Influence of Dietary Andrographis paniculata and Zingiber cassumunar Mixture on Growth Performance and Small Intestinal Morphology in Broiler Chickens




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Influence of Dietary Andrographis paniculata and Zingiber cassumunar Mixture on Growth Performance and Small Intestinal Morphology in Broiler Chickens

Danet Laing1,*, Sawitree wongtangtintharn2,#, Bundit Tungjarernkul3

Srisuda Sirilaophaisan4 and Jowaman Khajarern2

1 M.Sc program, Department of Animal Science, Faculty of Agriculture, Khon Kaen University 40002, Khon Kaen, Thailand

2 Department of Animal Science, Faculty of Agriculture, Khon Kaen University 40002, Khon Kaen, Thailand

3 Faculty of Veterinary medicines, Khon Kaen University 40002, Khon Kaen, Thailand

4 Faculty of Natural resources, Rajamangala University of Technology Isan Sakon Nakhon Campus, Thailand

* e-mail: laingdanet@yahoo.com, # e-mail: sawiwo@kku.ac.th


Abstract

The objective of this experiment was to detemine the effects of dietary inclusion of Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®) on growth performance and morphology of duodenum, jejunum, and ileum. 50 seven-day-old broiler chicks (Arber Acres) of both sexes were used for 42 days in battery cage. The chicks were randomly allocated to five treatments (T) with two replications, five chicks per replication according to completely randomized design and assigned to receive one of five dietary treatments including T1 (Control), T2 (0.50 kg/ton), T3 (1.00 kg/ton), T4 (1.50 kg/ton), and T5 (2.00 kg/ton). The results rvealed that final body weight, body weight gain, feed intake, feed conversion ratio and productive index were improved (P>0.05) in four graded levels of Mu-Plus® fed birds over the control group. The economic benefit returns in term of return on investment (ROI) on salable bird return from feed cost showed higher when compare to the control group. The survival rate of broilers was not significantly different (P>0.05) between treatment. There was a highly significant (P<0.01) increase villi height, crypt depth and villi height: crypt depth ratio of duodenum, jejunum and ileum of four graded levels of Mu-Plus® fed birds when compare to the control group. Additionally, dietary inclusion of Mu-Plus® at 2.00 kg/ton diet showed to improve growth performance, economic benefit return and villi height of broilers.


Key words: Duodenum, Jejunum, Ileum, Andrographis paniculata, Zingiber cassumunar
Introduction

Nowadays, Andrographis paniculata and Zingiber cassumunar are widely used as an animal growth promotion in the poultry industry and production. The combination mixture of herbs are being actively research and showed better efficacy response to improve feed consumption, feed efficiency, body weight gain, and survival rate [1, 2, 3, 4, 5]. Both herbs feed additive can be considered by their important function cover on pathogen resistance within antimicrobial, antibacterial and antifungal activity, antioxidant activity in control of metabolic auto-oxidation, hepatoprotective activity within mycotixin tolerance and liver function, anti-inflammatory activity by prevent small intestinal enterocyte cell and crypt of lieberkuhn, and digestive donor within stimulation of endogenous enzyme activity and nitrogen absorption as well as beneficial physiological and morphological effects [1, 6, 7, 8, 9]. Intestinal histomorphology correlates markedly affected by dietary feed components and the histomorphological changes in the intestine with intestinal function [10, 11, 12]. Assessment of intestinal villi morphology is important because the small intestine is a primary site for nutrient digestion and absorption [13]. Measuring villi height, crypt depth and villi height: crypt depth ratio are the way of investigating intestinal morphology and function [14]. Therefore, the study was investigated the effects of dietary Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®) on growth performance and morphology of duodenum, jejunum and ileum in broiler chickens.



Materials and Methods

A total of 50 seven-day-old birds (Arber Acres) were weighted and separated from the floor-pen open house to battery cage under a natural tropical condition with the same housing and general management practices. Birds were randomly allotted to five dietary treatments, two replicatications and each replication consist five birds.



The Basal diet was supplemented with Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®). Diet was divided to five treatments (T), T1 (control group), T2, T3, T4 and T5 with Mu-Plus® at 0.50, 1.00, 1.50 and 2.00 kg/ton diet, respectively. Moreover, diets in mash form were separated into three periods; starter (CP = 25.51 % and ME = 4,468.82 kcal/kg), grower (CP = 22.87% and ME = 4,471.55 kcal/kg) and finisher (CP = 19.36 % and ME = 4,572.64 kcal) (Table 1) consisting of the same ingredient in the diet and it meets the bird requirement [15]. All birds were allowed ad libitum access to water and feed throughout the experimental periods. Each diet was analyzed as described method in [16].


Table 1 Composition of the basal experimental diets of broiler chickens

Ingredients (%)

Starter

Grower

Finisher

Yellow corn (7.8 % CP)

47.1

53.24

59.17

Soybean meal (44% CP)

28

19.7

14

Full-fat soy bean (36 % CP)

18

20

20

Monocalcium phosphate (P21)

2.9

2.9

2.38

Limestone

1.4

1.4

1.3

DL-methionine

0.3

0.22

0.17

L-lysine

0.15

0.21

0.15

Crude rice bran oil

1.3

1.5

2

Salt

0.4

0.4

0.4

Choline Chloride 60%

0.1

0.08

0.08

Vitamin-mineral mixes a

0.35

0.35

0.35

Nutrient composition (calculated)

Crude protein (%)

25.51

22.87

19.36

ME (kcal/kg)

4,468.82

4,471.55

4,572.64

Calcium (%)

1.12

1.13

0.98

Avai. phosphorus (%)

0.52

0.53

0.45

Lysine (%)

1.42

1.30

1.12

Methionine (%)

0.67

0.56

0.49

Methionine and cystine (%)

1.06

0.91

0.81

Tryptophan (%)

0.29

0.25

0.22

Threonine (%)

0.90

0.80

0.72

a Vitamin-mineral mix supplied per kilogram of diet: vitamin A, 10,000 IU; vitamin D3, 2,500 IU; vitamin E, 10 IU; vitamin K3, 2.50 mg; thiamine, 2.0 mg; riboflavin, 7.0 mg; pyridoxine, 5.0 mg; vitamin B12, 30 mcg; d-pantothenic acid, 12 mg; niacin, 50 mg; choline, 1200 mg; folic acid, 1 mg; biotin, 15 mcg; ethoxyquin, 125 mg; Mn, 60 mg; Zn, 40 mg; Fe, 80 mg; Cu, 8 mg; I, 0.35 mg; Se, 0.15 mg.

Body weight, body weight gain, feed intake and feed conversion ratio were weighed and calculated for evaluating the productive performance at period intervals. Mortality was daily recoeded while occurrence. At termination 42 days of age, one-male and one-female chickens from each treatment were randomly sampled and serial euthanized immediately after the killing, the whole of small intestine was separated and various parts of small intestine including duodenum (from the gizzard outlet at the end of pancreas loop), jejunum (segment between pancreatic loop and the Meckel’s diverticulum) and ileum (segment between Meckel’s diverticulum and ileocaecal junction) removed. About 3.5 cm tissue segment, from midpoint of duodenum, midpoint of jejunum, and midpoint of ileum for measure villi height and crypt depth, were removed and immediately flushed with physical saline and fixed into a 10 % neutral buffer formalin solution until further processing. After embedding the samples in paraffin, 5µm tissue section of each sample placed on a glass slide and then stained with hematoxylin and eosin. A total of 25 villi and 25 crypts per sample were measured by using a light microscope. Villi height from the tip of the villi to the villi crypt and crypt depth defined as the depth of the invagination between adjacent villi.



Statistic Analysis

The experiment data were subjected to analyze with the ANOVA procedure of SAS program and significant differences between the treatments mean were determined to Duncan’s multiple range test (DMRT) [17]. The level of significance was considerate at P<0.05.



Results and discussion

Chemical composition of dietary treatment

The feed ingredients used in the experiment are same among groups, but it is different only the level of Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®). The percent chemical composition analyzed of the dietary treatments are similar in all treatments (Table 1).



Produvtive performance of broilers

Growth performance on body weight (BW), body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), survival ratio (SR), productive index (PI), feed cost per gain (FCG) of broilers were computed and analyzed for three cycle periods (day 7-21, day 22-35 and day 36-42) and overall period (day 7-42) are presented in Table 2. Broilers fed with Mu-Plus® showed both greater (P>0.05) body weight and body weight gain in all periods. Similar results were found by [2], [3], [18], [19] and [20] who reported that Andrographis paniculata fed broilers improved to increase in live weight of broilers. [4] and [21] also reported that Zingiber cassumunar fed broilers had a greater in body weight gain. [22] also reported that Andrographis paniculata fed sows showed a great daily gain when compared to the control group. There was improve (P>0.05) to increase feed efficiency after fed with four graded levels of Mu-Plus® for three cycle periods and overall periods when compared with the control no added Mu-Plus® group. However, in finisher period, there was significantly different (P<0.01) by level of concentrate 0.5 kg/ton diet leaded to the lower productive index within the higher feed cost per gain when compared to the control group. [23], [24], [25] and [26] reported that animals fed with Andrographis paniculata, Zingiber cassumunar and Curcuma longa mixture diet was decrease feed intake and feed conversion ratio. Otherwise, [18], [19] and [20] also reported that Andrographis paniculata fed broilers showed to decrease feed consumption and feed conversion ratio. [27] reported that feeding Andrographis paniculata was give positive response to decrease feed consumption of laying hens.

Feeding Mu-Plus® with four levels in control diet showed higher economic benefit returns (Table 3). The results from this study revealed that the economic benefit returns on salable bird return from feed cost showed higher when compared with the control no added group. Additionally, four graded levels of Mu-Plus® showed higher return on investment (ROI) when compared to the control group and showed the highest of ROI when added at the level of 2.00 kg/ton diet.

Table 2 Effects of Mu-Plus® in the diet of broilers on growth performance, productive index and feed cost per gain (FCG)

Treatment

Mu-Plus®

(kg/ton)


BW

(g)


BWG

(g)


FI

(g)


FCR


SR

(%)


PI1

FCG2

Baht/kgBW



7-21days

T1


T2

T3

T4



T5

SEM


Contrast

Lin


Quad

Cubic


Quar

-

0.50



1.00

1.50


2.00

742.00


774.00

749.75


750.00

766.63


28.244
NS

NS
NS

NS

618.90


647.50

622.45


625.10

642.13


26.956
NS

NS

NS



NS

923.88


915.93

876.75


923.88

937.35


46.909
NS

NS

NS


NS

1.49


1.41

1.41


1.48

1.46


0.100
NS

NS

NS



NS

100


100

100


100

100


-
-

-

-



-

296.67


328.21

315.81


302.63

315.38


31.754
NS

NS

NS



NS

21.21


20.38

20.58


21.90

21.91


1.443
NS

NS

NS



NS

22-35 days

T1


T2

T3

T4



T5

SEM


Contrast

Lin


Quad

Cubic


Quar

-

0.50



1.00

1.50


2.00

1795.50


1866.00

1843.70


1844.50

1860.00


124.906
NS

NS
NS

NS

1053.00


1091.50

1093.70


1094.50

1093.00


117.706
NS

NS

NS



NS

2120.80


2022.50

1909.00


2003.80

2042.00


159.762
NS

NS
NS

NS

2.02


1.89

1.74


1.83

1.87


0.188
NS

NS

NS



NS

100


100

100


100

100


-
-

-

-



-

373.52


430.04

448.38


427.45

417.99


80.709
NS

NS

NS



NS

28.27


26.86

25.09


26.79

27.65


2.690
NS

NS

NS



NS

36-42 days

T1


T2

T3

T4



T5

SEM


Contrast

Lin


Quad

Cubic


Quar

-

0.50



1.00

1.50


2.00

2330.80


2237.00

2349.50


2330.50

2371.80


165.917
NS

NS

NS



NS

535.25a

371.00b

505.84ab

486.00ab

511.75ab

54.925
NS

NS

*



NS

1283.80


1178.50

1205.00


1155.30

1199.00


100.870
NS

NS

NS



NS

2.41b

3.18a

2.39b

2.38b

2.34b

0.110
*

*

**



**

100


100

100


100

100


-
-

-

-



-

319.72a

166.85b

303.83a

292.12a

312.24a

43.832
NS

NS

*



NS

33.18b

44.41a

33.91b

34.20b

34.07b

1.544
*

*

**



**

7-42 days

T1


T2

T3

T4



T5

SEM


Contrast

Lin


Quad

Cubic


Quar

-

0.50



1.00

1.50


2.00

2330.80


2237.00

2349.50


2330.50

2371.80


165.917
NS

NS

NS



NS

2207.70


2110.50

2222.20


2205.60

2247.30


169.102
NS

NS

NS



NS

4328.50


4116.90

3990.80


4083.00

4178.40


250.896
NS

NS

NS



NS

1.96


1.95

1.79


1.85

1.86


0.08
NS

NS

NS



NS

100


100

100


100

100


-
-

-

-



-

321.73


310.37

353.58


340.65

345.44


35.936
NS

NS

NS



NS

27.44


27.79

25.80


27.04

27.46


1.254
NS

NS

NS



NS

a,b Values in the same column with a common letter are significantly different at P<0.05

* = P<0.05, ** = P<0.01, NS = non significant



1 Productive index (PI) = (BWG x Survival rate) ÷(Age x FCR),

2 Feed cost per gain (FCG) = (( FI x feed cost) ÷ (Survival x BWG)) x 100



Table 3 Effects of Mu-Plus® in the diet of broilers on economic benefits returns for overall period (42days of testing)


Treatments

Mu-Plus®

Kg/ton


FCG1

Baht/bird



FCG2

Baht/kgBW



SBR

Baht/bird



NPR1

Baht/bird



NPR2

Baht/kgBW



ROI1

Baht/bird



ROI2

Baht/BW


T1

T2

T3



T4

T5

SEM



Contrast

Lin


Quad

Cubic


Quar

-

0.50


1.00

1.50


2.00

60.55

58.42


57.38

59.54


61.73

3.582
NS

NS

NS

NS



27.44

27.79


25.80

27.04


27.46

1.254
NS

NS

NS

NS



88.57

85.00


89.28

88.56


90.12

6.304
NS

NS

NS

NS



28.01

26.58


31.89

29.02


28.40

3.928
NS

NS

NS

NS



12.01

11.80


13.59

12.41


11.97

1.095
NS

NS

NS

NS



-

-1.43


+3.88

+1.00


+0.38

-
-


-

-

-



-

-0.22


+1.58

+0.40


-0.04

-
-


-

-

-



Feed cost per gain (FCG1) = ((FI x feed cost, FC) ÷ Survival)x 100

Feed cost per gain (FCG2) = FCG1 ÷ BWG

Salable bird return (SBR) = Price of live chicken (38 Baht) x BW

Net profits return per bird (NPR)

NPR1 = SBR – FCG1

NPR2 = NPR1 ÷ BW

Return on investment by comparing with the control group(ROI)

ROI1 = NPR1 (added Mu-Plus®) – NPR1 (control)

ROI2 = NPR2 (added Mu-Plus®) – NPR2 (control)


Small intestinal morphology

Villi height, crypt depth and villi height: crypt depth ratio of duodenum, jejunum, and ileum at termination of age at day 42 are presented in Table 4. The results from this study reveal that the duodenal, jujenal and ileal villi height was significantly different (P<0.01) between treatment. In addition, four graded levels of Mu-Plus® showed higher in villi height when compared to the control group and showed the highest of villi height when supplemented at a level of 2.00 kg/ton diet. This finding is in agreement with [28] who reported that broilers fed with Andrographis paniculata, Curcumin longa, Momordica charantia and Zingiber cassumunar mixture diets led to increase (P<0.01) the villi height of duodenum and jejunum when compared to the control group. On one hand, the duodenal, jejunal and ileal crypt depth were significantly different (P<0.05) between treatment. These results are contrasted from a report made by [24] who reported that Mu-Plus® supplementation had no any effect on duodenal, jejunal and ileal villi height of weaning pigs when compared to the control group. [25] also reported that weaning pigs that receive Andrographis paniculata, Zingiber cassumunar, Momordica charantia and Curcuma longa mixture diet did not show any effect on the villi height of duodenum, jejunum and ileum when compare to the control group. However, the results from this trial showed that in all small intestinal segments of four graded levels of Mu-Plus® showed higher in crypt depth and villi height: crypt depth ratio when compared to the control group. It had mentioned that a crypt depth is indicative of a faster tissue turnover and a higher demand for new tissue. Furthermore, It also considered that a villi height: crypt depth ratio is associate with well-developed intestinal mucosa with high digestion and absorption [14, 29, 30, 31]. The response of intestinal microstructure to dietary or lumen factors may be affected by battery cage stress, dietary Mu-Plus® level inclusion and age of bird when taking tissue samples for histomorphology. Therefore, it is necessary to determine the optimum dietary inclusion level of Mu-Plus® in poultry diets.





Table 4 Effects of Mu-Plus® in the diet of broilers on the villi and crypt of duodenum, jejunum and ileum at 42 days of age

Treatment

Duodenum

Jejunum

Ileum

Mu-Plus® (Kg/ton)

VH

CD

VH:CD

VH

CD

VH:CD

VH

CD

VH:CD

T1 -

T2 0.50


T3 1.00

T4 1.50


T5 2.00

SEM


Contrast

Lin


Quad

Cubic


Quar

1,012.53bc

925.22c

1,037.77bc

1,222.43ab

1,371.4a

210.23
**

NS

NS

NS



90.70

94.10


86.83

94.48

89.99


16.34
NS

NS

NS



NS

11.20b

10.65b

12.08b

13.08ab

15.47a

2.84
**

NS

NS

NS



810.57c

543.17d

594.23d

1,314.61a

1,049.48b

149.39
**

**

**

**



85.32b

73.90c

78.92bc

86.43ab

97.01a

8.98
**

**

NS

NS



9.54bc

7.41c

7.68c

15.33a

10.99b

2.11
**

NS

**

**



491.56c

408.29d

673.37a

590.56b

499.04c

72.09
*

**

**

**



79.75a

65.79c

76.05ab

76.86ab

70.56bc

5.86
NS

NS

**

NS



6.14c

6.39c

8.87a

7.96ab

7.23bc

1.11
**

**

NS

*



a, b, c, d Values in the same column means with different superscripts differ significantly at P <0.05.

* = P<0.05, ** = P<0.01, NS = non significant, VH = Villi height, CD = Crypt depth




Conclusions

The results of this study clearly showed that dietary inclusion four graded level of Mu-Plus® in the diet had improved growth performance, productive index and economic benefit returns in term of return on investment. Dietary supplemtation four graded levels of Mu-Plus® had influenced on intestinal duodenal, jejunal and ileal villi morphology. Additionally, dietary inclusion Mu-Plus® at 2.00 kg/ton diet level showed potential to enhance broilers growth performance small intestinal morphology and economic benefit returns.



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Acknowledgements

The authors express their appreciation to Lily FoodAnSci LTD, Thailand for providing Mu-Plus® and funds for this project.


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