Dear Label_Name,

Greetings! I hope you are well.

I am writing on behalf of an international publishing house, Science and Education Publishing, USA (http://www.sciepub.com).

Science and Education Publishing (SciEP) is a multidisciplinary open access publisher of Journals covering the fields of Science, Technology and Medicine. All SciEP publications are subject to high-quality peer review, editorial and production processes and are freely available online.

Welcome to the April, 2013 edition of SciEP newsletter, providing monthly summaries of articles on science & education related topics of interest to scientist and professionals. The mission of our journals is to provide an open access forum for publishing new findings on issues pertinent to scientific and academic field. We also invite you to submit original research papers, review articles, short reports, or case studies are considered for publication in our Multidisciplinary Journals.

American Journal of Public Health Research
DOI: 10.12691/ajphr-1-1-3

The Implementation of a Tele-Homecare System with Service Oriented Architecture and HL7 Message Transmission Standard
Ching-Sung Wang

American Journal of Medical and Biological Research
DOI: 10.12691/ajmbr-1-1-2

The Influence of Ethanol on Pyruvate Kinases Activity in Vivo, in Vitro, in Silico
Lelevich Sergey Vladimirovich, Khrustalev Vladislav Victorovich, Barkovsky Eugene Victorovich, Shedogubova Tatyana Aleksandrovna

American Journal of Medical Sciences and Medicine
DOI: 10.12691/ajmsm-1-1-1

Clinical and Histological Features in Wegener Granulomatosis
Li Cavoli Gioacchino, Passantino Rita, Bono Luisa, Tortorici Calogera, Ferrantelli Angelo, Giammarresi Carlo, Rotolo Ugo

American Journal of Infectious Diseases and Microbiology
DOI: 10.12691/ajidm-1-1-2

The Curse of Lymphatic Filariasis: Would the Continual Use of Diethylcarbamazine Eliminate this Scourge in Papua New Guinea?
Anne Lanham, Lillian Mwanri

We have released issues of some journals on February 2013.
American Journal of Public Health Research
American Journal of Medical Sciences and Medicine
American Journal of Medical and Biological Research
American Journal of Pharmacological Sciences
American Journal of Clinical Medicine Research
American Journal of Nursing Research
American Journal of Cancer Prevention
American Journal of Medicine Studies
American Journal of Infectious Diseases and Microbiology
American Journal of Cardiovascular Disease Research
American Journal of Biomedical Research
American Journal of Mechanical Engineering
American Journal of Microbiological Research
Journal of Preventive Medicine
American Journal of Food and Nutrition
American Journal of Hypertension Research
American Journal of Sports Science and Medicine
Journal of Cancer Research and Treatment
American Journal of Zoological Research
Journal of Biomedical Engineering and Technology
American Journal of Microbiological Research
Biomedical Science and Engineering

We are pleased to announce that submissions are now being accepted for peer review for all the journals across all our subject areas in SciEP.

Please submit your manuscript via our online submission system:
http://www.sciepub.com/author/signin

To unsubscribe from our mailing list, please click on the following link: unsubscribe

I am looking forward to hearing from you.

Kind regards,

David Rain
Email:join@sciepub.com
Science and Education Publishing (http://www.sciepub.com)

Individual and Combined Effects of Moringa Leaf and Garlic Powder on Growth and Plasma Biochemical Indices of Clarias gariepinus Juveniles

Comfort Adetutu Adeniji*, Dorcas Wusu, Elizabeth Ololade Falana

School of Agriculture, Lagos State University, Ojo, Lagos State, Nigeria

Abstract: The study was designed to compare the growth promoting and some plasma biochemical indices of moringa leaf powder (MLP) and garlic powder (GP) each at 15.00g/kg; treatments 1 and 2, treatments 3 and 4 contained a combination of 7.50g and 15.00g/kg each of the phytoaddivites respectively and a control diet without additive. These five diets were fed to juveniles of Clarias gariepinus (46.45±0.12g). Relative to the juveniles in control diet, growth and nutrient utilization indices, (except survival percentage) of juveniles in phytoadditive diets increased. Plasma transaminase enzymes reduced significantly (p< 0.05) except for juveniles in treatment 4, likewise there were significant (p<0.05) reduction in ALP activities, except for juveniles in treatments 1 and 3. Also, there was significant (p<0.05) reduction in plasma cholesterol of juveniles in the phytoadditive diets. However, juveniles in treatments 1 and 2 recorded significant (p<0.05) increase in plasma phospholipids and triglycerides concentration compared to juveniles in control diet. Likewise, all juveniles in phytoadditive diets had significant (p<0.05) increase concentration of total protein, creatinine increased in treatment 2 and bilirubin increased in juveniles in treatment 3 as well as an increase in urea concentration with juveniles in treatments 1 and 3. Contrarily, creatinine concentration of juveniles in treatments 1, 3 and 4, albumin of juveniles in treatment 3 and bilirubin of juveniles in treatments 1 and 4 significantly (p<0.05) reduced compared to the values obtained with juveniles fed the control diet. While the bilirubin of juveniles in treatment 3 increased significantly (p<0.05). The results of this study showed improvement in the performance of juveniles fed phytoadditive diets when compared to those in the control diet. However, the inclusion of garlic powder at 15.g/kg produced the best growth, nutrient utilization and improved plasma biochemical indices of Clarias gariepinus juveniles.

Keywords: Moringa leaf, garlic, growth performance, feed utilization, plasma biochemical indices

Received April 04, 2019; Revised May 17, 2019; Accepted June 20, 2019

Citation: Comfort Adetutu Adeniji, Dorcas Wusu, Elizabeth Ololade Falana, Individual and Combined Effects of Moringa Leaf and Garlic Powder on Growth and Plasma Biochemical Indices of Clarias gariepinus Juveniles. American Journal of Food Science and Technology, 7 (5). 137-145. January 2019. doi:10.12691/ajfst-7-5-1

1. Introduction

Globally, there is increasing emphasis on the ban on the use of chemically synthesized antimicrobial drugs and growth enhancers. This emanated from the need to reduce environmental pollution, eliminate the emergence of antimicrobial resistance diseases and the quest for safe food. Hence, nutritionists are employing the use of natural alternatives, thus the emergence of natural feed additives. Natural feed additives or phytoadditives comprise of plant materials which possess growth promoting, antimicrobial, immune modulating and anti-oxidative properties among others.

Garlic (Allium sativa) is a plant species in the onion family. It has been used in food flavouring, traditional medicine and recently in animal nutrition. Garlic has antioxidant effects and useful in the control of pathogens; particularly bacteria and fungi. The allicin found in garlic has a broad range of chemical and biological properties [1]. Garlic also contains minerals (phosphorus and calcium), vitamins (A, C and B complexes), linolenic acid, carbohydrates [2] and many valuable compounds such as iodine, silicates and sulphur which have positive effects on circulatory, skeletal, cholesterolemia and control of liver diseases [3]. While Moringa oleifera is the most widely cultivated species of the genus Moringa. Its leaves possess nutritional and medicinal values. Nutritionally, it has a high exact protein content, an excellent source of B-vitamins, vitamin C and provitamin A as beta-carotene, vitamin k, minerals; manganese, iron and other essential nutrients. Medicinally, moringa has antioxidant, antibacterial and antifungal activities [4,5]. The leaves are free from antinutrients except for phenols that are much below the toxic threshold for animals and saponins which are inactive as far as haemolytic properties are concerned [6]

The growth promoting activities of garlic powder has been reported in Clarias gariepinus fingerlings [7,8] and Oreochromis niloticus [9] fingerlings. Likewise, there are reports on the growth enhancing potentials of moringa leaf meal in the diet of C. gariepinus juveniles [10] and the fingerlings [11]. Few studies suggested a potential synergistic effect of garlic with other phytoadditives [12,13,14], this suggestion needs further investigation before its adoption in practice. Consequently, this study was designed to compare the growth promoting, liver and kidney as well as lipid enhancing activities of garlic and moringa leaf powder in the diets of Clarias gariepinus juveniles when fed individually or in combination. Thus, growth performance, nutrient utilization, survival and some blood plasma chemistry of the juveniles fed diets containing these phytoadditives were assessed.

2. Materials and Methods

2.1. Materials and Feed Preparation

Two phytoadditives were used for the feeding trial. Flash dry moringa leaf powder (MLP) was purchased from the producer Julie Manny products limited Sagamu, Ogun State, Nigeria and garlic powder (GP) was purchased from a reputable Supermarket in Lagos, Nigeria. Five isoproteinous (35% crude protein) and isolipidic diets (2% palm oil) were formulated. The control contained no additive, treatment 1 contained 15.00g of MLP, treatment 2 contained 15.00g of GP, treatment 3 contained a mixture of 7.50g each of MLP and GP, while treatment 4 contained a mixture of 15.00g each of MLP and GP. Percentage gross composition of experimental diets is shown in Table 1. The feed preparation was done at a reputable commercial fish feed manufacturer where all other feed ingredients were purchased. The ingredients were weighed, ground into fine powder, thoroughly mixed and pelleted into 2mm with a 200kg/hr. table-top pelleting machine. The pellets were stored in air tight containers until when needed.

2.2. Fish Feeding and Management

The experiment was conducted at the indoor of Lagos State University Hatchery, Ojo Campus, Ojo, Lagos, Nigeria. Fifteen plastic aquaria tanks with 80L water capacity and 0.5m depth were filled with borehole water to three quarter capacity. The water in the experimental tanks was aerated by an electric air pump (Shining model; horsepower 50Hz). One hundred and fifty (150) Clarias gariepinus juveniles with a mean weight of 46.45±0.12g were obtained from a reputable commercial farm in Iba, Lagos, Nigeria. The fish were transported early in the morning between 6.30 and 7.00 am to the hatchery and acclimatized for three days before the start of the experiment. The fish were randomly distributed to 15 plastic aquaria at the stocking rate of 30 juveniles per experimental treatment and 10 per replicate. The fish were starved for 72hours to eliminate residual feed from the gut. Daily 50% of water in each tank was gently siphoned in exchange for fresh water. This was done to get rid of left over feed and faecal matter. Fish were fed to satiation twice daily between 7.00-8.00 and 17.00-18.00hours for a period of 12weeks. Feeding was done by hand and spread evenly across water surface of the aquarium.

2.3. Water Quality Analysis

Daily water temperature was measured using a mercury in-glass thermometer, hydrogen ions (pH) concentration measured using pH meter (Jenway Model 9060) and weekly dissolved oxygen (DO) concentration was measured by oxygen meter (Hanna Model HI-9142). Ammonia concentration was determined according to [15].

2.4. Growth Performance and Feed Utilization

Batch weighing of fish in each replicate aquarium was done at the beginning of the feeding trial, subsequently biweekly, using a MettleR 20110 top-loading balance. The evaluation of experimental diets for growth and feed utilization was according to [16].

2.5. Blood Collection

Blood samples were collected in triplicate following the procedure of [17,18]. The blood was collected using 5ml heparinise K3 vacuum tubes; the needle was run quite deep as much as possible through a middle line just below the sex organ in a dorsocranial direction till it strikes the vertebra column. The blood was immediately kept in the iced container after each collection. It was later taken to the laboratory and centrifuged for 10minutes. After that, the clear fluid which is the plasma was collected using a syringe and discharged into clean and sterilized tubes and kept in the freezer as described by [19] further analysis.

2.6. Biochemical Blood Analysis

2.6.1. Measurement of Plasma Liver Function Test

The activities of both aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were determined according to the methods before [20]. While, alkaline phosphatase (ALP) activity was assayed using the method of [21]. The albumin analysis was performed using the method of [22], bilirubin analysis was performed using the method of [23], urea analysis was performed using the method of [24], creatinine analysis was performed using the method of [25]. Triglycerides, cholesterol and phospholipids analyses were performed using the method of [26],

2.7. Statistical Analysis

All data collected were subjected to statistical analysis using GraphPad.

Table 1. The composition of the experimental diets (g/kg)

3. Results

The water quality and the summary of results of the growth performance, feed utilization and percentage survival of Clarias gariepinus juveniles fed diets fortified with flash dry moringa leaf powder, garlic powder and a combination of the two are presented in Table 2 and Table 3 respectively. While the results of plasma blood biomarkers measured are presented in Figure 1 to Figure 3.

3.1. Water Quality Analysis.

The water quality parameters of Clarias gariepinus juveniles fed the untreated and diets containing MLP and GP (treatments 1-4) individually or combined MLP and GP (treatments 1-4) were not significantly (p>0.05) different.

Table 2. Water quality analysis of Clarias gariepinus juveniles fed the experimental diets

Table 3. Growth, feed utilization and percentage survival of Clarias gariepinus juveniles fed the experimental diets

Figure 1. Liver Biomakers

Figure 2. Kidney Biomarkers

Figure 3. Lipid Profile

3.2. Growth Performance and Nutrient Utilization

The addition of flash dry moringa leaf powder (MLP) and garlic powder (GP) individually or in combination to the diets of Clarias gariepinus juveniles did not significantly (p>0.05) affect feed conversion ratio, protein efficiency ratio and the survival rate throughout the 12 weeks of the experiment.

However, Clarias gariepinus juveniles fed the control diet had significantly (p<0.05) lowest final weight (165.34±2.80) and weight gain (118.93±2.77), juveniles on diet containing 15.0 0g/kg GP (treatment 2) had significantly (p<0.05) highest final weight (210.93±14.75) and weight gain (164.25±14.88) while the final weight and weight gain of juveniles in other treatments did not differ (p>0.05) significantly from each other. Also, the lowest specific growth rate (0.76±0.01) was observed among juveniles in the control, while the significant (p<0.05) highest specific growth rate (0.90±0.40) with significant difference (p<0.05) significantly from values obtained with juveniles in other phytoadditives diets.

Feeding juveniles of African catfish (Clarias gariepinus) with 15.0g/kg GP (treatment 2) appeared to promote best growth performance and nutrient utilization. However, the mixture of MLP and GL did not improve growth performance and feed utilization of juveniles beyond the values obtained with juveniles fed either 15.00g/kg MLP or GP.

3.3. Plasma Biochemical Analysis

Blood plasma biochemical analysis of Clarias gariepinus juveniles fed the experimental diets for 12 weeks are presented in Figure 1 to Figure 3.

3.4. Liver Biomarkers

The inclusion of the phytoadditives to the diets of Clarias gariepinus juveniles resulted in significant (p<0.05) reduction in the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in juveniles fed treatments 1, 2 and 3 and alkaline phosphate (ALP) activities were also significantly (p<0.05) reduced in the plasma of juveniles fed treatments 1 and 3. However, the addition of 15.00g/kg⁻¹ each of MLP and GP (treatment 4) increased significantly (p<0.05) ALT activities of juveniles of Clarias gariepinus fed with this diet. Feeding of Clarias gariepinus juveniles with treatment 3 (7.50g/kg -1 each of MLP and GP) resulted in significant (p<0.05) reduction in albumin concentrations compared with values obtained from juveniles on the control and other diets. Likewise, significant reduction (p<0.5) was observed with bilirubin concentration of juveniles fed with treatments 1 and 4 (15.00 MLP and 15.00g/kg -1 of MLP and GP respectively). Contrarily, fortification of the juveniles’ diets with the phytoadditives resulted in significantly (p<0.05) increase in total protein concentration (though, lower value was obtained with juveniles fed with treatment 4) as well as in bilirubin concentration of juveniles fed with treatment 3 (7.50g/kg-1 each of MLP and GP).

3.5. Kidney Biomarkers

Feeding juveniles of African catfish with treatment 2 (15.00g/kg⁻¹ of GP) resulted in significant (p<0.05) increase in creatinine concentration when compared with juveniles fed the control diet. While significant (p<0.05) reduction in creatinine concentrations were observed with Clarias gariepinus juveniles fed other treatments when compared with the values obtained with the juveniles in control diet

3.6. Lipid Profile

The combination of MLP and GP at 7.50 and 15.00g/kg⁻¹ each (treatments 3 and 4) in the diets of Clarias gariepinus juveniles significantly (p<0.05) reduced the plasma cholesterol when compared with the values obtained from other treatment diets and the control. However, the HDL-Cholesterol, HDL-phospholipids and triglycerides concentrations of juveniles on treatments 1 and 2 were significantly (p<0.05) higher than values obtained from juveniles in the control. Also, juveniles fed treatment 1 (15.00g/kg⁻¹ MLP) had significant (p<0.05) higher HDL-Cholesterol among the phytoadditive fortified diets. In contract, HDL triglycerides were not significantly (p>0.05) affected by the inclusion of the phytoadditives.

4. Discussion

4.1. Growth Performance and Nutrient Utilization

Despite the global shift to the use of phytoadditives, there still exist dearth of information on the comparative use of moringa leaf powder and garlic powder individually or their combination as additives in the diets of African catfish; Clarias gariepinus juveniles. The results of parallel use of moringa, garlic and combination of MLP and GP showed significant improvement in growth performance and nutrient utilization of the Clarias gariepinus juveniles. Earlier work has reported the individual abilities of moringa and garlic powder to improve growth, efficiencies of feed and protein utilization, [9,27,28] in the diets of fish. The results of this study further attest to this claim, as juveniles on the phytoadditives showed better performance in feed utilization and growth. However, the improvement observed in feed utilization and growth performance parameters measured was highest when 15.00g/kg of garlic powder (treatment 2) was included in the diets of African catfish juveniles. Juveniles fed this diet consumed the highest amount of feed and protein, which were translated to best feed and protein efficiencies, cumulating in highest final weight and weight gain. This result agrees with the findings of [29,30,31] as well as that of [32] when garlic powder was included in the diets of Sterlet surgeon fingerlings, Dicentrarchus labrax fry, Tilapia zillii and Dicentrachus labrax juveniles respectively. The improvement in growth and utilization of nutrients of African catfish juvenile induced by garlic inclusion over the performance of juveniles fed 15.00gkg⁻¹ moringa could have emanated from the presence of allicin and other biological organo-sulphur compounds found in garlic. Allicin is known to promote feed intake, the performance of intestinal flora, improve digestion and enhances energy utilization [14,33]. These could have boosted nutrient utilization, resulting in higher weight gain than juveniles in treatment 1 (15.00g/kg⁻¹ MLP).

The lower performance observed with juveniles fed with diets containing the mixture of the two phytoadditives; moringa and garlic powder seems not to support the synergetic potentials of garlic with other phytoadditives suggested by previous studies [12,13,14,34]. Rather the results of this study showed lower growth performance, feed and protein intake when garlic and moringa leaf power mixture was added to the diets of C. gariepinus juveniles. The low performance was more pronounced at higher inclusion (15.00g/kg each of MLP and GP) treatment 4. Consequently, there might be the need to adjust the quantity of inclusion of moringa leaf and garlic powder, when used individually and when combined with other phytoadditives. Also, there is need to consider the intended fish to be cultured. This suggestion is borne out of the fact that the two mixtures (7.50 and 15.00g/kg⁻¹ each of MLP and GP, though with similar ratios; 1:1 in the diets of the C. gariepinus juveniles) had different feed utilization and growth performance. The discrepancy observed in this study may have resulted from the quantity and type of phytoadditives (among other factors) accompanying garlic in the diet used [32]. Hence, there might be the need to establish moringa leaf powder and garlic powder ratio when combined with the diets of Clarias gariepinus juveniles. This suggestion is buttressed by the findings of [13] who concluded that favourable health status of Cyprinus carpio L was only observed when garlic powder was used at 10g/kg diet or 7.5g and 2.5g/kg⁻¹ of garlic and ginger respectively as against other inclusion mixtures.

4.2. Plasma Biochemical Indices

Physiological stress indication such as biochemical variables can be useful in assessing nutritional effects of phytoadditives in fish. They are also useful biomarkers of liver and kidney physiological status.

4.3. Liver Biomarkers

ALT, AST and ALP are non-plasma specific enzymes that are localized in tissue cells of liver, kidney, heart, gills, muscles and other organs. Their presence in the blood may give specific information about organ dysfunction [35]. Transamination represents one of the pathways for the synthesis and deamination of amino acids. Thus, allowing interplay between carbohydrate and protein metabolism during fluctuating energy demands in various adaptive situations. It is considered necessary in assessing the state of the liver, kidney, heart and other organs. Therefore, attention has been focused on changes in these enzymes activities, their levels have been proposed to be useful indicator of extreme stress and provide information on organ dysfunction [36,37].

The decrease activities of the amino transaminases enzymes of phytoadditives fortified diets compared with the values obtained with juveniles on control diet may be attributed to moringa and garlic; which may have stabilized cell membrane and protect the liver and other organs [14,38] against deleterious agents and free radicals. This agreed with the findings of many studies when garlic was included in the diets of Nile tilapia and common carp [9,13,39]. ALT plays a vital role in synthesis and deamination of amino acids during stress-imposed conditions for meeting high energy demand of organisms [40,41,42]. Therefore, the elevated ALT activities at 15.00g each of MLP and GP (treatment 4) may suggest liver damage, which may be due to a disturbance in normal physiological and biochemical processes of C. gariepinus juveniles as fed in this study. This suggestion is borne out of the submission of [9] who reported that an alteration in a variety of chemical, biological and physiological factors or a disturbance in Krebb’s cycle might alter ALT activities. Consequently, the increased ALT activities may have arisen from any of these factors, which may have imposed some stress on the liver of the C. gariepinus juveniles in treatment 4. Alkaline phosphate is membrane-bound primarily found in liver and bone, to a lesser amount in cell lining of the intestine, placenta and kidney. Whatever is measured in the blood is the total amount of ALP released from these tissues, which indicate the possibility of bone or liver diseases [43]. The level of ALP is a reliable biomarker for other fish organs but not in plasma and muscles of Clarias gariepinus [42]. The reduced levels of ALP in treatments 1 and 3 compared with levels recorded with fish on other treatments and control diet may indicate an interruption in the secretion and flow of bile in the liver as reported by [44].

The elevated total protein concentration of juveniles of Clarias gariepinus fed the phytoadditive fortified diets as against the control may have arisen from the high feed and protein intake of juveniles fed with these diets [45]. Also, the additives may have repairing effects on the liver due to their nutritional properties [46,47]. Albumin is used as an indicator of liver function, reduced absorption or protein loss [48]. There were reductions in the concentrations of albumin in treatments 3 and 4 as well as in bilirubin concentration of juveniles in treatments 4. Conversely, higher bilirubin levels were observed with juveniles fed with treatment 3. Albumin and bilirubin are some of the biochemical indices for monitoring liver function in the blood. Abnormal levels of these proteins have been associated with liver damage [49]. Consequently, the reduced albumin concentrations observed in treatments 3 and 4 seem to indicate that the livers of the juveniles fed with these treatments were negatively affected. Also, the higher bilirubin concentration of juveniles on treatment 3 seems to lend credence to liver damage. The results observed seem to point to the probability of an inappropriate combination of moringa leaf meal and garlic powder, or perhaps improper mixing ratios of moringa leaf powder and garlic (7.50 and 15.00g/kg each) for the juveniles of Clarias gariepinus in this study. This could have exerted some level of stress on the liver resulting perhaps in reduced protein absorption by juveniles fed these treatment diets when compared with juveniles on other phytoadditive diets. These may have been responsible for the lower weight gained by juveniles of Clarias gariepinus fed with treatments 3 and 4 when compared with juvenile in other phytoadditive diets (treatments 1 and 2). However, the reduced bilirubin concentration recorded with Clarias gariepinus juveniles on 15.00g/kg MLP may be attributed to the inappropriate level of inclusion (rather than improper use) of the phytoadditives in the diet of the juveniles of Clarias gariepinus. This is because moringa leaf powder is known to possess nutritional qualities that enhance liver function [5,46] and protein absorption, which is evident in higher growth performance next to the performance recorded with juveniles on 15.00g/kg garlic diet. Hence, suggesting the need to further investigate this abnormality or establish their level of inclusion in the diet of Clarias gariepinus juveniles.

4.4. Kidney Biomarkers

The high creatinine concentration of juveniles fed treatment 2 (15.0g of garlic) may have resulted from the high muscle mass of juveniles in this diet [47]. While the reduced levels of creatinine observed with juveniles in other phytoadditives diets (treatments 1, 3 and 4) relative to juveniles in the control diet may be suggestive of the ability of the kidney in filtering the metabolic end products of the diets [50,51], rather than low muscle mass. This is because juveniles in these diets gained higher body weights when compared with juveniles fed the control diet. As earlier explained, the increased urea concentrations of juveniles fed treatments 1 and 3 (15.00g MLP, 7.50g each of MLP and GP) may have resulted from increased activity of the kidney [47,51].

4.5. Lipids Profile

Contrary to earlier reports the inclusion of moringa and garlic powder at 15.00g/kg each in treatments 1 and 2 did not have a hypolipidemic effect on Clarias gariepinus juveniles as fed in this study. Likewise, the increased plasma concentration of phospholipids and triglycerides found with all moringa and garlic supplemented diets. The discrepancy observed in this study may have emanated from the types and preparation adopted for the phytoadditives, time and dosage of application, species and stage of fish development as reported by many authours [32,52,53]. Also, since the diets fed were not high-fat diets, the hypolipidemic effect might not come into play [52,53]. Also relative to control and other phytoadditives diets, the inclusion of a mixture of MLP and GP at 7.5 and 15.0g/kg each (treatments 3 and 4) reduced plasma cholesterol concentration in the diets of African catfish juveniles. It can be inferred that the reduced cholesterol concentration observed with the juveniles of African catfish tend to suggest that the mixture of the phytoadditives as used in this study possess a synergetic ability that tend to enhance their hypolipidemic properties.

However, of importance in the moringa and garlic fortified diets are the increased plasma concentrations of HDL-cholesterol and HDL-phospholipids in the plasma of juveniles fed with these diets. These compounds are known to transfer fat away from cells, arteries and tissues. Increasing HDL particles are associated with decreasing accumulation of atherosclerosis, thus of significance in the production of fish for human consumption.

5. Conclusion

In conclusion, judging by the growth performance, nutrient utilization and plasma biochemical parameters (which are indices of the state of liver and kidney’s health status) assessed in this study, the inclusion of garlic at 15.00g/kg⁻¹ in the diets of Clarias gariepinus juveniles produced best results. While the mixture of moringa and garlic did not improve the performance observed with the individual use of either of the phytoadditives in the diets of Clarias gariepinus juveniles. Consequently, the potential synergic effect of garlic with other phytoadditives appear not to hold for moringa leaf powder as used in the diets of Clarias gariepinus juveniles in this study. There might also be the need to further investigate the level of the phytoadditives inclusion individually or in combination to establish their level (s) of inclusion in the diet of juveniles of Clarias gariepinus. However, of importance is the reduced tendency of atherosclerosis in man consuming catfish fed with these phytoadditives diets.

Statement of Competing Interest

The authours have no competing interests.

References

[1]   | Borlinghaus, J., Albrecht, F., Gruhike, M.C.H., Nwachukwu, I.D., Slusarenko, A.J. (2014). Allicin: Chemistry and biological properties. Molecules, 19; 12591-618.
[2]   | Dragon, S., Gergen, I., Socaciu, C. (2008). Alimentatia Functionala cu componente bioactive natural in sindromul metabolic Ed. Timisoara Eurostampa, p. 200-202, 160-161, 314.
[3]   | Corzo-Martinez, M., Corzo, N., Villamiel, M. (2007). Biological properties of ions and garlic. Trends in Food Science Technology, 18: 609-625.
[4]   | Anwar, F., Latif, S., Ashraf, M., Gilani, A.H. (2007). A food plant with multiple medicinal uses. Phototherapy Research, 2007, 21: 17-25.
[5]   | Gopalakrishnan, L., Doriya, K., Kumar, D.S. (2016). Moringa oleifera: A review on nutritive importance and application. Food Science and Human Wellness. 20(5): 249-566.
[6]   | Makkar, H.P.S., Becker, K. (1997). Nutrients and antiquality factors in different morphorlogical parts of the Moringa oleifera tree. Journal of Agricultural Science, 128: 311-322.
[7]   | Nwabuze, .A.A. (2012). The effect of garlic (allium sativum) on growth and haematological parameters of Clarias gariepinus (Burchell,1822). Sustainable Agricultural Research, 1: 222-228.
[8]   | Agbebi, O.T., Ogunmuyiwa, T.G., Herbert, S.M. (2013). Effect of dietary garlic source on feed utilization, growth and histopathology of the African catfish (Clarias gariepinus). Journal of Agriculture Science, 5:26-34.
[9]   | Shalab Y.A.M., Khattab, Y.A., Abdel-Rahman, A.M. (2006). Effects of garlic (Allium sa9tiva) and chloramphenicol on growth performance, physiological parameters and survival of Nile tilapia (Oreochromis niloticus). Journal of Venom. Toxins Incl. Tropical. Disease, 12: 172-201.
[10]   | Ayoola, S.O., Kuton, M.P., Shokefun, O. O. (2013). An evaluation of nutritional quality and haemat26zological parameters of (Moringa oleifera) Lam leaves in the diets of African catfish (Claria gariepinus). Agrosearch, 13:1-15.
[11]   | Bello, N.O., Nzeh, G.C. (2013). Effects of varying levels of Moringa oleifera leaf meal on growth performance, hematological indices and biochemical enzymes of African catfish Clarias gariepinus (Burchell 1822). Elixir Aquaculture, 59A: 1445-14466.
[12]   | Elias, N.S., El-Ghany, N.A.A. (2008). Monitoring the effect of garlic (Allium sativum) and black seed (Nigella sativa) on fusarium monifom infection in fish with emphasis on fecundity. Global Veterinary, 2: 242-249.
[13]   | Ajeel, S.G., Al-Faragi, J.K. (2013). Effect of ginger (Zingiber officinate) and garlic (Allium sativum) to enhance health of common carp (Cyprinus carpio L). The Iraqi Journal of Veterinary Medicine, 37: 59-62.

[14]  
Abu-Elala, N. M. Galal, M.K. Abd-Elsalam, R. M., Mohey-Elsaeed, O. Ragaa, N.M. (2016). Effects of dietary supplementation of Spirulina platensis and garlic on the growth performance and expression levels of immune-related genes in Nile tilapia (Oreochromis niloticus). Journal of Aquaculture Research and Development, (7): 433.

[15]   | Boyd, C.E. 1979. The chemical oxygen demand of water and biological materials fom pond. Journal of Environmental Quality, 8: 423-429.
[16]   | Olvera-Novoa, M.A., Campos, G.S., Sabido, M.G., Palacios, C.A.M. (1990). The use of alfalfa protein concentrates as a protein source in diets for tilapia (Oreochromis mossambicus) Aquaculture, 90: 291-302.
[17]   | Klontz, G.W., Smith, L.S. (1968). Methods of using fish as a biological research subjects. In: GREY, W. J. Methods of Animal Experimentation. New York: Academic Press, 3: 323-383.
[18]   | Wedemeyer, G.A., Yasutake, W.T. (1977). Clinical methods for the assessment of effects of environmental stress on fish health. United States Fish and Wildlife Service Technical Papers, Pp. 89.
[19]   | Ogbu, S.I., Okechukwu, F.I. (2001). The effect of storage temperature prior to separation on plasma and serum potassium. Journal of Medical Laboratory Science, 10: 1-4.
[20]   | Reitman, S., Frankel, S.A. 1(957). Colometric method for the determination of glutamic- ozaloacetic and glutamic-pyruvic transaminase. American Journal of Clinical Pathology, 33: 1-13.
[21]   | Englehardt, A. (1970). Measurement of alkaline phosphatase. Aerdti or aerzti Labor. 16:42-43.
[22]   | Grant, G. Het 1987. Amino acids and proteins. In: TIETZ N. W. (Eds), Fundamentals of Clinical Chemistry, 3rd ed., Philadelphia, WB Saunders Company, Pp. 328-329. 200-202.
[23]   | Tietz, N.W. (1995). Clinical guide to laboratory tests. 3rd ed., Philadelphia PA, W.B. Saunders Company, Pp. 518-519.
[24]   | Fawcett, J.K., Scott, J.E. (1960). A Rapid and Precise Method for the Determination of Urea. Clinical Pathotology, (13) 156-159.
[25]   | Bartel, S.H., Bohme, M. (1972). Creatinine. Clinical Chemical Acta, (37): 193.
[26]   | Kaplan A. Et AL 1984. Triglycerides. Clinical Chemistry, The C.V. Mosby Co. St Louis, Toronto, Princeton, Pp. 437 and Lipids Pp.1194-1206.
[27]   | Ali, S. M., El-Nadi,K., Mohamed, K.K. (2015). Effect of dietary supplementation with (Moringa oleifera) leaves on growth performance parameters on Nile tilapia (Oreochromis niloticus). Research Journal of Fish Hydrobiology, (10): 10-14.
[28]   | Irkin, L.C., Yigit, M., Yilman, S., Maita, M. (2014). Toxicology evaluation of dietary (Allium sativum) powder in European Sea Brass (Dicentrarchus labrax) juveniles. Food and Nutrition Sciences, 5: 989-996.
[29]   | Lee, D. H., Lim, S., Han, J.J., Lee, S.W., Ra, C.S., Kim, J.D. (2014). Effects of dietary garlic powder on growth, feed utilization and wholebody composition changes in fingerling sterlet sturgeon, Acipenser ruthenus. Asian-Australasian Journal of Animal. Science, (27): 1303-1310.
[30]   | Saleh, N.E., Micheal, F.R., Toutou, M.M. (2015). Evaluation of garlic and onion powder as phyto-additives in the diet of sea bass (Dicentrarcus labrax). Egytian Journal of Aquacultural Research, 41 (2): 211-217.
[31]   | Ajiboye, O.O., Quari, R. (2016). Short –term evaluation of graded levels of dietary garlic powder (Allium sativum L.) as growth promoter on growth, survival and feed utilization of Redbelly Tilapia, Tilapia zillii reared in glass aquaria tanks. International Journal of Marine Science, 6:.34.
[32]   | Irkin, L.C., Yigit, M. (2016). The use of garlic (Allium sativum) meal as a natural feed supplement in diets for European seabass (Dicentrarchus labrax) juveniles. Journal of Aquaculture, Engineering and Fish Research, 2:28-141.
[33]   | Khalil, R.H., Nadia, B.M., Soliman, M.K. (2001). Effects of biogen and evamisol HCL on the immune response of cultured Oreochromis niloticus to Aeromonas hydropila vaccine e. Beni-Suef css, Veterinary Medical Journal of Egypt, 11: 381-392.
[34]   | Kanani, H. G., Nobahar, Z., Kakoolaki, S., Jafarian, H., Gholipour, K.H. (2014). Effect of ginger and garlic-supplemented diet on growth performance, some hematological parameters and immune responses in juvenile Huso huso. Fish Physiology and Biochemistry, 40: 481-490.
[35]   | Gabriel, U., George, A.D 2005. Plasma enzymes in Clarias gariepinus exposed to chronic levels of roundup (glyphosphate). Environmental Ecology, 25: 271- 276.
[36]   | Hadi, A., Shoker, A., Alwan., S. (2009). Effect of aluminium on the biochemical parameters of fresh water fish Tilapia zillii. Journal of Science Application. 3: 33- 41.
[37]   | Nkpondion, N. N., Ugwuba, O.A., Esenowo, I.K. (2016). The toxicity effect of detergent on enzymatic and protein activities of African mud fish (Clarias gariepinus). Journal of Environmental and Analytical Toxicology, 6:361.
[38]   | Hussein. E. E. S. M. 2016. Effect of Moringa leaves, Moringa oleifera meal as feed additive in the performance of Nile tilapia, Oreochromis niloticus. International Journal of Aquaculture, 7: 1-8.
[39]   | Naeji, N., Shahsavani, D., Baghishani, H. (2013). Effects of dietary supplementation on lipid peroxidation and protein oxidation biomarkers of tissues as well as some serum biochemical parameters in common carp (Cyprinus carpio). Fish Science, 79 (4): 699-705.
[40]   | Svoboda, M. (2001). Stress in fish: A Review. In: Research Institute of Fish Culture and Hydrobiology. Vertebratae. 37: 169-191.
[41]   | Hukkeri, V.I., Jaiprakash, B., Lavhale, M.S., Karadi, R.V., Kuppast, I.J. (2002). Hepatoprotective activity of Ailanthus excelsa Roxbi leaf extracts in experimental liver damage in rats. Journal of Pharmacognosis, 11: 120-128.
[42]   | Wala, C., Bawa-Allah, A., Adams, A. (2014). Alkaline phosphate (ALP) activities in selected tissues and organs of Clarias gariepinus exposed to different levels of Paraquat. Journal of Biology, Agriculture and Healthcare, 2014, vol. 4 (21) 8-10.
[43]   | Raimi, O.G., Hammed, A.M., Adeola, S.A., Seriki, A.G. (2012). Isolation and characterization of ALP (E.C.3.1.3.1) from cultured and wild African mud catfish. Journal of Cell Tissue Research, 12: 3113-3116.
[44]   | Gabriel, U.U., Akinrotimi, A.O., Ariweriokuma, V.S. (2012). Changes in metabolic enzymes activities in selected organs and tissues of Clarias gariepinus exposed to cypermethrin. Journal of Chemical Engineering, 1: 25-30.

[45]  
Mlay, P.S., Seth, M., Balthazary, S.T., Chinbunda, R.T., Phiri, E.C.J.H., Balemba, O.B. (2007). Total plasma proteins and hemoglobin levels as affected by worm burden in freshwater fish in Morogoro, Tanzania. Livestock Resea rch and Rural Development, 19, Article ≠19. http;//www.Irrdl19/2/mlay 190919.htm

[46]   | Ekam, V.S., Johnson, J.T., Dasofunjo, K.., Odey, M.O., Anyahara, S.E., Annals of Biological Research, 3: 5590-5594.
[47]   | Ujah O.F., Ujah, I.R., Johnson, J.T., Oka, V.O. (2013). Effect of ethanolic extract of moringa oleifera leaf on haematological and biochemical parameters of wistar rats. Journal of Natural Plant Resources, 3 (2):10-14.
[48]   | Sacher, R.A., Mcpherson, R.A., Campos, J.M., Widmann, F.K. (2000). Widmann’s clinical interpretation of laboratory tests (ed. 11), Philadelphia, Davis Company.
[49]   | Islam, M.S., Lucy, N.S., Islam, M.R., Ahad, A., Das, R.R., Raham, M.M., Siddiul, M.S. (2004). Haematological parameters of Fayoumi, Assil and local chickens reared in Sylhet region in Bangladesh. International Journal of Poultry Science, 3: (2), p. 144-147
[50]   | Helmy, S.A., Morsy, N.F.S., Elaby, S.M., Ghaly, M.A.A. (2017). Hypolipidemic effect of Moringa oleifera Lam leaf powder and its extract in diet- induced hypercholesterolemic rats. Journal of Medicinal Food, 20: 755
[51]   | Halaby, M. S., Elmetwaly, E.M., Omar, A.A.A. (2013). Effect of Moringa oleifera on serum lipids and kidney function of hyperlipidemic rats. Journal of Applied Science and Research, 9: 5189-5198.
[52]   | Johnson, C., Banerji, A., Oommen, O.O. (2003). A comparative study of allitin and garlic on lipid turnover in a teleost, Anabas testudineus (Bloch). Indian Journal of Experimental Biology, 41: 242- 247.
[53]   | Fereidouni, M.S., Akbary, P., Soltanian, S. 2015. Survival rate and E biochemical parameters in Mugil cephalus (Linnaeus, 1758) larvae fed garlic (Allium sativum L.) extract. American Journal of Molecular Biology, 5: 7-15.