Polyherbal Aqueous Extracts in Comparison with Antibiotic on Growth Performance and Haematological Paramete...

Introduction

Antibiotics are utilized as growth promoters and for disease prevention and treatment. The beneficial effects of antibiotics in combating bacteria problems and as growth promoters are well known. Medication of water using antibiotics helps birds to recover from diseases (of bacterial origin). However, there may be problems associated with antibiotics such as drugs toxicity, residual effects and development of bacteria resistance. The negative impact on consumers of meat or poultry products due to residual effects has led to the ban on the use of antibiotics as growth promoters since 2006 by the European Union. Animal scientists and veterinarians are now turning attention to safe and natural alternatives such as plants (phytobiotic) to replace antibiotics. Plants contain phytonutrients and phytochemicals (such as saponins, tannins, oxalates, phytates, trypsin inhibitors and cyanogenic glycosides), which are referred to as secondary metabolites. Secondary metabolites are applied in nutrition and as pharmacologically active agents ¹ . Plants are also known to have high amounts of essential nutrients, vitamins, minerals, fatty acids and fibre² . There are reports on the presence of certain bioactive chemicals in plants or herbs and mushroom which have nutritional and medicinal benefits ^3,4,5,6 . Proximate and phytochemical analysis showed they contained essential nutrients and bioactive compounds (or phytochemicals) that play a role in nutrition (as feed supplement) and as medicines for the treatment of certain diseases ^6,7 .

Plants like Moringa oleifera are in high demand for their nutritional and medicinal value. Moringa leaves and seeds are used by humans as a good source of vitamins (B and C) and amino acids ^8,9 . Moringa oleifera was also claimed to boost immune systems ^9,10 . It has relatively high crude protein, low anti-nutritional factors and antimicrobial activity^11,12 . Gum Arabic is a secondary metabolite or natural secretion from trees (such as Acacia nilotica and Acacia Senegal), which is widely used as an additive in food materials. It is an edible coating used to increase stability and shelf-life and to enhance microbial safety of food ¹³ . Plant substances that are foods are of little or no side effects. Most of the prescribed medicines today (about 25%) are substances derived from plants ¹⁴ .

In developing countries (like Nigeria), modern medicines (or use of antibiotics) are increasingly unaffordable. There are concerns being raised on the adverse effects of antibiotics and drugs residues in meat and poultry products. Information on the utilization of plant products alone and in combination in form of phytobiotic or phytomedicine as alternative to reduce overdependence on antibiotics (as growth promoters or for treatment) in poultry production is scanty in Nigeria.

The objectives of this research work were: i. To determine the nutritional composition and the phytochemical constituents of Moringa oleifera, wild Ganoderma lucidum and Gum Arabic. ii. To evaluate the effects of polyherbal aqueous extracts from Moringa oleifera, wild Ganoderma lucidum and Gum Arabic in comparison with antibiotic on feed intake, growth performance and haematological parameters of broiler chickens.

Material and Methods

Study Area: The research was conducted at the Poultry Unit of the Livestock Teaching and Research Farm, Faculty of Agriculture, Nasarawa State University, Keffi, Nigeria. It is located in the Guinea savanna zone of North Central Nigeria and found on latitude 08° 35 N and longitude 08° 33 E.

Collection and processing of Moringa leaves, Gum Arabic and wild Ganoderma lucidum:Moringa oleifera leaves, Gum Arabic and wild Ganoderma lucidum were collected at different locations in the morning hours of the day. Moringa leaves were collected during flowering stage in Lafia, Nasarawa State. Stems were cut from Moringa trees and spread out under the shade to dry in the sun at 35° C for 3-5 days. The leaves were removed manually by hand and milled into powder form using a locally made miller machine (unbranded). Wild Ganoderma lucidum with red open caps were collected from decaying logs of woods and tree stumps during rainy season (June - September) in Vom, Plateau State. The wild mushroom was washed with water from the borehole and spread out in the sun to dry before milled into powder using a locally made mortar and pestle (unbranded). The milled mushroom powder was again dried in the sun for about 3 hours and then stored in plastic polythene bags at room temperature (28° C). Dried solid Gum Arabic granules were procured from the open market in Kaduna, Kaduna State, and milled into powder form using a locally made miller machine (unbranded).

Proximate study of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: The proximate study (crude protein, crude fibre, nitrogen, fat and moisture) was determined according to the methods recommended by the Association of Official Analytical Chemists ¹⁵ . All the determinations were done in duplicates. The proximate values were reported in percentage.

Determination of carbonhydrate and fatty acid: Total soluble carbohydrate was determined by the difference of the sum of all the proximate composition from 100%. The calorific (energy) value was obtained according to the methods of Akinyeye et al. ^16,17 . This was done by multiplying the value of carbohydrate, protein and crude fat by the Atwater factors of 17, 17 and 37 respectively. Crude fat was converted into fatty acid by multiplying with conversion factor of 0.80, as described by Akinyeye et al. ¹⁷ .

Mineral analysis of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: Mineral elements (calcium, magnesium, potassium, sodium, iron, zinc, manganese and copper) in Moringa oleifera leaves, Gum Arabic and wild Ganoderma lucidum were determined using Atomic Absorption Spectrophotometer (AAS-Buck 205 model). Phosphorus was determined colorimetrically. Flame photometer was used for the determination of sodium and potassium. All determinations were done in duplicates. The values of calcium, magnesium and potassium were reported in percentage (%), while sodium, iron, zinc, phosphorus, manganese and copper were reported in parts per million (ppm).

Phytochemical analysis and anti-nutrients of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: The quantitative phytochemical and anti-nutrients analysis of Moringa oleifera leaves; Gum Arabic and wild Ganoderma lucidum were carried out according to the methods of Sofowora ¹⁸ . All the determinations were done in duplicates.

Experimental birds and housing: Eighty day (80) old Arboracre commercial broiler chicks were procured from a hatchery at Ibadan, Nigeria. All the chicks were first brooded together on deep litter system of housing for one week in order to acclimatize. At two weeks of age, they were randomly distributed into five groups of treatments: Group A (Moringa oleifera + Gum Arabic + Ganoderma lucidum), B (Moringa oleifera + Ganoderma lucidum), C (Gum Arabic + Ganoderma lucidum), D (Moringa oleifera + Gum Arabic), E (Antibiotic only, as control for comparison). Each group was replicated (in duplicates) made of 8 chicks kept in separate wire-mesh compartments supported with wooden frame-work and the dimension measured about 200 cm x 100 cm. The chicks in all the groups (A-E) were fed on broiler starter mash containing 22% CP and 2800kcal ME from 1-4 weeks of age, followed by broiler finisher mash containing 20% CP and 2649kcal ME from 5-8 weeks of age, ad libitum. All the chicks in all the groups were given routine vaccinations against infectious bursal disease (first and second dose) at 2 and 5 weeks of age, and Newcastle LaSota vaccination at 3 weeks of age. Vitamins were administered to all the chicks in their drinking water for 3 days only after each vaccination.

Preparation of polyherbal aqueous extracts and administration to broilers: The Procedure involved; i. Different levels of aqueous extract were prepared (5, 10 and 20% w/v) using milled (powder) samples of Moringa leaves, Gum Arabic and wild Ganoderma in boiling water at 100 0 C for 12 h. Each solution was sieved and filtrate collected. ii. The filtrate from each extract was mixed at equal volume to form a polyherbal extract (1:1): Moringa leaves + Gum Arabic + wild Ganoderma (group A), Moringa leaves + wild Ganoderma (B), Gum Arabic + wild Ganoderma (C), Moringa leaves + Gum Arabic (D). Each polyherbal extract was labeled A – D, according to the groups of the broilers. iii. 5% w/v polyherbal aqueous extract was administered orally to the broilers at 2 weeks of age, 10% w/v at 4 weeks, and 20% w/v was administered at 6 weeks of age in each group.

Collection of data: Feed intake by the broilers in each group was monitored on daily basis. Growth performance in terms of body weight (gain) was determined on weekly basis and feed conversion efficiency was calculated by dividing the total feed intake by the weight gain of the broilers in each group. Blood samples were collected (4 birds/group) on weekly interval through jugular veni-puncture (2 ml/bird) using sterile syroinges and needles and the blood was immediately transferred into testtubes containing disodium salt of ethylene diamine tetra-acetic acid (EDTA). Blood samples were labeled according to the groups for haematology. Microhaematocrit method was used to determine packed cell volume (PCV) and cyanmethaemoglobin method was used to determine haemoglobin (Hb). Red blood cell (RBC) and white blood cell (WBC) counts were determined using Neubauer haemocytometer¹⁹ . At 9 weeks of age, carcass weights were determined from six randomly selected birds per group after slaughter (by severing the carotid arteries and jugular veins using a sharp knife) and carcass dressed (evisceration). The birds were first fasted for 12 hours (no access to feed except water) before slaughter. The organs and carcass in each group were separated into different parts and weighed: head, neck, shank, wing, thigh, breast, backbone and rib-cage. The organs: Liver, heart, gizzard and spleen were also weighed and expressed as percentage of the live weight.

Statistical analysis: Statistical analysis of data (nutrients, minerals phytochemicals, feed intake, weight gain, carcass and organ weights and haematological values) were all subjected to analysis of variance²⁰ . Duncan’s multiple range tests was used for the mean separation ²¹ . All the determinations were carried out in duplicates and results presented in mean ± standard deviations.

Result and Discussion

Proximate composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: The results of proximate study of Moringa oleifera leaves, Gum Arabic and wild Ganoderma lucidum are presented in Table 1. Moringa leaves contained appreciable amount of nutrients: crude protein (17.01% CP), carbohydrate (63.11% CHO) and energy (1440.11kcal/kg). Gum Arabic also contained appreciable amount of crude protein (15.38% CP), carbohydrate (62.26% CHO) and energy (1387.591kcal/kg). Wild Ganoderma lucidum also contained valuable amount of crude protein (16.79% CP), carbohydrate (63.27% CHO) and energy (1417.26kcal/kg). A higher crude protein value, crude fibre, fat and ash content in Moringa oleifera leaves was reported by Mutayoba et al. ²² . The crude protein observed in wild Ganoderma lucidum was higher (16.79 ±0.13%) than the crude protein value (13.3%) reported earlier by us. The crude protein, fat and fatty acids values obtained in this study were significantly different (P<0.05). These differences may not be unconnected to variations in the geographical location and stage of maturity of the plants and the wild mushroom. The presence of these important nutrients (carbohydrate, low fat and fatty acids) implies that Moringa oleifera leaves, gum arabic and wild Ganodema lucidum could be a nutritionally valuable and healthy ingredient for poultry. Low fat foods are known to reduce cholesterol level²³ .

Mineral composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: Table 2 showed that Moringa oleifera leaves contained minerals, Ca (1.91% ±0.08), Mg (0.38% ±0.01), K (0.97% ±0.01), Na (192 ±4.48), Fe (107.48 ±8.81), Zn (60.06 ±0.30), P (30.15 ±0.47), Mn (81.65 ±2.31) and Cu (6.10 ±0.20) in parts per million (ppm). Gum Arabic also contained minerals, Ca (2.10% ±0.13), Mg (0.42% ±0.10), K (1.30% ±0.04), Na (259.85 ±1.78), Fe (98.42 ±1.55), Zn (47.77 ±1.06), P (30.11 ±0.20), Mn (58.83 ±0.54) and Cu (5.94 ±0.30). Wild Ganoderma lucidum also contained minerals, Ca (1.99% ±0.04), Mg (0.34% ±0.01), K (1.11% ±0.04), Na (229.85 ±0.34), Fe (121.37 ±1.82), Zn (51.49 ±2.16), P (30.17 ±1.29), Mn (71.06 ±1.56) and Cu (7.43 ±0.13). These minerals, Mn (81.65, 58.83 and 71.06), Zn (60.06, 47.77 and 51.49) and Cu (6.10, 5.94 and 7.43), observed in Moringa leaves, gum arabic and wild Ganoderma lucidum, respectively were higher than those reported by Mutayoba et al ²² . Higher values of Fe (318.81), K (1.63%) and Mg (1.03%) were reported by Mutayoba et al ²² . Lower values of Ca (0.40%) and P (0.30) in wild Ganoderma lucidum were reported by Ogbe et al ⁵ . The values of Mg, K, Na, Fe, Zn, P, Mn and Cu obtained from Moringa leaves, gum arabic and wild Ganoderma lucidum were significantly different (P<0.05). The differences in the composition of minerals may be due to variations in the locality of growth and stage of maturity. The presence of these essential nutrients and minerals in Moringa leaves, gum arabic and wild Ganoderma mushroom would promote normal growth and health status of broiler chickens.

Phytochemical composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum: Table 3 showed that Moringa oleifera leaves contained phytates (2.59%), oxalate (0.45%), saponins (1.60%), tannins (21.19%), trypsin (3.00%) and hydrogen cyanide (0.10%). Gum Arabic also contained phytates (2.05%), oxalate (0.41%), saponins (1.47%), tannins (15.15%), trypsin (2.01%) and hydrogen cyanide (0.06%). Wild Ganoderma lucidum also contained phytates (2.43%), oxalate (0.57%), saponins (1.26%), tannins (18.27%), trypsin (2.39%) and hydrogen cyanide (0.08%). The levels of phytochemicals and anti-nutrients were low (less than 5%). The cyanide content of Moringa leaves, gum arabic and the wild mushroom was very low. Tannins appeared high in Moringa leaves (21.19%), followed by the wild Ganoderma mushroom (18.27%) and gum arabic (15.15%). The level of oxalates, saponins, tannins and trypsin in Moringa leaves, gum arabic and wild Ganoderma are significantly different (P<0.05). Some phytochemicals (such as tannins) were reported as phytoconstituents of plants ²⁴ . Phytoconstituents (which include saponins and other bioactive compounds) are antibiotic principles of plants ²⁴ .

Table – 1. Proximate composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum

Table – 2. Mineral composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum

Table – 3. Phytochemical composition of Moringa leaves, Gum Arabic and wild Ganoderma lucidum

Effect of polyherbal aqueous extracts (Moringa oleifera, Gum Arabic and wild Ganoderma lucidum) on feed intake and weight gain (kg/bird) of broilers: Table 4 showed the effect of polyherbal extracts from Moringa oleifera leaves, Gum Arabic and wild Ganoderma lucidum on feed intake of broilers. Group A (Moringa leaves + Gum Arabic + wild Ganoderma) recorded an increase in feed intake from 0.37 to 1.67kg/bird, and from 1-8 weeks of age the total feed intake was 6.89kg, which cost N822.34. Group B (Moringa leaves + wild Ganoderma) also recorded an increase in mean feed intake from 0.38 to 1.69kg, and the total feed intake from 1-8 weeks was 7.03kg, which cost N839.07. Group C (Gum Arabic + wild Ganoderma) also recorded an increase in feed intake from 0.37 to 1.65kg/bird. However, at higher concentration of polyherbal extract (20%w/v), there was slight decline in feed intake from 7 to 8 weeks of age (table 4). The total mean feed intake recorded in this group was 6.95kg and the total cost of feed consumed was N830.11. Group D (Moringa leaves + Gum Arabic) also recorded an increase in feed intake from 1 to 7 weeks of age (0.36kg/bird to 1.69kg/bird), which reduced to 1.65kg/bird at 8 weeks, and the total feed intake in this group was 7.18kg and the cost of feed consumed was N857.58. Group E (control) also recorded an increase from 0.40kg/bird to 1.67kg/bird, and the total feed intake was 7.18kg and the cost of feed consumed was N857.580. In general, when the broiler chickens were administered with aqueous extracts from Moringa leaves, gum arabic and wild Ganoderma (at 5% w/v), there was no significant difference (P>0.05) in all the groups when compared with that of antibiotic. However, at higher concentration (10 to 20% w/v), there was slight reduction in feed intake by the broiler chickens in group A. This difference in feed intake was significantly different (P<0.05) at 7 weeks of age (table 4). At 8 weeks of age, the broiler chickens in group C and E also showed a significant decrease in feed intake (P<0.05), but this was attributed to infection of the birds with intestinal coccidiosis (as revealed by the clinical signs of bloody diarrhea and postmortem). In general, group D and E had the highest feed intake (7.18 ±0.2, each), followed by B (7.03 ±0.13) and C (6.95 ±0.2). Group A had the lowest feed intake (6.89 ±0.2).

When the broilers were administered with polyherbal extract at higher concentration (10 to 20%w/v), group A (1.51kg) had the least weight gain (table 5). The feed cost per weight gain of the broilers in this group (A) was higher (N79.37), followed by C (N75.86), B (N74.09), D (N72.63) and E (N68.46) had the least cost. The feed cost per kg live weight was also higher in group A (N74.42), followed by C (N71.33), B (N69.73), D (N68.46) and E (N64.74) had the least cost. The feed to gain ratio was also high in group A (4.6). The reduction in feed intake and body weight in group A and C was significant (P<0.05). The combined effects of tannins in the polyherbal aqueous extracts may be responsible for reduction in feed intake and growth performance (weight gain) of the broilers (figure 1). Tannins are plant polyphenols, which (in excess) have ability to form complex with metal ions and this can reduce feed efficiency and weight gain ^25,26 . High level of saponnins and phytates can also affect feed intake due to reduction of protein digestibility and bioavailability of minerals ^27,28 .

Table – 4 Group weekly mean feed intake and weight gain (kg/bird) of broilers administered polyherbal aqueous extracts (Moringa oleifera, Gum Arabic and wild Ganoderma lucidum)

Effect of Polyherbal Aqueous Extracts (Moringa oleifera, Gum arabic and wild Ganoderma lucidum) in Comparison with Antibiotic on Growth Performance and Haematological Parameters of Broiler Chickens - Image 4

Effect of polyherbal aqueous extracts (Moringa oleifera, Gum Arabic and wild Ganoderma lucidum) on carcass and organs weight (%) and haematological parameters of broilers: The carcass analysis showed that pre-slaughter mean weight and dressed (carcass) weight of broilers in group E, D and B was 2.23, 2.17 and 2.16kg, respectively at the end of the experiment (9 weeks of age). Group A and C (2.07kg and 2.09) had the least mean weight (table 5). The carcass weight in Group E, D and B were heavier (1.65, 1.58 and 1.56kg), respectively. Group A and C had the least (1.49 and 1.54kg) mean weight. Although, the pre-slaughter and dressed (carcass) weights did not show any significant difference (P>0.05), but the increase in weights in these groups (E, D and B) may be attributed to the beneficial effect of the polyherbal extracts and antibiotic. High value of live body weight was said to attract high value of dressed (carcass) weight ²⁹ . The result also showed that dressing percentage was higher in group E and C (73.90% and 73.41%) than the others (table 5). In terms of carcass weight, the breast cut was heavier in weight in group A (28.53%). Group E had the least breast weight (25.51%). The thigh was also heavier in weight in group A (31.22%). The high values or weights of the cut carcass parts (thigh, breast and wings) in all the groups imply better growth performance.

Generally, the carcass and organ weight analysis in all the groups did not show appreciable variations. There was no significant difference (P>0.05) in the organ weights in all the groups. However, the weight of the gizzard in group C was high (10.64 %,) possibly due to the response of the broilers in this group to the combined effect of the high fibre content of the Gum Arabic and the mushroom (table 1). Also, when the broilers were administered with either polyherbal extracts or antibiotic, all the haematological parameters appeared within the normal range values (table 6): PCV = 29-44%; HB = 9.1- 13.9g/dl; RBC = 2.01 x 10 ¹² /L – 3.41 x 10¹² L; WBC = 2.35 x 10 ⁹ /L – 2.82 x 10 ⁹ /L. Wide ranges of normal haematological values were reported by Ogbe et al. ^19,30. Fluctuations in haematological values of avian blood are normal phenomenon and may be associated with the physiological status of the birds ^19,30 . Earlier studies have found Moringa (and other plants products) to be non-toxic and recommended for use in developing countries ³¹ . Aweng et al ³² reported that clean drinking water has become scarce nowadays due to poor land use management and pollution by sewage or waste dumping site. This polluted water will have to go through treatment process before it can be circulated for drinking. Possibly, Moringa oleifera and other plants products may also be useful for poultry drinking water treatment.

Figure – 1 Phytochemicals and anti-nutrients in Moringa leaves, Gum Arabic and wild Ganoderma lucidum

Table – 5 Carcass and organ body weights (%) of broilers administered polyherbal aqueous extracts from Moringa oleifera, Gum Arabic and wild Ganoderma lucidum

Table – 6 Mean (±SD) haematological values of broilers administered polyherbal aqueous extracts from Moringa oleifera, Gum Arabic and wild Ganoderma lucidum

Conclusion

In conclusion, the results of this study showed that Moringa oleifera leaves, gum arabic and wild Ganoderma lucidum contained appreciable amount of crude protein, dietary fibre, fatty acids and minerals, which are nutritional requirements of broiler chickens. These plants (products) and the wild Ganoderma mushroom contained valuable phytochemical constituents that improve growth performance and health of broiler chickens. Anti-nutritional factors in Moringa oleifera leaves, gum arabic and wild Ganoderma lucidum were low. However, the combined effects of anti-nutritional factors (like tannins) as a result of the combination of two or more aqueous extracts from these plants (products) could affect feed intake and weight gain of broilers. It was also concluded that the polyherbal extracts had no adverse effects on heamatological parameters of broilers. The heamatological values observed in this present study were all within the normal range. However, further trials on single doses of aqueous extract from Moringa oleifera leaves and seeds, gum arabic and wild Ganoderma lucidum and in feed are recommended.

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