Growth performance, haematological indices and carcass characteristics of finisher broilers fed Ocimum gratissimum leaf extract as a supplemmeantary source of vitamins and minerals

Ezenwosu C, Anizoba N W, Osita C O, Onodugo M O, Nwoga C C, Emezie U C, Onyimonyi A E

Department of Animal Science, University of Nigeria, Nsukka, Nigeria
celestine.ezenwosu@unn.edu.ng

Abstract

A 4 weeks study was carried to investigate the growth performance, haematological indices and organ characteristics of finisher broilers fed Ocimum gratissimum leaf extract (OGLE) as a supplementary source of vitamins and minerals. A total of hundred and forty (140) Anak broiler strains of mixed sex were used for the experiment. The birds after 4 weeks of brooding, were weighed and randomly assigned to four dietary treatments in a completely randomized experimental design (CRD) with twenty (35) birds per treatment. Each treatment was replicated 5 times with seven (7) birds per replicate. OGLE0=(control) received only water, =vitalyte + water, OGLE30, 30ml OGLE + water and OGLE6 = 60ml OGLE + water. Results showed significant (P < 0.05) differences among the treatments means in total feed intake, average daily feed intake, total weight gain, average daily weight gain, final body weight and feed conversion ratio (FCR). Haematoligical analysis showed significant (P>0.05) differences among the treatments in WBC, RBC, HB and PVC values. However values of organ characteristics among the treatments were not significant (p>0.05). In conclusion, treatment groups had better growth performance than the control group and the better performance of birds on OGLE suggests that OGLE stands the chance of becoming possible replacement for costly synthetic commercial vitamins and mineral premixes that are used in broiler production.

Introduction

According to recent projections, the world human population will likely attain eight billion. Also, according to United Nations Population Division and United Nations Department of Economic and Social Affairs (2010) the global human population by 2042 will be expected to rise to nine billion. As of Wednesday, October 23, 2019, Nigeria population was about 202,559,797million (World meters, 2019).

This escalating human population might need a doubling and sustainable source of farm animal protein and provision of sustainable source of farm animal protein that will meet the protein need of the rising global human population calls f or the production of all classes of poultry meat such as chicken and turkey. However, investing meaningfully in poultry production will help to reduce hunger and poverty which are currently ravaging the increasing human population globally. FAO (2010) revealed that poultry sector is among the top sectors that provide various sources of animal proteins for human utilization. Poultry are most inexpensive source of farm animal protein, contributing meaningfully to the rising demand for animal food products globally (Farrell, 2013). During the last two decades, poultry industry was the greatest dynamic meat sector, displaying the utmost growth of the entire meat sectors as mirrored in world consumption. According to Adene (1989) and Moore et al. (1996) majority of organizations and individuals globally depends on poultry sector for considerable portion of their revenue and protein which is low in cholesterol

Poultry, despite its role in provision of meat for global protein demand, poultry production has been set back due the shortage and elevated cost of feed ingredients. Therefore, there is a need to look for a non-conventional and less competitive plant feed sources that can serve as alternative in poultry production. Such an alternative is leaf extract made from various tropical herbs and plants. It is beneficial to substitute limited and costly commercial vitalytes with locally available plant sources. However, inclusion of leaf extract as a alternative for scarce and high cost of synthetic commercial vitamin premix used in broiler production can be a good option when we want to reduce cost of production. According to Ikepeme et al (2012) leaf plants and vegetables are recognized to be loaded in indispensable protein, fatty acids, vitamins, minerals and also possess anti-microbial (anti-viral, bacteria, fungal and parasitic) effects and as such they can be used in the nutrition of poultry to improve production.

However, one of the widely cultivated vegetables in the tropics from which extract can be obtained is Ocimum gratissimum. In the southern part of Nigeria, it is called “Nchuanwu”. Photochemical evaluation of this plant has shown that it is rich in alkaloid, tannins, phytates, flavonoids and oligosaccharides (Ijeh et al 2004), cinnamate and camphor (Matasyoh et al 2007), saponins and phenolic compounds (Victor et al 2009). It also contains xanthones, terpenes and lactones (Ezekwesili et al 2004). The proximate composition shows a crude protein of 4.7%, 10.8% crude fibre and ash content of 12.24% (Mensah and Okoli 2008). In the Eastern part of Nigeria, Ocimum gratissimum is a plant whose extract (stem, leaves and roots) have been found to be beneficial in both animal and human beings. The extract of form Basil leaves (Ocimum gratissimum) which is commonly called scent leaves when crushed is used in the treatment of convulsion, stomach pain and catarrh and oil from the leaves have been found to possess antiseptics, antibacterial and antifungal activities (Sofowora 1992).

The utilization of diverse plant extracts in the production of broiler chickens is well documented (Nworgu et al 2007; Essien et al 2007; Galib and Noor 2010). According to Anaso and Onochie (1992) plant leaves extract has been utilized as bitter spice of flavoring agents in many traditional dishes. Rural farmers use it in the treatment of common respiratory diseases in poultry (Esien et al 2007). Leaf meal extract have been revealed to posses anti-oxidative properties and are being utilized in the management of sugar disease and other diseases in the tropics (Agbo et al 2005; Ugochukwu et al 2003). According to Gamaniel and Akah (1996) leaf meal extracts contains 5 pyhytochemical compounds including flavonoids, saponis, alkaloids, tannis, and glycosides and proposed possible variation in pharmacological effects. In another research work conducted by Ladokun et al (2016) oral administration of leaf extract such as Telfairia occidentalis leaf extract to laying birds enhanced haematological indices, hen day and internal egg qualities. Addition of pumpkin leaf extract to the drinking water of broiler finisher birds significantly enhanced their average daily weight gain and feed conversion ratio (Alabi et al 2017). Also, Agbo et al (2005) identified crops to be high in zinc, iron, vitamins, proteins and amino acids and thus could complete the inadequacies of these substances in feed. Unfortunately, its beneficial inclusion as a source of vitamins and mineral in broiler production is still at a much reduced ebb tide. This study was designed to investigate:

This study was designed to investigate the growth performance, haematological indices and carcass characteristics of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamins and minerals.

Materials and methods

Location and duration of the study and duration

The study was carried out at the Poultry Unit of the Department of Animal Science Teaching and Experimental Farm, University of Nigeria Nsukka Enugu State. It is located on longitude 6⁰25¹ N and latitude 7⁰24¹ E at an altitude of 430m above sea level (Breinholt et al 1981). The climate this study area is humid tropical with average maximum ambient temperature ranges from 33 ⁰C and 37⁰C (Okonkwo and Akubuo 2007). The annual rainfall ranges from 1567mm-1846.98mm (Metrological Center, Crop Science Department, University of Nigeria, Nsukka Enugu State). The study lasted for 8weeks

Collection and preparation of Ocimum gratissimum leaf extract

Ocimum gratissimum leaves were harvested fresh without the stalk from the Crop Science Research farm of University of Nigeria, Nsukka. The leaves were washed in a clean water to get rid of dirt and sands and under room temperature the leaves were air dried for 10 days. The dried leaves were crushed into miniature particles and later were soaked in water at the rate of 1.25kg in 5 litres. At 5⁰C, the supernatant was stored in a refrigerator and used on daily basis as O gratissimum leaf exract (OGLE). Other feed ingredients were purchased from Chidera feedmill limited, Nsukka, Enugu Nigeria.

Experimental diet

Experimental diet was formulated for finisher diets. The percentage composition of experimental diet and its proximate compositions are presented in Table 1. The diet was formulated to contain minimum of 20.00% crude protein and 2900.00 Met.Energy Kcal.kg^-1 . Proximate compositions of experimental diet were determined using standard methods (AOAC 1990).

Experimental layout

The trial was done using completely randomized design (CRD). The experiment involved four treatments with 35 birds per treatment. Each treatment was replicated 5 times with 7 birds per replicate.

Experimental procedure and management

The experiment was carried out in accordance with the provisions of the Ethical Committee on the use of animals and human for biomedical research of the University of Nigeria Nsukka (2006). One Hundred and forty (140) day old Anak broiler strains were purchased from Fidan Breeders, Ibadan, Nigeria. The chicks were brooded together for 4 week (28days) before they were weighed and randomly assigned to four dietary treatments with 35 birds per treatment in a completely randomized design (CRD). Each treatment had 5 replicates with 7 birds per replicate. The experimental treatments were as follows:

OGLE0: Water only

Vitalyte: Water +vitalyte

OGLE30: Water +30ml OGLE

OGLE60: Water+60ml OGLE

The vitalyte was administered in water according to the manufacturer’s recommendation of 5g per litre of drinking water. Birds on treatment OGLE30 and OGLE60 were provided drinking water having 30ml and 60ml OGLE per litre of water respectively. Water and feed were served ad libitum during the experimental periods. The research lasted for 4 weeks. Prior to the arrival of the birds from the hatchery, the brooding house was cleaned with soap and disinfected with strong disinfectant after which wood shavings were spread. The pen was pre-heated few hours before the arrival of the birds with charcoal pot. The pre-heating was to achieve a nice brooding environment that would enhance bird’s activities. Feeding troughs and drinkers were also procured, disinfected and strategically positioned. Clean drinking water and feed were made ready before the arrival of the birds. Timely vaccination and drugs were administered appropriately. The diets were prepared to meet the nutrients of broiler birds.

Parameters measured

Initial body weight and body weight gain

At the beginning of each phase of this experiment, the birds were weighed and also weighed on weekly basis throughout the experimental periods. At the end of the previous week, weight measured was subtracted from that of the present week in order to get the body weight gained for the week. A box on a top pan balance was used to weigh the birds and it was done in batches.

Feed intake (g)

On each day throughout the experimental periods, feed was weighed before being given to the birds. Then, the difference between the feed provided the preceding day and left over feed in the feeding trough the next morning was divided with the number of birds in each replicate in order to get daily feed intake per bird for each replicate.

Average daily feed intake (ADFI)

This was obtained by dividing the total feed intake of birds with the number of days the feeding trial lasted.

Average daily weight gain ( ADWG)

This was obtained by dividing total weight gained per bird per replicate with the number of days the feeding trial lasted.

Final body weight (FBW)

Weights of the birds at end of trial periods.

Feed conversion ratio (FCR) = Feed consumed (g) / Weight gain (g)

Haematological determination

At the eight (8) week of the study, a veterinarian was invited from the Faculty of Veterinary Medicine, University of Nigeria, Nsukka for haematological determination. Three birds were randomly selected from each replicate. Blood samples were collected from the wing vein using a 5ml syringe and placed in sterilized bottles containing Ethylene Diamine Tetra-acetic Acid (EDTA) anticoagulant for determination of the hematological values (Ritcheie et al, 1994). The samples was cooled at 4⁰c, using ice packs and transferred to the laboratory of Faculty of Veterinary Medicine, University of Nigeria, Nsukka within 12 hours of collection.

The following hematological parameters were determined.

Packed Cell Volume (PCV)-The packed cell volume (PCV) was determined by the microhaematochrit method of (Thrall and Weiser, 2002).

Red Blood Cell Count (RBC)-Haemocytometer method of (Thrall and Weiser, 2002) was used

White Blood Cell Count (WBC)- Haemocytometer method of (Thrall and Weiser, 2002) was used.

Haemoglobin Concentration (HbC)- The haemoglobin concentration of the blood samples was determined by the cyanomethaemoglobin method (Higgins et al 2008).

Mean corpuscular values

Mean Corpuscular Volume (MCV) = PCV(%) x 10/RBCcounts(million)

Mean Corpuscular Haemoglobin (MCH) = Hb (g/dl) x 100/RBCcounts(millions)

Mean Corpuscular Haemoglobin Concentration (MCHC) = Hb(g/d) x 100/PCV

Organ evaluation

At the end of the experiment, two (3) birds were randomly selected from each treatment. They were weighed and slaughtered in accordance with the provisions of the Ethical Committee on the use of animals and human for biomedical research of the University of Nigeria Nsukka (2006) in a research abattoir at the Department of Animal Science, University of Nigeria. The carcasses were de-feathered, eviscerated and cleaned. The weights of organs were determined using a manual and electronic weighing scale.

Experimental design

The experiment was executed using Completely Randomized Design (CRD).The experimental model of the Completely Randomized Design:

Xij=µ+T₁ +∑ij

Where, Xij = any observation or measurement taken

µ = population mean

T₁ = treatment effect

∑ij =experimental error

i= number of treatments

j=number of replicates

Statistical analysis

Data generated were subjected to the analysis of variance (ANOVA) in CRD using statistical package (SPSS, 2003) Windows version 8.0. Mean differences were separated using Duncan^’s New Multiple Range Test (Duncan, 1955) as outlined by Obi (2002).

Results and discussion

The results of the growth performance, haematological indices and carcass characteristics of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamin and minerals are shown in table 5, 6 and 7 respectively.

Table 4. Growth performance of finisher broilers fed Ocimum gratissimum leaf extract as a supplementary source of vitamin and mineral
Parameters (g)	Treatments
	Vitalyte	OGLE0	OGLE30	OGLE60	SEM	p
Initial body weight	1727.33	1713.67	1741.67	1709.00	14.41	0.890
Total feed intake	3856.67b	3450.00c	4120.00b	4733.33a	146.73	0.000
Average daily feed intake	137.73b	124.40a	147.14b	167.97a	5.02	0.000
Total weight gain	1366.67c	1090.00d	1476.67b	1950.00a	94.67	0.000
Final body weight	3094.00b	2787.00c	3215.00b	3659.00a	96.73	0.000
Average daily weight gain	48.80c	38.92d	52.73b	69.63a	3.38	0.000
Feed conversion ratio	2.82b	3.13a	2.78b	2.42c	0.08	0.000
abcd means on the same row with different superscript are significantly (P<0.05) different OGLE (ml)= Ocimum gratissimum leaf extract, SEM= Standard error of mean

 

Figure 1. Effect increse level on Ocimum gratissimum OGLE leaf extrac on feed intake

Figure 2. Effect increse level on Ocimum gratissimum OGLE leaf extrac on live weight gain	Figure 3. Effect increse level on Ocimum gratissimum OGLE leaf extrac on FCR

Table 5. The haematological indices of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamin and mineral
Parameters	Treatments
	Vitalyte	OGLE0	OGLE30	OGLE60	SEM	p
PCV (%)	31.00b	31.50b	33.00a	33.50a	0.35	0.001
RBC (X106/µ/l)	10.22b	10.11b	10.22b	10.69a	0.45	0.000
WBC (x103/ µ/l)	10710c	10700c	11200b	11600a	124.33	0.002
HB (g/dl)	10.95b	9.03c	11.50a	11.05a	0.11	0.000
MCV	10.39	10.40	10.51	10.61	0.12	0.062
MCH	35.34	34.63	34.82	35.69	0.46	0.142
MCHC	12.91	12.16	12.33	12.98	0.11	0.123
abc= means on the same row with different superscript are significantly different (p < 0.05). PCV = packed cell volume, RBC=red blood cell, WBC = white blood cell count, Hb = haemoglobin concentration, MCV= mean corpuscular volume, MCH= mean corpuscular haemoglobin, MCHC= mean corpuscular haemoglobin concentration. OGLE= Ocimum gratissimum leaf extract, SEM = Standard error of mean

Table 6. The organ characteristics of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamin and minerals.
Parameters (% LW)	Treatments
	Vitalyte	OGLE0	OGLE30	OGLE60	SEM	p
Gizzard	2.32	2.24	2.27	2.29	0.03	0.061
Liver	2.46	2.39	2.38	2.41	0.11	0.150
Kidney	0.13	0.14	0.15	0.16	0.03	0.140
abc= means on the same row with different superscript are significantly different (p<0.05). OGLE= Ocimum gratissimum leaf extract, SEM = Standard error of mean

Table 4. shows the growth performance of finisher broilers fed Ocimum gratissimum leaf extract as a supplementary source of vitamins and minerals. Results showed that administration of vitalyte and OGLE to finisher broilers caused significant (P<0.01) differences in the mean values for growth parameters. Daily feed intakes and weight gain of birds on vitalyte and OGLE (30ml and 60ml) were higher (P<0.05) compared to those on control, while feed intake of birds on vitalyte and those on OGLE were the same (P<0.05), but weight gain of those on OGLE were the highest among the treatments. This is an indication that finisher broilers can tolerate up to 60ml concentration of OGLE in water with positive effects on their growth performance. It may be concluded that the organs of the birds are already well developed and equipped to tolerate any anti-nutritive factors inherent in OGLE extract. Nworgu et al (2007) gave similar suggestions for broiler finisher birds fed Fluted pumpkin (Telfaria occidentalis) leaves extract.

However, increase (P<0.05) in the mean values for feed intake and weight gain in favour of birds on vitalytes and OGLE can be attributed to the increased availability of useful vitamins and minerals in the vitalyte and those inherent in the extract. Oluyemi and Roberts (2000) reports affirmed that the inclusion of both macro and micro nutrients in poultry ration improves intake of feed and utilization. It may be significant to conclude that the inclusion of Vitalyte® and OGLE increased the ability of the birds to utilize available nutrients in the feed they consumed compared to control birds.

Despite the facts that birds on vitalytes and OGLE had higher (P<0.05) values for feed intake, but body weight gain value of birds on increased OGLE60 intake were the highest (Table 5) among the treatments. This was an indication that the level of OGLE used in this study had nutritional benefits to the birds probably because of its content of active vitamins and minerals, phenols and flavonoids (Ijeh et al 2004; Victor et al 2009) which are very beneficial to the growth of the birds. Active ingredients like vitamin E and C, beta carotenes, phenols, lycopene, alkaloids, haemagluttinins, and flavonoids have been reported to have anti-oxidative properties (Ugochukwu et al 2003; Agbo et al 2005) and possible pharmacological effects (Okafor 2005). Alabi and Chime (2007) reported that scent leaf has high mineral and vitamin content and can increase the digestibility of diets and consequently increase the utilization of the feed as indicated by the better body weight gain in favor of birds on OGLE. Similarly, El-Deek et al (2003) and Hassan et al (2004) have reported the efficiency of spices in improving the performance of poultry species. Vitamins and minerals enhance feed digestion, nutrient absorption and utilization in form of co-enzymes and co-factors in the body of animals. Increase in nutrient absorption and utilization culminates into better performance. This may have led to the highest growth performance observed for birds on OGLE among the treatments. The proximate composition of Ocimum gratissimum shows a crude protein of 4.7%, 10.8% crude fibre and ash content of 12.24% (Mensah and Okoli, 2008). Also better growth performance observed for birds on OGLE can attributed to its antimicrobial properties and impact on gut function. Intake of herbs and spices or their products affects the gastrointestinal microflora composition and population (Leung and Foster, 1996) and controlling potential pathogens (Roth and Kirchgessner, 1998). Improvement in gut micro flora will enhance better feed digestion and utilization.

The feed conversion ratio of birds on vitalytes, (2.82), 30ml OGLE+water (2.78) and 60ml OGLE (2.42) were lower compared with 3.13 observed for the control birds that were on water only. Birds on vitalyte and, 30ml OGLE had similar feed conversion ratio, while birds on 60ml OGLE had the lowest value for feed conversion ratio among the treatments. The comparable performance of birds on vitalytes with those on 30ml and on OGLE suggests that OGLE stands the chance of possibly replacing the use of costly synthetic commercial vitamins and mineral premixes as a major source of these nutrients in poultry nutrition.

Generally, growth performance of birds can vary due to breed/genotype, age, nutrition, duration of experiment, ambient temperature, disease as well as management.

Table 5. shows the haematological indices of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamin and minerals. From the results, birds on the OGLE had the highest (P<0.05) immunity since they recorded the highest mean values for PVC, RBC,WBC and HB. Adamu et al (2006) observed that nutrition had significant effect on haematological values like PCV, HB and RBC. The white blood cells (leucocytes) are body defiance cells that prevent the entering of infections into the body system. Dietary inclusion of O. Gratissimum leaf extract caused an increase in the erythrocyte count. The high production on RBC in birds OGLE could be as a result of the fact that O. gratissimum contains erythropoietin-like agent(s) which is/are responsible for the increased production of erythrocytes.

Table 6. shows the organ characteristics of finisher broilers fed Ocimum gratissimum leaf extract as supplementary source of vitamins and minerals. There were no differences in the relative weights of the gizzard, liver, kidney and lungs of the birds among the treatments (Table 7). OGLE is not toxic to the liver and because it has the ability to reduce the level of liver enzymes in the blood, its role is rather protective and not destructive to the liver. Perrissoud (1986) reported that flavonoids exert a membrane-stabilizing action that protects organs like the liver cells from injury.

Conclusion

• Birds on vitalyte and OGLE had better growth performance than those on control, While birds on OGLE had the highest performance among the treatments.
• Since Ocimum gratissimum are high in abundant in rural communities, the extract made from it can conveniently replace the use of some conventional sources of vitamins and mineral supplements used in poultry production in rural villages where these premixes are not readily accessible and affordable due to their exorbitant prizes.

References

Adene D F 1989 An appraisal of the health management problems of rural poultry stock in Nigeria. Proceedings of the International Workshop on Rural Poultry in Africa. November 13-16, 1989, IleIfe,Nigeria, pp: 989- 999.

Agbo C U, Baiyeri K P and Obi I U 2005 Indigenous knowledge and utilization of Gongronema latifolia Benth: A case study of women in University of Nigeria, Nsukka. Bio-Research Journal 3(2) 66-69.

Alabi O, Ayoola M and Akinoso O 2017 Performance characteristics and physiological response of broiler chickens at finisher stage to oral supplementation with fluted pumpkin, Telfairia occidentalis leaf extract. Journal of Central European Agriculture, 18: 646-656.

Anaso H U and Onochie C C 1999 A comparative study of nutrients in Gongronema latifolium, Piper Guidneense and Piper nigrum, to testify high acceptability in local dishes. Journal of Science Engineering and Technology 6 (2): 1321 –1832.

AOAC 1990 Association of Official Analytical Chemist. Official Methods of Analysis 15 ^th edn., Washington D.C.

Breinholt K A L, Gowen F A and Nwosu C C 1981 Influence of environmental and animal factors on day and night grazing activity of imported Holstein-Friesian cows in the humid lowland tropics of Nigeria. Tropical Animal Production 6 (4):300-307 http://www.fao.org/Ag/aga/agap/frg/tap64/64_300.pdf

El-Deek A A, Attia YA, Hannfy M M 2003 Effect of anise (Pimpinella anisum), ginger (Zingiber officinale roscoe) and fennel (Foeniculum vulgare) and their mixture on performance of broilers. Archiv Für Geflügelkunde; 67(2):92-96.

Essien J P, Ebong G A and Akpan E J 2007 Antioxidant and antitussive Properties of Gongronema latifolium leaves used locally for the treatment of Fowl Cough in Nigeria . Journal of Applied Science Environment and Management 11(4) 47 – 50 http://www.bioline.org.br/pdf?ja07092

Ezekwesili C N, Obiora K A and Ugwu O P 2004 Evaluation of Anti-Diarrhoeal Property of Crude Aqueous Extract of Ocimum gratissimum L. (Labiatae) In Rats. Biokemistri. 16(2):122-131.

FAO 2010 Chicken Genetic Resources Used in Small holder Production Systems andOpportunities for their Development. FAO Small holder Poultry Production Paper, No. 5.Rome: Food and AgricultureOrganization.

Farrell D 2013 The role of poultry in human nutrition.http://www.fao.org/docrep/013/al709e/al709e00.pdf Accessed 10 April 2015.

Galib A M A and Noor M W 2010 Comparative study on diet supplementation with a mixture of herbal plants dandelion as a source of prebiotics on the performance of broilers Pakistan Journal of Nutrition 9(1):67. http://www.pjbs.org/pjnonline/fin1620.pdf

Gamaniel K S and Akah P A 1996 Analysis of the gastrointestinal relaxing effect of the stem extract of Gongronema latifolia. Phytomedicine 2(4): 293-296.

Hassan W N, Saed A M 2012 Protective effect of anise fruit ( Pimpinella anisum) against osteoporosis in rat model. American Journal of Biomedical Sciences 3(1):49-56.

Ijeh I I, Njoku O U, Ekenze E C 2004 Medicinal evaluation of xylopia aethiopica and ocimum gratissimum. J. Med. Aromatic Sci. 26(1): 44-47.

Ikpeme E V et al. 2012 Phytochemical screening and haematological evaluation of Parkia biglobosa and Gonglonema latifolium. J. Basic Appl. Res., 2 (3): 2599-2606.

Ladokun AO, Obe A A, Oso, A O, Oke O E and Abiona J A 2016 Performance and egg quality characteristics of egg-type chickens as influenced by fluted pumpkin (Telfaria occidentalis) leaf extract. Nigerian Journal of Animal Science, 18: 42-48

Leung A Y and Steven F 1996 Encyclopaedia of common natural ingredients used in food, drugs and cosmetics. 2^nd Ed. Pp. 649.

Matasyoh L G, Matasyoh, J C, Wachira F N, Kinyua M G, Muigai Thairu A W and Mukiama T K 2007 Chemical composition and antimicrobial activity of the essential oil ofOcimum gratissimum L. growing in Eastern Kenya. Afr J Biotechnol. 6(6):760–765.

Mensah J K, Okolie R I, Ohaju-Obodo J O and Eifedi K 2008 Phytochemical, nutritional and medical properties of some leafy vegetables consumed by Edo people of Nigeria. Afr. J. Biotechnol., 7, 2304-2309

Moore Jr P A, T C Daniel, D R Edwards and D M Miller 1996 Evaluation of chemicalamendments to reduce ammonia volatilization from poultry litter. Poult. Sci., 75: 315-320.

Nworgu F C, Ogungbenro S A and Solesi K S 2007 Performance and some blood chemistry indices of broiler chicken served Fluted Pumpkin (Telfaria occidentalis) leaves extract supplement. American-Eurasian Journal of Agriculture & Environmental Sciences, 2 (1): 90-98. http://ajol.info/index.php/ajb/article/viewFile/56911/45319

Obi I U 2002 Statistical methods of detecting differences between treatments means a research methodology issues in laboratory and field experiments. 2ndEd.Express Pub.Ltd.Enug u.Pp13-21.

Ofomata G E K 1975 Soil Erosion. In: Nigeria in Maps. Eastern States. Benin Ethiope Publishing House.pp:43-45

Okonkwo, W I and Akubuo C O 2007 Trombe wall system for poultry brooding. Int. J. Poultry Sci .6ː125-130.

Oluyemi J A and Roberts F A 2000 Poultry Production in Warm Climate. Spectrum Books Limited. 2^nd Rev. Edn., pp: 140-153.

Perrisoud D 1986 The development of cianidanol and 3-palmitoyl-9(+)-Catechin as drugs for the treatment of liver diseases. In: Plant Flavanoids in Biology and medicine: Biochemical, Pharmacological and structure- Activity Relationships, Cody, V., E. Middleton and J.B. Harborne (Eds.). Alan R. Liss, Inc. New York, pp: 559-569.

Sofowora A 1992 Medicinal Plants and Traditional Medicine in Africa. 24th Edn. John Wiley and Sons, New York, USA., Pages: 8.

SPSS for Windows 2001 Standard Version (8), SPSS INC. London

Ugochukwu N H, Babady N E, Cobourne M and Gasset S R 2003 The effect of Gongronema latifolia leaf extract on serum lipid profile and oxidative stress of hepatocytes of diabetic rats. Journal of Biological Sciences, 28:1-5. Received 1 August 2010; Accepted 11 January 2011; Published 1 April 2011

United Nations 2017 Resolution adopted by General Assembly on 6 ^th ,July,2017,work of the Statistical Commission pertaining to the 2030 Agenda for Sustainable Develpoment(A/RES/71/313-Archived, 28 November 2020 at the Wayback Machine).

United Nations Development Programme 2006 Socio-economic Impact of Avian Influenza inNigeria, Abuja.

Victor N O and ChidI O 2009 Phytochemical constituents of some selected medicinal plants, African Journal of Pure and Applied Chemistry, 3(11), 228-233

World meters RTS algorithm 2019 Elaboration of the latest United Nations data