± 0.13c	7.88± 0.12c	30.45± 0.13c	23.41± 0.05d	125.61± 2.70c	44.57± 0.90c
Group V	25	2.38± 0.00c	7.27± 0.03cd	57.31± 0.07d	7.11± 0.11e	24.50± 0.61e	22.14± 0.03e	94.85± 2.98e	33.26± 1.05e
Group VI	30	2.32± 0.03c	7.17± 0.01d	53.18± 1.04e	6.95± 0.07e	21.65± 0.54f	21.91± 0.06f	86.93± 1.80f	29.09± 0.10f

 

RBC, red blood cell; WBC, white blood cell; PLT, platelet; Hb, hemoglobin; MCHC, mean corpuscular hemoglobin concentration; PCV, packed cell volume; MCV, mean corpuscular volume; MCH, mean corpuscular hemoglobin. Means within rows having different superscripts are significantly different (p<0.05). Data are the mean of three replicate.

 

Table V. Whole body mineralization C. catla fingerlings fed on various replacement levels of MOLM based diets.

Experimental diets	MOLM (%)	Zn (μg/g)	P (%)	Ca (%)	Na (mg/g)	Mg (%)	Fe (μg/g)	Mn (μg/g)	K (%)	Cu (μg/g)	Se (mg/g)
Group I (Control)	0	3.43± 0.03d	1.05± 0.01c	0.87± 0.02ab	5.11± 0.02d	2.69± 0.04d	47.55± 0.19d	8.21± 0.02d	7.77± 0.01d	3.33± 0.02d	0.36± 0.01b
Group II	10	4.13± 0.02a	1.24± 0.03a	1.00± 0.03a	5.83± 0.04a	3.27± 0.01a	61.44± 0.10a	12.15± 0.01a	8.68± 0.02a	4.08± 0.02a	0.32± 0.02b
Group III	15	3.98± 0.01b	1.16± 0.01b	0.97± 0.02cd	5.69± 0.02b	3.05± 0.02b	54.63± 0.06b	11.09± 0.01b	8.19± 0.01b	3.92± 0.02b	0.24± 0.01c
Group IV	20	3.77± 0.04c	1.11± 0.02bc	0.92± 0.01bc	5.41± 0.02c	2.87± 0.02c	51.64± 0.25c	9.33± 0.02c	7.96± 0.02c	3.62± 0.04c	0.23± 0.01c
Group V	25	3.01± 0.02e	0.97± 0.03d	0.81± 0.01de	3.98± 0.01e	2.37± 0.02e	37.43± 0.13e	7.79± 0.02e	6.91± 0.02e	2.87± 0.01e	0.38± 0.02b
Group VI	30	2.23± 0.04f	0.89± 0.02e	0.76± 0.02e	3.16± 0.01f	1.97± 0.02f	33.59± 0.06f	7.49± 0.02f	6.09± 0.05f	2.16± 0.01f	0.49± 0.03a

 

Means within rows having different superscripts are significantly different at (p< 0.05). Data are means of three replicates.

 

group II having 10% inclusion of MOLM based diet while lowest value of PLT (53.18) was recorded in the group VI having 30% MOLM based diet. Maximum value of Hb (8.76 g/100ml) was found in the fingerlings when fed with 10% MOLM based diet while poorest value of Hb (6.95 g/100ml) was indicated by fingerlings fed 30% MOLM based diet. Best values of PCV, MCHC, MCH and MCV (26.63%, 34.81%, 53.92 pg and 178.27fl, respectively) were indicated in the fingerlings fed 10% inclusion of MOLM based diet while minimum values of PCV, MCHC, MCH and MCV (21.91%, 21.65%, 29.09 pg and 86.93 fl, respectively) were recorded in the fingerlings having 30% MOLM based diets. These findings of current study, suggested that inclusion of 10% MOLM in the diet improved hematological indices of fingerlings.

Body mineralization

The body mineralization of C. catla treated on various levels of MOLM-based diet is indicated in Table V. Body mineral contents of the fingerlings revealed that there were significant differences (p<0.05) between them. Best values of Zn, P, Ca, Na, Mg, Fe, Mn, K and Cu (4.13 μg/g, 1.24%, 1.00%, 5.83 mg/g, 3.27%, 61.44 μg/g, 12.15 μg/g, 8.68% and 4.08 μg/g) were deposited in the fingerlings body recorded at group II having 10% replacement of MOLM based diet and these values were significantly different (p<0.05) from the fingerlings fed on other experimental diets. While minimum values of Zn, P, Ca, Na, Mg, Fe, Mn, K and Cu (2.23 μg/g, 0.89%, 0.76%, 3.16 mg/g, 1.97%, 33.59 μg/g, 7.49 μg/g, 6.09 % and 2.16 μg/g) were observed in the fingerlings body fed 30% replacement of MOLM based diet. Maximum value of Se (0.49 mg/g) was found in the fingerlings body at group VI having 30% inclusion of MOLM based diet while lowest value (0.23 mg/g) of Se was indicated in the group IV.

DISCUSSION

Our research aimed to examine the effects of six various levels of MOLM based diet on growth performance, body composition, hematological indices and body mineralization in C. catla fingerlings. Current study revealed that feeding C. catla fingerlings with 10% MOLM based diet in the replacement of fishmeal improved the overall growth performance. The growth rate of Nile tilapia improved by MOLM based diet, according to a recent Egyptian study (Elabd et al., 2019). Findings of this study agreed with results of Doctolero and Bartolome (2019) who used MOLM in the diets of Oreochromis niloticus and indicated that up to 20% replacement of MOLM improved the growth performance of fish. In the same way, findings of this study are also similar with results of Elabd et al. (2019) who used diet based on MOLM for Nile tilapia and found that growth of fish was significantly increased. Moreover, he concluded that supplementing Nile tilapia with moringa based diets had beneficial effects as a growth stimulant. Results of Hussain et al. (2018) also showed that 10-20% inclusion of MOLM in the diet of Labeo rohita significantly increased the weight gain of fish while further increase in the concentration of MOLM had not satisfactory effect on growth performance of fish. Current study results were also in line with the findings of Idowu et al. (2017) who revealed that 15% inclusion of MOLM in the diets of Clarias gariepinus post-fingerlings significantly enhanced the weight gain of fish while 25% inclusion of MOLM in the diet, lower the mean weight gain of fish. Tabassum (2017) also recorded that 10% inclusion of MOLM in diet of Mozambique tilapia improved the growth performance of fingerlings. MOLM was proved as a fish meal alternative in the diet of C. gariepinus up to 10% without impairing growth, according to Ezekiel et al. (2016). Our results were dissimilar with outcomes of Khetran et al. (2018) who observed that addition of 30% moringa meals in diets of L. rohita can boost the growth. The higher growth rates could be linked to the high antioxidant contents of moringa leaves, such as vitamin C and carotene, which are favorable for fish health (Vergara-Jimenez et al., 2017). This research also concluded that 30% inclusion of MOLM in the diet didn’t show significant results, because the increasing MOLM in the diet contains lots of fiber.

The findings of current study match with those of Arsalan et al. (2016) who reported that 10% replacement level had the greatest CP when compared to a control group while 40% incorporation had the lowest CP. It was also shown that adding 10% MOLM to fish feed is a significant protein substitution. Similarly, Ganzon-Naret (2014) studied the carcass composition of Asian sea bass at various inclusion levels of MOLM based diets and indicated maximum CP value at 10% which was statistically significant than other MOLM based diets. Hence, according to current study, an excellent alternate source of protein for fingerlings’ diet is 10% replacement level of MOLM.

The growth and health of fish are now reflected by hematological parameters, which are key factors used in aquaculture (Fazio, 2019). Present study findings also similar to Tabassum et al. (2023), those who indicated that MOLM increased in the diet of Cirrhinus mrigala showed decreased improvement in hematological studies of fish and suggested that 10% replacement of MOLM in the diet had beneficial effects on the hematology of fish. In the same way, Billah et al. (2020) also observed that increasing the MOLM in the diet of fish declined the improvement of RBCs and WBCs in O. niloticus. The results of Arsalan et al. (2016), indicated parallel results with current study, those who showed that 10% inclusion of MOLM had positive effects on the hematology of L. rohita but hematological analysis revealed that MOLM inclusion of more than 20% in the diets caused hematological disturbance. The findings of the current investigation were in conflict with those of El-Gawad et al. (2020), who noted a significant increase in WBCs when fish fed with 1.5% moringa leaf powder whereas, Hb and RBCs amounts in the O. niloticus not significantly improved. It might be due to size of fish, concentration of dose, variety of fish and health condition of fish. According to the findings of current research, 10% MOLM based diet had improved the hematological indices of fish in earthen ponds.

Minerals are naturally occurring inorganic substances that are required for normal functioning of the fish body. In the current study, results indicated that highest values of minerals such as Zn, P, Ca, Na, Mg, Fe, Mn, K and Cu were observed in the body of fingerlings fed 10% inclusion of MOLM based diet while lowest values of minerals as like Ca, Na, P, Mg, Fe, Mn, Zn, K and Cu were found in group VI having 30% replacement of MOLM based diet. Whereas highest value of Se was found in the group VI having 30% addition of MOLM based diet but lowest value of Se was observed in the fingerlings body fed 20% MOSM based diet. Our results, were similar to Shahzad et al. (2021), who found that addition of MOLM in diet of common carp had improved the mineral contents. Present research work, revealed that inclusion of 10% MOLM in the diet of fish significantly boosted the mineral contents of C. catla. While increasing the concentration of MOLM in the diet of fish lowered the body mineralization of fish.

CONCLUSION

It was deduced that 10% replacement of fish meal with MOLM in the diet causes considerable improvement on overall response of fingerlings while increasing the levels of MOLM in the diet didn’t show significant effects. Additionally, it was confirmed that MOLM could replace costly FM by up to 10–20% without negatively affecting the overall response of C. catla fingerlings.

AKNOWLEDGEMENT

The authors are thankful to HEC Pakistan for funding Projects No. 20-4892/NRPU/RandD/HEC/14/1145 and 5649/Punjab/NRPU/RandD/HEC/2016 at Department of Zoology, Government College University, Faisalabad for provision of facilities for this research.

Funding

The funds for this project were provided by HEC project no. 20-4892/NRPU/RandD/HEC/14/1145 and 5649/Punjab/NRPU/RandD/HEC/2016.

Ethical statement

All applicable international, national, and/orinstitutional guidelines for the care and use of animals were followed.

Statement of conflict of interest

The authors have declared no conflict of interest.

REFERENCES

Abdulkadir, A.R., Zawawi, D.D. and Jahan, M.S., 2016. Proximate and phytochemical screening of different parts of Moringa oleifera. Russ. Agric. Sci., 42: 34-36. https://doi.org/10.3103/S106836741601002X

Adebayo, I.A., Akin-Obasola, B.J. and Abe, B.A., 2020. Growth and haematological responses of Clarias gariepinus juveniles fed diets containing varying digestible lipids of plant origin. Int. J. Fish. Aquacult. Res., 6: 10-23.

AOAC, 2005. Official methods of analysis, 18th ed. Association of Official Analytical Chemists, Washington, D.C. USA, pp. 1094.

Arsalan, M.Z.H., Hussain, S.M., Asrar, M., Anwar, H., Rehan, M.M.H., Shahzad, M.M. and Wahab, N., 2016. Effects of Moringa oleifera leaf meal (MOLM) based diets on carcass composition and hematology of Labeo rohita fingerlings. J. Biodivers. environ. Sci., 9: 214-223.

Aslam, S., Abbas, S., Kalhoro, M.A. and Shoaib, A., 2016. Anchor worms (lernaeid parasites), Lernaea polymorpha yü and Lernaea cyprinacea (Copépode: Lernaeidae) on major carps at different fish farms in Punjab, Pakistan. Pak. Sci. Int., 28: 295-298.

Ayotunde-Ezekiel, O., Ada-Fidelis, B. and Udeh-Grace, N., 2016. Effect of partial replacement of fishmeal with Moringa oleifera leaf meal on the haematology, carcass composition and growth performance of Clarias gariepinus (Burchell 1822) fingerlings. Int. J. Fish. aquat. Stud., 4: 307-311.

Billah, M.B., Haque, M.E., Sarkar, S., Hossain, M.M. and Dey, S.K., 2020. Growth performance, hematological disorder and bacterial challenge on Nile tilapia (Oreochromis niloticus) using Moringa oleifera plant leaf as feed supplement. Bangladesh J. Zool., 48: 151-166. https://doi.org/10.3329/bjz.v48i1.47884

Blaxhall, P.C. and Daisley, K.W., 1973. Routine hematological methods for use with fish blood. J. Fish Biol., 5: 771-781. https://doi.org/10.1111/j.1095-8649.1973.tb04510.x

Brown, B.A., 1988. Routine hematology procedures. In: Hematology: Principle and procedures, pp. 7-122.

Dawood, M.A., Koshio, S., Ishikawa, M. and Yokoyama, S., 2015. Effects of partial substitution of fish meal by soybean meal with or without heat-killed Lactobacillus plantarum (LP20) on growth performance, digestibility, and immune response of amberjack, Seriola dumerili juveniles. BioMed Res. Int., 2015: Article ID 514196. https://doi.org/10.1155/2015/514196

Doctolero, J.S. and Bartolome, R.M., 2019. Utilization of horseradish (Moringa oleifera) as an alternative protein-source feed ingredient on the diet of red Nile tilapia (Oreochromis niloticus). Int. J. Fish. aquat. Stud., 7: 94-97.

Elabd, H., Soror, E., El-Asely, A., Abd El-Gawad, E. and Abbass, A., 2019. Dietary supplementation of Moringa leaf meal for Nile tilapia (Oreochromis niloticus): Effect on growth and stress indices. Egypt. J. aquat. Res., 45: 265-271. https://doi.org/10.1016/j.ejar.2019.05.009

El-Gawad, A., Eman, A., El-Asely, A.M., Soror, E.I., Abbass, A.A. and Austin, B., 2020. Effect of dietary Moringa oleifera leaf on the immune response and control of Aeromonas hydrophila infection in Nile tilapia (Oreochromis niloticus) fry. Aquacult. Int., 28: 389-402. https://doi.org/10.1007/s10499-019-00469-0

Elumalai, P., Kurian, A., Lakshmi, S., Faggio, C., Esteban, M. A. and Ringø, E., 2020. Herbal immunomodulators in aquaculture. Rev. Fish. Sci. Aquac., 29: 33-57. https://doi.org/10.1080/23308249.2020.1779651

Ezekiel, M.O., Salaudeen, M.M., Olusola, A.O. and Adeyemi, R.S., 2016. Yield and proximate composition of farmed african fresh water catfish (Clarias gariepinus). Garden City 2021, pp. 554.

Falowo, A.B., Mukumbo, F.E., Idamokoro, E.M., Lorenzo, J.M., Afolayan, A.J. and Muchenje, V., 2018. Multi-functional application of Moringa oleifera Lam. in nutrition and animal food products: A review. Fd. Res. Int., 106: 317-334. https://doi.org/10.1016/j.foodres.2017.12.079

FAO, 2008. Fishery statistics (Aquaculture production). Food and agriculture Organization of the United Nations, pp. 99.

FAO, 2010. The state of World fisheries and aquaculture. Food and Agriculture organization of United Nations, Rome. https://www.fao.org/3/i1820e/i1820e.pdf

FAO, 2014. Food and agriculture organization of the United Nations, Rome, https://www.fao.org/3/i4040e/i4040e.pdf

FAO, 2020. Sustainability in action. State of World fisheries and aquaculture, Rome, pp. 200.

Fazio, F., 2019. Fish hematology analysis as an important tool of aquaculture: A review. Aquaculture, 500: 237-242. https://doi.org/10.1016/j.aquaculture.2018.10.030

Ganzon-Naret, E.S., 2014. Utilization of Moringa oleifera leaf meals as plant protein sources at different inclusion levels in fish meal-based diets fed to Lates calcarifer. Anim. Biol. Anim. Husband., 6: 158-167.

Gopalakrishnan, L., Doriya, K. and Kumar, D.S., 2016. Moringa oleifera: A review on nutritive importance and its medicinal application. Fd. Sci. Hum. Wellness, 5: 49-56. https://doi.org/10.1016/j.fshw.2016.04.001

Hardy, R.W., 2010. Utilization of plant proteins in fish diets: effects of global demand and supplies of fishmeal. Aquac. Res., 41: 770-776. https://doi.org/10.1111/j.1365-2109.2009.02349.x

Hussain, S.M., Javid, A., Hussain, A.I., Aslam, N., Ali, Q., Hussain, M. and Riaz, D., 2018. Replacement of fish meal with Moringa oleifera leaf meal (MOLM) and its effect on growth performance and nutrient digestibility in Labeo rohita fingerlings. Pakistan J. Zool., 50: 1815-1823. https://doi.org/10.17582/journal.pjz/2018.50.5.1815.1823

Hussain, S.M., Shahzad, M.M., Afzal, M., Javid, A., Mubarik, M.S., Shah, S.Z.H., Hussain M., Ahmad, S., Arsalan, M.Z.H., Manzoor, R. and Riaz, D., 2015a. Efficacy of phytase enzyme for increasing mineral digestibility of Cirrhinus mrigala fingerlings fed on soybean meal-based diet. Pakistan J. Zool., 47: 1807-1816.

Idowu, E., Adewumi, A., Oso, J., Edward, J. and Obaronbi, G., 2017. Effects of varying levels of Moringa oleifera on growth performance and nutrient utilization of Clarias gariepinus post-fingerlings. Am. Sci. Res. J. Eng. Technol. Sci., 32: 79-95.

Khetran, Z., Noor, Z., Rasool, F., Aslam, N., Rahujo, Z., Mehdi, H., Khetran, A., Khetran, R., Khan, K., Ahmad, M. and Noor, M., 2018. Effect of Moringa oleifera leaves on the growth and enzymatic activities of Labeo rohita by replacing with fish meal. Int. J. Biosci., 12: 200-205. https://doi.org/10.12692/ijb/12.1.200-205

Leone, A., Spada, A., Battezzati, A., Schiraldi, A., Aristil, J. and Bertoli, S., 2015. Cultivation, genetic, ethnopharmacology, phytochemistry and pharmacology of Moringa oleifera leaves: An overview. Int. J. mol. Sci., 16: 12791-12835. https://doi.org/10.3390/ijms160612791

Lovell, T., 1989. Nutrition and feeding of fish. Van Nostrand Reinhold, New York. Vol. 260. https://doi.org/10.1007/978-1-4757-1174-5

Nasr, M.A., Reda, R.M., Ismail, T.A. and Moustafa, A., 2021. Growth, hemato-biochemical parameters, body composition, and myostatin gene expression of Clarias gariepinus fed by replacing fishmeal with plant protein. Animals, 11: 889. https://doi.org/10.3390/ani11030889

Naylor, R.L., Goldburg, R.J., Primavera, J.H., Kautsky, N., Beveridge, M.C.M., Clay, J., Folke, C., Lubchenco, J., Mooney, H. and Troell, M., 2000. Effect of aquaculture on world fish supplies. Nature, 405: 1017-1024. https://doi.org/10.1038/35016500

Puycha, K., Yuangsoi, B., Charoenwattanasak, S., Wongmaneeprateep, S., Niamphithak, P. and Wiriyapattanasub, P., 2017. Effect of moringa (Moringa oleifera) leaf supplementation on growth performance and feed utilization of Bocourti’s catfish (Pangasius bocourti). Agric. Natl. Resour., 51: 286-291. https://doi.org/10.1016/j.anres.2017.10.001

Rowland, S.J. and Ingram, B.A., 1991. Diseases of Australian native fishes. Fisheries Bulletin 4 NSW Fisheries, Sydney, NSW, Australia.

Saini, R.K., Sivanesan, I. and Keum, Y.S., 2016. Phytochemicals of Moringa oleifera: A review of their nutritional, therapeutic and industrial significance. Biotech, 6: 1-14. https://doi.org/10.1007/s13205-016-0526-3

Shahzad, M.M., Bashir, S., Hussain, S.M., Javid, A., Hussain, M., Ahmed, N. and Khalid, F., 2021. Effectiveness of phytase pre-treatment on growth performance, nutrient digestibility and mineral status of common carp (Cyprinus carpio) juveniles fed Moringa by product based diet. Saudi J. biol. Sci., 28: 1944-1953. https://doi.org/10.1016/j.sjbs.2020.12.046

Sherif, A.H., Prince, A., Adel, S.A., Saad, S.M., Eldessouki, E.A. and Harfoush, M.A., 2022. Moringa oleifera mitigates oxytetracycline stress in Oreochromis niloticus. Aquacult. Res., 53: 1790-1799. https://doi.org/10.1111/are.15707

Snedecor, G.W. and Cochran, W.G., 1991. Statistical methods 8th Ed. Iowa State University Press, USA, pp. 503.

Soliva, C.R., Kreuzer, M., Foidl, N., Foidl, G., Machmüller, A. and Hess, H.D., 2005. Feeding value of whole and extracted Moringa oleifera leaves for ruminants and their effects on ruminal fermentation in vitro. Anim. Feed Sci. Technol., 118: 47-62. https://doi.org/10.1016/j.anifeedsci.2004.10.005

Steel, R.G.D., Torrie, J.H. and Dickey, D.A., 1996. Principles and procedures of statistics (3rd ed.). McGraw Hill International Book Co. Inc., New York. USA, pp. 336-352.

Tabassum, S., 2017. Effect of Moringa oleifera leaves on growth performance and survival rate of Mozambique tilapia fingerlings (Doctoral dissertation).

Tabassum, S., Hussain, S.M., Ali, S., Arsalan, M.Z.H., Ahmad, B., Asrar, M. and Sharif, A., 2023. Partial replacement of fish meal with Moringa oleifera leaf meal in practical diets of Cirrhinus mrigala fingerlings. Braz. J. Biol., 83: e246333. https://doi.org/10.1590/1519-6984.246333

Tacon, A.G.J. and Jackson, A.J., 1985. Utilization of conventional and unconventional protein sources in practical fish feeds. In: Nutrition and feeding in fish (eds. C.B. Cowey, A.M. Mackie and J.G. Bell). Academic Press, London, pp. 119–145.

Utilization of conventional and unconventional protein sources in practical fish feeds. Nutr. Feed. Fish, pp. 119-145.

Vergara-Jimenez, M., Almatrafi, M.M. and Fernandez, M.L., 2017. Bioactive components in Moringa oleifera leaves protect against chronic disease. Antioxidants, 6: 91. https://doi.org/10.3390/antiox6040091

Wedemeyer, G.A. and Yasutake, W.T., 1977. Clinical methods for the assessment of the effects of environmental stress on fish health. Department of the Interior, Fish and Wildlife Service. Vol. 89.

Wee, K.L. and Wang, S.S., 1987. Nutritive value of Leucaena leaf meal in pelleted feed for Nile tilapia. Aquaculture, 62: 97-108. https://doi.org/10.1016/0044-8486(87)90314-0