Effect of Megathyrsus maximus (Guinea Grass) leaf meal on performance indices, carcass characteristics and blood profiles of broiler chickens

Gafotor J M^1,2, Sarkwa F O³, Ayizanga R A¹, Adjei-Mensah B¹ and Obese F Y¹

¹ Department of Animal Science, University of Ghana, Legon, Ghana
sarkwafelix@yahoo.com
² University College of Agriculture and Environmental Studies, Bunso, Ghana
³ Livestock and Poultry Research Centre, University of Ghana, Legon, Ghana

Abstract

This study investigated the effects of substituting wheat bran with varying levels of Megathyrsus maximus leaf meal (MLM) on the growth indices, haematological and serum biochemical parameters and carcass characteristics of broiler chickens production. A total of 240 unsexed day-old Cobb-500 broiler chicks (average initial weight of 42.43 g) were used for the experiment comprising four dietary treatments with four replicates per treatment for eight weeks. The experimental treatments included diets in which wheat bran was substituted with varying levels of MLM [0% (MLM 0), 50 % (MLM50), 75 % (MLM75) and 100 % (MLM100)]. Parameters of interest included feed intake, weight gain, feed conversion ratio, carcass characteristics haematological, and serum biochemical indices. The results showed that substitution of wheat bran with MLM in broiler chicken diets up to 75 % did not have any (p> 0.05) adverse effect on growth performance. However, beyond 75 % feed intake (p = 0.044) and weight gain (p = 0.023) declined, and feed conversion ratio (p< 0.002) was poorer. With respect to blood indices, dietary inclusion of MLM in broiler diets up to 100 % had no adverse effects (p> 0.05) on haematological and serum biochemical characteristics of broiler chickens. There were no differences (p> 0.001) among dietary treatments (MLM0, MLM50, MLM75 and MLM100) with regards to % bleeding, heart weight, neck weight, and abdominal fat weight. In conclusion, wheat bran can be substituted with MLM up to 75 % without adversely influencing the growth performance, haematological and biochemical parameters in broiler chicken production.

Key words: feed intake, haematology, serum biochemistry, weight gain, wheat bran

Introduction

Agriculture plays a pivotal role in improving food security and alleviating poverty in most economies (Karolina and Malgorzata 2020). The cost of livestock feed represents, approximately 70 to 80 percent, of the overall production cost (Flake and Ashitey 2008; Davis et al 2013). In Ghana, the cost of animal feed has been on a consistent upward trajectory in recent years, leading to a major negative impact on the country's livestock industry, particularly the poultry sector (Flake and Ashitey 2008) influencing both economic viability and production efficiency. In order to maintain profitability in this industry, it is therefore crucial to effectively manage animal feed costs. The seasonality and frequent scarcity of raw materials especially conventional feed ingredients such as maize and soyabean used in feed production account for the rising prices of livestock feed (Apantaku et al 2006; Aruwayo et al 2019). Currently, there is a growing search for easily available and cheaper non-conventional feed resources, and a focus on the development of innovative feeding approaches aimed at promoting healthier gut function to enhance the efficiency of utilization of nutrients for an improved broiler production at a cheaper cost (Attia et al 2020).

Wheat bran is the predominant source of dietary fibre used in poultry feed in Ghana. This by-product of the milling process of wheat is notably abundant in insoluble fibre, and to a lesser extent, contains cellulose and β-glucans (Kamal-Eldin et al 2009). Wheat bran as source of fibre regulate gastrointestinal functions, including gastric emptying time and intestinal transit rate which can subsequently impact digestive processes (de Mora 2015). It also enhances gut health, particularly in terms of gut microbiota, which in turn can improve digestive characteristics (Akhtar et al 2012).

The abundant forage resources in Ghana have not been extensively studied as potential local feed sources for poultry production, even though they are believed to contain valuable nutrients. Given the high cost and shortages associated with conventional feeds, it is imperative to explore alternatives that are easily available to lower production cost. Megathyrsus maximus (Guinea grass) is an important grass used for feeding ruminants in tropical regions, known for its high palatability to ruminant animals (Lawal-Adebowale 2012). Guinea grass contains a combination of fibre and protein, as well as a variety of vitamins and minerals (Quang et al 2015). These attributes make it a potential substitute for wheat bran in broiler diets. It was hypothesised that substituting wheat bran with MLM would not have any negative impact on the overall performance of the birds. Thus, this study was initiated to assess the effect of substituting wheat bran with Megathyrsus maximus meal (MLM) on growth performance, blood profiles and carcass traits of broiler chickens production.

Materials and methods

Study location

The experiment was conducted at the University College of Agriculture and Environmental Studies (UCAES) Poultry Farm at Bunso (6.2892⁰N, 0.4679⁰W), located in the East Akim Municipal of the Eastern Region within the semi-deciduous forest zone of Ghana. The area has a bi-modal rainfall pattern. The major rainy season typically occurs from April to June, followed by the minor rainy season from September to October. November to March are the dry months. The annual rainfall ranges between 125 and 175 cm and temperatures fluctuate between 26^oC and 30^o C. Relative humidity levels are generally high throughout the year ranging between 75 % and 80 % (Amissah 2014).

Ingredients used in formulating experimental diets

The experimental diets consist of various components, including maize, fishmeal, soyabean meal, wheat bran, iodated salt, oyster shell, dicalcium phosphate, premix (containing vitamins and minerals), lysine, methionine and dried MLM. All of these ingredients were sourced from the local market in Koforidua except the MLM. With the exception of the vitamin, all other feed ingredients were milled to a size of 5 mm as described by Svihus et al (2024). These ingredients were then carefully blended in accordance with the specific treatment requirements to form the standard diets.

Megathyrsus maximus meal (MLM) preparation

The Megathyrsus maximus grass was harvested from the local bushes surrounding the UCAES. The harvested herbage was chopped into pieces and dried in the shade till a moisture level of 18-20 % was achieved as recommended for hay (Nascimento et al 2020). During the drying process, tedding was done daily to prevent the potential growth of molds. Once completely dried, the grass was ground using a commercial grinding mill (2A Corn Mill, Rajasthan, India) to produce the MLM. The meal was then incorporated into standard diets by replacing varying proportions of wheat bran.

Experimental design

The experimental design was a Complete Randomized Design (CRD) comprising 4 dietary treatment groups, with 4 replicates per treatment. Each replicate was made up of 15 birds. Thus, a total number of 240 Cobb – 500 broiler chicks (initial average weight of 42.43 g) were used for the experiment with each dietary treatment comprising of 60 birds.

Experimental diet

The composition of experimental diet is shown in Table 1.

Experimental birds and their management

Brooding

Prior to the arrival of the chicks, thorough cleaning and disinfection of the brooder house was carried out. Upon arrival of the chicks, they were weighed and given water infused with Glucovit as an anti-stressor to provide energy. Five 100 watts filament bulbs were used as source of heat during the brooding period. The chicks remained in the brooder house and then were fed commercial starter diet ad libitum with same measured quantities of diet comprising MLM for 2 weeks, to accustom them to the experimental feed.

Bird distribution after brooding

A dip litter grower pen was partitioned with bamboo into 16 pens, each measuring 7.0 x 8.0 ft. These pens were thoroughly cleaned and disinfected with a solution of potassium permanganate and 37 % formalin prior to the arrival of the chicks. Each pen was equipped with feeders and waterers (nipples). Wood shavings of about 5 cm deep were used as bedding material. Following the adaptation period, 240 chicks were randomly selected, individually weighed using a digital balance, and then transferred into the various experimental pens.

Feeding and Medication

Throughout the experimental period, the designated feed was provided to the chickens three times daily, at 6:30 am, 12:30 pm and 5:30 pm. Fresh water was provided ad libitum. Vaccinations and medication for the chickens were administered according to a vaccination schedule obtained from the Eastern Regional Veterinary office in Koforidua.

Determination of chemical composition of MLM, wheat bran and formulated diets

Samples of MLM, wheat bran and the experimental diets were ground to pass through a 1 mm sieve. The dry matter (DM), crude fibre (CF), crude protein (CP), and ash contents were analyzed according to the standard procedure of AOAC (2005). The neutral detergent fibre (NDF) and acid detergent fibre (ADF) of MLM, wheat bran and experimental diets were determined according to the method of Van Soest et al (1991). The metabolizable energy (ME) of the wheat bran, MLM and experimental diets were determined by the use of formula given by Wiseman (1987) as:

ME (kcal/kg DM) = 3951 + 54.4 EE – 88.7CF – 40.8 ash

Determination of growth performance and feed utilization efficiency

Using a Wadfow Electronic Scale, the birds were weighed at the start of the experiment and then once every week. In addition, daily records of mortalities were kept during the experiment, along with weekly weighing of the amount of feed leftovers and the amount that was offered each time. The following formulae were used to calculate average weekly weight gain (AWG), average daily weight gain (ADG), and average daily feed intake (ADFI) based on the data. Feed intake (FI) divided by weight gain was used to calculate the feed conversion ratio (FCR), which was done for each phase of the trial across the whole duration.

Slaughtering procedure and carcass dissection

The birds were starved the night before they were euthanized. A total of 48 birds (12 birds per treatment) were chosen at random and weighed and then the carotid arteries were gently severed. The birds were fully bled and dressed to enable determination of carcass characteristics, including dressed weight, heart weight, liver weight, gizzard weight, drumstick weight, thighs weight, wings weight, breast muscle weight, abdominal fat weight, and small intestine length.

Blood sampling and determination of blood parameters

On the 57th day of the trial (end of experimental period), blood samples were obtained from two birds per replicate making up a total of 32 samples, for the investigation of haematological and blood biochemical variables. By using a needle and syringes to puncture the brachial left wing, 4mls of blood was collected from each bird into vacutainer tubes containing ethylenediamine tetraacetic acid (EDTA) as an anticoagulant for the purpose of determining the haematological parameters. Additionally, 4 ml of blood sample was collected into vacutainer tubes containing a gel separator for the determination of serum biochemical parameters. The gel separator kept the serum and the blood's cellular components apart, making separation easier for analysis. Following collection, the blood samples for the haematological and blood biochemical indices were kept on ice and then sent to the laboratory for further analysis. Blood samples in the gel separator tubes were centrifuged at 1,500 rpm for 5 minutes at room temperature (25 ^oC) at 4 ^oC after a thirty-minute clotting interval. The sera were removed, and the blood biochemical indices were determined.

An automatic Prokan PE-7080 (Shenzhen Prokan Electronics Inc, China) analyser was used to determine levels of haematological indices namely Hb, PCV, RBCcount, MCV, MCH, MCHC, WBC count, heterophils, lymphocytes, monocytes, eosinophils, basophils and blood platelets. The BA-88A Mindray semi-auto chemistry analyser (Guangzhou Medsinlong Medical Equipment Co. Ltd.) was used to determine the concentrations of serum biochemical parameters namely total protein, albumin, urea, total cholesterol, high density lipoproteins, low density lipoproteins, very low- density lipoproteins, creatinine. The difference between total protein and albumin was used to estimate the concentration of globulin.

Statistical Analysis

Data on growth performance (ADG, FI, FCR), carcass characteristics and blood parameters were analysed using the general analysis of variance procedure of GenStat 12^th Edition software (VSN 2009).

The model used was;

Y _іlk = µ + T _j + R _j + ԑ _іϳk

Where:

Y _ijk = Response variable (feed intake, weight gain, FCR, etc)

µ = Overall mean

T_j= Effect due to peculiarities of experimental diets

R_j = Effect due to replication

ԑ_ij= Random error term

Significant differences between treatment means were separated using the Student-Newman Keuls (SNK) test at 5 % level of significance.

Results

Chemical composition of Megathyrsus maximus leaf meal (MLM) and wheat bran

The chemical composition of MLM and wheat bran (Table 2) showed that, MLM had comparable dry matter content to wheat bran (85.4 versus 83.9 %), but higher ash (8.62 versus 6.30 %), neutral detergent fibre (69.9 versus 44.7 %) and acid detergent fibre (38.9 versus 13.6 %) content than wheat bran. Wheat bran however, had higher crude protein (14.9 versus 9.20 %) and metabolizable energy (2104 versus 2089 kcal/kg) content than MLM.

Feed intake and growth performance of broiler chickens

The influence of substitution of wheat bran with varying levels of MLM in the diet of broilers on growth performance is shown in Table 3. The average daily feed intake (ADFI) of birds on diets MLM0, MLM50 and MLM75 were higher (p=0.044) than that of diet MLM100. The values for diets MLM0 (119 g), MLM50 (120 g), and MLM75 (119 g) were similar but higher (p<0.044) than for diet MLM100 (111.02 g). Likewise, average daily gain (ADG) for birds on MLM0 (60.10 g), MLM50 (60.49 g) and MLM75 (60.5 g) were higher (p=0.032) than their counterparts on MLM100 (51.9 g). The feed conversion ratio (FCR) followed a similar trend being better (p<0.001) in birds on diet MLM0 (1.98), MLM50 (1.98) and MLM75 (1.96) than those on diet MLM100 (2.14). Furthermore, birds on diet MLM0, MLM50 and MLM75 had high (p<0.021) weight gain than those on MLM100. However, no mortalities were recorded for birds on any of the dietary treatments (Table 3). Figure 1 shows that, as MLM levels increased from MLM0 to MLM50, ADG slightly increased but decreased for MLM inclusion rate beyond 50 %. With respect to feed conversion ratio, increasing levels of MLM resulted in slight increase in FCR as indicated in Figure 2.

Figure 1. Relationship between ADG and varying levels of MLM	Figure 2. Relationship between FCR and varying levels of MLM

Live and organ weights, and carcass characteristics of broiler chickens fed diets with graded levels of Megathyrsus maximus leaf meal

The live weight, organ weight and carcass characteristics of broiler chicken fed diets containing various levels of MLM are presented in Table 4. Birds on diet MLM0 (0 % MLM substitution), diet MLM50 (50 % MLM substitution) and diet MLM75 (75 % MLM substitution) had similar (p>0.001) live weights, bled weight, % defeathering, drumstick weight, thigh weight, wing weight, and breast weight but were higher (p< 0.001) than those on diet MLM100 (100 % MLM substitution). Dressed weight was higher (p<0.001) in birds on MLM75 (2437 g) than MLM0 (2342 g) and MLM50 (2358 g) which did not differ (p>0.001) and MLM100 (1762 g) was the lowest (p<0.001). Full gizzard weight and empty gizzard weight were higher (p<0.001) in birds on diet MLM100 than those on MLM0, MLM50 and MLM75 which had similar (p>0.001) weights. Birds on diet MLM75 and MLM0 had longer length of the intestine (p<0.001) than those on diet MLM50 and MLM100. Liver weight was higher (p<0.001) in birds on diet MLM75 (47.3 g) than those on diet MLM0 (40.7 g). There were no differences (p> 0.001) among dietary treatments (MLM0, MLM50, MLM75 and MLM100) with regards to % bleeding, heart weight, neck weight and abdominal fat weight.

Effect of MLM on haematological parameters of broiler chickens

The result on haematological parameters (Table 5) indicated that the substitution of wheat bran with graded levels of MLM had effect (p=0.042) on MCHC levels. MCHC levels in birds on diet MLM50 (48.4 g/dL) was higher (p=0.042) than those on diet MLM0 (46.3 g/dL) and diet MLM100 (48.0 g/dL). There were no differences (p>0.05) among dietary treatments with all other haematological parameters determined.

Effect of MLM on serum biochemical parameters

The results in Table 6 indicate that the substitution of wheat bran with graded levels of MLM had no effect (p>0.05) on all serum biochemical variables except creatinine and HDL concentrations. Creatinine concentrations were higher (p=0.042) in birds on diet MLM0 (0 % MLM substitution) and diet MLM50 (50 % MLM substitution) than those on diet MLM75 (75 % MLM substitution) and MLM100 (100 % MLM substitution). However, birds on diet MLM0 and MLM50 had similar (p>0.05) creatinine concentrations. Birds on diet MLM0 had lower (p< 0.05) HDL concentrations than those on diet MLM75 and MLM100.

Discussion

Chemical composition of MLM and wheat bran

The dry matter (83.9 %) and crude protein (14.9 %) contents of wheat bran obtained in this study fell within the reported range of 83.6 to 90.3 % and 14.1 to 20.5 % respectively, reported by Bach - Knudsen and Canibe (2000). Also, the value of 44.7 % and 13.6 % for neutral detergent and acid detergent fibre were within the range of 32.4 to 56.5 % and 8.4 to 17.6 % respectively, reported for wheat bran (Gendley et al 2002). In the present study, the value for dry matter (85.37 %), acid detergent fibre (38.9 %) and ash (8.62 %) contents of MLM determined fell within the range of 83.4 to 93.4 %, 36.9 to 53.7 % and 6.6 to 14.2 % reported for dry matter, neutral detergent and ash respectively (Babayemi et al 2009). Similarly, the crude protein (9.20 %) and neutral detergent fibre (69.7 %) contents were within the range of 4.0 to 13.8 % and 68.3 to 81.5 % obtained by Aganga and Tshwenyane (2004)

Effect of wheat bran substituted with varying levels of MLM in broiler diet on feed intake and growth performance of broiler chickens

Feed intake (FI) is the main factor influencing the pace of broiler growth (Abdollahi et al 2018). The lower ADFI in birds placed on diet MLM100 (100 % MLM substitution) compared to birds on the other diets MLM0 (0 % MLM substitution), MLM50 (50 % MLM substitution) and MLM75 (75 % MLM substitution) may be due to the higher NDF content of the diet. Neutral detergent fibre concentration is inversely related with voluntary feed intake (Sarkwa et al 2020; Diatta et al 2021; Adogla-Bessa et al 2022). According to Massuquetto et al (2020) NDF raises intestinal viscosity and slows down feed passage, which lowers feed intake and nutrient absorption rate and may impact chicken growth performance. Likewise, Tesfaye et al (2013) reported that higher fibre content of rations containing higher levels of leaf meal may have a negative effect on broiler growth performance. Higher amount of fibre especially in poultry make the gastrointestinal tract bulkier decreasing feed intake (Buragohain 2016). This bulkiness can lead to a reduction in nutrient digestibility and subsequently impact growth performance, though the specific effects can vary significantly based on the fibre type and concentration (Singh and Kim 2021). The lower ADG of birds on MLM100 (100 % MLM substitution) was due to their reduced ADFI. Consequently, the lower ADFI coupled with lower ADG in the birds on diet MLM100 resulted in their poorer (high value) FCR in the current study.

The dietary treatments with varying amounts of Megathyrsus maximus leaf meal had no effect on the mortality of broilers. Manjaniq et al (2017) and Valdez et al (2024) have also reported that adding cassava leaf meal at 3 to 5 % and Moringa oleifera leaf meal at 3 % respectively had no effect on the mortality of broilers.

Live weight, organ weights and carcass characteristics of broiler chickens fed diets with graded levels of MLM

Carcass evaluation is an important process in assessing an animal value and meat quality during slaughter (Delgado-Pando et al 2021). The lower live weight, bled weight, intestine length, thigh weight, drumstick weight, wing weight, and breast weight of birds on diet MLM100 (100 % MLM substitution) compared to those on diet MLM0 (0 % MLM substitution), MLM50 (50 % MLM substitution) and MLM75 (75 % MLM substitution) may be due to their lower feed intake and utilization that hampered growth and carcass development. Some authors have also observed poor growth performance in chickens supplemented with high doses of leaf meals (Modisaojang-Mojanaja et al 2019; Manyelo et al 2022).

The gizzard is expected to grow in size and mass as more fibrous material is included in the diet of poultry due to its workload and this may account for the higher full and empty gizzard weights of the birds on diet MLM100 (100 % MLM substitution) which had higher fibre content compared to their counterparts on diet MLM0 (0 % MLM substitution), MLM50 (50 % MLM substitution) and MLM75 (75 % MLM substitution). Voemesse et al (2018) added graded levels of Moringa oleifera leaf meal to broiler diet and had similar result. The authors ascribe the large weight of the gizzard to have resulted from its muscles having to work harder to grind the high fibre content of feed that were high in the Moringa oleifera leaf meal.

The length of the gut has a major impact on the digestion and absorption of nutrients. Longer intestines enable longer digesta retention in the gastrointestinal tract, extending the duration of time that substrates and digestive products, as well as intestinal mucosa and products of digestion, can come into contact with one another (Ravindran and Abdollahi 2021). This prolonged contact time subsequently maximises the enzymatic activity and absorption surface, thereby improving the overall nutrient absorption efficiency (Farago et al 2020). The increased exposure facilitates more comprehensive breakdown of complex dietary components and greater uptake of liberated nutrients, particularly in species adapted to diets requiring extensive digestive processing (Duque-Correa et al 2022). Thus, the highest intestinal length of birds on diet MLM75 and MLM0 than those on diet MLM50 and MLM100 may partly account for their higher feed utilization and growth. The similar % bleeding, dressed %, heart weight, neck weight, and abdominal fat weight of birds among dietary treatments in the present study suggest no adverse effect on these parameters.

Effect of MLM on haematological parameters of broiler chickens

Blood metabolites are useful indicators of nutritional, physiologic, metabolic and health status of livestock (Onasanya et al 2015) and hence essential in evaluating the suitability of introduced feed resources. The similar levels of haematological parameters determined in the current study suggest similar ability of the dietary treatments in enhancing the production of haemoglobin (Hb) for normal synthesis of RBCs for effective transport of gases. It also indicates adequate synthesis of WBCs to defend the body against infections. The level of haematological parameters measured were within the normal physiological ranges reported for poultry (Bounous and Stedman 2000; Baudouin et al 2021), suggesting that substitution of wheat bran with MLM in diets of broilers did not adversely influence their haematological parameters. Substitution of wheat bran in the diet of broilers with MLM increased MCHC levels than the control. All the levels of MCHC were higher than the normal (26 – 35 g/dL) in the birds on the control diet MLM0 and the birds on MLM50, MLM75 and MLM100. Although higher than normal MCHC level may lead to anaemia, birds on the various dietary treatment diets did not display any sign of anaemia. Moreover, the Hb and RBC concentrations of birds on the various treatments were within the normal physiological range of 7 – 13 g/dL and 2.5 – 3.5 x 10¹²/L respectively reported for birds (Bounous and Stedman 2000).

Effect of MLM on serum biochemical profile of broiler chickens

In the present study, the similar concentrations of total protein, albumen, globulin, urea, total cholesterol, triglyceride, low density and very low- density lipoproteins (LDL and VLDL) among dietary treatments and which were within the normal physiological range of values reported for poultry (Harr 2002; Bueno et al 2017) suggest that substituting wheat bran with MLM in the diets of broilers did not adversely affect the health and physiology of the birds. The diets provided adequate protein and lipids for protein and lipid metabolism and efficient maintenance of osmotic pressure and transport of various substances in the serum in addition to proper immune response, fat metabolism liver and kidney functions.

The lower creatinine concentration of birds on diets MLM75 and MLM100 compared to their counterparts on diets MLM0 and MLM50 may be due to the relatively lower protein levels which could have reduced protein intake and hence lower production of creatinine. Although the concentrations of creatinine in the birds on all the dietary treatments were above the normal physiological range of 8.84 to 35.36 µmol/L which may suggest impaired kidney function or reduced clearance (Benzo et al 1986; Scanes 2022), the birds did not display any signs of kidney malfunction.

The high - density lipoprotein concentrations increased with increase in concentration of MLM in the diet of the birds and were within the normal physiological range of 1.44 – 2.26 mmol/L (Bueno et al 2017) suggesting protection against cardiovascular diseases (Scanes 2022).

Conclusion

The results of this current study show that;

· Substitution of wheat bran with MLM in broiler chicken diets up to 75 % did not negatively affect growth performance (ADFI, ADG and FCR), but rather enhanced growth performance.

· Inclusion of MLM in broiler chicken diets up to 75 % has no detrimental effect on carcass characteristics.

· Dietary inclusion of MLM in broiler diets up to 100 % had no adverse effects on broiler chicken physiology and health status.

Acknowledgement

Authors are grateful to Leventis Foundation London for the financial support.

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