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. 2022 Dec 1;8(12):e11958. doi: 10.1016/j.heliyon.2022.e11958

Intake, digestibility, growth performance and blood profile of rams fed sugarcane bagasse or rice husk treated with Trichoderma viride and effective microorganisms

Regasa Begna Roba a,∗, Mengistu Urge Letta b, Tegene Negesse Aychiluhim c, Getachew Animut Minneeneh d
PMCID: PMC9720023  PMID: 36478836

Abstract

Sugarcane bagasse and rice husk are lignocellulose rich agricultural by-products designated as an alternative feed for ruminant. However, their nutritive value is relatively low due to low digestibility, high fiber content, low protein content and low palatability. Therefore, the nutritional value of these by-products can be improved with biological treatments. The objective of this study to evaluate the feed intake, digestibility, growth performance and blood profile of Hararghe highland sheep fed biologically treated rice husk (RH) or sugarcane bagasse (SCB) in concentrate mixtures. Thirty six rams with an average age of 10 months and an average initial weight of 18 ± 1.8kg were assigned in randomized complete block design (RCBD) in 2 × 3 factorial arrangements, with two feeds (SCB and RH) and three treatment groups (Control, Trichoderma viride and effective microorganism (EM)). 5% EM1 was used to prepare the activated EM solution and 5% Trichoderma viride (Tv) solution was used to prepare inaculant. Sheep were fed ad libitum for 90 days at 20% refusal rate followed by10 days of digestibility trial. Nutrient intake and digestibility of sheep were significantly (p < 0.05) affected by the interaction. Sheep fed either treated or untreated rice husk based diets had significantly higher dry matter (DM g/d), organic matter (OM g/d), crude protein (CP g/d) and metabolizable energy (ME MJ/d) intake than a diet based on untreated sugarcane bagasse or treated with Trichoderma viride. Sheep fed diets containing either treated or untreated rice husks had a higher average daily weight gain (ADG) and feed conversion efficiency than sugarcane bagasse containing diets. Sheep fed diets containing biologically treated roughages had significantly higher (p < 0.05) average daily weight gain and feed conversion efficiency. Conversely, no significant differences were observed between rams fed diets containing EM and Trichoderma viride (Tv) treated rice husk or sugarcane bagasse in average daily weight gain (ADG) and feed conversion efficiency (FCE). Values of some blood hematology (red blood cell, haemoglobin and packed cell volume) and total protein, urea, glucose and cholesterol of serum metabolites were significantly (p < 0.05) higher for rams fed diet containing rice husk. In conclusion, the results show that the biological treatment of feed with the effective microorganism or Trichoderma viride improved nutrient utilization and average daily gain in the sheep fattening diet.

Keywords: Average daily gain, Biological treatment, Blood hematology, Feed conversion efficiency, Sheep

Average daily gain; Biological treatment; Blood hematology; Feed conversion efficiency; Sheep.

1. Introduction

Small ruminants are important components of livestock raised in various agro-ecology and production systems in Ethiopia. A recent report shows that Ethiopia has largest sheep and goat population (91 millions) among the African countries (CSA, 2020). They are primarily kept for meat productions. Sheep are particularly very important to in the large rural areas in terms of income sources, food and non-food, socioeconomic and cultural functions. Moreover, they contribute more to the national economy in the form of exports of live animal, meat and processed and semi-processed hides and skin (Adane and Girma, 2008). Despite their importance, the current production level of sheep is far below their potential because of shortage of feeds, poor genetic potential of indigenous breeds, inadequate veterinary services and poor animal managements. Among these feed shortages is considered as the most important one, because sheep are fed on natural pastures and crop residues which are characterized by a low nutritional value. Although, concentrate feeding and improving the nutritional value of the existing feed resources may reduce the problem of feed shortage, feeding high proportion of concentrate to ruminants can increase the total cost of production. Thus, this situation attracted the attention of many researchers to use agro-industrial by-products rich in lignocellulose as unconventional feeds.

Lignocellulose rich agro-industrial by-products are generated in large amount during the industrial processing of agricultural products. Sugarcane bagasse (SCB) and rice husk (RH) are the most abundant lignocellulose by-products produced by the sugar and milling industries respectively. The annual global productions of these by-products were estimated at about 1287 million tons (Melati et al., 2017) and 150 million tons (Aulakh, 2018) for sugarcane bagasses and rice husk respectively. In Ethiopia, about 4 million tons (Tena et al., 2018) and 27,200 tons (CSA, 2016) of sugarcane bagasses and rice husk, respectively were annually produced. However, the use of these by-products as animal feed is very restricted due to their low nutritional value. Sugarcane bagases contain high cellulose (47.8%), hemicellulose (25.13%), lignin (15.12%) and low protein (2.61%) which lower digestibility and palatability of feed (Begna et al., 2019). Rice husks also contain high cellulose (30.3%), hemicellulose (21.4%), lignin (14%) and low protein (7.9%) (Begna et al., 2019). The nutritional value of poor quality roughage can be improved through mechanical, chemical or biological and combined biological and chemical treatments (Sarnklong et al., 2010; Salman et al., 2011). However, the use of chemical treatments may produce toxic substances that can directly affect the animals through interfering with microbial fermentation or animals and man through feces and urine polluting soils and water sources (Shi et al., 2008). Because of these facts, biological treatments have concerned interest due to its possible rewards over physical treatments such as better substrate and response specificity, minimum energy requirements, lower pollution generation, and higher yield of desired products (Fadel and El-Ghonemy, 2015).

The use of biologically treated lignocellulose rich agricultural by-products as ruminant feed has been reviewed and documented by Mahesh and Mohini (2013). Moreover, Abdel-Azim et al. (2011) reported that ram fed crop residues treated with Trichoderma viride with concentrate had been showed higher feed intake (1399 g/d vs. 1459 g/d), fiber digestibility (40% vs. 45%), growth performance (128 g/d vs. 149 g/d) and better economic efficiency (73 % vs. 88%). Smilarly, Shazad et al. (2016) reported that feed intake (2.88 kg/d vs. 2.95 kg/d), dry matter digestibility (65% vs. 69%), growth performance (0.52 kg/d vs. 0.56 kg/d) and better economic gain (1.38 vs. 1.44) of buffalo calves fed on fungal treated wheat straw in total mixed ration was enhanced as compared to buffalo calves fed on untreated wheat straw. Huyen et al. (2019) also reported that dry matter digestibility (60% vs. 70%), nitrogen retention (23% vs. 35%) and microbial protein synthesis (4.3 g/d vs. 6.0 g/d) were significantly improved for sheep consumed fungal treated rice straw. Feeding of effective microorganism (EM) treated grass hay supplemented with noug cake improved dry matter digestibility (52% vs. 78%), average daily gain (6.6 g/d vs. 61.5 g/d) and better economic gain (Fikre et al., 2019). Similarly, Mulgeta (2015) and Girma and Alemayhu (2018) reported higher dry matter and nutrient intake in lactating dairy cow fed EM treated barley straw as compared to untreated. However, the use of fungus as biological treatment to improve the nutritive value of roughage feed is almost nil in Ethiopia, but limited studies have been conducted on the use of effective microorganisms and the application of the technology has been limted to research stations and universities.

We hypothesized that feeding biologically treated sugarcane bagasse and rice husk gave difference response in the intake and digestibility, which help us to make decision to identify which feed yields the best response in terms of nutrition and production performance of growing ram. Therefore, the objective this study was to determine how biological treatments with Trichoderma viride and effective microorganism (EM) of sugarcane bagasse and rice husk with a concentrate mixture affected yearling Hararghe highland rams feed intake, digestibility, growth performance, and blood profile.

2. Materials and methods

2.1. Experimental site

The study was conducted at Haramaya University goat farm. Haramaya University is located at 9° 26′N latitude and 42°3′ E longitude. The site is situated at about 1980 m above sea level (m.a.s.l) and located at 520 km east from the capital city, Addis Ababa. The average annual rainfall of the study area is about 870 mm with a range of 560–1260 mm, and the average maximum and minimum temperatures are 23.4 °C and 8.25 °C, respectively (Mishra et al., 2004).

2.2. Treatment preparation and experimental design

Trichodermaviride (Tv) was obtained from plant protection section of the school of plant sciences from Haramaya University, Ethiopia. The sample of Tv maintained below 4 °C were collected and grown on Petridish to activate the microorganisms. Three days old slant culture samples of Tv were inoculated on petridish (9 cm) containing 25 ml potato dextrose agar (PDA) for seven days. The seven days old cultures were used to inoculate plastic bottles containing the following substances per litre of distilled water: 4% molasses, 0.4% urea, 0.2% KH2PO4 and 0.03% MgSO4(7H2O) per one litre of water and the inoculate was kept for seven days under room temperature. This solution was contained 653∗ 108 colony forming units (CFU) per ml. The rice husks and sugarcane bagasse were inoculated at a ratio of one liter solution to 20 kg feed dry matter to prepare spawn. Next, either untreated sugarcane bagasse or untreated rice husk was moistened with water at 55% and inoculated by 10 % the respective spawn, as spawn fungi (Abdel-Azim et al., 2011). The treatments were conducted in a clean room padded with plastic sheet disinfected by Dettol and Formalin. The materials were covered with plastic sheet and incubated for 21 days under room temperature. At the end of the incubation, the treated materials were exposed to sunlight until the moisture content reached approximately 10%, then packed and stored until use in the feeding trials.

Effective micro-organism (EM-1) packed in 20 L of plastic bottles were purchased from Weljijie PLC (Debrezeit). The EM used in this trail was developed from three major microbes; namely photosynthetic bacteria, lactic acid bacteria and yeasts (Higa, 1994). Then EM1 was mixed with molasses and chlorine free water. Water, molasses and EM1 were mixed at a ratio of 18:1:1, respectively (Higa and Wididana, 1991) to generate activated EM. After stirring, the solution was stored in 200 L capacity barrel for seven days to activate the EM solution. Next, the activated EM solution was evenly sprayed on to the sugarcane bagasse and rice husks at a proportion of one litre EM solution to one kg of dry matter feed, mixed well and packed in an airtight plastic bag of 100 kg capacity and stored in a large barrel of 200–250 kg capacity, covered with plastic sheet and kept at room temperature for 21 days for feeding.

Six experimental rations were prepared from a concentrate feed mixture (CM) and the two roughages (Rice husks and Sugarcane bagasse). The treatments had a 50:50 ratios of the roughages to concentrate as shown in Table 2.

Table 2.

Ingredients (%) and chemical compositions (% DM basis) of the experimental Treatments.

Ingredients (%) Treatments
Rice Husks (RH)
 Sugarcane Bagasse (SCB)
U EM Tv. U EM Tv.
Untreated RH 50 - - - - -
EM treated RH - 50 - - - -
Tv treated RH - - 50 - -
Untreated SCB - - - 50 - -
EM treated SCB - - - - 50 -
Tv treated SCB - - 50
Maize grain 11 11 11 11 11 11
Wheat bran 11 11 11 11 11 11
Noug seed cake 23.5 23.5 23.5 23.5 23.5 23.5
Molasses 3 3 3 3 3 3
Salt 1 1 1 1 1 1
Pre.mix 0.5 0.5 0.5 0.5 0.5 0.5
Chemical compositions of diet (% DM basis)
DM 89.6 66.6 89.5 89.8 61.1 89.9
OM 85.8 85.1 84.6 92.5 91.5 90.5
CP 13.7 13.8 15.6 11.4 11.9 13.2
NDF 50.3 45.7 43.7 60.2 57.6 55.7
ADF 30.0 25.8 24.0 38.1 35.6 35.7
ADL 9.7 9.5 8.8 10.2 9.6 9.3
ME MJ/kg DM 10.59 11.37 12.05 10.4 11.20 11.14
Feed cost birr/kg 4.64 5.14 5.14 4.14 4.64 4.64
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U=Untreated; EM = effective microorganism; Tv = Trichoderma viride; RH = Rice husk; SCB = Sugarcane bagase; DM = Dry matter; OM = Organic matter; CP = Crude protein; NDF = Neutral detergent fiber; ADF = Acid detergent fiber; ADL = Acid detergent lignin; ME = Metabolizable energy; premix contained: 500000IU vitamin A, 100000IU vitamin D3,100mg vitamin E, 190g Ca, 90 g Mg, 50g Na, 2g Mn, 3g Fe, 0.3g Cu,3gZn, 0.1g Co and 1 mg Se.

Experimental rations were randomly assigned to individual housed thirty six yearlings Hararghe highland rams in a randomized complete block design (RCBD) in a 2 × 3 factorial arrangement (2 feeds and 3 treatments). Six replications were performed for each treatment. The treated or untreated materials were mixed with the concentrate prior to feeding to form the total mixed ration (TMR). All total mixed diets were prepared according to the National Research Council recommendations (NRC, 2007) for protein (12%) and energy requirements (10 MJ/kg) of growing ram at least to guarantee 50 g/d average daily weight gain. The total mixed diets were offered ad libitum with a 20% refusal rate adjusted at four days intervals based on individual animal intake on the previous day. Daily offerings were made over two meals at 8:00 AM and 2:00 PM. The amount of feed offered and refused was recorded daily for each animal and the daily intake for the individual animal was determined as the difference between the two. Samples of feed prepared were taken in batches, whereas the refusal responses were taken daily from each animal, recorded, and pooled per animal. Wet samples of feed offered and refused were kept in a deep freezer at-20oc until analysis was required. Feed offer and feed refusal samples were collected over the experimental period and sub-sampled to determine chemical composition. All animals had access to clean tap water at all times during the entire study.

2.3. Animals and management

A total of thirty six yearling intact Hararghe highland sheep were purchased from Chelenko and Kulubi markets. The age of the animals was determined based on dentition and information provided by the owner. Animals were transported to Haramaya University, quarantined for three weeks, dewormed with Albendazole (15 mg/kg) to treat against internal parasites, injected with Ivermecin (0.2 mg/kg) and sprayed with Diazinine (1 ml/L of water) for treating ecto-parasite. Sheep were ear-tagged for easy identification. The initial body weight of experimental ram was 18 ± 1.8kg, which was taken as the average weight of the rams after two consecutive days of measurment. Sheep were kept in an individual pens equipped with feeding and watering trough.

2.4. Growth trial and economic efficiency

Growth studies lasted for 90 days. A digital balance with a sensitivity of 0.02 kg was used to record the amount of total mixed ration (TMR) offered and refused daily for the duration the study. The animal was weighed at 10 days interval after an overnight fast and before the daily feeding using a hanging scale graduated at 0.2 kg intervals. Total weight gain (TWG) was calculated as the difference between final and initial weight. Weight versus atime regression was used to determine the average daily weight gain (ADG) of rams. Feed conversion efficiency (FCE) of the animal was determined as the ratio of daily weight gain to the daily DM intake. The total cost of total mixed ration was determined based on the cost of feed ingredients and biological treatments according to existing price from 2017 and 2018. The selling market price of sheep was 120 Birr per kg live weight. Feed cost (FC) per kg of weight gain was determined as a proportion of consumed feed cost to total weight gain of sheep fed experimental ration. Total returns (TR) per animal was estimated by multiplying the total weight gain by the selling price in Birr per kg live weight of sheep. Net profit (NP) was calculated as difference between total return and feed cost.

2.5. Digestibility trial

All animals were fitted with fecal collection bags and acclimated to wearing the bag for three days. The total amount of faeces that each animal excreted into the bag was emptied into a plastic bucket every 24 h, weighed and recorded. It was thoroughly mixed in the bucket and approximately 20% of each daily collection was taken and stored in a deep freezer at -20 °C until required for analysis (McDonald et al., 2010). Frozen fecal samples were thawed, bulked across the collection period, thoroughly mixed, adequate amount of samples were taken, a portion of it was oven dried at 105 °C for 24 h to obtain DM content, and the remaining portion was dried at 60 °C for 72 h in forced draft oven, ground to pass through a 1mm sieve in a Wiley mill for use in chemical assays. Feed offer and refusal samples were taken daily, collected per animal, and finally subsampled. The daily feed intake of the experimental sheep was calculated on the DM basis as the difference between the DM of the offered feed and the DM of the refused feed. Apparent digestibility of DM and nutrients was determined as the difference between the intake and faecal excretions divided by intake multiplied by 100.

2.6. Blood sampling

Blood was collected from the jugular vein of experimental animal on the last days of the trials. A blood sample of about 10 ml was taken from all animals via the jugular vein in to heparinized vaccutainer tube (5ml) for determination of blood hematology and an additional 5 ml in to plain tubes for biochemical analysis of the blood. Hematological parameters of blood such as packed cell volume (PCV), haemoglobin (Hgb), mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), red blood cells (RBC) and total white blood cells (WBC) were measured from whole blood using an electronic cell counter (Hematology Analyzer, 2.5 Release, Germany). Total protein was determined from serum by the biuret method, urea by the enzymatic method (GLDH method), creatinine by the photometric colorimetric assay for kinetic measurements (Jaffe’-reaction) and glucose by the enzymatic colorimetric test (GOD-PAP method), cholesterol levels were determined using an enzymatic colorimetric method with lipid clearing factor (CHOD-PAP method) using the reagents developed for each parameter test and an instrument of Germany Huma analyzer 3000.

2.7. Animal care and ethical issues

Animal care and ethical issues were carefully evaluated, and the experiment was approved by the Ethics Committee of Haramaya University College of Agriculture and Environmental Sciences (1956ET-12/2017). European Union Directive 2010/63/EU (2010) guideline on handling and use of animals for research and development purposes was implemented.

2.8. Chemical analysis

Chemical analysis of the samples was perfomed at the Haramaya University Laboratory of Animal Nutrition. Representative samples of feed, refusals, and faeces were dried at 60 °C for 72 h and the ground to pass 1 mm sieve using a laboratory grinder. Dry matter (DM), and ash contents of the feed and faecal samples were determined according to the methods of AOAC (1990). Van Soest and Robertson (1985) methods were used to determine acid detergent fiber (ADF), neutral detergent fiber (NDF) and acid detergent lignin (ADL) contents of feed and faecal samples. The N content of the samples was determined using the Micro-Kjeldahl method and CP was calculated as Nx6.25 (AOAC, 1990). Metabolize energy (ME) of the experimental feed (MJ/kg DM) was estimated from digestible organic matter (DOM) by using the formula ME = DOM x 0.016 (McDonald et al., 2010). Where: DOM = digestible organic matter per kg of DM.

2.9. Data analysis

Collected data on feed intake, digestibility, live weight gain and blood profile for each treatment diet in sheep were analysed using SAS software 9.1 version according to the Guide for Statistical Analysis Systems (SAS, 2008). If there was a significant difference between means, Tukey honestly significance test was used to adjust the mean separation. The model for the analysis was:

  • Yijkl = μ +ai + bj + cl + b∗cij+ εijkl

  • Where:Yijkl = the dependent variables,

  • μ = overall mean,

  • ai = Theithblock

  • bj = The jth feed type,

  • cl = The lth treatment method,

  • c∗b = The jlth interaction (between feed type and treatment method)

  • εijkl = random error.

3. Results

3.1. Chemical compositions of diets

Rice husk based diets have higher CP as compared to sugarcane bagasse (Tables 1 and 2). CP values were higher in roughage treated with EM and Tv than in untreated. Dietary fiber components (ADF and NDF) were higher in sugarcane bagasse base diets than in rice husk. Biological treatment of sugarcane bagasse and rice husks decreased OM, NDF, and ADF and increased dietary CP and ash content.

Table 1.

Chemical compositions (% DM basis) of feed ingredients.

Feed Ingredients DM (%) Nutrients (%DM)
OM CP NDF ADF ADL
Maize grain 89.0 98.4 9.2 13.7 1.0 1.7
Noug seed cake 92.1 89.6 31.6 38.5 20.1 8.5
Wheat bran 90.8 94.5 15.3 48.7 15.4 3.2
Rice husks (untreated) 90.3 82.4 6.8 69.3 47.8 14.4
Rice husks (RH-EM) 45.4 80.9 7.2 59.8 38.6 14.4
Rice husks (RH-Tv) 90.5 79.1 10.6 55.7 35.4 12.7
Sugarcane bagasse (untreated) 90.7 94.7 2.4 88.6 63.8 15.7
Sugarcane bagasse (SCB-EM) 36.5 92.9 3.4 83.9 58.7 14.4
Sugarcane bagasse (SCB-Tv) 90.2 91.2 5.9 79.8 58.5 13.6
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DM = Dry matter; OM = Organic matter; CP = Crude protein; NDF = Neutral detergent fiber; ADF = Acid detergent fiber; RHEM = Rice husk treated with effective microorganism; RHTv = Rice husk treated with Trichoderma viride; SCBEM = Sugarcane bagasse effective microorganism; SCBTv = Sugarcane bagasse treated with Trichoderma viride.

3.2. Nutrient intake and digestibility

There were significant (p < 0.05) feed by treatment interaction for nutrient intake and digestibility (Table 3). Sheep consumed either treated or untreated RH based diets had higher DM, OM, CP and ME intake than a diet based on untreated SCB or treated with Tv. Treating RH and SCB with EM enhanced feed intake and there was no significant difference between these treatments. Except significantly (p < 0.05) higher metabolizable energy intake observed in sheep consumed diet contained RH treated with Tv, other nutrient intake were similar amoung groups fed ration contain untreated or RH treated with EM and Tv. Conversely, significantly (p < 0.05) higher DM and nutrient intake were recorded for sheep fed diet contained SCB treated with EM than SCB treated with Tv and untreated. Likewise, DM and nutrient digestibility of RH based diet was better improved when treated with Tv than EM as compared to untreated RH and all treatment combination of SCB, although EM displayed similar effect on the digestibility of the diets. Significantly (p < 0.05) higher CP digestibility observed for diet contained RH treated with Tv as compared to untreated. However, significantly (p < 0.05) higher NDF and ADF digestibility was observed in sheep fed diet containing Tv and EM treated SCB as compared to sheep fed diet contained untreated SCB, while higher CP digestibility observed in sheep fed diet contained SCB treated with EM.

Table 3.

Nutrient intake and digestibility of Hararghe highland sheep fed diet containing rice husk or sugarcane bagasse treated with Trichodermaviride and effective microbes.

Item Rice Husk (RH)
 Sugarcane Bagasse (SCB)
 SEM p-value
U EM Tv. U EM Tv. Feed Treatment F x T
Intake g/h/day
DM 942.2ab 962.4ab 1051.9a 554.7d 819.0bc 661.9cd 38.7 <0.001 0.003 0.004
OM 805.1ab 811.7ab 884.1a 504c 735.3b 588.3c 33.1 <0.001 0.004 0.002
CP 130b 133.8b 163a 69.6d 101.1c 92.8c 4.8 <0.001 <0.001 0.003
NDF 470.4a 444.1ab 460.4a 321.8c 467.8a 366.4bc 18.7 <0.001 0.01 0.003
ADF 278.2a 246.1abc 251.4ab 202.2c 278.8a 228.8bc 11.3 <0.001 0.09 0.002
MEI,MJ/day 10.00b 10.94ab 12.67a 5.74d 9.17bc 7.40cd 0.44 <0.001 <0.001 0.001
Digestibility, %
DM 67.4bc 69.5b 74.6a 66.5c 68.2bc 67.3bc 0.6 0.001 <0.001 <0.001
OM 70.5bc 72.4b 76.8a 69.9bc 71.4b 71.0b 0.62 <0.001 <0.001 0.001
CP 66.8bc 68.7ab 69.8a 63.2d 67.7abc 65.2cd 0.7 <0.001 0.002 0.03
NDF 63.7de 68.9b 72.7a 60.8e 67.7bc 64.5cd 0.8 <0.001 <0.001 <0.001
ADF 41.5cd 45.4b 51.5a 39.9d 46.6b 44.1bc 0.64 <0.001 <0.001 <0.001
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a-dMeans bearing different superscripts within factor across column are significantly different at P < 0.05; SEM = Standard Error Mean; RH = Rice husk; SCB = Sugarcane bagasse; U=Untreated; EM = effective microorganism; Tv = Trichodermaviride; DM = Dry matter; OM = Organic matter; CP = Crude protein; NDF = Neutral detergent fiber; ADF = Acid detergent fiber; MEI = Metabolizable energy intake.

3.3. Growth performance and economic efficiency

Significantly (p < 0.001) higher final body weight (FBW), total body weight gain (TWG), average daily gain (ADG), and feed conversion efficiency (FCE) were observed for sheep fed diet containing either treated or untreated rice husks was observed in compared with the SCB based diet consumed (Table 4). Biological treatments also affect growth performance of animals. Numerically higher final body weights, total body weight gain, average daily gains, and feed conversion efficiencies were observed in sheep fed diets containing biologically treated rice husks compared to sheep fed diets containing untreated rice husks, while significantly (p < 0.05) higher values of these parameters were recorded for groups fed diet contained biologically treated sugarcane bagasse (SCB) as compared to sheep consumed diet contained untreated SCB. However, differences between EM and Tv was not significant, though biologically treated sugarcane bagasse had better feed conversion efficiency than untreated sugarcane bagasse. An economic analysis showed that significantly (p < 0.05) lower feed costs per kg of weight gain for sheep fed diets containing either treated or untreated RH compared to sheep fed diets containing untreated SCB and higher net profit per head were observed for these groups. Correspondingly, lower feed cost/kg gain and higher net profit per animal were observed in sheep fed diet contained biologically (Tv and EM) treated SCB in comparison to untreated. However, there was no significant changes were observed among the group fed the diet contained rice husk.

Table 4.

The growth performance and economic efficiency of Hararghe highland sheep fed TMR contained rice husks or sugarcane bagasse treated with Trichoderma viride and effective microbes.

Parameters Rice Husk (RH)
 Sugarcane Bagasse (SCB)
 SEM p-value
U EM Tv. U EM Tv. Feed Treatment F x T
IBW(Kg) 18.2 18 18.1 18.1 18.1 18.2 0.91 0.98 0.99 0.99
FBW(Kg) 26.7a 27.9a 28.3a 20.9c 24.4b 23.2b 0.94 <0.001 0.04 0.49
TWG (Kg) 8.5a 9.9a 10.2a 2.9c 6.3b 5.0b 0.48 <0.001 <0.001 0.1
ADG (g/d) 93.9a 110.3a 113.5a 32c 70.5b 55.5b 5.4 <0.001 <0.001 0.1
FCE 0.1a 0.114a 0.11a 0.06c 0.083b 0.09b 0.01 <0.001 0.001 0.25
FC (Birr) 46.71b 46.08b 47.71b 79.11a 53.83b 56.28b 4.34 <0.001 0.012 0.01
NP(Birr) 620.53ab 746.40a 739.38a 130.47d 419.38bc 323.60c 51.2 <0.001 0.001 0.3
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a-dMeans of each parameter with different superscripts within in the same column are significantly different at P < 0.05; SEM = Standard Error Mean; U=Untreated; EM = effective microorganism; Tv = Trichoderma viride; IBW = Initial body weight; FBW = Final body weight; TWG = Total weight gain; ADG = Average Daily gain; FCE = Feed conversion efficiency (g ADG/g DMI/d), FC = Feed cost/kg gain, NP = Net profit per animal.

3.4. Blood hematology and chemistry

Red blood cells (RBC), haemoglobin (Hgh), and packed cell valume (PCV) were significantly (p < 0.05) higher in the sheep fed diets containing RH (Table 5). White blood cell (WBC) counts were significantly (p < 0.05) higher in the sheep fed diets containing roughages (RH and SCB) treated with EM as compared to Tv. Sheep fed diet containing roughages (RH and SCB) treated with Tv had lower RBC and PCV compared to diet containing roughages treated with EM. MCH values were higher in sheep fed diets containing sugarcane bagasse treated with Tv as compared to EM. Total protein, albumin, globulin, urea, cholesterol and glucose were significantly (p < 0.05) higher in sheep fed diet containing RH (Table 5). Values of total protein, serum urea and albumin were significant (p < 0.05) higher in sheep fed diet containing roughage treated with Tv and EM, whereas other parameters were unaffected by biological treatment.

Table 5.

Blood hematology and metabolites of Hararghe highland sheep fed diet contained rice husk or sugarcane bagasse treated with Trichoderma viride and effective microbes.

Variables Rice husk (RH)
 Sugarcane bagasse (SCB)
 SEM p-value
U EM Tv U EM Tv Feed Treatment FxT
Blood hematology
Red blood cell (103/μl) 13.28ab 13.71a 12.23b 11.68bc 12.45ab 10.48c 0.40 <0.001 0.01 0.8
White blood cell (103/μl) 12.42 ab 12.93a 12.05b 12.19ab 12.53a 11.47b 0.31 0.12 0.01 0.85
Haemoglobin (g/dl) 12.36ab 12.88a 11.50ab 11.63ab 11.00b 10.77b 0.39 0.001 0.06 0.24
Packed cell volume (PCV) (%) 41.10ab 42.87a 36.62bc 37.28bc 38.50ab 36.23c 1.07 0.01 0.001 0.15
MCV (fl) 31.00 31.41 30.03 32.04 31.1 34.66 1.12 0.06 0.6 0.08
MCHCH (g/dl) 30.11 30.07 31.48 31.20 28.57 29.75 0.78 0.27 0.17 0.15
MCH (pg) 9.30ab 9.40ab 9.42ab 9.97ab 8.88b 10.29a 0.27 0.13 0.03 0.06
Blood chemistry
Total protein (g/dl) 7.03bc 7.30ab 7.61a 6.65c 7.10ab 7.01bc 0.13 0.001 0.003 0.34
Albumin (g/dl) 3.63bc 3.83ab 3.99a 3.45c 3.80ab 3.80ab 0.07 0.03 <0.001 0.49
Globulin (g/dl) 3.40ab 3.47ab 3.62a 3.20b 3.30ab 3.22ab 0.1 0.03 0.45 0.42
Urea (mg/dl) 32.33b 34.32a 35.33a 30.46c 31.37bc 32.00bc 0.40 <0.001 <0.001 0.19
Cholesterol (mg/dl) 63.30a 64.17a 64.50a 52.50b 54.32b 54.31b 0.66 <0.001 0.06 0.75
Glucose (mg/dl) 77.38a 76.00a 72.30ab 66.67bc 67.50bc 65.08c 1.44 <0.001 0.052 0.48
Creatine (mg/dl) 1.13 1.13 1.07 1.10 1.12 1.13 0.08 0.9 0.95 0.83
Alanine transferase (ALT) u/l 26.02 26.78 27.97 24.69 25.57 27.33 3.05 0.67 0.75 0.99
Aspartate transferase (AST) u/l 130.83 148.17 158.83 137.94 148.00 143.33 15.21 0.82 0.51 0.75
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a-cMeans within in the same column with different superscripts are significantly different at P < 0.05; SEM = Standard Error Mean; U=Untreated EM = effective microorganism; Tv = Trichoderma viride, RBC = red blood cell, WBC = white blood cell, PCV = packed cell volume, MCV = mean corpuscular volume; MCH mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; ALT = Alanintransferase; AST = Aspartate transferase.

4. Discussion

4.1. Chemical compositions of diets

The observed difference between diets in chemical compositions is due to biological treatments and variation in chemical composition of the feed ingredients. The CP content in sugarcane bagasse is below minimum value required to support optimum rumen function (7–7.5% CP), whereas the CP content of rice husks is in the range of 7–10% (Van Soest, 1994). The higher CP and lower OM of the diet based on RH is the reflection of higher CP and ash content of rice husk as compared to sugarcane bagasse. The observed higher ADF and NDF in the SCB containing diet is due to the higher ADF and NDF in sugarcane bagasse. However, lower fiber fractions (ADF and NDF) and higher CP were observed in the diet containing biologically treated rice husk and sugarcane bagasse. The increased crude protein (CP) and decreased fiber components (ADF and NDF) of by-products indicate that biological treatment improves their nutritional value. These results are in line with various studies (Salman et al., 2011; Gado, 2012; Khattab et al., 2013; Hassan et al., 2015; El-Bordeny et al., 2015), who observed that biological treatments of poor quality roughages increased their nutritional value through increasing the concentration of CP and reducing fiber fractions (NDF, ADF and ADL). The reduction of fiber fractions (ADF and NDF) and increased CP content in biologically treated rice husk and sugarcanes bagasse may be due to carbohydrate utilization by microorganisms as a carbon sources for the growth and formation of microbial proteins (Maharous et al., 2021).

4.2. Nutrient intake and digestibility

Animal dry matter intake is inversely proportional to the concentration of cell wall components (NDF and ADF) and directly proportional to the dietary CP content (Bosa et al., 2012; De Carvalho et al., 2017). Therefore, higher dry matter and nutrient intakes were observed in sheep fed diets containing RH compared to SCB, which could be related to higher CP and lower fiber fraction of diets based on RH. The lack of effect on nutrient intake of sheep fed diets containing either biologically treated or untreated rice husks was smiler to the findings of Huyen et al. (2019), who reported insignificant difference in nutrient intake of sheep fed diets containing fungal treated rice straw. However, sheep fed diets containing sugarcane bagasse trated with effective microorganism had showed signficantly higher nutrient intake in comparsion to untreated sugarcane bagasse or treated with Trichoderma viride. This result agrees with Mulgeta (2015) and Girma and Alemayhu (2018), who obtained significantly higher DM and nutrient intake in lactating dairy cow fed EM treated barley straw as compared to untreated. Also Maharous et al. (2019) and Maharous et al. (2021) reported higher DM intake in growing ram fed ration contained olive tree by-products or date palm leaves treated with EM compared to untreated roughages.

Improved NDF and ADF digestibility of biologically treated rice husk and sugarcane bagasse are associated with improved rumen fermentation efficiency and microbial biomass production, increased number and/or activity of rumen microbiota, enhanced rumen microbial colonization and attachment to the digesta (Wangi et al., 2001; Olafadehan, 2013; Maharous et al., 2021). Abo-Donia et al. (2014) reported that higher nutrient (OM, CP, NDF and NFC) digestibility for sheep fed fungal (Trichoderma viride) treated peanut hulls, compared to sheep fed untreated. Likewise, Abdel-Azim et al. (2011) reported that treatment of rice straw (RS) and corn stalks (CS) with Trichoderma viride improve the digestibility of CP, CF, NDF, ADF, ADL and cellulose with a ration containing biologically treated by-products than untreated. Mahrous et al. (2019) stated that the digestibility of dry matter, organic matter, crude protein, nutreal detergent fiber and acid detergent fiber were significantly higher in sheep fed ration containing olive tree by-products treated with EM than untreated. Correspondingly, Fikre et al. (2019) reported that supplementing growing rams with EM treated hay significantly improved the digestibility of nutrients as compared to sheep fed control diet. Supplementing the growing rams with probiotics also improve the digestibility of DM, OM and CP, compared to the rams fed only concentrate diets (Saleem et al., 2017). The higher nutrient digestibility observed for sheep fed biological treated feed may be due to microbial solubilization or increased biodegradability of cell wall components (Salman et al., 2011).

4.3. Growth performance and economic efficiency

The Differences in growth performance among treatments reflect the varations in feed conversion efficiency. The higher weight gain and feed conversion efficiency (FCE) observed with the sheep fed RH based diet than SCB based diets are an attributes of higher crude protein and metabolizable energy intake, allowing the growth of microbial population which encourages efficient nutrient utilization. These results are consistent with Ayele et al. (2017), who reported higher body weight gain and feed conservesion efficiency for different Ethiopian sheep breeds consumed high crude protein in g/day and metabolizable energy in MJ/day as compared to those consumed lower crude protein and metabolizable energy. The average daily weight gain recorded for the RH diet group was higher than the values 32.2 g/d-63.2 g/d (Hirut et al., 2011) and 30 g/d- 65 g/d (Gebeyew et al., 2011) reported for the same breed fattened with diets based on different basal diets and at different levels of concentrate supplementation, these could be due to lower CP and ME intake by these group. The higher body weight gain and feed conversion efficiency (FCE) of the biologically treated by-products as compared to untreated is the result of improved feed utilization by treatments. Smiler trend was obtained by Fouda (2008) and El-Bordeny et al. (2015) who reported that sheep fed diets containing biologically treated sugarcane bagasse or rice straw had higher body weight gain and feed conversion efficiency as compared to sheep feed untreated roughages. Moreover, Mahrous et al. (2019) and Maharous et al. (2021) reported higher body weight gain and feed conversion efficiency for sheep fed diets containing olive tree by-products or date palm leaves treated with EM. Sheep fed diets containing ethier treated or untreated rice husks resulted in lower feed costs per kg of weight gain and higher net profit per head as compared to sheep fed diets containing sugarcane bagasse, this could be either due to lower fiber and higher crude protein content of the diets leads to better feed conversion efficiency. Significantly lower feed costs per kg weight gain and higher net profit per animal were observed in sheep fed diet containing biological treated by-products as compared to untreated by-products. The finding is in agreement with Fouda (2008) who reported a lower feed costs per kg of weight gain and higher net profit per animal in lambs fed fungal treated diets compared to lambs fed untreated diet. Similarly, lower feed cost per kg gain and higher net revenue were reported by Mahrous et al. (2019) and Maharous et al. (2021) for sheep fed diet containing olive tree by-products or date palm leaves treated with EM.

4.4. Blood hematology and chemistry

The higher red blood cell (RBC) counts, packed cell volume (PCV) and haemoglobin (Hgb) obtained from sheep fed diet containing rice husk as compared to diet containing sugarcane bagasse might be a result of high protein and energy content of the rice husk based diets (Radostis et al., 2006). However, all blood hematology results in this study were 9–15 × 106/μl, 27–45%, 9–15 g/dl, 4–12 × 103/μl, 28-40fl, 8–12pg and 31–34 g/dl for RBC, PCV, haemoglobin, WBC, MCV, MCH and MCHC respectively, with in the normal physiological ranges reported for clinical healthy sheep (Jackson and Cockcroft, 2002; Radostits et al., 2006). The reduction in red blood cell (RBC) counts observed in sheep fed diet containing by-products treated with Trichoderma viride was not below the normal RBC counts reported in healthy sheep, indicated that no haemolytic anaemia and depression of erythrogenesis (Olafadehan, 2011; Ibhaze et al., 2021). Packed cell volume (PCV) concentrations below the normal range may indicate anaemia and poor protein quality in the diet (Radostis et al., 2006). Nevertheless, the lower packed cell volume values obtained from the Trichoderma viride treatment by-products fed to the sheep were within the normal physiological range, indicating that the sheep were not anaemic (Ibhaze et al., 2021). Furthermore, the blood haemoglobin concentrations observed in this study were within the normal physiological range, indicate that the sheep had sufficient blood pigment for proper oxygen transport, thus prevent microcytic and hypochromic anaemia caused by iron deficiency (Olafadehan, 2011). White blood cells above the normal range are known to indicate infection in sheep (Radostits et al., 2006). The normal values obtained in this study indicate that the animal immunity was not compromised by feed type or biological treatments. Normal values for mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH) and mean corpuscular haemoglobin concentration (MCHC), and lack of effect of feed type and biological treatment explained that the sheep were not anaemic (Olafadehan, 2011; Ibhaze et al., 2021).

All serum metabolite values obtained in this study were within the range of values reported by Radostits et al. (2006) and Kaneko et al. (2008) for healthy sheep. Higher serum total protein, albumin, urea and cholesterol from sheep fed diets containing rice husk and biologically treated by-products may be attributed to increased nutrients intake and digestibility, especially protein and organic matter (Yang et al., 1999). Blood total protein concentrations, albumin and serum urea reflect the nutritional status of the animals and are positively correlated with dietary protein intake (Kaneko et al., 2008). Kaneko et al. (2008) reported that normal serum total protein, albumin and urea in the healthy animals ranged from 6-7.9 g/dl, 2.4–3 g/dl and 10–35 mg/dl, respectively, but altered during any liver and kidney diseases. Therefore, the values recorded in this study were within the normal range, indicated that the diets supplied satisfactory amount of protein needed to maintain normal serum protein levels and organ function of the animals. The results of this study agree with Maharous et al. (2021), Abd El-Razik et al. (2012), Okab et al. (2012) and Fouda (2008) who found that lamb fed diets containing biological treated date palm leaves, rice straw, sugar beet pulp and sugarcane bagasse respectively, had higher values of blood serum urea, total protein and albumin. The higher serum total protein and albumin in sheep fed biological treated ration also indicated the better utilization of dietary protein and ruminal true protein-N through the digestive tract (Recce, 1991). The observed value of cholesterol levels ranged from 55 to 63 mg/dl was similar with the values reported by Diriba et al. (2015) for the same breed and falls within in the range 43–103 mg/dl of normal blood cholesterol concentration of sheep various ages and breeds (Radostits et al., 2006). Sheep fed diets containing RH had higher glucose concentration, possibly due to the animals higher dietary energy intake (Ayele et al., 2017). Blood serum creatinine, Alanintransferase (ALT) and Aspartate transferase (AST) were unaffected by feed type and biological treatment. Normal values for serum creatinine, Alanintransferase and Aspartate transferase and no effects of feed type and biological treatment indicate normal liver activity and function (Maharous et al., 2021).

5. Conclusion

The results of the present study indicate that sheep fed diets containing either treated or untreated rice husk were performed well in feed intake, apparent digestibility, live weight changes and economic efficiency as compared to untreated sugarcane bagasse. Feeding biologically treated sugarcane bagasse with effective microorganism and Trichoderma viride had better improved growth performance and economic efficiency as compared to untreated sugarcane bagasse. Blood hematology and metabolites were also found in the normal physiological range of healthy sheep. Hence, it can be concluded that both feeds are treated with Trichoderma viride and effective microorganism can be incorporated into the ration of a ruminant. However, further investigation will be needed for sugarcane bagasse to determine the optimum level of inclusion to get maximum performances.

Declarations

Author contribution statement

Regasa Begna: Conceived and designed the experiments; Performed the experiments; Analysed and interpreted the data; Wrote the paper.

Mengistu Urge: Conceived and designed the experiments; Performed the experiments; Analysed and interpreted the data; Contributed reagents, materials, analysis tools or data; Wrote the paper.

Tegene Negesse and Getechew Animut: Conceived and designed the experiments; Contributed reagents, materials, analysis tools or data; Wrote the paper.

Funding statement

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for profit sectors.

Data availability statement

The authors are unable or have chosen not to specify which data has been used.

Declaration of interest's statement

The authors declare no conflict of interest.

Additional information

No additional information is available for this paper.

Acknowledgements

The authors would like to acknowledge all the participants and peer reviewers.

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