The effect of Acacia eriolaba leaf meal inclusion on growth performance, blood parameters and methane gas emission in lambs fed diets containing ammoniated maize stover.

doi:10.21203/rs.3.rs-3905536/v1

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The effect of Acacia eriolaba leaf meal inclusion on growth performance, blood parameters and methane gas emission in lambs fed diets containing ammoniated maize stover.

https://doi.org/10.21203/rs.3.rs-3905536/v1

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published 03 Oct, 2024

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The study evaluated the effect of Acacia erioloba leaf meal in diets containing ammoniated maize stove on growth performance, methane emission and heath of growing lambs. Thirty-two female lambs were allocated to the following four dietary treatments: total mixed ration (TMR, control), 20% inclusion of untreated maize stover (UMS), 20% inclusion of ammoniated maize stover (AMS), and combined inclusion of 10% ammoniated maize stover and 10% Acacia erioloba leaves (AMSAL). Each treatment was replicated 8 times and a lamb in an individual pen was regarded as an experimental unit in a completely randomized design. Feed intake was higher (P < 0.05) in lambs fed the AMS and AMSAL diets compared to those fed UMS. Final body weights were higher in lambs fed the AMS and AMSAL diets. Both ADG and FCR were not affected (P > 0.05) by diet. In comparison with the AMS and AMSAL diets, the lambs fed the UMS diet had the highest (P < 0.05) Methane emission. Overall, lambs fed the control diets had the lowest methane gas emission. Hematological values were affected by diet with the AMSAL fed lambs having the highest (P < 0.05) MPV and PCT values. Furthermore, total albumin, amylase and total bilirubin were highest (P < 0.05) in lambs fed on the AMSAL diet. Lambs fed on AMS diet had the highest (P < 0.05) serum urea levels. It can be concluded that combined inclusion of ammoniated maize stover and Acacia leaves improved feed value and lamb performance when compared to the individual inclusion of both UMS and AMS.

lambs

growth performance

haematology

methane gas emission

serum biochemistry

Maize stover is the major crop residue that can significantly contribute to the feed resource base for livestock. However, its utilisation by ruminants is often limited by high levels of lignified matter that reduce its digestibility (Lynch et al., 2014). Coupled with reduced digestibility, its utilisation may result in the production of high amounts of absolute methane (CH₄) emission that contribute towards global warming (Aluwong et al., 2011). According to Aluwong et al. (2011) and Eckard et al. (2010) ruminants fed on grass or crop residues produce more CH₄ than ruminants fed on legumes forage. Energy loss through CH₄ production represents a significant loss of dietary energy that could potentially be redirected towards the meat production (Eckard et al., 2010). Therefore, treatment of residues, including ammoniation and inclusion of leaf meals in crop residues based diets could be employed as strategies to improve the nutritive value, digestibility and ultimately utilisation of the poor quality maize stover.

Previous studies have shown that physical, biological and chemical treatment improve the chemical composition and digestibility of the stover (Arora et al., 2011; Buthane et al., 2021; Wan Zahari et al., 2003). The ammoniation of crop residues is one of the chemical strategies to improve utilisations of poor-quality residues. Previous studies have shown that ammoniation of crop residues Ammonia treatment increased non-protein nitrogen (NPN) (Huang et al., 2022; Kraiem et al., 1991), that in turn improved reticulo-ruminal microbial activity, coupled with increased digestibility, suppression of mould growth, inhibition of bacterial proliferation ultimately improving animal performance (Belanche et al., 2021; Huang et al., 2022; Spanghero et al., 2017). Moreover, the alkalinising properties of ammonia are often associated with a decline in the rate of ruminal proton accumulation subsequently reducing the risk of reticulo-ruminal acidosis (Humer and Zebeli, 2017; Huang et al., 2022). In addition, the inclusion of protein-rich fodder tree leaves has also been shown to improve the performance of some ruminants (Liu, et al. 2001; Marume et al., 2012; Mnisi and Mlambo, 2017). The use of fodder trees such as Acacia sp which are abundantly available during dry seasons in animal feeding systems has the potential to ameliorate problems of feed shortages for ruminants (Mlambo et al., 2008). Natural fodder trees such as Acacia erioloba require limited agronomic inputs to grow and they are rich in readily fermentable nitrogen and energy (Kabir et al., 2004). Besides improving nutritive values of maize stover, the inclusion of leaf meal may help reduce methane gas emission. Methane (CH₄) gas emission from ruminants is a global concern.

Globally it has been estimated that ruminants produce 80 million tons of CH₄ annually, which accounts for 28% of anthropogenic emissions (Beauchemin et al., 2008; Metawi et al., 2017). This accounts for about 6–10% of the gross available energy from feed consumed by the animal. Several attempts have been made to reduce methane emissions, including supplementation with various feed stuff, antibiotics and probiotics (Benchaar et al, 2001; Metawi, et al., 2017; Ningrat et al., 2018). According to Ningrat et al. (2018) tannins are naturally occurring compounds that can act as anti-methanogenic agents in the rumen, therefore, have the ability to reduced methane production by ruminants. It is therefore hypothesized that the inclusion of Acacia erioloba leaf meal in diets improves nutritive value and performance in lambs and reduces methane gas emission in lambs. Therefore, this study aimed at examining the effects of Acacia erioloba foliage leaves inclusion in diets on the growth performance, blood parameters and greenhouse gas emission in lambs fed ammoniated maize stover based diets.

2.1. Study site and ethical considerations

The study was conducted at North-West University (NWU) Mafikeng Campus, South Africa. The NorthWest Province is situated between 25° and 28° south latitude and between 22° and 28° east longitude, 1500m above sea level. The experimental procedures were approved by the NWU-ANIMPROD Research Ethics Committee of North-West University following the guidelines for use and care of animals in research and the following ethics number was granted: NWU-00238-18-A5.

2.2. Ammoniation process

Two tons of stover obtained from the local commercial farms, were placed in airtight polyvinyl chloride plastic bags using garden forks., and subsequently tightly closed. Ammonia gas was then introduced through a pipe that pressed through the polyvinyl chloride into the middle of the sack, at a rate of 3.5% of the total dry matter. The opening left in the polyvinyl chloride bags after the withdrawal of the pipe was sealed with tape. The treated material was incubated for 4 weeks, in accordance with ammonia treatment procedures by Sundstoel et al. (1978). At the end of incubation period, samples of treated maize stover together with the untreated stover were obtained for the analysis of dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF), and total nitrogen (N) at the North-West University Farm (Molelwane) in the Animal Science Research laboratory. The chemical composition of untreated maize stover (UMS) and ammoniated maize stover (AMS) is presented in Table 1.

Table 1

Chemical composition (g/kg DM) of Maize Stover (MS) and Ammoniated Maize Stover (AMS)
Parameter	MS	AMS
DM	60	58
CP	3.59	9.23
CF	37.28	31.77
EE	1.45	1.72
NDF	79.10	71.52
ADF	56.78	45.28
DM = dry matter, CP = crude protein, CF = Crude fibre, EE = Effective energy, NDF = neutral detergent fibre ADF = acid detergent fibre.

2.3. A. erioloba leaf harvesting and processing.

Acacia erioloba leaves were collected at the North-West University farm, Molelwane. The leaves were collected at an advanced maturity stage during the rainy season. The branches were clipped from the growing point; placed on polyethylene sheets and air-dried for 3 days by placement under a shade to prevent nutrient heat damage of heat-sensitive nutrients and turned regularly to prevent a build-up of moulds. The leaf meal was obtained by carefully shaking the branches and passed through a 2mm sieve to eliminate thorns and pods and subsequently milled. Milled leaf samples of Acacia erioloba species were analysed for dry matter (DM), organic matter (OM), neutral detergent fibre (NDF), acid detergent fibre (ADF), and total nitrogen (N) at the North-West University Farm (Molelwane) in the Animal Science Research laboratory. The chemical composition of harvested Acacia erioloba leaf is represented in Table 2.

Table 2

*Acacia erioloba* leaf meal chemical composition
Parameters	Chemical composition (g/kg)
DM	95.0
ASH	3.8
CP	13.0
ADF	41.3
NDF	55.0
DM: dry matter; ash; CP: crude protein; ADF: acid detergent fibre; NDF = neutral detergent fibre

2.4. Dietary treatments

Four isnonitrogenous and isoenergetice diets were formulated to meet the requirements for growing lambs with the inclusion of ammoniated maize stover and A. erioloba leaf meal. A conventional lamb finisher diet (total mixed ration, TMR) was used as the control diet (Control). The fibre and protein component of the total mixed ration was replaced with untreated maize stover (UMS), ammonia-treated maize stover (AMS) and a combination ammonia-treated maize and Acacia leaves (AMSAL) as follow:

(a) TMR: Conventional lamb finisher diet (TMR) (Control)

(b) UMS: TMR + 20% untreated maize stover

(d) AMSAL: TMR + 10% treated maize stover: + 10% Acacia leaves

The dietary formulation and chemical compositions are shown in Tables 3 and 4.

Table 3

Proportion of ingredients composition (%) of the experimental diets
Ingredients (kg/DM)	Control	UMS	AMS	AMSAL
Lucerne	10	0	0	0
Hay	10	0	0	0
Maize stover	0	20	0	0
Amonniated maize stover	0	0	20	10
Acacia erioloba leaves	0	0	0	10
Maize/Chop	52.3	49.1	45.2	51
Wheat bran	10	10	15	12
Molasses meal	10	10	10	8.6
Soybean OCM	5	8	7	6
Feed grade urea	1	1.3	1.1	1.2
salt	0.5	0.5	0.5	0.5
Mono Ca:P	0.5	0.5	0.5	0.5
Feed limestone	0.7	1.1	1.2	0.7
Vitamin/mineral premix	1	1	1	1
Experimental diets: Diet 1 (Control, TMR), Diet 2 (20% maize stover), Diet 3 (20% ammoniated maize stover) and Diet 4 (10% ammoniated maize stove: 10% Acacia leaves). The diets were formulated to meet the nutritional requirements of growing fattening lambs (NRC, 2007). The diets were formulated to be iso-caloric and iso-nitrogenous, using Format®.

Table 4

Chemical composition of the diets (g/kg)
¹ Dietary treatments
Parameters	control	UMS	AMS	AMSAL
DM	89.45	90.01	86.65	89.00
ME MJ/kg	11.71	11.46	11.43	11.73
EE	1.21.55	2.80	2.67	3.33
CP	15.28	15.14	15.44	15.63
CF	9.07	9.95	11.29	9.56
NDF	20.93	26.04	26.43	22.70
Ca	0.69	0.63	0.66	0.56
P	0.35	0.38	0.38	0.32
Ca:P K S	1.97 0.95 0.20	1.65 1.12 0.17	1.73 1.15 0.17	1.75 1.01 0.16
Na	0.24	0.23	0.23	0.23
Cl	0.52	0.49	0.50	0.47
¹Dietary Treatments: Refer to Table 1. DM = dry matter, ME = Metabolism energy, EE = Ether extract, CP = crude protein, CF = Crude Fiber, NDF = neutral detergent fiber, Ca = calcium, P = phosphate Ca: P = ratio of calcium to phosphate, K = Potassium, S = Sulfur, Na = Sodium, Cl = Chloride.

2.5. Animal management and experimental design

Thirty-two (32) female lambs (aged 5–6 months) were used in this study. The lambs were randomly assigned to four (4) dietary treatments in a complete randomized design. Each treatment had 8 replications with the lambs housed individually as experimental units. Feed and water were provided ad libitum throughout the experimental period. Seven days for acclimatization was allowed for the lambs to adapt to the diets and the experiment was run over 63 days. All the experimental protocols were approved by the NWU-ANIMPROD Research Ethics Committee, and the following approval number was granted: NWU-00238-18-A5.

2.6. Feed intake and Growth performance measurements

Feed intake was measured daily by subtracting the weight of feed leftovers from that of the feed offered, and thereafter, average daily feed intake (ADFI) was calculated as follows.

$$ADFI= \frac{\varvec{f}\varvec{e}\varvec{e}\varvec{d} \varvec{o}\varvec{f}\varvec{f}\varvec{e}\varvec{r}\varvec{e}\varvec{d}-\varvec{f}\varvec{e}\varvec{e}\varvec{d} \varvec{r}\varvec{e}\varvec{f}\varvec{u}\varvec{s}\varvec{a}\varvec{l}\varvec{s}}{\varvec{N}\varvec{o}. \varvec{o}\varvec{f} \varvec{d}\varvec{a}\varvec{y}\varvec{s}}$$

The initial weight of the lambs was recorded at the start of the study. Thereafter weekly weights were measured in the morning before feeding at 08H00 for two months. The average daily weight gain (ADWG) was calculated as:

ADWG =$\frac{\varvec{F}\varvec{i}\varvec{n}\varvec{i}\varvec{s}\varvec{h} \varvec{w}\varvec{e}\varvec{i}\varvec{g}\varvec{h}\varvec{t} \left(\varvec{k}\varvec{g}\right)-\varvec{S}\varvec{t}\varvec{a}\varvec{r}\varvec{t} \varvec{w}\varvec{e}\varvec{i}\varvec{g}\varvec{h}\varvec{t}\left(\varvec{k}\varvec{g}\right) }{\varvec{N}\varvec{o}. \varvec{o}\varvec{f} \varvec{d}\varvec{a}\varvec{y}\varvec{s} }$

Where: t0 = initial time, T = final time, W (T) = final body weight, and W (t0) = initial body weight.

Feed conversion ratio (FCR) per animal was also determined using the following formula:

$$FCR=\frac{Feed intake \left(kg\right)}{weight gain \left(kg\right)}$$

2.7. Methane emission measurement

The methane production was measured using a portable intelligent gas detector (IGD) (Tokyo Gas Engineering Co., Ltd. Anritsu Devices Co., Ltd., Tokyo, Japan) in ppm/ minute. The IGD is a hand-held open path laser measuring device.

2.8. Haematological analyses

Blood samples were collected from the jugular vein using bleeding needles in a purple top vacutainer tube containing EDTA as an anticoagulant a day before the termination of the feeding trial. Blood was analyzed for hematology (erythrocyte count, hematocrit, hemoglobin, neutrophils, lymphocytes, monocytes, eosinophils, reticulocytes, white blood cells (WBC), and basophils) using an automated IDEXX LaserCyte Haematology Analyzer.

2.9. Serum biochemical analysis

About 8 mL of whole blood samples were collected in red top vacutainer tubes that allow for clotting. The blood samples were then centrifuged at 2000 × g for 10 min at 15°C within 4 h after collection to obtain serum, which was immediately stored in Eppendorf tubes in multiple aliquots at − 20°C until analysis. The serum samples were analysed for enzyme activity, Calcium (Ca), cholesterol (Chol), creatinine (Crea), globulin (Glob), Glucose (Glu), Lipase(Lipa), Phosphorus (Phos), Total Bilirubin (Tbil), total protein (TP) and serum urea nitrogen (SUN), were analysed using an automated IDEXX Vet Test Chemistry Analyser (IDEXX Laboratories, Inc.).

2.10. Statistical analysis

The general linear model's procedure (PROC GLM) of SAS (2012) was used to analyze the effects of diet on growth performance, methane emission and blood parameters while the mixed model’s procedure for repeated measures of SAS (2012 was used to analyze the effect of diet on cumulating weight gain considering the effect of a week of measurement and diet. The following models were used:

Yij = μ + D_i + ε_ij

Where Yij = individual observation of the i-th Diet, µ = overall mean, D_i = effect of diet, εij = random error.

Y_ijk = µ + D_i+ W_j + (D * W)_ij + ε_ijk

Where: Y_ijk = observation (Cumulative weight gain parameter), µ = population mean constant common to the observation, D_i = effect of diet, Wj = effect of week of measurement, D*W_ij = effect of diet interacting with week and ε_ij = random error term. For all tests, the level of significance was set at (P < 0.05).

3.1. Growth performance measures and methane emission of lambs

The effects of diet treatment on growth performance measures and methane emission of lambs are presented in Table 5. The results showed that while there were no differences in initial weights, the lambs fed the AMS and AMSAL diets had higher (P < 0.05) final body weights. Feed intake was, however, highest (P < 0.05) in lambs fed the control diet followed by those fed on AMSAL (P < 0.05) them those on AMS diet. The lambs fed untreated maize stover had the lowest feed intake compared to all other diets. Inclusion of AL had a considerable impact on feed intake. Feed intake in AMSAL fed lambs was significantly higher than that of the AMS fed lambs. No effect of diet was observed on ADG and FCR although the values for ADG were slightly higher in lambs fed the AMS and AMSAL diets while having slight slower FCR. Lambs fed a diet with UMS had the highest (P < 0.05) methane gas emissions while the lowest methane gas emission was observed in lambs fed the control diet. Generally, there was a decline (P < 0.05) in methane gas emission with the inclusion of treated maize stover and the leaf meal. The lambs fed the AMSAL diet had lower (P < 0.05) methane gas emissions in comparison with those fed the AMS diet. In all the dietary treatments, cumulative weight gain (CWG) increased linearly from week 1 to week 9 (Fig. 1). The lambs fed the UMS diet had the lowest CWG compared to all the other treatments throughout the trial. Lambs fed AMSAL appeared to have a similar growth trend to that of those fed the control. The final weight was generally high in lambs fed on AMS followed by those fed the AMSAL diet with the lambs fed the UMS diet having the lowest end weight.

Table 5

Growth performance and methane emission of lambs’ fed on the inclusion of untreated maize stover, ammoniated Maize and inclusion of ammoniated maize stover and *Acacia* leaves in a complete ration
Parameters	AMS0	UMS	AMS	AMSAL	SEM
Initial body weight (kg)	20.55	22.79	21.95	22.13	3.514
Final body weight (kg)	34.08^a	35.34^a	37.50^b	36.93^b	3.421
ADFI (kg)	1.39^a	1.21^d	1.27^c	1.32^b	0.043
ADG (kg)	0.22	0.20	0.24	0.24	0.046
FCR (kg/kg)	6.54	6.41	5.36	5.94	1.291
Methane emission (CH₄) (ppm)	75.38^c	200.57^a	148.13^b	126.88^b	32.740
^a,b Means in the same row with different superscripts differ (P < 0.05) are significantly different. ADFI = Average daily feed intake, ADG = Average daily gain, FCR = Feed conversion ratio. ²Experimental diets: AMS0 (control), UMS (20% untreated maize stover), AMS (20% ammoniated maize stover) and AMSAL (10% ammoniated maize stover: 10% Acacia leaves).

3.2. Haematological parameters

The results on hematological parameters are presented in Table 6. Hematological blood parameters were affected (P < 0.05) by the inclusion of AMSAL in diets. Red blood cell count (RBC), hematocrit value and basophil granulocytes (BASO) were lower (P < 0.05) in lambs fed the AMSAL diet. Nevertheless, AMSAL fed lambs had higher (P < 0.05) mean platelet volume (MVP) and procalcitonin (PCT) compared all other lambs. No significant differences were observed in other hematological parameters across treatments.

Table 6

Effect of feeding experimental rations on lambs’ hematological parameters
Parameters	Control	UMS	AMS	AMSAL	SEM
Red blood cell (X10^12/L)	10.69^a	10.82^a	12.39^a	8.57^b	3.416
Hematocrit value (%)	19.85^ab	19.75^ab	23.49^a	16.49^b	6.329
Hemoglobin concentration (g/dL)	12.15	11.80	11.91	11.64	0.950
Mean corpuscular volume (fL)	18.55	18.28	19.04	19.41	1.128
Mean corpuscular hemoglobin (pg)	13.76	11.25	10.70	15.83	5.531
Red cell distribution width (%)	22.79	22.09	22.09	22.46	2.262
White blood cells (x10^9/L)	6.79	7.45	6.17	7.56	2.040
Neutrophil granulocyte (%)	28.96	33.06	32.31	31.16	8.965
Lymphocyte (%)	58.76	54.21	54.30	56.51	9.569
Monocytes (%)	6.85	6.48	6.75	7.59	1.674
Eosinophil granulocytes (%)	7.33	5.43	5.60	3.99	4.121
Basophil granulocyte (%)	0.93^ab	0.89^ab	1.06^a	0.79^b	0.241
Platelets (k/µL)	289.88	240.75	258.00	310.13	8.490
Mean platelet volume (fL)	6.54^b	5.96^b	5.96^b	9.04^a	2.411
Platelet distribution width (%)	23.96	24.53	23.06	22.28	4.324
Procalcitonin (%)	0.18^ab	0.13^b	0.15^b	0.25^a	0.088
^ab Means in the same row with different superscripts (P < 0.05) are significantly different. SEM: Standard error of the mean. Experimental diets: Control (TMR), UMS (20% untreated maize stover), AMS (20% ammoniated maize stover) and AMSAL (10% ammoniated maize stover: 10 Acacia leaves)

3.3. Serum biochemical profile

The results on serum biochemical parameters are presented in Table 7. Diet had no effect on liver enzyme activity (ALT and ALP) irrespective of treatments. However, amylase activity was observed to be higher (P < 0.05) in lambs fed on AMSAL as compared to those fed the control diet. Total bilirubin (TBIL) was higher (P > 0.05) in lambs fed AMSAL diet while urea concentration was highest in the lambs fed the AMS diet. The lambs fed the control diet had the lowest values for TBIL and urea concentrations. The level of serum glucose, phosphorus, cholesterol, creatinine, calcium and total protein were similar (P < 0.05) across the four diets. Total albumin was also no affected by diet although the values were observed to be lower in lambs fed on UMS diet.

Table 7

Effect of feeding experimental rations on lambs’ serum biochemical parameters
Parameters	Control	UMS	AMS	AMSAL	SEM
Albumin (g/L)	26.13^ab	24.00^b	26.29^ab	28.25^a	2.980
Alkaline phosphatase (U/L)	297.50	292.63	297.43	319.25	97.271
Alanine transaminase (U/L)	19.88	24.63	21.43	21.88	12.211
Amylase (U/L)	16.75^b	20.98^ab	22.43^ab	25.50^a	5.712
Lipase (U/L)	179.75	170.50	140.00	175.63	63.410
Calcium (mmol/L)	2.04	1.70	1.81	1.93	0.505
Cholesterol (mmol/L)	1.20	1.77	0.92	0.92	9.520
Urea (mmol/L)	7.90^bc	9.51^ab	11.20^a	6.83^c	1.230
Glucose (mmol/L)	2.53	2.43	2.53	2.43	0.554
PHOS (mmol/L)	3.589	4.21	3.81	3.59	10.666
Total bilirabin (µmol/L)	2.75^b	3.10^b	3.57^b	6.25^a	2.336
Creanine (µmol/L)	49.50	51.50	59.00	52.00	9.989
Total Protein (g/L)	68.25	56.63	65.86	68.63	13.306
Globulin (g/L)	48.00	45.43	45.43	51.75	11.398
^abMeans with different superscript letters within treatments and sex in the same row, differ significantly (P < 0.05). SEM: Standard error of the mean. Experimental diets: Control (TMR), UMS (20% untreated maize stover), AMS (20% ammoniated maize stover) and AMSAL (10% ammoniated maize stover: 10 Acacia leaves)

The major challenges observed when ammoniated stover has been used in ruminant feeding systems are the feed intake depression and toxicity, which are related to several factors such as total daily ingestion of ammonia, dietary CP and proportion of the CP supplied as non-nitrogen protein. In order to prevent these adverse effects, inclusion level of 20% AMS was used in the current study as recommended in previous studies (Dayani et al., 2011). The increase in feed intake in lambs fed the AMSAL diet can be attributed to the effect of Acacia leaf meal could have improved rumen fermentation and nutrient availability as observed in other studies (Abdul et al., 2012; Huang et al., 2022; Yadete. 2014;). The improved intake in the AMSAL fed lambs could also be due to the diluent effect of the leaf meal on the high ADF and NDF that characterize maize stover in comparison with the normal fibre source in the total mixed ration as observed elsewhere (Undi et al., 2001). Although ADFI for lambs fed on AMSAL diet was slightly lower than that of the control diet, observation from the results showed that the inclusion of the leaf meal substantially increased the feeding value of the diet and the feed intake when compared to the AMS and UMS respectively. The differences in ADFI between lambs fed on UMS and AMS, shows that ammonia treatment also improved the quality of the stover and ultimately the utility of the treated maize stover as reported elsewhere (Huang et al., 2022; Mahesh and Mohini, 2013; Souzza et al., 2010; Undi et al., 2001). This is because ammoniation exposes rumen degradable protein that is locked between the lignin bonds in untreated maize stover.

Although ADG and FCR were statistically not affected by diet, numerically, lambs fed on a diet containing AMS and AMSAL had marginally higher ADG and FCR as compared to the control and the UMS diets. The lower body weights obtained in lambs fed the UMS diet confirms the fact that untreated maize stover on its own is unable to release sufficient nutrients to meet the ruminant’s requirements even if it had similar CP to all dietary treatments, owing to the effect of the fibrous nature of untreated maize stover on supply parameters such as voluntary feed intake. This is consistent with reports from other studies which showed poor growth and productivity in ruminants fed only on maize stover-based diet (Barde et al., 2010; Ngongoni et al., 2009). Improvement in growth performance of lambs fed on combined inclusion of AMS and leaf meal was expected based on their nutritive value and a probable increase in digestibility of the diets. Metawi et al. (2017) also confirm improved performance in goats supplemented foliage leaves.

Maize stover is known to have lower digestibility, and as the digestibility of the diet decreases, inefficient fermentation occurs resulting in its energy being converted to methane gas (Metawi et al., 2017). Lambs fed the UMS diet had the highest methane gas emission compared to all other dietary treatments, with the control diet producing the least. The high methane production in lambs fed the UMS inclusion diet may be due to maize stover having a slow passage rate, prolonged residency time in the rumen, and inefficient fermentation while a comparative reduction in CH₄ emission in AMSAL fed lambs could be due to the increase feeding value and quality of the diet with the inclusion the leaf meal. Inclusion of AMS alone decreased CH₄ emission by 26%. However, the combination of AMS and Acacia leaf meal inclusion led to a 36% decrease in CH₄ emission compared to the UMS diet. Therefore, treatment of stover and inclusion of A. erioloba leaf meal in diets has the potential to improve feeding value and nutritive quality of diets, which consequently improves digestibility, whilst reducing methane emission. This is in line with findings by Zhang et al. (2019) who reported a gradual decrease in methane emission when feeding urea-treated rice straw on a small ruminant. Acacia leaf meal inclusion in highly fibrous diets is known to have the ability to lower methanogenesis indirectly through inhibition of the growth of archaea methanogens (Tavendale et al., 2005). The effect of Acacia leaf inclusion on CH₄ emission reduction is also linked to the presence of some secondary plant metabolites (Benchaar et al., 2008; Metawi et al., 2017; Montoya-Flores et al., 2020).

Blood profiles provide a significant basis for the assessment of the health and nutritional status of animals. They also provide a good indication of the effects of different extraneous factors on health, immune response, and normal physiological functioning of the body (Beigh et al., 2018). The hematological values obtained in the current study were within the expected ranges (Beigh et al., 2018) for all diets. The observed reduction in RBC in AMSAL fed lambs is consistent with the decrease in the hematocrit value. Hematocrit values indicate the adequacy of the supply of healthy red blood cells, which, when deficient may result in anemia. The Acacia species contain secondary plant compounds such as tannins. Intake of condensed tannins (CT) may precipitate proteins that inhibit the absorption of iron and reduce the bioavailability of iron (Rani et al., 2018) hence the observed results. In contrast Paswan et al. (2016) and Brown et al. (2016) reported no effect on RBC with the inclusion of Acacia leaves. Lower basophil count may be due to the suppressive effect of CT contained in the Acacia leaves. This was in agreement with Brown et al. (2016), Olafadehan, (2011) and Solaiman et al. (2010) who reported a reduction in the WBC component when the animals were fed on CT rich diet.

Serum biochemical parameters are also bio-indicators of the nutritional and general health status of the animals. The results of the current study show an increase in albumin (ALB), amylase (AMYL), and total bilirubin (TBIL) values with the inclusion of Acacia leaves in a diet. The observed increase in ALB indicates an increased amount of available metabolic protein to the animal contributed by the Acacia leaf meal as observed in other studies (Zhao et al., 2019). The observed increase in ALB and AMYL concentrations in the AMSAL fed lambs compared to other diets may reflect better protein and energy digestion and utilization for that diet. TBIL was also significantly higher in lambs fed on AMSAL inclusion diet, however, their values remained within the reference range of 0.88 to 8.84 µmol/L (Borjesson, et al. 2000). Serum AMYL activity was significantly higher in lamb fed on a diet with combined inclusion of AMS and AL. This may be attributed to higher particles involved in fibre digestion of the stover and nutrient availability in Acacia leaves. An increase in AMYL activity in lamb fed on combined inclusion of AMS and AL was expected as the AMYLY activity depends mostly on improved fibre digestion in the rumen (Raghuvansi et al., 2007).

Calcium (Ca), Alkaline transferase (ALT), cholesterol (CHOL), creatinine (CREA), globulin (GLOB), glucose, lipase (LIPA), phosphorus (PHOS) and total protein (TP) were not affected in all the dietary treatments. Similarly, Solaiman et al. (2010) reported no effect Ca, ALT, AMYL, CHOL, CREA, GLOB, GLU, LIPA, PHOS and TP of goats consuming CT containing forage. In contrast to the results of the current study other studies confirm a decrease in TP with the inclusion/supplementation of acacia sp leaves (Zhao et al., 2019). Sallam et al. (2017) also observed reduced TP with partial substitution of clover hay by acacia sp. The current study shows no significant effect of diet on TP across all the dietary treatments, however, numerically lambs fed on a diet containing Acacia leaves (AMSAL) had higher serum TP and more comparable to control diet-fed lambs, this could be enhanced by the interaction of Acacia leaves and ammoniated stover as observed in as study by Sallam et al. (2017).

Surprisingly, with no effect on the serum TP, the serum urea nitrogen was significantly high in lambs fed on AMS with values that were above the normal range of 2.8–7.1 mol/L as outlined in the Merck Manual (2012) for healthy lambs. The possible reason for an increase in serum urea could be as a result of rumen undegradable protein supplied through ammoniation. The increase in serum urea due to ammoniation agrees with the findings reported by Kraidees, (2005). Blood urea is an indirect indicator of the protein composition of feed associated with greater ruminal degradation of protein with a concurrent increase in ammonia production (Broderick and Clayton, 1997; Soul et al., 2010). As expected, the serum urea level was reduced with the inclusion of Acacia leaves. Sallam et al. (2017) reported a reduction in blood urea in goats fed acacia saligna leaves. Waghorn et al. (1994) also concluded that Acacia supplementation has the potential to lower serum urea in sheep. Both hematological and serum biochemical parameters observed were all within normal ranges for healthy growing lambs (Merck manual, 2012), except for serum urea in lambs fed on the AMS diet. This implied that the test diets were able to supply an adequate amount of nutrients needed to maintain normal lamb’s blood metabolites except for AMS based diet on serum urea.

Inclusion of Acacia erioloba leaf meal in AMS based diet significantly increased feed intake and reduced CH₄ emissions, nutritional and health status of the lambs, as it had the potential to increase the serum albumin, which is the main component indicator of the nutritional and health status of the animal. The desirable blood metabolite levels observed in the current study indicate the potential of the combined effect of AMSAL inclusion in improving the general health status of lambs. Use of ammoniated maize stover with Acacia leaves could be implemented in lamb finisher feeding without detrimental effects on growth performance and health status of the animals but rather improve growth performance and health status of the lambs.

Acknowledgements

We would like to thank the Dept of Animal Science (NWU) and PureGas, SA (Pvt, Ltd), for providing resources and funds for the study.

Author contributions

Getrude M Chelopo: Conceptualization, Methodology, Data curation, Formal analysis, Investigation, Writing-Original draft preparation; Upenyu Marume: Conceptualization, Methodology, Formal analysis Project administration, Editing, Supervision, Funding acquisition

Data Availability

The data sets used in the current study are available from the corresponding author on request.

Funding

We are grateful to the Puregas, South Africa (PTY) LTD for provision of research funds and ammonia gas and the NWU for providing the infrastructure and the research animals.

Declaration of conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethical approval

The experimental procedures were approved by the NWU-ANIMPROD Research Ethics Committee of North-West University following the guidelines for use and care of animals in research and the following ethics number was granted: NWU-00238-18-A5.

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The effect of Acacia eriolaba leaf meal inclusion on growth performance, blood parameters and methane gas emission in lambs fed diets containing ammoniated maize stover.

Status:

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Version 1

Abstract

Figures

1. Introduction

2. Materials and methods

2.1. Study site and ethical considerations

2.2. Ammoniation process

2.3. A. erioloba leaf harvesting and processing.

2.4. Dietary treatments

2.5. Animal management and experimental design

2.6. Feed intake and Growth performance measurements

2.7. Methane emission measurement

2.8. Haematological analyses

2.9. Serum biochemical analysis

2.10. Statistical analysis

3. Results

3.1. Growth performance measures and methane emission of lambs

3.2. Haematological parameters

3.3. Serum biochemical profile

4. Discussion

5. Conclusion

Declarations

References

Status:

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