Food & Feed Research

Influence of mistletoe (Viscum album) leaf meal on growth performance, carcass characteristics and biochemical profile of broiler chickens

DOI: UDK:
636.52/.58:[636.087.8:582.728.4
JOURNAL No:
2017-2
PAGES
163-172
KEYWORDS
antibiotics, broiler chickens, feeding trial, Viscum album
TOOLS Creative Commons License
Anthony D. Ologhobo1, Isaac Oluseun Adejumo*2, Temitope Owoeye1, Akangbe Esther1

1University of Ibadan, Department of Animal Science, Animal Biochemistry and Nutrition Research Unit, Ibadan, Nigeria
2Federal University Gashua, Department of Animal Science, Animal Nutrition, Biotechnology and Food Safety Laboratory, Gashua, Nigeria

ABSTRACT

The quest for alternatives to antibiotics has resulted in the discovery of prebiotics. The search for the alterative antibiotics is on-going. Therefore, this study was carried out to investigate possible prebiotic potentials inherent in mistletoe (Viscum album) leaf meal with the aim of developing prebiotics as an alternative to antibiotics thus optimizing animal performance, carcass characteristics and a healthy blood profile as indicators of systemic conditions. Five experimental diets were formulated and mistletoe leaf meal (AMLM) was incorporated into the diets at different concentrations (0% with 0.05% antibiotics (positive control), 2.5%, 5.0%, 7.5% without antibiotics (negative control)). The values of body weight were not significantly different across the treatments. Live weight, bled weight, wings, drumsticks, thighs, breasts and heads were not significantly different across the treatments. Aspartate aminotransferase, globulin and creatinine contents were not significantly different across the treatments. Meanwhile, birds on AMLM-supplemented diets obtained significantly (p <0.05) higher values of alanine aminotransferase than those on negative control diet (3.75 IU/l). Urea and glucose contents followed a similar pattern. The AMLM could be used as alternative antibiotics in broiler production, although further studies are required to ascertain this.

Introduction

With the intensification of the livestock production in Nigeria, came an increase in clinical and sub-clinical enteric diseases, thus animals became vulnerable to harmful bacterial such as E. coli, Salmonella and Clostridium perfringens, resulting in reduced productivity, increased mortality and the associated contamination of meat, meat products and eggs for human consumption (EFSA BIOHAZ panel, 2013). In response to these problems, antibiotics have been used as a growth promoter (AGP) to promote good health and enhance feed efficiency, growth and production performance in farm animals.

Antibiotics are naturally occurring, semi-synthetic and synthetic compounds with anti-microbial activity that can be administered orally, parentally or topically and also be used as growth promoters at sub-therapeutic levels. However, the use of antibiotics has not been without side effects. These include increase in populations of resistant pathogens and commensal bacteria in the animal given antibiotics. Leaf meals have been incorporated into poultry diet for several positive reasons. The beneficial effects of leaf meals in poultry production have been reported (Egbenwade and Olorede, 2003; Murthy et al., 2006). Leaf meals have been reported to provide antioxidants (Cross et al., 2007), antimicrobial (Manzanilla et al., 2004), immunity (Ko et al., 2008) and growth promoting effects (Lee et al., 2009). In the livestock industry, herbs and other plant extracts improve feed intake, digestibility and reinforce immunity (Wenk, 2003).

The search for the alterative antibiotics is on-going. Ologhobo et al. (2017) earlier reported that Viscum album did not have any significant on packed cell volume, haemoglobin, red blood cell counts, monocytes, eosinophils, basophils, platelets, MCV, MCH and MCHC of broiler chickens fed with diets supplemented with Viscum album. Therefore, this study was carried out to investigate possible prebiotic potentials inherent in mistletoe (Viscum album) leaf meal with the aim of developing prebiotics as an alternative to antibiotics thus optimizing animal performance, carcass characteristics and a healthy blood profile as indicators of systemic conditions.

MATERIAL AND METHODS

The study was carried out at the Poultry Unit of the Department of Animal Science, Teaching and Research Farm, University of Ibadan, Nigeria. The experimental pens were thoroughly cleaned, washed and disinfected. Fresh leaves of Viscum album from Citrus spp. (orange) trees were harvested, washed and air dried for about two weeks. The dried leaves were separately micronized with a hammer mill into a fine powder, known as African Mistletoe Leaf Meal (AMLM), weighed and kept in sterile containers for use later.

Table 1. Gross composition of experimental diets for birds (%) (as fed basis)

Ingredients

(%)

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

Maize

53.30

53.30

53.30

53.30

53.30

Soy bean meal

35.65

34.76

33.87

29.55

35.65

Fish meal

3.00

3.00

3.00

3.00

3.00

Wheat offal

2.85

2.85

2.95

2.95

2.95

Oyster shell

2.00

2.00

2.00

2.00

2.00

DCP*

2.00

2.00

2.00

2.00

2.00

Lysine

0.25

0.25

0.25

0.25

0.25

Methionine

0.35

0.35

0.35

0.35

0.35

Premix**

0.35

0.35

0.35

0.35

0.35

Salt

0.25

0.25

0.25

0.25

0.25

AMLM

0.00

0.95

1.75

2.55

0.00

Antibiotics***

0.05

0.00

0.00

0.00

0.00

Calculated analysis

Crude protein

23.25

23.11

23.18

23.10

23.24

Crude fibre

3.00

3.34

3.69

3.98

3.00

Ether extract

3.65

3.66

3.64

3.68

3.66

Calcium

1.45

1.47

1.49

1.52

1.45

Phosphorus

0.72

0.74

0.73

0.75

0.72

Lysine

0.93

0.93

0.95

0.97

0.93

Methionine

0.32

0.34

0.34

0.35

0.32

ME (Kcal/kg)

3199.00

3227.00

3263.00

3279.00

3200.00

*DCP – Dicalcium Phosphate

**Composition of Vitamin Premix per kg of diet: Vitamin A, 12500IU; Vitamin D3, 2500IU; Vitamin E, 40mg; Vitamin K, 3.2mg; Vitamin B1, 3mg; Vitamin B2, 5.5mg; Calcium pantothenate, 11.5mg; Vitamin B6, 5mg; Vitamin B12, 0.025mg; Choline Chloride, 500mg; Folic Acid, 1mg; Biotin, 0.08mg; Manganese, 120mg; Iron, 100mg; Zinc, 80mg; Copper, 8.5mg; Iodine, 1.5mg; Cobalt, 0.3mg; Selenium, 0.12mg; Antioxidant, 120mg.

***Antibiotics contained tetracycline hydrochloride 500 mg

AMLM – African Mistletoe Leaf Meal

Experimental diets

A total of five experimental diets were formulated to meet the NRC (1994) nutrient requirements for broilers. Mistletoe leaf meal (AMLM) was incorporated into the diets at different concentrations (0%, 2.5%, 5.0% and 7.5%). Treatments were: T1 (standard diet + 0.05% antibiotic – positive control), T2 (standard diet + 2.5% AMLM), T3 (standard diet + 5.0% AMLM), T4 (standard diet + 7.5% AMLM) and T5 (standard diet – negative control). The gross composition of experimental diet is shown in Table 1.

Determination of the chemical composition of the diets

Triplicate samples of mistletoe leaf meal (AMLM) were subjected to chemical analysis according to the method of AOAC (2000). Nitrogen free extract (NFE) was determined by difference between 100 and the sum of moisture, protein, crude fibre, fat and ash values (Table 2).

The spectrophotometric method of Akinmutimi (2006) was used for the determination of saponin, tannin, oxalate and phytates in AMLM. One gram of AMLM was dissolved in 50 ml of butanol in a 25 ml beaker, the mixture was left for 5 h and then shaken to have a homogenous mixture. The mixture was filtered through a What man filter paper into a 100 ml beaker and 20 ml of 40% saturated solution of magnesium carbonate (MgCO3) was added to the filtrate. The saturated solution of magnesium carbonate obtained was again filtered using Whatmanfiltter paper to obtain a clean colourless solution. 1 ml of the colorless solution was pipetted into a 50 ml volumetric flask and 2 ml of 5% FeCl3 solution added. It was made to the mark with distilled water and allowed to stand for 30 min. The absorbance of the solution was read on an Agilent spectrophotometer at a wavelength of 380 nm.

Tannin was quantified by taking 2 g of each of AMLM in a conical flask and 10 ml of distilled water was added. The solution was left to stand for 30 min after which 2.5 ml of the supernatant was taken into a 5 ml volumetric flask and 1 ml of Folin–Denis’ reagent was added. This was followed by the addition of 2.5 ml of saturated Na2CO3 and diluted to 50 ml in a volumetric flask with distilled water. It was allowed to stay for 90 min after which the absorbance was read at 250 nm on a spectrophotometer.

Oxalate was determined by dissolving 2g of the AMLM in 100 ml of distilled water in a 500 ml volumetric flask, followed by addition of 10 ml 6 M HCL. It was boiled for 1 h, cooled and filtered. The content was made up to 300 ml with distilled water. Duplicate portions of the filtrate (125 ml) were taken into 5 different beakers and drops of methyl red indicator were added, followed by concentrated NH4OH solution drop wise until the test solution changed from pink to faint yellow colour.

Phytate was determined by extracting 5 g of AMLM with 0.2 N HCl; 0.5 ml of the extract was pipetted into a test tube and heated in boiling water bath for 30 min. The test tube was cooled in ice for 15 min and allowed to reach the room temperature. The content of the tube was mixed and centrifuged for 30 min at 3000 rpm. 1 ml of the supernatant was transferred to another test tube and 1.5 ml of HCl solution was added before the absorbance was read at 514 nm in a spectrophotometer. All determinations were carried out in triplicates.

Management of experimental animals

A total of two hundred unsexed day-old Cobb broiler chicks were used for the study. They were weighed for their initial weights and randomly allotted into five dietary treatments with forty (40) chicks per treatment. Each group had five replicates with eight (8) chicks per replicate in a completely randomized design (CRD). The birds were placed on conventional feeds for the first week after which they were randomly assigned to dietary treatments

The brooding pens were thoroughly cleaned, disinfected and allowed to rest for a period of two weeks before the arrival of the chicks. During this period, the pens were sealed up with polythene bags and fumigated in preparation for brooding, feeders and drinkers with other brooding materials were thoroughly washed and disinfected. Wood shavings used as litter materials were spread on the floor of the pen and a warm temperature was maintained within the pen with 100 watt electric bulbs before the arrival of the chicks. On arrival, the chicks were carefully unboxed, weighed and brooded for a period of one week before they were randomly allotted into treatments. Fresh cool water and feed were provided ad-libitum to the birds throughout the period of the experiment and routine medication (vaccination and drugs) were administered at appropriate times to birds on the positive control only.

Data collection and analyses

Feed consumption for each animal was measured daily as the difference between the daily feed supplied and refusal, and live-weight changes of the animals were taken weekly throughout the experimental period.

Carcass characteristics

The carcass characteristics were determined at the end of the experiment by selecting randomly, three birds from each replicate. The selected birds were starved of feed and water over night. Before slaughtering, the individual weight of the birds was recorded. Thereafter, the birds were slaughtered by cutting the jugular vein around the neck. The birds were immediately scalded in warm water and the feathers were manually removed. Thereafter, the fully dressed weights of the carcasses were taken and recorded. The carcasses were then separated into breast, back, upper back, thigh, shank, neck, arm, wing, drumstick, head and the internal organs (viscera). The parts were individually weighed and the weights were expressed as percentage of the live weight of the carcass. In addition, the length of the intestine of each carcass was taken and recorded. The dressing percentage and percentage weight of body in relation to the live weights of the birds were calculated by this formula:

Haematological parameters

At the end of the feeding trial, blood samples were collected from the jugular vein of animals from each group into two sets of Monoject® vacutainers. One set containing ethylenediaminetetraacetic acid vacutainers (EDTA) for haematology, while the other set without EDTA was covered and centrifuged, the serum decanted and deep-frozen for serum biochemical and enzymological analyses.

Statistical analysis

Data obtained from the experiment were subjected to analysis of variance (ANOVA) (SPSS 17.0). The variations in means were separated using the Duncan’s Multiple Range Test (Duncan, 1995).

Results and Discussion

The result of the proximate composition of the tested ingredient (African mistletoe leaf meal) is shown in Table 2. The leaf was rich in phytates (22.75%) and oxalates (15.80%) while the proximate composition of the feed samples is shown in Table 3.

The growth performance, carcass characteristics, organ weights and serum biochemical profile of broiler chickens fed with AMLM are presented in Tables 4, 5, 6 and 7 respectively. The values of body weight were not significantly different across the treatments. Feed intake was not significant during weeks 4 and 5, while on positive control (T1) had the highest feed intake during the second week closely followed by those on negative control (T5). The feed conversion ratios (FCR) were statistically similar across the treatment during weeks 2 and 5, while the values were not significant during weeks 3 and 4. Birds on control diets had the highest rate of mortality when compared with those on 5.00% and 7.50% AMLM supplemented diets.

Table 2. Chemical composition of African mistletoe leaf meal (AMLM)

Parameters

Proportion (%)

Moisture content

7.70

Crude protein*

3.50

Ash*

11.21

Ether extract*

7.11

Crude fibre*

8.90

Saponins*

3.25

Tannins*

9.90

Oxalate*

15.80

Oxalate*

22.75

*Determined on dry matter basis

Table 3. Proximate composition of feed samples

Samples

(%)

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

Dry matter

92.10

92.18

92.49

92.27

92.40

Crude protein

23.00

23.50

23.13

22.77

24.10

Crude fibre

10.0

9.96

9.61

9.01

9.40

Ether extract

7.12

6.90

7.02

6.80

7.00

Ash

10.89

11.30

11.50

11.30

12.00

AMLM – African mistletoe leaf meal


Table 4. Growth performance of broiler chickens fed with graded levels of mistletoe leaf meal

Parameters

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

SEM

Body weight gain (g/bird)

Week 2

2.67ns

2.25 ns

2.25ns

2.74ns

2.68 ns

0.96

Week 3

3.57ns

2.68 ns

2.98ns

2.36ns

3.08 ns

4.33

Week 4

6.63ns

7.14ns

7.36ns

8.05ns

9.05ns

0.63

Week 5

2.21ns

2.43ns

1.96ns

3.07ns

2.69ns

4.10

Feed intake (g/bird)

Week 2

59.64a

52.14c

54.54cd

51.61c

54.54ab

0.85

Week 3

111.00bc

111.61bc

104.82b

118.75bc

123.21a

2.47

Week 4

118.39ns

117.50ns

116.43ns

128.87ns

138.71ns

3.76

Week 5

117.11ns

117.14ns

114.12ns

137.02ns

140.21ns

4.40

FCR

Week 2

22.63ab

20.21ab

26.33a

18.91b

21.32ab

0.08

Week 3

31.54ns

43.22ns

46.52ns

60.65ns

42.16ns

0.19

Week 4

19.01ns

16.78ns

15.37ns

16.49ns

15.37ns

0.35

Week 5

46.99ab

38.63b

67.83a

47.52ab

59.50ab

0.25

Mortality (%)

1.80ab

0.40bc

0.20c

0.20c

2.80a

0.30

a,b,… Means with different superscripts in the same row differ significantly (p< 0.05) different, ns = non-significant, SEM = Standard Error of Mean

AMLM – African mistletoe leaf meal

Table 5. Carcass characteristics of broiler chickens fed with graded levels of mistletoe leaf meal

Parameters (g)

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

SEM

Live weight

954.00ns

936.00ns

960.00ns

1030.00ns

900.00ns

19.12

Bled weight

867.20ns

913.60ns

908.20ns

962.00ns

879.00ns

14.86

Defeathered weight

834.80c

847.00bc

865.00abc

930.00a

914.80ab

12.33

Dressed weight

631.60b

657.00ab

633.00b

722.00a

666.00ab

11.32

Wings

75.56ns

71.00ns

78.20ns

82.54ns

77.84ns

3.24

Drumstick

76.98ns

85.00ns

77.70ns

74.00ns

83.44ns

1.87

Thigh

88.00ns

88.80ns

92.98ns

95.42ns

94.84ns

1.85

Breast

156.54ns

173.70ns

125.46ns

164.78ns

147.04ns

4.00

Back

102.08b

125.46a

108.26b

126.32b

105.760a

3.12

Neck

40.54abc

38.28bc

36.00c

47.00a

44.00ab

1.18

Shank

37.74ab

35.90b

63.64a

46.32ab

41.10ab

3.93

Head

27.54ns

28.84ns

28.50ns

30.04ns

29.06ns

4.73

a b,.. Means with different superscripts in the same row differ significantly (p< 0.05) different, ns = non-significant

SEM - Standard Error of Mean

AMLM – African mistletoe leaf meal

Table 6. Organ weight of broiler chickens fed with graded levels of mistletoe leaf meal

Parameters (g)

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

SEM

Heart

4.16b

4.78ab

5.66ab

4.82ab

7.86a

0.50

Liver

34.68a

25.98b

29.18ab

29.32ab

29.08ab

1.11

Empty gizzard

26.28ab

21.52b

29.22a

26.96ab

25.04ab

0.50

Whole gizzard

43.20ns

35.52ns

42.82ns

39.14ns

39.44ns

1.13

Spleen

1.00ab

0.90b

0.68b

1.50a

0.76b

0.98

Lungs

80.30b

92.36ab

87.10ab

84.64ab

100.44a

2.54

Abdominal fat

5.64a

2.34b

0.80c

5.24a

0.90c

0.55

a,b,… Means with different superscripts in the same row differ significantly (p< 0.05) different, ns = non-significant. SEM - Standard Error of Mean

AMLM – African mistletoe leaf meal

Table 7. Serum profile of broiler chickens fed with graded levels of mistletoe leaf meal

Parameters (g)

T1-positive control

(0% AMLM, 0.05% antibiotics)

T2

(2.5% AMLM, 0% antibiotics)

T3

(5% AMLM, 0% antibiotics)

T4

(7.5% AMLM, 0% antibiotics)

T5-negative control

(0% AMLM, 0% antibiotics)

SEM

AST (IU/l)

197.16ns

213.83ns

191.24ns

192.97ns

198.44ns

285.86

ALT (IU /l)

3.75d

4.87c

8.08a

6.07b

8.30a

0.56

TP (g/dl)

3.75b

4.85a

4.38ab

4.19ab

4.74a

0.29

Globulin (g/dl)

0.76ns

1.04ns

1.00ns

0.75ns

1.18ns

0.28

Creatinine (mg/dl)

0.28ns

0.44ns

0.33ns

0.39ns

0.45ns

0.03

Urea (mg/dl)

6.30c

7.75b

9.83a

10.11a

10.24a

0.30

Glucose (mg/dl)

174.32a

184.51b

192.22b

198.15b

250.00b

795.77

a,b,… Means with different superscripts in the same row differ significantly (p = 0.05) different, ns = non-significant. TP-total protein, SEM-Standard Error of Mean, AMLM – African mistletoe leaf meal, AST – Aspartate amino transferase, ALT - Alanine Amino transferase

Live weight, bled weight, wings, drumsticks, thighs, breasts and heads were not significantly different across the treatments. Birds on 7.50% AMLM-supplemented diet (930.00 g) obtained higher defeathered weights than those on positive control (834.80 g). Dressed weight followed a similar trend. The mean values of whole gizzard were not significantly different across the treatments. However, weights of hearts, empty gizzards and lungs were statistically similar to those of the control diets.

Aspartate aminotransferase, globulin and creatinine contents were not significantly different across the treatments. Meanwhile, birds on AMLM-supplemented diets obtained significantly (p< 0.05) higher values of alanine aminotransferase than those on negative control diet (3.75 IU/l). Urea and glucose contents followed a similar pattern.

Traditional herbal practice in many parts of the world involves prescribing combinations of herbs with a wide range of actions that concurrently cover several treatment strategies. Rather than focusing on a specific disease pathology, Herbal practitioners treat holistically with individualized herbal formulae (Williamson, 2001). Combinations provide multiple active constituents working together which may produce additive, or synergistic interactions.

The result of the proximate composition of the test diet showed that the crude protein, metabolizable energy, and ether extract are adequate. Even though, there was variation in the analysed crude protein composition, this did not affect the performance of the birds. The reason would perhaps be due to the fact that all the diets had crude protein that was well above the least recommendation. In this study, feed intake became not significant as the experiment progressed. Also, Mottaghitalab (2000) earlier observed no differences in body weight gain of broiler chickens given diets supplemented with different natural feed additives as alternatives to antibiotic growth promoters, which is similar to the findings of the present study, although Guoet al. (2004) reported a significant positive effect on broiler body weight gain exposed to herbs and herbal products. Also, Ocaketal.  (2008) and Sarkeret al. (2010b) reported no significant difference for body weight gain and feed intake. The variations in the results could be explained by the type and dosage of herbs and herbal products used. Plant extracts contain different molecules that have intrinsic bioactivities affecting animal physiology and metabolism. Some of these compounds have been reported to improve animal performance due to their stimulating effect on salivation and pancreatic enzyme secretions or by having a direct bactericidal effect on gut microflora (Hardy, 2002).

Previous studies disagree on the effects of herbs on carcass characteristics of animals. Hassan et al. (2004) reported an increase in dressing and liver percentages for broiler chicks fed the supplemented herbal feed additives while Sarkeret al. (2010a) reported that herbal plants had no influence on organ weight. The AMLM did not influence most of carcass parts and such results are not strange since it did not influence body weight gain of the experimental animals.

The non-influence of AMLM on creatinine content of the experimental animals is in harmony with the findings of Hossainet al. (2012) who earlier reported that water plantain, mistletoe and antibiotics had no influence on the creatinine content of birds. The observed ALT content was higher for AMLM-supplemented diets but fell within the range reported by Mitruka and Rawnsley (1977). Elevated serum activities could be an indication of heart, kidney and or liver damage owing to cellular destructions caused by toxins (Ewuola and Egbunike, 2008). More studies may be required to ascertain the hematological, immunological, and antimicrobial activities of these plants.


Conclusions

It is concluded from the results of this study that AMLM did not influence body weight gain, feed intake and some parts of the carcass characteristics of the experimental animals. The AMLM did not seem to pose a threat on biochemical profile of the experimental animals. However, further studies may be required to ascertain the effects of AMLM on biochemical profile of animals, antibacterial  properties and effects on immune response of animals.

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