Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken

Two hundred and forty day old broiler chicks were used to investigate the effect of aqueous Moringa oleifera leaf extracts (AMOLE) on growth performance and carcass characteristics of broiler chicken. The birds were randomly allocated into six treatments with four replicates, and each replicate containing 10 broiler chicks; the CRD was used. The treatments contained AMOLE 0+ (positive control with antibiotic treatment), AMOLE 0-(negative control with ordinary water), AMOLE 60 (60 ml/l), AMOLE 90 (90 ml/l), AMOLE 120 (120 ml/l) and AMOLE 150 (150 ml/l) inclusion levels of AMOLE, respectively. Birds on positive control had the highest final body weight and growth rate (2392.00 g and 53.61 g respectively) and the ones on 150 ml/l of AMOLE had the least (2042.00 g and 45.37 g respectively). Results of feed intake showed that birds on positive control had the highest (84.70 g) and the ones on 90 ml/litre of AMOLE had the lowest (73.19 g); while the results of feed conversion ratio indicated that birds on AMOLE 90 and AMOLE 120 performed better than the positive control treatment. Birds on the AMOLE had similar dressing percentages though that of positive control was highest (94.93 %); while those on AMOLE 60 and AMOLE 150 had the highest large intestine and lung weights respectively. Aqueous Moringa oleifera leaf extract can be included up to 90 ml/litre in the drinking water of broiler chicken for reduced feed intake (12.83 %) and improved feed conversion efficiency (9.11) thus, AMOLE can be used to replace synthetic antibiotics as growth promoter.


INTRODUCTION
Poultry production remains the most wide spread of all livestock enterprises; it constitutes an important pillar of food security improvement as well as socio-cultural and economic development in most countries (Alders, 2005;Dieye et al., 2010).Broiler production is a source of income, it is a good source of protein and quick returns on investment (Kekocha, 1994).However, the industry in the developing countries is facing some challenges; these challenges include high feed to gain ratio and increase in the cost of feed because of high prices of feed ingredients (Abbas, 2013).Numerous attempts have been made to overcome these challenges, and one of them involves the use of antibiotics in feed.Antibiotics have been utilized as growth promoters and to prevent outbreak of diseases (Thomke & Elwinger, 1998;Phillips et al., 2004).Furthermore, medication in water using antibiotics helps birds to recover from diseases (Khalafalla et al., 2010).However, the bene t of the use of antibiotics as growth promoters has some disadvantages; these include drug toxicity, residual effects and development of bacteria resistance (Ogbe & John, 2012).Studies have shown that usage of chloramphenicol resulted into bacteria of the

Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken
genus Salmonella developing resistance to the drug (Gassner & Wuethrich, 1994).The use of Avilamycin as a growth promoter resulted in an occurrence of avilamycin resistant Enterococcus faecium in broiler farms (Aarestrup, Bager and Andersen, 2000).These problems have led to the ban on the use of antibiotics as growth promoters by the European Union (Butaye et al., 2000;Catalá-Gregori et al., 2008).Thus, attention to safe and natural alternatives such as plants to replace antibiotics (phytobiotics) is on the increase.
Moringa oleifera leaves are reported to have potential prebiotic effects and potentially antioxidant phytochemicals, such as chlorogenic acid and caffeic acid (Siddhuraju and Becker, 2003).Moringa oleifera leaf meal, widely available in many tropical countries, is also a good source of antioxidant compounds such as ascorbic acid, avonoids, phenolics and carotenoids (Teixeira et al., 2014).The underlying effects of the bioactive compounds in M. oleifera leaves are not clear.However, they are believed to induce prebiotic effects, bacterial and immune-stimulant activities (Ghazalah & Ali, 2008) resulting in increased productivity of broiler chickens.Similar effects have been observed in the presence of antibiotic growth promoters (Khalafalla et al., 2010).However, data on the effects of M. oleifera leaf meal (MOLM) extract inclusion in the diet on growth performance and carcass characteristics of broiler chickens is limited and contradictory.Gakuya et al. (2014) reported a decrease in intake but an increase in feed conversion ratio.Olugbemi et al. (2010) observed a reduction in growth performance of broiler chickens when Moringa oleifera leaf meal was beyond 5% level of inclusion in the feed.Generally, Kakengi et al. (2007), Olugbemi et al. (2010, andAbou-Elezz et al. (2011) agreed that the use of Moringa oleifera leaf meal up to a level of 10% had no negative effect on the productive performance of broiler chicken, however, levels above 10 % produce adverse effects.This might be because the pure leaf meal are not extracted, tinctured, or cooked as a leaf extract, thus, the birds depend solely on the digestive system to extract the medicinal chemicals from the plant.Furthermore, the low performance at higher inclusion level could be due to high level of anti-nutritional factors and dustiness of MOLM and low digestibility of the bre, energy and protein presence in the raw leaf (Abba, 2013).Safa & El-Tazi (2014) observed signi cant in uence of MOLM on all the carcass parameters measured except the thigh weight.There are limited studies of the effect of Moringa oleifera leaf extracts (MOLE) growth performance and carcass characteristics of broiler chickens.Kachik et al. (1992) reported that the presence of phytate and other anti-nutrients can reduce the bioavailability of certain nutrients and processing can be done for maximum utilization of required nutrients from the leaves.Fuglie (1999) reported 53.77, 30.06 and 16.18 % decreased in extracted NDF, ADF and ADL, respectively of MOLE.Makkar & Becker (1997) reported that signi cant quantity of anti-nutritional factors, particularly saponins can be removed through solvent and aqueous extractions.Information on this extract is limited.The objective of this study was, therefore, to determine the effect of aqueous Moringa oleifera leaf extracts (AMOLE) on growth performance and carcass characteristics of broiler chickens.

Study location
This study was carried out at the Animal Production Teaching and Research Farm of the Federal University of Technology, Minna, Niger State in Nigeria.Minna is located between latitude 9º37' North and longitude 6º33' East.It is located in the Southern Guinea Savanna vegetation zone of North Central Nigeria.The mean monthly minimum and maximum temperatures are 38ºC and 42ºC respectively.The mean annual rainfall is between 1200 mm -1300 mm while the mean monthly relative humidity is 65 % (Climatemp, 2011).

Source of the test ingredients and preparation of the extracts
Fresh Moringa oleifera leaves were purchased between May and June from farmers in Minna.The leaves were air-dried in a laboratory for ve days and ground into ne particles using a simple hammer mill.60 g of the ground particles were then soaked in one litre of water for 24 hours, and this was done daily.The preparation were then ltered using a muslin cloth to separate the debris from the ltrate, and the extracts were placed in clean containers and diluted using borehole water (volume/volume) to form 0, 60, 90, 120 and 150 ml/1000 ml water for Treatments 2 to 5, respectively.This procedure was carried out daily and the ltrate served to the experimental birds in their drinking water.

Source of the experimental birds, experimental diets and experimental design
A total number of 240 day old Hubbard broiler chicks were purchased from Bnot Harel Hatchery, Oluyole Extension, Ring Road, Ibadan, Oyo State in Nigeria.The birds were randomly allocated to six treatments Obaje P, Ojo BK

Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken
of aqueous Moringa oleifera leaf extracts (AMOLE) in a completely randomized design experimental model.Each of the treatments had four replicates with ten birds per replicate.The birds were acclimatized for one week before being given the experimental treatments.Treatment 1 was the Control of which antibiotics (GENDOX ® ) at 1.25g/litre was used; Treatment 2, 3,  4, 5 and 6 were given 0, 60, 90, 120 and 150 ml per litre of aqueous Moringa oleifera leaf extracts (AMOLE) respectively (Table 1).

Management of the experimental birds
Twenty four (24) pen units, with an area of a square meter each that could accommodate ten broilers were constructed.The walls and oors of the pens were disinfected with Germicide (IZAL ®) after washing with detergent and water.Old newspapers spread on wood shavings as litter materials were used for the rst one week of the chicks' life.Clean and disinfected feeders and drinkers were set in a place accessible to the birds.Each pen unit was properly labeled for easy identi cation of each treatment group.In addition, a traditional charcoal pot was placed at a strategic area of each pen unit.The distance of the charcoal pot was adjusted based on the response of chicks to weather condition and rate of feather growth.The charcoal pot was removed during the third week when feathers were fully grown.Super starter mash from TOP FEED ® containing a crude protein (CP) content of 26.60 % and metabolizable energy (ME) of 2985 kcal/ kg were given to the birds during the rst two weeks, and starter mash (containing 24.85 % CP and 3013 kcal/kg ME) during the third and fourth week.Finisher feed pellets containing 22.05 % CP and 3026 kcal/ kg ME were given during the fth week of age till the sixth week.Feeds were given ad libitum and shifting from one form of feeds to another was done gradually to avoid digestive disorder.Medications and proper vaccinations were given to the birds based on the recommendations of the Nigerian Veterinary Medical Association (NVMA) for this region.

Data collection
The following data were collected and determined over a period of six weeks.

Water intake (ml)
The drinking water given to the birds in each treatment were measured daily and the left over water were also measured.Water intakes were determined by calculating the difference between the left over and the initial quantity of water given.

Feed intake (g)
The feed given to the birds in each treatment were weighed daily and the left over feed also weighed.The daily feeds consumed were determined by nding the differences between the left over and the initial quantity of feed given.The weekly records of average feed consumed were obtained for each of the replicate by dividing the total quantity of feed consumed by the total number of chickens in each replicate.

Body weight gain (g)
The body weight gains for each week were determined by subtracting the previous week's body weight from the current week's body weight.The initial weights of the birds were taken at the commencement of the study.The record obtained was used to calculate the average body weight gain per replicate (the weight for each replicate was added per replicate and then divided by the number of birds in each replicate).

Feed conversion ratio
The feed conversion ratio (FCR) were determined from the average weight gained and average feed consumed by the birds in each treatment.

Feed conversion ratio =
average feed intake (g) average body weight gain (g)

Carcass quality examination
At the end of the experiment, two birds from each replicate were randomly selected and fasted for 18 hours.Live body weight were recorded prior to slaughter.The birds were slaughtered using most humane method "killing cone".After evisceration, the data on hot carcass weights and organ weights were recorded and expressed as a percentage of the live body weights.The eviscerated carcass were chilled at 1-4 ºC for 24 hours.Then cold carcass weights were determined.The carcasses were partitioned and the breast, wing, thigh, drumsticks and feet yields were

Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken
weighed and expressed as percentage of the cold carcass weight according to the procedure of Hassan et al. (2004).

Chemical analysis
Proximate composition of the ne powder of Moringa oleifera leaf meal and the phytochemical composition of the leaf meal and the aqueous leaf extract were determined using the procedures of AOAC ( 2006).

Statistical analysis
Data collected were subjected to one way analysis of variance (ANOVA) based on the Completely Randomized Design model, using Statistical Analysis System (SAS, 2012).Where differences occurred at 5 % (p<0.05),they were separated using Duncan's Multiple Range Test (SAS, 2012).

RESULTS AND DISCUSSION
The results of the proximate composition of Moringa oleifera leaf is presented in Table 2. Results obtained in this study show that Moringa oleifera leaf meal has high crude protein content (23.80 %).This is similar to the result of Nuhu (2010) who reported values of crude protein, ether extract, crude bre, ash and nitrogen free extracts to be 29.55,22.30, 19.50 and 7.13 % respectively.It is also similar to the results of Oduro et al. (2008) who reported that Moringa oleifera leaf meal contains 27.51 % CP, 19.25 % CF, 22.3 % EE, 7.13 % ash and 76.53 % dry matter.Furthermore, crude protein results of 23.80 % obtained in this study is within the 20 to 33 % reported by Foidl & Paull (2008).However minor differences in the compositions could be due to environmental factors which play a minor role in determining the nutritive value of Moringa oleifera leaf meal.Birds on the AMOLE 0+ treatment had the highest nal body weight (FBW) and daily body weight gain (DBWG).Furthermore, the FBW and DBWG of birds on the aqueous Moringa oleifera leaf treatments increased until 120 ml and then declined; this implies that the optimal dosage must have been reached at about 120 ml before it declined.The increment in the FBW and DBWG might be as a result of the pharmacological chemical compounds (carbohydrates, saponins, cardiacglycosides, terpenes, steroids, avonoids and alkaloids) present in the extract as reported by Ambali & Furo (2012).This result is similar to the report of Zanu et al. (2012) who observed that the FBW and DBWG increased with increase in dosage level until 10 % and then signi cantly declined with increasing level of Moringa oleifera leaf meal.Unlike the results from previous researchers (Portugaliza & Fernandez, 2012, John & Kenaleone, 2014and Ashong & Brown, 2011), birds on the AMOLE treatments had signi cantly (p<0.05)lower FBW than the control.This might be because Moringa oleifera leaf meal was incorporated in the experimental diet as opposed to the aqueous extracts used in the present study.The antimicrobial (lipophilic compounds) and antioxidant (polyphenols, tannins, anthocyanin, glycosides compound) present in MOLE may attach to the cytoplasmic membrane and remove free radicals, activate antioxidant enzymes, and inhibit oxidases thus, making this elements more available for the birds to use (Jabaeen et al., 2008 andLuqman et al., 2012).Furthermore, the synergy between individual bioactive compounds in MOLM extract may be an important feature of their action which may affect broad aspects of physiology, such as nutrient absorption and processing, red-ox state, or immunity (Wallace et al., 2010;Mbikay, 2012).Results of feed intake showed that birds on the control treatment had the highest value (84.70 g/day), which is an indication that AMOLE inclusion depressed feed intake which might be as a result of nutrient satisfaction.This nding is similar to those of Ashong & Brown (2011) who reported that the control group had higher feed intake compared to the treatment groups when Moringa oleifera leaf meals were fed to White Leghorn type of chickens.The decrease in feed intake in birds on the AMOLE treatments is also in line with those of Portugaliza & Fernandez (2012) who observed that Moringa oleifera aqueous leaf extracts in drinking

Effect of Aqueous Moringa Oleifera (Lam) Leaf Extracts on Growth Performance and Carcass Characteristics of Hubbard Broiler Chicken
water signi cantly decreased feed intake of broilers as the concentration increased.This could be as a result of improved digestion and metabolism activities of Moringa oleifera (Ghazalah & Ali, 2008), thus, meeting the nutrients requirements at lower feed intake.
Furthermore, leaves of Moraga oleifera are rich in carotenoids, vitamins, minerals, amino acids, alkaloids, and avonoids (Siddhuraju & Becker, 2003).They have rare combination of phenolic compounds (zeatin, quercetin, kaempferol, apigenin), the combination of these compounds are essential for growth and they reduce disease infestation in the GIT (Teixeira et al., 2014) and, hence, improving the food utilization and leading to less feed needed to meet the requirements for maintenance and production of the birds.
The results of the FCR indicate that the birds on the AMOLE treatments at an inclusion of 90ml and 120ml/l (1.46 and 1.50 respectively) performed better than the control (1.58).This implies that the AMOLE treatments at these levels can be used to replace the antibiotic growth promoter.This might be because of the presence of bioceutical agents in Moringa oleifera plant as reported by Lannaon (2007) and bacterial and immune-stimulant activities of Moringa oleifera plant (Ghazalah & Ali, 2008).Furthermore, the AMOLE 90 and AMOLE 120 birds gave better FCR than the control, which means better returns on investment.This assertion is supported by David et al. (2012), Safa & El-Tazi (2012) and Ebenebe et al. (2012) who reported better feed conversion ratio for birds on M. Oleifera diets as compared to the control diets.
There was a signi cant (p<0.05)effect of aqueous Moringa oleifera leaf extracts on the dressing percentage of broiler chicken.This is in line with the results of Aderinola et al. (2013) who studied the effect of Moringa oleifera leaf meal on broiler chicken.Safa & El-Tazi (2012) also reported a positive in uence of Moringa oleifera treatments on rabbit.However, Ayssiwede et al. (2011) and Ochi et al. (2015) who studied the effect of Moringa oleifera seed powder on broiler chickens did not observe signi cant differences in the dressing percentage among the treatments.The variation could be attributed to the difference in the supplementation form of Moringa oleifera leaf meal.Inclusion of aqueous Moringa oleifera leaf extract showed no signi cant (p>0.05) in uence on the weight of the breast meat, thighs, wings and drumsticks.This is in line with the results of Ayssiwede et al. (2011).There were signi cant differences (p>0.05) in the weight of the large intestine and lungs.The reason for this is not known as AMOLE treatments had no in uence on the weight of the other organs in the broiler birds.Similar results were reported by Zanu et al. (2012) and Aderinola et al. (2013).

CONCLUSION
Aqueous Moringa oleifera leaf extract can be included up to 90 ml/litre in the drinking water of broiler chicken for reduced feed intake (12.83 %) and improved feed conversion ef ciency (9.11 %) when compared with the control.There was similarity in the thigh weight up to 90 ml/litre inclusion level and there were no signi cant differences in all the other carcass characteristics measured.It could, therefore, be concluded that Aqueous Moringa oleifera leaf extract can be used to replace synthetic antibiotic as growth promoter.An inclusion level of up to 90 ml/litre is recommended for improved feed conversion.

Table 2 -
Proximate composition of Moringa oleifera leaf meal

Table 3 -
Phytochemical composition of Moringa oleifera leaf meal and aqueous leaf extracts

Table 4 -
Growth performance of broiler chicken administered different levels of aqueous Moringa oleifera leaf extracts in their drinking water for the period of forty two days

Table 5 -
Effect of aqueous Moringa oleifera leaf extracts on carcass characteristics and carcass cut-up parts of Hubbard broilers