Effects of Andrographis Paniculata and Zingiber Cassumunar Mixture in the Diets on Productive...

EFFECTS OF ANDROGRAPHIS PANICULATA AND ZINGIBER

CASSUMUNAR MIXTURE IN THE DIETS ON PRODUCTIVE

PERFORMANCE, CARCASS QUALITY, NUTRIENTS

DIGESTIBILITY AND INTESTINAL

HISTOMORPHOLOGY IN

BROILER CHICKENS

MR. DANET LAING

A THESIS FOR THE DEGREE OF MASTER OF SCIENCE

KHON KAEN UNIVERSITY

2014





HISTOMORPHOLOGY IN

BROILER CHICKENS

MR. DANET LAING

A THESIS FOR THE DEGREE OF MASTER OF SCIENCE


2014





HISTOMORPHOLOGY IN

BROILER CHICKENS

MR. DANET LAING

A THESIS SUBMITTED IN PARTIAL FULFILLMENT OF THE REQUIREMENTS FOR THE DEGREE OF MASTER OF SCIENCE

IN ANIMAL SCIENCE GRADUATE SCHOOL KHON KAEN UNIVERSITY

2014

THESIS APPROVAL


FOR

MASTER OF SCIENCE

IN ANIMAL SCIENCE

Thesis Title: Effects of Andrographis paniculata and Zingiber cassumunar mixture

in the diets on productive performance, carcass quality, nutrients

digestibility and intestinal histomorphology in broiler chickens

Author: Mr. Danet Laing

Thesis Examination Committee: Asst. Prof. Dr. Winai Jaikan

Assoc. Prof. Dr. Supaporn Isariyodom Member

Dr. Sawitree Wongtangtintharn Member

Assoc. Prof. Dr. Jowaman Khajarern Member

Assoc. Prof. Dr. Bundit Tengjaroenkul Member

Thesis Advisors:

……………………………………… Advisor

(Dr. Sawitree Wongtangtintharn)

……………………………………… Co-Advisor

(Assoc. Prof. Dr. Jowaman Khajarern)

……………………………………… Co-Advisor

(Assoc. Prof. Dr. Bundit Tengjaroenkul)

………………………………… … ……………………………………..

(Assoc. Prof. Dr. Lampang Manmart) (Assoc. Prof. Dr. Monchai Duangjinda)

Dean, Graduate School Dean, Faculty of Agriculture

Copyright of Khon Kaen University

Chairperson

ดาเนตร ลง. 2557. ประสทธภาพการเสรมฟาทะลายโจรรวมกบไพล ในอาหารตอสมรรถนะการ

ผลต คณภาพซาก การยอยไดของโภชนะ และสณฐานวทยาของลาไสในไกเน-อ. วทยานพนธปรญญาวทยาศาสตรมหาบณฑต สาขาวชาสตวศาสตร บณฑตวทยาลย มหาวทยาลยขอน

อาจารยท1ปรกษาวทยานพนธ: ดร. สาวตร วงศต งถ"นฐาน, รศ. ดร. เยาวมาลย คาเจรญ, รศ. ดร. บณฑตย เตงเจรญกล

บทคดยอ

การศกษาคร งน มวตถประสงคเพ"อศกษาการเสรมสมนไพรฟาทะลายโจร (Andrographis paniculata) รวมกบไพล (Zingiber cassumunar) (Mu-Plus®) ในอาหารตอสมรรถนะการผลต (growth performance), คณภาพซาก (carcass quality) การยอยไดของโภชนะ (nutrients digestibility) และสญฐานวทยาของลาไส (intestinal histomorphology) ในไกเน อ โดยแบงการทดลองออกเปน 3 การทดลอง ดงน

การทดลองท" 1 ศกษาผลของการเสรมสมนไพรฟาทะลายโจรรวมกบไพล (Mu-Plus®) ในอาหารท"ระดบ 0.00 (T1, กลมควบคม), 0.50 (T2) 1.00 (T3), 1.50 (T4) และ 2.00 กก./ตน (T5) ตอสมรรถนะการผลต คณภาพของซาก และองคประกอบของเน ออก (breast) และตบ (liver) ของไกเน อโดยใชลกไกพนธ อารเบอร เอเคอร (Arbor Acres) คละเพศ อาย 1 วน 480 ตว แบงเปน 5 กลม กลมละ 4 ซ า ซ าละ 24 ตว เพศผ 12 ตว เพศเมย 12 ตว วางแผนการทดลองแบบสมสมบรณ (Completely Randomized Design: CRD) แบงระยะการเล ยงเปน 3 ระยะตามสตรอาหาร ไกเลก อาย 1-21 วน (starter) มปรมาณ CP = 22.47 % และ ME = 3,102 kcal/kg;ไกรน อาย 22-35 วน (grower) มปรมาณ CP = 20.02 % และ ME = 3,155 kcal/kg และไกใหญ อาย 36-42 วน (finisher) มปรมาณ CP = 17.98 % และ ME = 3,228 kcal/kg ในดานสมรรถนะการผลต พบวา น าหนกตวสดทาย (final body weight, FBW) น าหนกตวท"เพ"มข น (body weight gain, BWG) ปรมาณการกนได (feed intake, FI) ประสทธภาพการใชอาหาร (feed conversion ratio, FCR) และดชนประสทธภาพการผลตไกเน อ (productive index, PI) ไมแตกตางกนทางสถต (P>0.05) ในแตละกลมการทดลอง แตในกลมเสรม Mu-Plus® ท ง 4 ระดบ พบวา สามารถปรบปรงน าหนกตวท"เพ"มข น ประสทธภาพการใชอาหาร และดชนประสทธภาพการผลตไกเน อเพ"มข น เม"อเปรยบเทยบกบกลมควบคม ในกลมเสรม Mu-Plus® ท"ระดบ 0.50 กก./ตน สงผลตอ น าหนกตวท"เพ"มข น ประสทธภาพการใชอาหาร อตราการเล ยงรอด และดชนประสทธภาพการผลตสง ในดานผลตอบแทนทาง

แกน.

ii

เศรษฐกจ พบวา สงผลตอผลตอบแทนเพ"มข น เม"อเปรยบเทยบกบกลมควบคม การเสรม Mu-Plus® ท ง 4 ระดบ พบวา ผลตอบแทนการลงทน (returns of investment, ROI) เพ"มข น เม"อเปรยบเทยบกบกลมควบคม นอกจากน ยงพบวา กลมเสรม Mu-Plus® ท"ระดบ 0.50 กก./ตน ในอาหาร สงผลตอผลตอบแทนการลงทนสงสด เม"อเปรยบเทยบกบกลมอ"น

สาหรบผลตอคณภาพซาก พบวา การเสรม Mu-Plus® ในอาหาร สงผลไมแตกตางกนทาง สถต (P>0.05) ตอน าหนกซาก (carcass weight) อวยวะภายใน (organ weights) มาม (spleen) ไทมส(thymus) และเบอรซา ออฟฟาบรเชยส (bursa of fabricius) เม"อเปรยบเทยบกบกลมควบคม อยางไรกตาม ในกลมเสรม Mu-Plus® ท"ระดบ 0.50 กก./ตน ในอาหาร พบวา สงผลตอเปอรเซนซาก (dressing percentage) และเน อเพ"อบรโภคท งหมด (total edible meat) เพ"มข น และสงผลตอ ปรมาณไขมนชองทอง (abdominal fat) ลดลง นอกจากน ยงพบวา การเสรม Mu-Plus® ท ง 4 ระดบ สงผลตอไขมนในเน ออก และตบลดลง เม"อเปรยบเทยบกบกลมควบคม

การทดลองท" 2 ศกษาผลของการเสรมสมนไพรฟาทะลายโจรรวมกบไพล (Mu-Plus®) ในอาหารท"ระดบ 0.00 (T1, กลมควบคม), 0.50 (T2) 1.00 (T3), 1.50 (T4) และ 2.00 กก./ตน (T5) ตอประสทธภาพการยอยไดของวตถแหง (dry matter) โปรตนหยาบ (crude protein) ไขมน (fat) และพลงงานรวม (gross energy) ในไกเน อ อาย 19-21 วน, 33-35 วน และ 40-42 วนโดยใชลกไก พนธ อารเบอร เอเคอร (Arbor Acres) อาย 7 วน 60 ตว แบงเปน 5 กลม กลมละ 4 ซ า ซ าละ 3 ตว วางแผนการทดลองแบบสมสมบรณ (Completely Randomized Design: CRD) โดยการเล ยงไกบนกรงแบตเตอร" ผลการทดลอง พบวา การยอยไดของวตถแหง โปรตนหยาบ ไขมน และพลงงาน รวม เพ"มข น (P<0.01) ในระยะไกรน และไกใหญ นอกจากน ยงพบวา การเสรม Mu-Plus® ท"ระดบ 0.50 กก./ตน ในอาหาร สงผลตอการยอยไดของโภชนะ (nutrients digestibility) สงสด โดยวดจาก การยอยไดของวตถแหงโปรตนหยาบ ไขมน และพลงงานรวม เม"อเปรยบเทยบกบกลมควบคม และกลมเสรม Mu-Plus® ท"ระดบแตกตางกน

การทดลองท" 3 ศกษาผลของการเสรมสมนไพรฟาทะลายโจรรวมกบไพล (Mu-Plus®) ในอาหารท"ระดบ 0.00 (T1, กลมควบคม), 0.50 (T2) 1. 00 (T3), 1.50 (T4) และ 2.00 กก./ตน (T5) ตอลกษณะทางสณฐานวทยาของลาไสสวนตน (duodenum) สวนกลาง (jejunum) และสวนปลาย (ileum) ในไกเน อ อาย 14, 21, 35 และ 42 วน โดยใชลกไกพนธ อารเบอร เอเคอร (Arbor Acres) คละเพศ อาย 7 วน 50 ตว แบงเปน 5 กลม กลมละ 2 ซ า ซ าละ 5 ตว วางแผนการ ทดลองแบบสมสมบรณ (Completely Randomized Design: CRD) โดยการเล ยงไกบนกรงแบตเตอร" ผลการทดลอง พบวาการเสรม Mu-Plus® ท ง 4 ระดบ สงผลตอความสงของวลไล (villi) ในลาไส สวนตน กลาง และปลาย เพ"มข นในทกชวงอาย เม"อเปรยบเทยบกบกลมควบคม ในสวนของ ความลกครปท (crypt

iii

depth) ในลาไสสวนตน กลาง และปลาย มผลแตกตางกนในทางสถต (P<0.01) นอกจากน ยงพบวา การเสรม Mu-Plus® สงผลตออตราสวนของความสงวลไลตอความลกครปท (villi height: crypt depth ratio) ในลาไสสวนตน กลาง และปลาย เพ"มข น (P<0.01) เม"อเปรยบเทยบ กบกลมควบคม

จากผลการศกษา พบวาการเสรม Mu-Plus® ในอาหารไกเน อมประสทธภาพในการเพ"ม สมรรถนะการผลต ประสทธภาพการใชอาหาร ประสทธภาพการผลต การยอยไดของโภชนะ ความสงวลไล และอตราการเล ยงรอดของไกเน อสงกวากลมควบคม โดยเฉพาะอตราการเล ยงรอดในระยะไกใหญมคาสงสด ซ" งอาจเปนผลเน"องจากสวนประกอบของเทอรพนอยด (terpenoids) ของสมนไพรท ง 2 ชนดชวยเพ"มการยอยไดของไขมน โดยชวยเพ"มการหล"งน าด และกระตนการหล"งการทางาน ของเอนไซม (endogenous enzymes) ในรางกาย ไดแก ไลเปส (lipase) อะไมเลส (amylase) และซเครส (sucrase) เปนตน และระดบการเสรม Mu-Plus® ท"เหมาะสมในอาหารไกเน อ คอ 0.50 กก./ตน

Danet Laing. 2014. Effects of Andrographis paniculata and Zingiber cassumunar

mixture in the diets on productive performance, carcass quality, nutrients

digestibility and intestinal histomorphology in broiler chickens. Master of

Science Thesis in Animal Science, Graduate School, Khon Kaen University.

Thesis Advisors: Dr. Sawitree Wongtangtintharn,

Assoc. Prof. Dr. Jowaman Khajarern,

Assoc. Prof. Dr. Bundit Tengjaroenkul

ABSTRACT

The purposes of the present study were to evaluate the effects of dietary

Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®) on productive

performance, carcass quality, nutrients digestibility and intestinal histomorphology in

broiler chickens. The study was divided into 3 experiments.

Experiment I. Aimed to determine the effects of Andrographis paniculata and

Zingiber cassumunar mixture at 0.00 (Control, T1), 0.50 (T2), 1.00 (T3), 1.50 (T4)

and 2.00 kg/ton diet (T5) on productive performance, carcass quality, and

composition of breast meat and liver of broilers. A total of 480 d-old commercial

Arbor Acres chicks were designed under completely randomized design (CRD), and

all of the birds were allotted to five dietary treatments with four replications and 24

birds (twelve males and twelve females) per each. The experimental diet was divided

to three phases: starter (1-21 d of age containing CP = 22.47% and ME = 3,102

kcal/kg), grower (22-35 d of age containing CP = 20.02% and ME = 3,155 kcal/kg)

and finisher (36-42 d of age containing CP = 17.98% and ME = 3,228 kcal/kg).

Growth performance on final body weight (FBW), body weight gain (BWG), feed

intake (FI), feed conversion ratio (FCR) and productive index (PI) were not

significantly different (P>0.05) among treatment groups. However, four graded levels

of Mu-Plus® showed improvement BWG, FCR and PI when compared with the

control group. Additionally, supplementation of Mu-plus® at 0.50 kg/ton diet showed

the highest BWG and PI with the lowest FCR. Survival rate (SR) was significant

increased (P<0.05) by increasing the levels of Mu-plus® supplementation in finisher

period, and the concentration of Mu-plus® at 0.50 kg/ton diet showed the highest SR.

v

The economic benefit returns on salable bird return from feed cost showed higher

when compared with the control group. Furthermore, four graded levels of Mu-Plus®

showed higher returns of investment (ROI) when compared to the control group and

showed the highest of ROI when added at the level of 0.50 kg/ton diet.

On carcass quality, Mu-plus® supplementation did not show any effects

(P>0.05) on carcass weights, organ weights, spleen, thymus and bursa of fabricius

when compared with the control group. However, Mu-plus® at 0.50 kg/ton diet trend

to give the higher in dressing percentage and total edible meat with the lowest in

abdominal fat and showed significant decrease (P<0.05) in the fat composition of

breast meat and liver of four graded levels of Mu-plus® fed broilers when compared

with the control group.

Experiment II. Aimed to determine the effects of Andrographis paniculata and


and 2.00 kg/ton diet (T5) on dry matter (DM), crude protein (CP), fat and gross

energy (GE) digestibility of broilers at d 19-21, d 33-35 and d 40-42. A total of 60

seven-d-old commercial Arbor Acres chicks were designed under completely

randomized design (CRD) in battery cages, and all of the birds were allotted to five

dietary treatments with four replications and three birds per each. The results

indicated that digestion of DM, CP, fat and GE were highly significant differences

(P<0.01) in grower and finisher periods. Additionally, Mu-Plus® at 0.50 kg/ton diet

showed the highest nutrients digestibility in terms of DM, CP, fat and GE when

compared to the control group and the other Mu-Plus® fed groups.

Experiment III. Aimed to determine the effects of Andrographis paniculata and


and 2.00 kg/ton diet (T5) on histomorphology of duodenum, jejunum and ileum of

broilers at d 14, d 21, d 35 and d 42. A total of 50 seven-d-old commercial Arbor

Acres chicks were designed under completely randomized design (CRD) in battery

cages, and all of the birds were allotted to five dietary treatments with two

replications and five birds per each. It indicated that the four graded levels of

Mu-Plus®-fed broilers had shown to increase (P<0.01) the villi height of duodenum,

jejunum and ileum in all periods when compared with the control group. Crypt depth

of duodenum, jejunum and ileum was a highly significant difference (P<0.01) among

vi

treatment groups. Moreover, there was also significant enhance (P<0.01) villi height:

crypt depth ratio of duodenum, jejunum and ileum higher than the control group.

The results from the present study suggested that dietary supplementation with

Mu-Plus® enhanced productive performance, FCR, PI, nutrients digestion, villi height

and SR of broilers over the control group. Especially, SR had the highest in finisher

period. This may be caused by terpenoids compound of both herbs enhance to digest

dietary fat including bile secretion and/or stimulate the endogenous enzyme activity

such as lipase, amylase, sucrase, etc. The optimum supplementing level of Mu-Plus®

was suggested at 0.50 kg/ton diet of broiler chickens.

The Present Thesis is Greatly Dedicated to Her Royal Highness

Princess Maha Chakri Sirindhorn, my Parents

and the Entire Teaching Staffs

ACKNOWLEDGEMENTS

Firstly, I would like to pay my highest respect and gratitude to Her Royal

Highness Princess Maha Chakri Sirindhorn, who awarded me the scholarship for

studying in M.Sc degree level at Khon Kaen University (KKU). Secondly and the

most is to my major advisor, Dr. Sawitree Wongtangtintharn, who I wish to express

my deepest and most personal sincere gratefulness. During my study, she has been

giving many useful continuous advices and supports throughout the course of my

study. Moreover, her encouragement, criticism, excellent skilled technique, assistance

and guidance were gratefully acknowledged. Next, I would like to express my

appreciation to my co-advisor, Assoc. Prof. Dr. Jowaman Khajarern and Assoc. Prof.

Dr. Bundit Tengjaroenkul, for their kind advices and comments during the course of

my study and in the preparation of the thesis. I also would like to say many thanks to

all examination committees for their suggestion and recommendation on this thesis

writing. I also wish to express my deepest gratitude to Her Royal Highness Princess

Maha Chakri Sirindhorn Project for providing full scholarship in my study and Lily

Food Animal Science Ltd for providing financial support to my research work.

Furthermore, I would like to thank Khon Kaen University and Department of Animal

Science, non-ruminant nutrition team for their facility supports both in course and

research work. In addition, I would like to extend words of thanks to all staff

members, M.Sc and Ph.D students under non-ruminant nutrition team for their great

help and encouragements. Furthermore, I would like to express my deepest gratitude

to my parents, brother, and sister for their understanding and moral support during my

study as well as those anonymous people whose names could not be appeared here,

but deeply in my heart, for their continuous encouragement and support.

Danet Laing

TABLE OF CONTENTS

Page

ABSTRACT (IN THAI) i

ABSTRACT (IN ENGLISH) iv

DEDICATION vii

ACKNOWLEDGEMENTS viii

LIST OF TABLES xiii

LIST OF FIGURES xv

LIST OF ABBREVIATIONS xvi

CHAPTER I INTRODUCTION 1

1.1 Background 1

1.2 Hypotheses 2

1.3 Objective of the research 3

1.4 Expected results 3

CHAPTER II LITERATURE REVIEW 4

2.1 Androgaphis paniculata 4

2.1.1 Botanical characteristics of Androgaphis paniculata 4

2.1.2 Utilization of Andrographis paniculata 5

2.1.3 Pharmacological action of Androgaphis paniculata 5

2.1.4 The chemical composition of Androgaphis paniculata 9

2.2 Zingiber cassumunar 11

2.2.1 Botanical characteristics of Zingiber cassumunar 11

2.2.2 Utilization of Zingiber cassumunar 12

2.2.3 Pharmacological action of Zingiber cassumunar 13

2.2.4 The chemical composition of Zingiber cassumunar 15

2.3 Utilization of Andrographis paniculata and Zingiber cassumunar

mixture in animal feed 17

2.4 Characteristics of small intestine of broiler chickens 20

x

TABLE OF CONTENTS (Cont.)

Page

CHAPTER III MATERIALS AND METHODS 24

3.1 Experiment I 24

3.1.1 Experimental unit 24

3.1.2 Experimental design 24

3.1.3 Research methodology 25

3.1.4 Data record 25

3.2 Experiment II 27


3.2.2 Diets formulation 27



3.3 Experiment III 28


3.3.2 Diets formulation 28



3.4 Analysis data 29

3.5 Location of research 29

CHAPTER IV RESULTS AND DISCUSSION 35

4.1 Experiment I 35

4.1.1 Nutritive values of dietary diet at 1-21 d,

22-35 d, 36-42 d of age 35

4.1.2 Effect of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on growth performance of

broilers 37

4.1.3 Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on feed cost and economic

benefit returns 47

xi


Page

4.1.4 Effect of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on carcass quality of

broilers 51

4.2 Experiment II 56

4.2.1 Effect of Andrographis paniculata and Zingiber

cassumunar mixture (Mu-Plus®) in the diets on dry matter

digestibility (DMD) of broiler chickens 56


cassumunar mixture (Mu-Plus®) in the diets on crude

protein digestibility (CPD) of broiler chickens 57


cassumunar mixture (Mu-Plus®) in the diets on ether

extract digestibility (EED) of broiler chickens 59


cassumunar mixture (Mu-Plus®) in the diets on gross

energy digestibility (GED) of broiler chickens 60

4.3 Experiment III 65


cassumunar mixture (Mu-Plus®) in the diets of broiler

chickens on the villi height 65



chickens on the villi width 68



chickens on crypt depth 70

xii


Page



chickens on the ratio of villi height: crypt of liberkhun depth 73

CHAPTER V CONCLUSION AND RECOMMENDATION 76

5.1 Conclusion 76

5.2 Recommendation and suggestion 77

REFERENCES 78

APPENDIX 90

CURRICULUM VITAE 103

LIST OF TABLES

Page

Table 2.1 Chemical constituents and bioactivities of Andrographis paniculata 10

Table 2.2 Small intestinal villi morphology of broiler chicken 22

Table 3.1 Composition of the experimental diets for starter period 30

Table 3.2 Composition of the experimental diets for grower period 31

Table 3.3 Composition of the experimental diets for finisher period 32

Table 3.4 Composition of vitamin-mineral mixes in diets of broilers

(vitamin, mineral /kg diet) 33

Table 3.5 Cost of feedstuff in experimental diets (January – February 2013) 34

Table 4.1 Analyzed nutrient composition of diets at 1-21d of age (proximate

analysis) 35


analysis) 36


analysis) 36

Table 4.4 Effect of Mu-Plus® in the diets on the body weight gain (BWG) at

different age of broiler chickens 38

Table 4.5 Effect of Mu-Plus® in the diets on feed intake (FI) at different

age of broilers 41

Table 4.6 Effect of Mu-Plus® in the diets on the feed conversion ratio (FCR) at

different age of broilers 43

Table 4.7 Effect of Mu-Plus® in the diets on the survival rate (SR) at


Table 4.8 Effects of Mu-Plus® in the diets of broilers on growth performance,

survival rate (SR) and productive index (PI) at 42 d of age 47

Table 4.9 Effect of Mu-Plus® in the diets on feed cost per gain (FCG) at


xiv

LIST OF TABLES (Cont.)

Page

Table 4.10 Effects of Mu-Plus® in the diets of broilers on economic benefit

returns for overall 42 d of testing 50

Table 4.11 Effect of Mu-Plus® in the diets on carcass quality of broilers at

termination (42 d of age) 52

Table 4.12 Effects of Mu-Plus® in the diets on organ weights and abdominal

fat of broilers at termination (42 d of age) 54

Table 4.13 Effects of Mu-Plus® in the diets on composition of breast meat

and liver of broilers at termination (42 d of age) 55

Table 4.14 Effect of Mu-Plus® in the diets on nutrients digestibility of

broilers at 1-21 d of age 62





Table 4.17 Effect of Mu-Plus® in the diets on the villi height of duodenum,

jejunum and ileum of broilers at different period of age 67

Table 4.18 Effect of Mu-Plus® in the diets on villi width of duodenum,


Table 4.19 Effect of Mu-Plus® in the diets on crypt depth of duodenum,


Table 4.20 Effect of Mu-Plus® in the diets on villi height: crypt depth ratio of

duodenum, jejunum and ileum of broilers at different period of age 75

LIST OF FIGURES

Page

Figure 2.1 Andrographis paniculata plant (A: stems, B: leaves, C: flowers) 5

Figure 2.2 Structures of the major diterpenoids in Andrographis paniculata 10

Figure 2.3 Zingiber cassumunar plant (A: stems, B: flowers, C: rhizomes) 12

Figure 2.4 Structures of the major constituent in Zingiber cassumunar 16

Figure 2.5 Layer of small intestine, villi and crypt of lieberkuhn 23

Figure 2.6 Morphology of small intestine 23

LIST OF ABBREVIATIONS

CD : Crypt depth

CPD : Crude protein digestibility

DMD : Dry matter digestibility

DMPBD : Dimethoxyphenylbutadiene

DMPRD : Division of Medicinal Plants Research and Development

EED : Ether extract digestibility

FCG : Feed cost per gain

GED : Gross energy digestibility

MPRI : Medicinal Plants Research Institute

NPR1 : Net profit returns per bird

NPR2 : Net profit return per kilogram body weight

ROI1 : Returns of investment per bird

ROI2 : Returns of investment per kilogram body weight

SBR : Salable bird returns

SCHRI : Sichuan Chinese Herb Research Institute

VH : Villi height

VW : Villi width

CHAPTER I

INTRODUCTION

1.1 Background

Livestock is the second most important sector within the agricultural economy

in terms of value added. The major animals in the livestock sector are poultry,

buffalo, cattle, and swine. Poultry is the smallest livestock investment that village

household can make. The poultry industry, in which chickens are the most important

products, has been consistently the largest in terms of value-added with an annual

growth rate of 5.2% (Nipon, 1985). During the past decade, there has been a

tremendous expansion of the commercial broiler industry. In the early 1970s, most

farmers grew a small number of indigenous chickens for on-farm-consumption and

there were no exports, but in 1981, Thailand became the eleventh largest chickens

producing country in terms of the number of broiler produced and the second largest

chicken exporter to Japan. This success can be attributed mostly to the initiative and

ability of a few private feed-mill companies. In fact, one of these farms has been so

successful that its market share in every activity within the broiler industry is more

than 50%. Poultry provides a major income-generating activity from the sale of birds

and eggs. Occasional consumption provides a valuable source of protein in the diets.

Poultry has also played an important socio-cultural role in many societies. For

developing countries’ smallholder farmers (especially in poor income, food-deficient

countries), family poultry represents one of the few opportunities for saving,

investment and security against risks. In several of these countries, family poultry

accounts for approximately 90 percent of the total poultry production (Sonaiya and

Swan, 2004).

Antibiotics play an important role in fighting diseases, but recently of undesired

antibiotic residues in poultry products and environment contamination has largely

added to the public concern regarding the use of antibiotics in the feed. As a

consequence of the above facts, European Union Scientific Steering Committee has

banned majority of the antibiotics used as growth promoters in monogastric animal

2

diets since 1999 in the European Union. That reason makes people and researchers

pay attention and come back to use herbals, prebiotics, probiotics, or/and botanicals to

enhance resistant and maintain animal health.

Commercial feed additives of plant origin like herbals, species and various plant

extracts have a growth promoting effect in animals on birds. Herbal is a plant or plant

part used for its scent, flavor, therapeutic properties, and medicinal products. Herbals

frequently take to improve health as dietary supplements. Both of the medicinal plants

seem promising were Andrographis paniculata and Zingiber cassumunar, a shrub

found throughout Southeast Asia. They were well-known as a medicinal plant that

commonly use in human as an immune system booster. They are very widely used in

China, for healing common colds, inflammations and diarrhea (MPRI, 1999).

Andrographis paniculata is used by some farmers as a feed additive to broilers

to avoid the use of drugs. Andrographis paniculata leaf supplementation reduced

mortality among broilers. It is one of the plants having antimicrobial and growth

promoting activity (Chopra et al., 1992). At 2g/kg of Andrographis paniculata fed

broilers had improved feed consumption, feed conversion ratio, live weight, and

decreased mortality rate (Mathivanan et al., 2006). In many Asian countries, Zingiber

cassumunar Roxb., (Zingiberaceae) is widely used in folklore remedies as a single

plant or as a component of herbal recipes. Zingiber cassumunar Roxb., has received

much attention since it can produce many complex compounds that are useful in food

as herbs and spices, flavoring, seasoning, and in the cosmetics and medical industries

such as antioxidant, antibacterial, antifungal, antimicrobial, and anti-inflammatory

agent.

Therefore, this study designed in broilers by inclusion different levels of

Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®) to determine

the growth performance, carcass characteristics, nutrient digestibility, and small

intestinal histomorphology in broiler chickens.

1.2 Hypotheses

1.2.1 Andrographis paniculata and Zingiber cassumunar powder as feed

additive for broilers reduce the mortality rate and lesion in the intestinal tract.

3

1.2.2 Andrographis paniculata and Zingiber cassumunar improve live weight,

feed consumption, feed conversion ratio in broilers, and digestive herbs.

1.2.3 Andrographis paniculata and Zingiber cassumunar help to stimulate the

immune system.

1.2.4 Compounds of Andrographis paniculata and Zingiber cassumunar are an

antibacterial and antifungal activity (against Salmonellae, E. coli and P. multocida).

1.2.5 Andrographis paniclata (andrographolide) is used as a stimulating agent

for liver enzymes, hepatoprotective agent and against liver damage.

1.3 Objectives of the research

1.3.1 To study the effects of different levels of Andrographis paniculata and

Zingiber cassumunar mixture (Mu-Plus®) in the diets on growth performance and

carcass quality of broiler chickens.


Zingiber cassumunar mixture (Mu-Plus®) in the diets on nutrients digestibility of

broiler chickens.


Zingiber cassumunar mixture (Mu-Plus®) in the diets on intestinal histomorphology

in broiler chickens.

1.4 Expected results

1.4.1 Results of Andrographis paniculata and Zingiber cassumunar mixture

(Mu-Plus®) supplemented diets will be improved growth performance and survival

rate (SR) of broiler chickens.


(Mu-Plus®) supplemented diets will be improved carcass quality.


(Mu-Plus®) supplemented diets will be improved nutrients digestibility.


(Mu-Plus®) supplemented diets will be further extended for use by smallholder

farmers and commercial farms in replacing antibiotic drug using in animal feed.

CHAPTER II

LITERATURE REVIEW

2.1 Androgaphis paniculata

2.1.1 Botanical characteristics of Androgaphis paniculata

Andrographis paniculata (Burm, f Nees.) is a scientific name, English

name is King of bitter and other names are Kalmegh, Chaun xinlian, Hempudu bumi,

Creyat root, Halviva, Green chirctta, Krent and Kirayat (Manjunath, 1948).

Andrographis paniculata is a 0.3 to 1.0 m height erects annual plant, which wildly

and abundantly grows in Asia country: India, Pakistan, Sri Lanka, Indonesia, China

and Thailand. Its stem is dark green, 2 to 6 mm in diameter, quadrangular with

longitudinal furrows and wings at angles of the younger parts. The leaves are

opposite, decussate, lanceolate, up to 8 cm long and broad, glabrous, margin entire,

and venation pinnate; the petiole is very short. The flowers are small with bilabial

corollas. The fruits are small 2-celled odorless capsules which taste intensely bitter.

Andrographis paniculata is found in every green, pine and deciduous forests and

along roadsides. It can grow in all types of soil and it is the only plant that grows on

what is called “serpentine soil” formations. This type of soil contains metals such as

aluminium, copper and zinc. Andrographis paniculata's ability to grow in such a

harsh environment may explain its wide distribution (DMPRD, 1990 and MPRI,

1999). Mostly the roots and leaves have been traditionally used over the countries for

different medicinal purposes in Asia and Europe.

Kingdom : Plantae

Division : Angiospermae

Class : Dicotyledoneae

Order : Tubiflorae

Family : Acanthaceae

Genus : Andrographis

Species : paniculata Nees

Source: Dhiman et al. (2012)

5

A B C

Figure 2.1 Andrographis paniculata plant (A: stems, B: leaves, C: flowers)

2.1.2 Utilization of Andrographis paniculata

Andrographis paniculata is extensively used as an anti-inflammatory and

antipyretic drug for the treatment of fever, cold, laryngitis, diarrhea, inflammation,

infections in the gastrointestinal tract, upper respiratory tract, and other chronic

infectious diseases (Wangboonskul et al., 2006 and Sheeja et al., 2006). Andrographis

paniculata has also been used for sluggish liver as an antidote in case of colic

dysentery and dyspepsia. It is then used as a bitter tonic, antispasmodic, antiperistaltic,

stomachic, and anthelmintic (Dhiman et al., 2012). Feeding 2g/kg of Andrographis

paniculata to broiler chickens improved feed consumption, feed conversion ratio, live

weight, and decreased mortality rate (Mathivanan et al., 2006).

2.1.3 Pharmacological action of Androgaphis paniculata

Andrographis paniculata is a bioactivities herb, plays role as an

antioxidant, anti-allergies, anti-inflammatory, stimulating agent for liver enzymes,

increase the bile flow and bile salt production, against infectious diseases, antipyretic

(Kanokwan et al., 2008; Ojha et al., 2009 and Dhiman et al., 2012). Mathivanan and

Kalaiarasi, (2007) also reported that Andrographis paniculata at level 2g/kg improved

immune status of broiler chickens.

2.1.3.1 Antioxidant activity

The role of free radical generated oxidative stress in isoproterenol-

induced myocardial ischemic injury is well established. A plenty amount of herbal

drugs possessing antioxidant activities has been demonstrated protective in the

6

isoproterenol-induced ischemic injury of the myocardium. On one hand, Ojha et al.

(2009) and Dhiman et al. (2012) postulated that a hydroalcoholic extract of

Andrographis paniculata prevented isoproterenol induced elevated lipid peroxidation

and elevated the activities of antioxidant enzymes, superoxide dismutase, calatase,

glutathione peroxidase and the level of reduced glutathione level in hearts.

Additionally, the extract also prevented leakage of lactate dehydrogenase from the

heart and salvaged it from isoproterenol induced myocardial ischemic injury.

2.1.3.2 Antipyretic activity

Andrographolide and andrographolide derivative at 4 mg/kg dose

showed antipyretic activity. The semisynthesis of isopropylidene-andrographolide and

14-deoxy-11, 12-didehydro-3, 19-dipalmitoylandrographolide were also shown a good

effect on antipyretic activity in mice (Suebsasana et al., 2009). Andrographis

paniculata is also used to prevent and treat the common cold (Research review,

1997). Intragastric administration of the major active components of Andrographolide

at 100 mg/kg body weight in mice decreased brewer’s yeast-induced pyrexia (Madav

et al., 1995). A similar previous study was conducted on deoxyandrographolide,

andrographolide, or 11, 12-didehydro-14-deoxyandrographolide at 100 mg/kg body

weight in mice, rats or rabbits were found to reduce pyrexia induced by 2, 4-

dinitrophenol or endotoxins (Chang and But, 1986 and Deng et al., 1982).

2.1.3.3 Anti-inflammatory activity

Inflammation is a response of vascularized tissue to sublethal

injury (Ballou and Kushner, 2000 and Sheeja et al., 2007). Andrographis paniculata is

also used as a folk medicine remedy for fever, pain reduction, and disorders of the

intestinal tract. The ability of Andrographis paniculata, andrographolide lowered the

fever produced by different fever-inducing agent such as bacterial endotoxins,

pneumococcus, and hemolytic streptococcus (Deng, 1978). The anti-inflammatory

activity of androgapholide is reduction of inducible nitric oxide synthase protein

expression (Chiou et al., 1998 and Kanokwan et al., 2008). Suebsasana et al. (2009)

also found that 4 mg/kg of andrographolide, isopropylidene andrographolide, or

7

14-deoxy-11, 12-didehydro-3, 19-dipalmitoylandrographolide had anti-inflammatory

activity by reduced inflammatory demyelinating disease. Dutta and Sukul, (1982)

reported that three substances of Andrographis paniculata leaf powder are

andrographolide, deoxyandrographolide, and neoandrographolide had an effect in

reducing inflammation.

2.1.3.4 Antibacterial activity

Andrographis paniculata is one plant that has antimicrobial and

growth promoting activity and be used as an alternative to antibiotics (Chopra et al.,

1992). Mathivanan and Edwin (2012) also mentioned that Andrographis paniculata

did not have direct antibacterial activity against organisms of broiler intestinal

contents. Stems and leaves blended into powder can be effective against the Shigella

bacteria, but it has no effect against cholera. An ethanol extract of Andrographis

paniculata has effects against Staphylococcus aureus and Escherichia coli (E. coli)

(Bunyapraphatsara, 2000). It has been believed that Andrographis paniculata was

affected against bacterial dysentery and diarrhea because of it has antibacterial

activities (Pleumjai, 1992 and Kanokwan et al., 2008). Sithisomwonges et al. (1989)

also conducted in in-vitro trial found that Andrographis paniculata extract with 70%

and 80% ethanol can kill bacteria that cause diarrhea. Thamaree et al. (1985) and

Sawasdimongkol et al. (1990) also conducted trial on mice found that 50% and 85%

alcohol extract of Andrographis paniculata leaves powder were affected in reducing

intestinal tract movements. Tipakorn (2002) also mentioned that 0.4% Andrographis

paniculata as feed additive in broiler feed can reduce the very serious impact of

coccidiosis and it can replace the use of antibiotic in poultry production.

2.1.3.5 Immunostimulatory activity

Puri et al. (1993) demonstrated that intragastric administration of

an ethanol extract of Andrographis paniculata leaves at 25 mg/kg body weight or the

purified andrographolide at 1 mg/kg body weight in mice stimulated antibody

production. The crude extract was found to be more effective than either

andrographolide or neoandrographolide alone indicating that the effects may be due to

the other active constituents involved in the immunestimulant response.

8

2.1.3.6 Gastrointestinal activity

Mixing Andrographis paniculata leaves into chicken diets reduce

mortality especially based on digestive tract diseases (Chaiwongkeart, 1997). Duke

and Ayensu (1985) had conducted an experiment in animal demonstrated that

Andrographis paniculata can prevent or stop diarrhea. It also has effects against the

diarrhea associated with E. coli infections. The active substances of Andrographis

paniculata against diarrhea are andrographolide and deoxyandrographolide.

Choudhury and Poddar (1985) have been reported that both Andrographis paniculata

leaves extract and andrographolide increased the intestinal digestion and absorption of

carbohydrate in a dose related and time dependent on characteristic activation of

brush-border membrane-bound hydrolysis.

Dhiman et al. (2012) also reported that ethanol extract tablets of

Andrographis paniculata able to cure 88.3% of acute bacillary dysentery and 91.3%

of acute gastroenteritis cases.

2.1.3.7 Hepatoprotective activity

In Ayurvedic medicine, there are 26 different remedies containing

Andrographis paniculata used to treat liver disorders, liver damage, and as a

stimulating agent for liver enzymes and hepatoprotective agent (Dhiman et al., 2012).

Administration of Andrographis paniculata prevented hexachlorocychohexance

induced increase in the activities of γ-glutamyl traspeptidase, glutathione-S-

transferase and lipid peroxidation in mouse liver, an indication potential antioxidant

and hepatoprotective effects of Andrographis paniculata (Trivedi et al., 2000 and

Trivedi et al., 2001). Leaf extract of Andrographis paniculata was affected in

preventing carbontetrachloride-induced liver damage in rats and mice (Choudhury

et al., 1984; Handa et al., 1990 and Rana et al., 1991). Andrographolide play role as a

hepatoprotective by reducing a lipid peroxidation product (Kapil et al., 1993). Chanda

et al. (1995) was noted that antihepatotoxic action of andrographolide against

plasmodium berghei K173-induced hepatic damage of mastomys natalensis. Shukla

et al. (1992) and Dhiman et al. (2012) reported that andrographolide also play a role

as a potent stimulator of gallbladder function because of produce significant increase

9

in bile flow, bile salts, and bile acid in conscious rats and anesthetized guinea pigs and

improved liver function after continuous treatment with Andrographis paniculata.

2.1.4 The chemical composition of Androgaphis paniculata

SCHRI (1973) postulated that Androgapholide was first isolated from

Andrographis paniculata by BOORSMA in 1896. It is a crystal compound with a

very bitter taste with colorless crystalline appearance, that compound as lactones.

Deng et al. (1982) also reported that there are four lactones in Andrographis

paniculata including deoxyandrographolide (Andrographis A), andrographolide

(Andrographis B), neoandrgraphplide (Andrographis C) and deoxydidehydro-

andrographolide (Andrographis D) which were also identified by Dhamma-Upakorn

et al. (1992).

Moreover, the chemical constituents of Androgaphis paniculata include

andrographolide, neoandrographolide, andrograpanoside, andrographanin, 14-deoxy-

12-methoxyandrographolide, 14-deoxyandrographis, 14-deoxy-11, 12-didehydro-

andrographolide and deoxyandrographolide (Kongkathip, 1995; Tipakorn, 2002 and

Dhiman et al., 2012). The aerial of the plant (leaves and stems) are used to extract the

active phytochemicals. The leaves contain the highest amount of andrographide

(2.39%) the medically most active phytochemical in the plant, which the stem and

seed contain the lowest amount (Research review, 1997).

10

Table 2.1 Chemical constituents and bioactivities of Andrographis paniculata

Chemical constituents Functions Sources

- Andrographolide Anticancer Ajaya et al., 2004

Immunostimulatory

activity

Ajaya et al., 2004

Hepatoprotective Mohamed, 2010

- Andrographide Liver cleaning and

Hepatitis

Dahanukar et al., 2000

- Andrpgraphiside Antioxidant Balachandranm et al., 2005

Antilipoperoxidant Balachandranm et al., 2005

Carcinogenic

detoxification

Surveswaran et al., 2007

- Neoadrographolide Anti-inflammatory Liu et al., 2007

- Kalmeghin Fever and Cold Koul et al., 1994

- 14-deoxy-11-

oxoandrographilide

Antiparasitic diseases

Lala et al., 2003

- 14-deoxy-11, 12-

didehydroandrographolide

Anticancer

Ajaya et al., 2004

Andrographolide Andrographide

Figure 2.2 Structures of the major diterpenoids in Andrographis paniculata

11

14-Deoxy-11, 12-didehydroandrographolide Neoandrogapholide

Deoxyandrographolide Dehydroandrographolide

Figure 2.2 Structures of the major diterpenoids in Andrographis paniculata

(Cont.) (Dhiman et al., 2012 and Yang et al., 2012)

2.2 Zingiber cassumunar

2.2.1 Botanical characteristics of Zingiber cassumunar

Zingiber cassumunar Roxb., is a scientific name, English name is Plai and

other names are Zingiber montanum, Theilade, and Zingiber purpureum. It is a short-

lived herbal plant in the family of Zingiberaceae found in many Asian countries and it

is an important medicinal plant in Southeast Asia and widely cultivated throughout

the tropics including Southeast Asia, Korea, India, Bangladesh, Malaysia, China, and

Thailand for its medicinal properties (Indian medicine plant, 1993). The essential oil

12

from the rhizome is used for reducing inflammation from injuries, sprains, muscles,

chest pain, swelling during wound healing of the skin and joint issues.

Kingdom : Plantae

Division : Angiospermae

Class : Monocots

Order : Zingiberales

Family : Zingiberaceae

Genus : Zingiber

Species : Cassumunar Roxb

Source: http://en.wikipedia.org/wiki/Zingiber_cassumunar

A B C

Figure 2.3 Zingiber cassumunar plant (A: stems, B: flowers, C: rhizomes)

2.2.2 Utilization of Zingiber cassumunar

Zingiber cassumunar Roxb., is extensively used in folk medicine to treat

inflammation, muscle pain, wounds, cough, fever, immunostimulant activity,

antibacterial activity, antifungal activity, antimicrobial activity, antihistaminic effect,

reliefs of sore muscles, antioxidant activity, antiplatelet aggregation activities, and

digestive herb. Plai oil has anti-inflammatory effect and exhibits antimicrobial

activity. The rhizome oil of plai was found to exhibit high activity against

dermatophytes and yeasts (Pithayanukul et al., 2007).

13

2.2.3 Pharmacological action of Zingiber cassumunar

2.2.3.1 Anti-inflammatory activity

Zingiber cassumunar Roxb., is a tropical ginger distributed in

Southeast Asia, has been widely used in traditional medicine for the treatment of

various conditions such as muscular or joint pain, rheumatism, asthma and

inflammation (Bhuiyan et al., 2008). Ozaki et al. (1991) has been reported that the

methanol extract obtained from the rhizomes of Zingiber cassumunar contains both

anti-inflammatory and analgesic activity.

Jeenapongsa et al. (2003) was conducted on rats and demonstrated

that (E)-1-(3, 4-Dimethoxyphenyl) butadiene (DMPBD) exerts a clear potent anti-

inflammatory effect. The difference in potency of DMPBD in inhibiting platelet

aggregation was induced by each inducer point out the specificity of DMPBD on the

pathways involved in platelet aggregation. It was also demonstrated that DMPBD is a

unique topical active anti-inflammatory agent. Possibly, it is a modulator of

arachidonic acid metabolism having activities on both cyclooxygenase and

lipoxygenase enzymes. It has a potential for local therapeutic applications in

inflammatory diseases.

Ong-chai et al. (2008) also reported that hyaluronan plays an

important role in the formation of some pathological conditions of connective tissue,

especially inflammatory reaction and edema during wound healing processes.

Consequently, the anti-inflammatory activity of Zingiber cassumunar extract may

partly be due to its ability to decrease hyaluronan synthesis.

Kaewchoothong et al. (2012) have been demonstrated that the

anti-inflammatory activity of phenylbutanoid-enriched Zingiber cassumunar extracts

via inhibition of nitric oxide production by murine macrophage-like RAW264.7 cells

was stronger than those of the four individual phenylbutanoids, the crude hexane

extract and the essential oil of Zingiber cassumunar.

2.2.3.2 Immunostimulant activity

Chairul et al. (2009) determined that the potential of phenylbu-

tenoid compounds isolated from Zingiber cassumunar: [(E)-4(3', 4'-dimethoxyphenyl)

but-3-en-1-ol)], [(E)-4(2', 4', 5'-trimethoxyphenyl) but-3-en-1-ol)], and [(E)-4(3', 4', 1-

14

dimethoxyphenyl) but-3-en-1-methoxy-1-ol] as immunostimulant through stimulation

mouse macrophage cells. The result showed that [(E)-4(3', 4'-dimethoxyphenyl) but-

3-en-1-ol)] compound had highest immunostimulant activity compared to other

compounds. Free radical and active oxygen is one of foreign bodies that suppress

body immune and antioxidant properties related to the immunostimulant properties.

2.2.3.3 Antibacterial activity

Bhuiyan et al. (2008) also reported that the high concentration of

sabinene and triquinacene, 1, 4-bis (methoxy) in the leaves and rhizome oil makes it

respectively potentially useful in the medicines because they exhibit antibacterial

activities. Bhusita et al. (2009) presented that essential oil of Zingiber cassumunar has

antibacterial effects against bacteria such as E. coli, Salmonella. Pithayanukul et al.

(2007) found that the antibacterial activity of plai oil against most gram-positive and

gram-negative bacteria tests at minimum bactericidal concentrate 0.62-2.5 Vol%. The

5% plai oil gel (pH 5.0) was more potent against dermatophytes and yeasts at much

lower concentrations than against gram-positive and gram-negative bacteria. The plai

oil gel (5%) can be considered as a very weak antibacterial (minimum bactericidal

concentrate 52–79 mg/ml). Golam et al. (2011) also demonstrated that a crude ethanol

extract of the Zingiberaceae (Zingiber zerumbet) has highest antibacterial activity (10

mm) against V. parahemo-lyticus.

2.2.3.4 Antifungal activity

Kishore et al. (1992) and Tripathi et al. (2008) had written that

Zingiber cassumunar essential oil found to exhibit absolute fungi toxic activity. Chen

et al. (2008) also demonstrated that many constituents of Zingiberaceae plants have

biological activity in antifungal, antioxidant activity. Pithayanukul et al. (2007) also

reported that plai oil could be considered as a weak antibacterial agent and a weak

antifungal agent in comparison with ketoconazole (minimum fungicidal concentrate

0.08-0.22 µg/ml) and Ampicillin (minimum bactericidal concentrate 0.24-0.32

µg/ml). Plai oil and the 5% plai oil gel (pH 5.0) should be more effective for the

treatment of fungi rather than bacteria. The plai oil gel (5%) can be considered as a

very weak antifungal (minimum bactericidal concentrate 13.8–39.5 mg/ml).

15

Moreover, Golam et al. (2011) also postulated that a crude ethanol extract of the

Zingiberaceae (Zingiber zerumbet) has good antifungal activity (9 mm) against all

pathogenic fungi.

2.2.3.5 Antimicrobial activity

Essential oil plant or/and their components are becoming

increasing popular as natural antimicrobial agents to be used for a wide variety of

purposes. Nevertheless, the potential use of these oils as natural antimicrobial agents

has been less explored. While some of the oils used on the basis of their repute

antimicrobial properties has well-documented in in-vitro activity (Cosentino et al.,

2003). The essential oil of Zingiber cassumunar has antimicrobial activity against to 7

strains of bacteria (Bhusita et al., 2009). A number of pure compounds isolated from

the plants have been shown to possess antimicrobial. Terpinen-4-ol (24-32%) and

sabinene (34-44%) were found as the major constituents of the rhizome oil of

Zingiber cassumunar and their antimicrobial activities were postulated in comparison

with the commercial terpinen-4-ol (Wasuwat et al., 1989) and exhibit a high activity

against yeasts (Bin et al., 2003). However, Wungsintaweekul et al. (2010) reported

that volatile oil and methanol extracts of Zingiber cassumunar did not have any

antimicrobial activity. Tg. Siti Amirah Tg. Kamazeri et al. (2012) also reported that

Zingiber cassumunar had very low or weak activity against the tested

microorganisms.

2.2.4 The chemical composition of Zingiber cassumunar

The compounds isolated from Zingiber cassumunar rhizome there are

four phenylbutanoids, (E)-4-(3, 4-dimethoxyphenyl) but-3-en-l-ol (I), (E)-4-(3, 4-

dimethoxyphenyl) but-3-en-l-yl acetate (II), (E)-1-(3, 4-dimethoxyphenyl) butadiene

(III) and (E)-3-(3, 4-dimethoxyphenyl)-4-[(E)-3, 4-dimethoxystyryl] cyclohex-1-ene

(IV), isolated from Zingiber cassumunar. It is used as standard markers for

quantitative determination and preparation of phenylbutanoid-enriched Zingiber

cassumunar extracts (PZEs) (Kaewchoothong et al., 2012). The plai oil obtained by

hexane extraction contained mainly: sabinene (24.05-39.11%), γ-terpinene (6.68-

7.74%), terpinen-4-ol (33.11-49.36%) and (E)-1-(3, 4-dimethoxyphenyl) butadiene

16

(DMPBD) (5.31-8.28%), whereas that obtained by hydro-distillation contained:

sabinene (36.71-53.50%), γ-terpinene (5.27-7.25%), terpinen-4-ol (21.85-29.96%)

and DMPBD (0.95-16.16%) (Udomlak et al., 2009). Han et al. (2005) reported that in

previous study, two phenylbutenoid dimers, (±)-trans-3-(3, 4-dimethoxyphenyl)-4-

[(E)-3, 4 dimethoxystyryl] cyclohex-1-ene (1) and (±)-trans-3-(4-hydroxy-3-methoxy

phenyl)-4-[(E)-3, 4-dimethoxystyryl] cyclohex-1-ene (2), and four phenylbutenoids,

4-(2, 4, 5-trimethoxyphenyl) but-1, 3-diene (3), 4-(3, 4-dimethoxyphenyl) but-1, 3-

diene (4), (E)-4-(3, 4-dimethoxyphenyl) but-3-en-1-ol (5), and (E)-4-(3, 4-dimethoxy-

phenyl) but-3-en-1-yl acetate (6) and (E)-4-(3, 4-dimethoxyphenyl) but-3-en-1-O-b-

D-glucopy-ranoside (7).

(E)-4-(3, 4-dimethoxyphenyl) but-3-en-l-ol (I) (E)-4-(3, 4-dimethoxyphenyl) but-3-en-l-ly-

acetate (II)

(E)-1-(3, 4-dimethoxyphenyl) butadiene (III) (E)-3-(3, 4-dimethoxyphenyl)-4-[(E)-3,4-

dimethoxystyryl] cyclhex-1-ene (IV)

Figure 2.4 Structures of the major constituent in Zingiber cassumunar

(Kaewchoothong et al., 2012)

17

2.3 Utilization of Andrographis paniculata and Zingiber cassumunar mixture in

animal feed

Many researches have been made to estimate the ability of animal to digest and

absorb the various nutrients in feed ingredients. It is important for the animal’s health

and well-being to provide sufficient amount of all critical nutrients. There are

persistent indications that age, breed or sex of a bird may influence the ability to

digest and use certain nutrients. Feed additives are considered to be natural products

that consumers might find acceptable. Phatarapheecha et al. (2011) have been

conducted experiment in piglets by supplement Andrographis paniculata, Curcuma

longa and Momardica charantia mixture (Herbatob-Mix®) in the diets at 0, 0.05 and

0.10% shown that Herbatob-Mix® can stimulate to increase daily milk yield (P<0.05)

when compared with the control group. Body weight gain has significantly different

(P<0.05) increase for group that treated Herbatob-Mix®. Even though there were no

significant difference (P>0.05) among groups, there was tendency for Herbatob-Mix®

group to have lower body fat and increase feed intake of sows and their piglets.

Nopparatmaitree (2008) have been conducted trial in weaning pigs with

supplement Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®)

at 1,000 and 2,000 ppm in the diets compared with the antibiotic group and indicated

that Mu-Plus® improved productive performance including feed intake, average daily

gain (P>0.05) and improved nutrient digestion such as organic dry matter, protein, fat,

and energy (P>0.05). Moreover, it had highly significant difference (P<0.01) increase

the duodenal villi height, jejunal villi height and crypt of duodenum.

Pratoomtong (2011) have been conducted experiment in piglets with

supplement Herbatob-Mix® (Andrographis paniculata, Curcuma longa and

Momardica charantia) and Mu-Plus® (Andrographis paniculata and Zingiber

cassumunar) in the diets compared to antibiotic at different level: 0, 500 ppm

(Herbatob-Mix®) and 500 ppm (Herbatob-Mix®) + 1,000 ppm (Mu-Plus®) shown that

final body weight, body weight gain, feed intake, feed conversion ratio, survival rate

has no effect (P>0.05) by Herbatob-Mix® or Herbatob-Mix® + Mu-Plus® concentrate

when compared with the control group. However, there was no effect among groups,

there was tendency for Herbatob-Mix® or Herbatob-Mix® + Mu-Plus® group to have a

18

higher final weight, body weight gain, feed intake and feed conversion ratio than

control.

Saengsophon et al. (2000) was conducted and allotted 3 experimental groups

(group I: feeding control group, group II: feeding with antibiotic 50 g/100 kg diet,

group III: feeding Andrographis paniculata 180 g/100 kg diet) under completely

randomize design. The results indicated that Andrographis paniculata was significant

affected the body weight and growth rate (P<0.05) at week 6 lower than control and

antibiotic group. However, it was not affecting (P>0.05) on feed consumption and

feed conversion. Both antibiotic and Andrographis paniculata feeding groups were

significant (P<0.05) lower dressing percentages than the control group.

Mathivanan et al. (2008) also conducted experiments with five dietary include

fed a basal diet (T1), basal diet with virginiarnycin 20 mg/kg (T2), basal diet with A.

paniculata 1.0 g/kg (T3), basal diet with A. paniculata 2.0 g/kg (T4) and basal diet

with A. paniculata 3.0 g/kg (T5) for a 42 days period. Reported that A. paniculata had

no effects on body weight and body weight gain at 6 weeks of age. However, feed

consumption was highly significant (P<0.01) in A. paniculata fed at 2.0 and 3.0 g/kg

of diet when compared with the control group. The feed conversion ratio was also

significantly different (P<0.05) better in A.paniculata fed at 2.0 and 3.0 g/kg at 6

weeks of age.

Tongwiti et al. (2012) was conducted to investigate the effects of herbal mixture

(Herbatob-mix®) at 0, 250, 500 and 1,000 ppm during 1-42 days and 0, 250, 500 and

1,000 ppm during 36-42 days. The results indicated that feed intake was not

significant (P>0.05) but shown the highest body weight gain and better feed

conversion ratio in all of the group fed with Herbatob-Mix® supplementation in the

diets. Feeding Herbatob-Mix® for the whole experiment had improved performance

and higher net profit returns per bird.

Watanasit et al. (2005) studied the effects of Andrographis paniculata at 0.1 and

0.2% diet on the growth performance and carcass quality compared with antibiotic

or/and control. The results shown that feed intake, feed conversion ratio, body weight

gain and mortality rate were not significantly different (P>0.05) among groups. In

addition, there were not affected (P>0.05) on breast and thigh among chicken fed

different diets.

19

Tummaruk and Limtrajitt (2010) made study the influence of Andrographis

paniculata compound 1,000 ppm in diets on average daily feed intake of lactating

sows and the litter weight gain of piglets. The results indicated that litter weight gain,

weaning weight, feed intake of Andrographis paniculata compound group were

highly significant difference (P<0.01) and average daily gain significantly different

(P<0.05) higher than the control group. It also concluded that 1,000 ppm of

Andrographis paniculata compound in post-partum sows improved average daily feed

intake of the sows and improved litter weight gain of piglets.

Mathivanan et al. (2006) had studied Andrographis paniculata as alternatives to

antibiotic growth promoter on broiler production and carcass characteristics. The

results of supplementation of 2 g/kg of Andrographis paniculata in the diets

compared with antibiotic (virginiamycin 20 mg/kg) or/and control shown that feed

consumption did not have any affect different among treatments. However, feed

conversion ratio was better (P<0.05) than antibiotic and control group. Nevertheless,

carcass quality did not different due to dietary supplementations.

Suci et al. (2012) studied dietary supplementation of Andrographis paniculata

meal at 0, 0.3, 0.6 and 0.9 g/kgBW on performance of laying hens. The results

indicated that laying hen fed 0.6 g/kgBW significant (P<0.05) increase hen day

production when compared with other groups. However, 0.3 and 0.9 g/kgBW had

lower feed consumption and hen day production but the feed conversion ratio was not

significant. It also concluded that dietary supplementation of Andrographis paniculata

meal at 0.6 g/kgBW was effective for increased egg production.

Khajarern and Siriloaphaisan (2007) investigated the effects of Herbatob-Mix®

(Andrographis paniculata, Curcuma longa and Momordica charantia) at 0, 0.05,

0.075 and 0.10% on egg performance and egg quality of laying hens. The results

revealed that it is trend to increased egg production, but there was not significant in

overall period. Feed intake and feed to egg ratio were not significant (P>0.05).

Testing at period 43-46 week of age, both feed intake and feed to egg ratio had a

linear response decrease (P<0.05) and better in feed efficiency while increasing level

of Herbatob-Mix® in the diet. It can save feed intake 0.73, 0.47, and 0.03% and better

in feed efficiency 0.42, 0.90 and 1.42% with Herbatob-Mix® in the diets at level 0.05,

20

0.075 and 0.10% respectively. Feeding laying hens with Herbatob-Mix® also

increased survival rate (P<0.05).

Chanjula et al. (2006) had studied the effects of Andrographis paniculata at 0.1,

0.2, 0.3, 0.5 and 0.6% diet compared to the antibiotic and the control group on growth

performance of chickens. The results revealed that Andrographis paniculata

supplement at level 0.1 to 0.3% diet were not significantly different (P>0.05) among

groups on feed intake, weight gain, feed conversion ratio, mortality and feed cost

when compared to both antibiotic and control group. Nevertheless, the supplement

level higher than 5% diet, it is trend to lower growth performance and feed intake and

feed cost also increased with the increasing Andrographis paniculata level in the diet.

Tangmutthapattharakul (2007) had studied the effect of Zingiber cassumunar at

level 0.2, 0.4 and 0.8% diet on the growth performance of broilers at 0-3 week of age

compared with the antibiotic (Avilamincin 2.5 mg/kg diet) and/or control group. The

results deal that feed intake, final body weight, body weight gain and feed conversion

ratio were not significantly different (P>0.05). However, at 0.4% of Zingiber

cassumunar in the diets, it trended to improve feed intake, feed conversion ratio and

final body weight when compared to control and/or antibiotic group.

Kungsoksomboun (2007) also studied the effect of Zingiber cassumunar at level

0.2, 0.4 and 0.8% diet on the carcass quality of broiler compared to the antibiotic

(Avilamincin 2.5 mg/kg diet) and/or control. The results indicated that hot carcass,

carcass percentage, breast, thigh, wings, abdominal fat, gizzard, liver and heart were

not significantly different (P>0.05) among the groups. Nevertheless, there was a

tendency to improved carcass quality for the group that received at 0.2 and 0.4% of

Zingiber cassumunar in the diets when compared with the antibiotic and/or control

group.

2.4 Characteristics of small intestine of broiler chickens

The main features of the small intestine include the three main sections such as

duodenum, jejunum and ileum. The small intestine is the part of the chicken’s

digestive tract where vitamins, mineral and nutrients are fully absorbed and digestion

into the blood stream.

21

The duodenum is the first part of the chicken’s small intestine. The duodenum is

largely responsible for the breakdown of food in the small intestine, using enzymes

that it receives digestive enzymes and bicarbonate from the pancreas and bile from the

liver. Digestion is completed here before the food move to lower small intestine for

nutrient absorption. The villi of the duodenum have a leafy-looking appearance, a

histologically identifiable structure. Brunner’s glands, which secrete mucus, are found

in the duodenum only. The duodenum wall is composed of a very thin layer of cells

that form the muscularis mucosae.

The jejunum is the middle section of the small intestine and it lies between the

duodenum and ileum. The jejunum area has a very large surface area. Villi also

increase the surface of the jejunum and it is responsible for absorbing nutrients. The

jejunum also secretes digestive enzymes that break down food into units that can be

absorbed by the intestine. The luminal surface of the jejunum is covered in finger like

projections of the mucosa, called villi, which increase the surface area of tissue

available to absorb nutrients from ingested foodstuffs. The epithelial cells that line

these villi have microvilli. The transport of nutrients across epithelial cells through the

jejunum and ileum includes the passive transport of sugar fructose and the active

transport of amino acids, small peptides, vitamins, and most glucose. The villi in the

jejunum are much longer than in the duodenum or ileum.

The ileum is the last segment of the small intestine. It is partially responsible for

absorbing nutrients into the blood stream. The ileum absorbs vitamin B12 and bile

salts and whatever products of digestion that were not absorbed by the jejunum. The

wall itself is made up of folds, each of which has many tiny finger-like projections

known as villi on its surface. In turn, the epithelial cells that line these villi possess

even larger numbers of microvilli. Therefore, the ileum has an extremely large surface

area both for the adsorption (attachment) of enzyme molecules and for the adsorption

of products of digestion. The villi contain large numbers of capillaries that take the

amino acids and glucose produced by digestion to the hepatic portal vein and the

liver. Lacteals are small lymph vessels and present in the villi. They absorb fatty acids

and glycerol, the products of fat digestion (figure 2.6).

Chicken’s small intestine consists of four layers included tunica serosa, tunica

muscularis extra, tunica submucosa and muscularis mucosa layer (figure 2.5).

22

Table 2.2 Small intestinal villi morphology of broiler chickens

Parameters Duodenum Jejunum Ileum Sources

Villi height 1,441.00 1,167.00 898.00 Pelicano et al., 2005

(µm) 1,635.70 986.07 731.69 Marchini et al., 2011

1072.50 544.91 434.10 Vaezi et al., 2011

2,777.80 1,595.70 957.20 Gordana et al., 2004

1,667.00 1,064.00 Awad et al., 2006

655.70 399.20 Kawalilak et al., 2010

1,640.00 614.00 Awad et al., 2009

2,090.00 1,070.00 Xiaolun, 2004

743.33 Mathivanan and Edwin, 2012

372.32 Chumpawadee et al., 2008

1,077.00 Nyamambi et al., 2007

Villi width 460.40 542.00 547.60 Vaezi et al., 2011

(µm 249.00 175.00 Awad et al., 2006



Crypt depth 247.00 200.00 203.00 Pelicano et al., 2005

(µm) 191.20 130.72 144.64 Marchini et al., 2011

143.00 105.20 101.10 Vaezi et al., 2011

534.10 322.30 175.20 Gordana et al., 2004

149.00 128.00 Awad et al., 2009

130.00 90.00 Xiaolun, 2004



173.00 Nyamambi et al., 2007

23

Figure 2.5 Layer of small intestine, villi and crypt of lieberkuhn

(http://www.vetmed.vt.edu/education/curriculum/vm8054/Labs/Lab19/

Lab19.htm/ April 8, 2014)

Figure 2.6 Morphology of small intestine (http://cikgurozaini.blogspot.com/

2011/07/digestive-system.html/ April 8, 2014)

CHAPTER III

MATERIALS AND METHODS

3.1 Experiment I. Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on productive performance and carcass quality

of broiler chickens.

3.1.1 Experimental unit

The experiment was used 480 d-old Arbor Acres chicks that bought from

local commercial hatching. Birds were randomly allotted to five treatments with four

replications and each replication consist 24 birds with equal sex.

3.1.2 Experimental design

The experiment was designed in completely randomized design (CRD),

and all of the birds were allotted to five dietary treatments with four replications and

24 birds (twelve males and twelve females) per each. Before the housing in the floor

pan, baby chicks were weighed body weight to separate as small, medium and large

size and then count the amount number of each size and then formulated the ratio of

chick size for the overall weight and weight range were similar for each replicate

group. Feed and water offered ad libitum throughout the experiment.

Treatment 1. Basal diet (Control group)

Treatment 2. Basal diet + Mu-Plus® at 0.50 kg/ton diet




Ingredients, nutrient compositions of experimental diets and feedstuff cost

are presented in Table 3.1, 3.2, 3.3 and 3.4. All of five formulations were agreement

with NRC (1994) and diets, every period, were collected to analyze to find out the

nutrients content according to AOAC (2000) procedure.

25

3.1.3 Research methodology

Birds were reared in floor pen open-side house with uniform floor, feeder

and waterer space for a period of six weeks. The broiler starter, grower, and finisher

diets were fed ad libitum to the birds from 1 to 21, 22 to 35 and 36 to 42 days of age,

respectively. Uniform management and vaccination schedules were followed for all

birds. Lighting program was provided according to the Arbor Acres broiler

management manual. All birds were vaccinated for Gumboro disease at 14 days of

age.

3.1.3.1 At the end of each period, the birds were weighed for calculating

body weight gain. The feed consumption also weighed in order to calculate to find out

feed intake and feed conversion ratio.

3.1.3.2 Week 6, four males and four females birds from each treatment

was randomly sampled with average body weight for serial euthanize to evaluate

carcass quality such as dressing percentage, percentage of breast, thigh, wings,

drumstick, edible meat, abdominal fat and also the percentage of liver, heart, thymus,

bursa, spleen and gizzard.

3.1.4 Data record

3.1.4.1 Productive performance

Initial weight, final body weight (FBW), body weight gain

(BWG), feed intake (FI), feed conversion ratio (FCR), mortality, survival rate (SR)

were recorded throughout the experiment.

- Body weight gain (BWG) (kg/bird)

Finished weight – start weight BWG =

Number of chickens

- Feed intake (FI) (kg/bird)

Offered feed – refused feed FI =

Number of chickens

- Feed conversion ratio (FCR)

Feed intake FCR =

Body weight gain

26

- Survival rate (SR) (%)

Survival chicken SR = x 100

Total chickens

- Productive index (PI)

BWG (Kg) x Survival rate PI = x 100

Age x FCR

3.1.4.2 Carcass characteristics

The selected chickens were weighed to record live body weight

and then euthanize, immediately, various body parts will be removed and weighed

(breast meat, drumstick, thigh, wings, liver, gizzard, spleen, heart, feet, abdominal fat,

bursa and thymus) for calculating percentage compare to live body weight basis.

New York dressing weight

- Dressing percentage (%) = x 100 Survival body weight Breast weight

- Breast (%) = x 100 Survival body weight

3.1.4.3 Economic efficiency

- Feed cost (FC) (Baht/bird)

FC = Feed intake (FI) x diet cost (Baht/kg)

- Feed cost per kilogram gain (FCG)

Feed cost (Baht/bird) FCG (Baht/bird) = x 100

Survival rate (%) Feed cost (Baht/bird)

FCG (Baht/kgBW) = x 100 Survival rate (%) x BW

- Salable bird returns (SBR) (Baht/bird)

SBR = Price of live chicken x BW

(Price 38 Baht/kgBW market price) (February 18-23, 2013)

27

- Net profit returns (NPR)

NPR1 (Baht/bird) = SBR – FCG

NPR2 (Baht/kgBW) = NPR1 ÷ BW

- Return of investment by comparing with the control group

(ROI)

ROI1 (Baht/bird) = NPR (added Mu-plus®) – NPR (control )

ROI2 (Baht/kg BW) = ROI1 ÷ BWG

3.2 Experiment II. Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on nutrients digestibility of broiler chickens.


A total of 60 seven-d-old Arbor Acres chicks were designed in completely

randomized design (CRD), and all of the birds were allotted to five dietary treatments

with four replications and three birds per each. Birds were moved from floor pens to

battery cages with similar average body weight.

3.2.2 Diet formulation

Birds were fed a diet that contains Cr2O3 0.20 % diet as indigestive

indicator daily during the last 5 days, but fecal sample collects daily during last 3 days

each period. Feed and water offered ad libitum throughout the experiment.







A total of 60 seven-d-old chicks were reared in battery cages and given

feed and water ad libitum throughout the experiment. Birds were fed feed Cr2O3 daily

during the last five days, but fecal sample collects only during the last 3 days.

Therefore, feed Cr2O3 and fecal sample were collected for analyzing nutrients and

28

chromium contain follow the AOAC (2000) procedure for nutrients digestibility of

broilers at different stage of age.

3.2.4 Data record: Nutrients digestibility

- Percentage of dry matter digestibility (DMD)

% indicator in fecal - % indicator in feed % DMD = x 100

% Indicator in fecal

- Percentage of nutrients digestibility (ND)

% indicator in feed nutrient in fecal % ND = 100 – 100 [ x ]

% indicator in fecal nutrient in feed

3.3 Experiment III. Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on small intestinal histomorphology in broiler

chickens.


A total of 50 seven-d-old Arbor Acres chicks were designed in completely

randomized design (CRD), and all of the birds were allotted to five dietary treatments

with two replications and five birds per each. Birds were moved from floor pans to

battery cages with similar average body weight.

3.3.2 Diet formulation

The experiment was divided to five dietary treatments within three periods

including starter, grower and finisher. Feed and water offered ad libitum throughout

the experiment.






29


A total of 50 seven-d-old chicks were reared in battery cages and given

feed and water ad libitum throughout the experiment. One-male and one-female

chickens each treatment was randomly selected to euthanize at day 14, day 21, day 35

and day 42 and collect segment of small intestine for measurement the height and

width of villi and crypt depth.

3.3.4 Data record: Small intestinal histomorphology

Supplementation of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the broiler diets were conducted to study the morphology of

the small intestine at various stages of ages during starter, grower and finishing. One-

male and one-female chickens from each treatment will be euthanized at 14, 21, 35

and 42 days of age to measure the villus height, villus width and crypt depth of

duodenum, jejunum and ileum. Segments of the small intestine were collected about

2.5 to 3.0 cm for each of all three parts; duodenum, jejunum and ileum and then

immediately fixed in formalin 10% until the process paraffin block.

3.4 Analysis data

All data were subjected to analysis of variance by using ANOVA procedure

follow completely randomize design, (CRD) and treatment mean difference were

determined by Duncan’s New Multiple Range Test (DMRT) procedure by the

Statistical Analysis System Institute (SAS, 1996). Difference treatment mean was

analyzed by orthogonal polynomial of Proc GLM.

3.5 Location of research

The experiments were conducted on the station at Poultry farm, Department of

Animal Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen, 40002,

Thailand.

30

Table 3.1 Composition of the experimental diets for starter period (1-21 d)

Ingredients (%) Levels of Mu-Plus (kg/ton)

0.00 0.50 1.00 1.50 2.00

Yellow corn (7.8 % CP) 47.10 47.05 47.00 46.95 46.90

Soybean meal (44% CP) 28.00 28.00 28.00 28.00 28.00

Full-fat soy bean (36 % CP) 18.00 18.00 18.00 18.00 18.00

Monodicalcium phosphate, P21 2.90 2.90 2.90 2.90 2.90

Limestone 1.40 1.40 1.40 1.40 1.40

DL-methionine 0.30 0.30 0.30 0.30 0.30

L-lysine 0.15 0.15 0.15 0.15 0.15

Crude rice bran oil 1.30 1.30 1.30 1.30 1.30

Salt 0.40 0.40 0.40 0.40 0.40

Choline chloride 60% 0.10 0.10 0.10 0.10 0.10

Vitamin-mineral mixes a 0.35 0.35 0.35 0.35 0.35

Mu-Plus® b 0.00 0.05 0.10 0.15 0.20

Total 100.00 100.00 100.00 100.00 100.00

Price per kg (Baht) 14.21 14.40 14.59 14.79 14.98

Chemical composition

CP, % 22.47 22.47 22.46 22.46 22.45

ME, kcal/kg 3,102 3,100 3,098 3,097 3,095 a presented in table 3.4, b presented in table 3.5

31

Table 3.2 Composition of the experimental diets for grower period (22-35 d)


0.00 0.50 1.00 1.50 2.00

Yellow corn (7.8 % CP) 53.24 53.19 53.14 53.09 53.04

Soybean meal (44% CP) 19.70 19.70 19.70 19.70 19.70



Limestone 1.40 1.40 1.40 1.40 1.40

DL-methionine 0.22 0.22 0.22 0.22 0.22

L-lysine 0.21 0.21 0.21 0.21 0.21


Salt 0.40 0.40 0.40 0.40 0.40



Mu-Plus® b 0.00 0.05 0.10 0.15 0.20

Total 100.00 100.00 100.00 100.00 100.00

Price per kg (Baht) 14.02 14.21 14.41 14.60 14.80


CP, % 20.02 20.01 20.01 20.00 20.00


32

Table 3.3 Composition of the experimental diets for finisher period (36-42 d)


0.00 0.50 1.00 1.50 2.00

Yellow corn (7.8 % CP) 59.17 59.12 59.07 59.02 58.97

Soybean meal (44% CP) 14.00 14.00 14.00 14.00 14.00



Limestone 1.30 1.30 1.30 1.30 1.30

DL-methionine 0.17 0.17 0.17 0.17 0.17

L-lysine 0.15 0.15 0.15 0.15 0.15


Salt 0.40 0.40 0.40 0.40 0.40



Mu-Plus® b 0.00 0.05 0.10 0.15 0.20

Total 100.00 100.00 100.00 100.00 100.00

Price per kg (Baht) 13.78 13.97 14.17 14.36 14.56


CP, % 17.97 17.97 17.96 17.96 17.96


33

Table 3.4 Composition of vitamin-mineral mixes in the diets of broilers (vitamin,

mineral /kg diet)

Composition Age of broilers (day)

1-21 22-35 36-42

Vitamin:

Vitamin A 10,000.00 IU 10,000.00 IU 10,000.00 IU

Vitamin D3 2.500.00 IU 2,500.00 IU 2,500.00 IU

Vitamin E 10.00 IU 10.00 IU 10.00 IU

Vitamin K3 2.50 mg 2.50 mg 2.50 mg

Thiamine 2.00 mg 2.00 mg 2.00 mg

Riboflavin 7.00 mg 7.00 mg 7.00 mg

Pyridoxine 5.00 mg 5.00 mg 5.00 mg

Vitamin B12 30.00 mcg 30.00 mcg 30.00 mcg

D-pantothenic acid 12.00 mg 12.00 mg 12.00 mg

Niacin 50.00 mg 50.00 mg 50.00 mg

Choline 1,200.00 mg 1,200.00 mg 1,200.00 mg

Folic acid 1.00 mg 1.00 mg 1.00 mg

Biotin 15.00 mcg 15.00 mcg 15.00 mcg

Ethoxyquin 125.00 mg 125.00 mg 125.00 mg

Mineral:

Mn 60.00 mg 60.00 mg 60.00 mg

Zn 40.00 mg 40.00 mg 40.00 mg

Fe 80.00 mg 80.00 mg 80.00 mg

Cu 8.00 mg 8.00 mg 8.00 mg

Co 2.00 mg 2.00 mg 2.00 mg

I 0.35mg 0.35 mg 0.35 mg

Se 0.15 mg 0.15 mg 0.15 mg

34

Table 3.5 Cost of feedstuff in experimental diets (January – February, 2013)

Feedstuffs Price (Baht/kg)

Yellow corn (7.8 %CP) 10.25

Soybean meal (44 % CP) 14.50

Full-fat soybean (36 % CP) 19.50

Monodicalcium phosphate, P21 18.00

Limestone 0.35

DL-methionine 125.00

L-lysine 72.00

Crude rice bran oil 35.00

Salt 3.50

Choline chloride 60 % 49.00

Vitamin-mineral mixes 80.00

Mu-Plus® 400.00

CHAPTER IV

RESULTS AND DISCUSSION

4.1 Experiment I. Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) on growth performance and carcass quality in broilers

4.1.1 Nutritive values of dietary diet at 1-21, 22-35, 36-42 d of age

Table 4.1, 4.2 and 4.3 indicates the analyzed nutritive composition of the

diets used in the experiment. Apparently, all diets had similar contents of moisture,

ash, calcium (Ca), phosphorus (P), crude protein (CP), ether extract (EE or crude fat),

crude fiber (CF), and gross energy (GE). The nutritive composition analyses of the

diets were reasonably agreed with the calculated values.

Table 4.1 Analyzed nutrient composition of diets at 1-21 d of age (proximate

analysis)

Nutrients Levels of Mu-Plus® (kg/ton)

0.00 0.50 1.00 1.50 2.00

Dry matter (%) 91.66 91.69 91.62 92.13 91.37

Crude protein (%) 23.39 23.66 23.38 23.19 23.00

Crude fiber (%) 2.52 3.02 2.02 2.01 2.52

Ether extract (%) 6.88 7.83 7.35 7.41 7.71

Ash (%) 7.39 8.04 7.70 8.22 7.85

Calcium (%) 1.16 1.13 1.20 1.11 1.17

Total phosphorus (%) 0.79 0.97 0.88 0.79 0.79

Gross energy (kcal/kg) 4469 4521 4433 4433 4448

36


analysis)


0.00 0.50 1.00 1.50 2.00

Dry matter (%) 91.95 91.95 91.97 91.85 92.00

Crude protein (%) 21.04 21.56 20.83 21.25 21.10

Crude fiber (%) 2.50 3.00 3.00 3.01 3.01

Ether extract (%) 6.84 6.90 7.12 7.02 7.25

Ash (%) 7.84 8.27 7.93 7.94 8.18

Calcium (%) 1.19 1.36 1.45 1.13 1.09




analysis)


0.00 0.50 1.00 1.50 2.00

Dry matter (%) 91.21 91.32 92.12 92.40 92.28

Crude protein (%) 18.01 18.56 18.54 18.17 18.33

Crude fiber (%) 2.02 2.02 2.00 1.99 1.99

Ether extract (%) 8.64 8.04 8.41 8.52 8.57

Ash (%) 6.99 6.71 6.56 6.93 6.80

Calcium (%) 1.11 1.03 1.09 1.17 1.20



37

4.1.2 Effect of Andrographis paniculata and Zingiber cassumunar mixture

(Mu-Plus®) in the diets on growth performance of broilers

4.1.2.1 Body weight gain (BWG)

The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and

2.00 kg/ton diet on BWG during 1-21 d, 22-35 d, and 36-42 d of broiler chickens are

presented in Table 4.5. During 1-21 d of age, the averages BWG of broilers are 813,

845, 815, 805 and 824 g/bird, respectively. The results found that supplementation of

Mu-Plus® at all levels showed to increase (P>0.05) BWG higher than the control

group. During 22-35 d of age, the averages BWG of broilers are 1,188, 1,214, 1,168,

1,212 and 1,231 g/bird, respectively. All groups that received dietary Mu-Plus®

showed to increase (P>0.05) BWG higher than the control group. During 36-42 d of

age, the averages BWG of broilers are 332, 329, 371, 340 and 383 g/bird,

respectively. During the overall period (1-42 d of age), the averages BWG of broilers

are 2,333, 2,388, 2,355, 2,357 and 2,448 g/bird, respectively. The results revealed that

BWG of all groups in all periods were not significantly different (P>0.05).

Nevertheless, all groups of broilers fed Mu-Plus® diet were improved to increase

BWG higher than the control group, but 2.00 kg/ton is the best level to provide the

best result. On the other hand, andrographolide and phenylbutenoid compounds active

substance caused growth regulator with increased appetite, which increase the

consumption of nutrients for growth performance. The active substance of both herbs

had the effect of immunostimulant, antibacterial, antifungal activity (Puri et al., 1993

and Chairul et al., 2009). This finding is in agreement with Tongwiti et al, (2012) who

reported that an herbal mixture of Herbatob-Mix® had no effect (P>0.05) on BWG,

but it had shown a greater BWG in all groups fed with Herbatob-Mix®

supplementation in diets. Similar to the results made by Mathivanan et al. (2006) who

reported that feeding 2g/kg of Andrographis paniculata to broilers improved live

weight of broilers. Tangmutthapattharakul (2007) also studied the effect of Zingiber

cassumunar in 0.2, 0.4 and 0.8% diets of broilers at 0-3 weeks of age on BWG

showed that it was not significantly different (P>0.05) among treatment groups.

However, groups fed Zingiber cassumunar at levels 0.2 and 0.4% diets had improved

the BWG higher than the control group. Arkan et al. (2012) also reported that broilers

fed diets with Zingiber officinale at 0.1 and 0.2% diets showed a better BWG when

38

compared with birds fed control diet for the overall period. Further, Tipakorn (2002)

reported that supplementation of Andrographis paniculata leaf powder at 0.1, 0.2, 0.3

and 0.4% diets of broilers compared with the control group indicated that all groups

that received Andrographis paniculata leaf powder diets showed improved BWG

higher than the control group even though it was not significantly different (P>0.05)

among treatment groups. These results contrast from a report made by

Nopparatmaitree (2008) who reported that supplementation of Mu-Plus® at 0.1 and

0.2% diets of weaning pigs had no effect (P>0.05) on BWG when compared to the

control group. Nonetheless, both levels of Mu-Plus® trend to decrease BWG lower

than the control group. Narkchamnarn et al. (2005) reported that supplement

Herbatob-Mix® 1,000 ppm + Mu-Plus® 2,000 ppm in broiler diets showed no effect

(P>0.05) on BWG among treatment groups, but herbal-fed broilers trend to have body

weight gain higher than control and antibiotic. Pratoomtong (2011) reported that

BWG of weaning

Table 4.4 Effect of Mu-Plus® in the diets on the body weight gain (BWG) at

different stage of age of broiler chickens

Treatments Mu-Plus®

(Kg/ton)

Body weight gain (g/bird)

1-21 d 22-35 d 36-42 d 1-42 d

T1 - 813 1,188 332 2,333

T2 0.50 845 1,214 329 2,388

T3 1.00 815 1,168 371 2,355

T4 1.50 805 1,212 340 2,357

T5 2.00 824 1,231 383 2,438

SEM 40.55 155.75 91.17 241.82

--------------------------- Probability --------------------------

Contrast

Lin NS NS NS NS

Quad NS NS NS NS

Cubic NS NS NS NS

Quar NS NS NS NS

39

pigs that fed Herbatob-Mix® (500 ppm) and Herbatob-Mix® + Mu-Plus® (500 ppm +

1,000 ppm) diets were not significantly different (P>0.05) among treatment groups.

Semakanit (2005) also reported that supplement different levels of andrographolide in

the diets of weaning pigs indicated that weaning pigs that received andrographolide

shown decreasing (P>0.05) BWG when compared to the control group.

4.1.2.2 Feed intake (FI)


2.00 kg/ton diet on feed intake during 1-21 d, 22-35 d, and 36-42 d of broiler chickens

are presented in Table 4.4. During 1-21 d of age, the averages FI of broilers are 1,098,

1,117, 1,075, 1,071 and 1,088 g/bird, respectively. The results revealed that

supplementation of Mu-Plus® at the levels 1.00, 1.50 and 2.00 kg/ton showed

decrease (P>0.05) FI lower than the control group. During 22-35 d of age, the

averages FI of broilers are 2,110, 2,126, 2,096, 2,122 and 2,175 g/bird, respectively.

The broilers that received dietary Mu-Plus® at 1.50 and 2.00 kg/ton diets showed

trend to have the highest (P>0.05) FI when compared to the control group. During 36-

42 d of age, the averages FI of broilers are 1,028, 978, 1,000, 996 and 1,086 g/bird,

respectively. The broilers that received dietary Mu-Plus® 2.00 kg/ton diet showed

trend to have the highest (P>0.05) FI when compared to the other groups.

Additionally, overall periods (1-42 d of age), different levels of Mu-Plus® in the diets

also show no effect on FI (P>0.05) and the averages of FI are 4,336, 4,221, 4,170,

4,190 and 4,349 g/bird, respectively. Even though there was not significant different

among groups and the group supplementation of Mu-Plus® at level 2.00 kg/ton

showed trend to have the highest feed intake when compared with control and other

groups. In addition, all groups in all periods, the results indicated that supplement

Mu-Plus® at 2.00 kg/ton diet showed improvement to increase FI higher than the

control group and other. However, bitter taste of Andrographis paniculata and bad

smell of Zingiber cassumunar were not influenced on feed consumption of broilers

because the physiology of broilers does not get the taste and smell of feed.

Nopparatmaitree (2008) investigated and reported the effect of supplementation of

Andrographis paniculata and Zingiber cassumunar mixture (Mu-Plus®) at level 0.1

and 0.2% diets of weaning pigs compared with the control group shown that both

40

levels of herbal mixture had no effect (P>0.05) on FI of weaning pigs. Similar data

also report by Tongwiti (2012) who conducted trials with herbal mixture

Andrographis paniculata, Curcuma longa and Momordica charantia (Herbatob-

Mix ®) at 250, 500 and 1,000 ppm in broiler diets indicated that all three levels of

Herbatob-Mix® had no effect (P>0.05) on FI of broilers in all periods. Pratoomtong

(2011) reported on supplementation of Herbatob-Mix® (500 ppm) and Herbatob-Mix®

+ Mu-Plus® (500 ppm + 1,000 ppm) in the diets of weaning pigs showed no

significantly different (P>0.05) in FI between treatments. Mathivanan et al. (2006)

also reported that feeding Andrographis paniculata at 2 g/kg diet to broilers improved

feed consumption. Previous study made by Tangmutthapattharakul (2007) who

conducted experiments by supplement Zingiber cassumunar at 0.2, 0.4 and 0.8% diets

in 0-3 week of age on FI showed that it was not significantly different (P>0.05)

among treatment groups, but group received Zingiber cassumunar at 0.2% diet shown

FI higher than other levels, antibiotic group and lower than the control group.

Narkchamnarn et al. (2005) also reported that supplement Herbatob-Mix® 1,000 ppm

+ Mu-Plus® 2,000 ppm in the diets of broilers had no effect (P>0.05) on FI, but

broilers that received herbal mixture diets trend to have FI higher than the control and

antibiotic group. Otherwise, Watanasit et al. (2005); Chanjula et al. (2007) and

Mathivanan et al. (2008) also reported that Andrographis paniculata fed broilers show

to decrease feed consumption and feed conversion ratio. Suci et al. (2012) reported

that Andrographis paniculata was using 3 g/kg was given a positive response to

decrease feed consumption of laying hens. Arkan et al. (2012) reported that feed

intake of broilers fed with Zingiber officinale at 0.1 and 0.2% diets did not differ

between treatments for the first period of the experiment, but the birds fed with

Zingiber officinale at 0.1 and 0.2% diets had less (P>0.05) FI for an overall period

when compared with the control birds. These results are in agreement with Semakanit

(2005) who reported that supplement different level of andrographolide in the diets of

weaning pigs indicated that weaning pigs that received andrographolide higher than 6

ppm shown to increase (P<0.05) FI when compared to the control group.

41

Table 4.5 Effect of Mu-Plus® in the diets on feed intake at different age of broilers


(Kg/ton)

Feed intake (g/bird)

1-21 d 22-35 d 36-42 d 1-42 d

T1 - 1,098 2,110 1,028 4,336

T2 0.50 1,117 2,126 978 4,221

T3 1.00 1,075 2,096 1,000 4,170

T4 1.50 1,071 2,122 996 4,190

T5 2.00 1,088 2,175 1,086 4,349

SEM 55.78 223.50 160.44 396.41

--------------------------- Probability --------------------------

Contrast

Lin NS NS NS NS

Quad NS NS NS NS

Cubic NS NS NS NS

Quar NS NS NS NS

4.1.2.3 Feed conversion ratio (FCR)


2.00 kg/ton diet on FCR during 1-21 d, 22-5 d, and 36-42 d of broiler chickens are

presented in Table 4.6. During 1-21 d of age, the averages FCR of broilers are 1.352,

1.322, 1.318, 1.331 and 1.321, respectively. All groups that fed dietary Mu-Plus® had

shown to decrease (P>0.05) FCR when compared with the control group. Moreover,

1.00 kg/ton diet is a good level to decrease the FCR in starter period. During 22-35 d

of age, the averages FCR of broilers are 1.780, 1.755, 1.797, 1.762 and 1.774,

respectively. Broilers fed Mu-Plus® at the levels 0.50, 1.50 and 2.00 kg/ton diets

showed lower (P>0.05) in FCR and the highest FCR in the group that fed 1.00 kg/ton

diet but no difference (P>0.05) when compared to the control group. During 36-42 d

of age, the averages FCR of broilers are 3.120, 2.984, 2.835, 3.035 and 2.854,

respectively. It found that all of the dietary Mu-Plus® had shown to decrease (P>0.05)

FCR and addition Mu-Plus® at 1.00 kg/ton diet showed best level to decrease the FCR

or the best feed efficiency in the finisher period when compared with the control

group. During the overall period (1-42 d of age), the averages FCR of broilers are

42

1.817, 1.767, 1.775, 1.780 and 1.784, respectively. The results found that it was

improved to decrease (P>0.05) FCR or improvement feed efficiency in all dietary

Mu-Plus® fed groups and added 1.00 kg/ton diet is a best level to decrease the FCR in

overall period. Andrographolide and phenylbutenoid compounds active substance

caused growth regulator with increased appetite, which lead to increase the

consumption of nutrients. The active substance of both herbs had the effect of

immunostimulant, antibacterial, antifungal activity (Puri et al., 1993 and Chairul et

al., 2009). A similar previous study of Nopparatmaitree (2008) who supplementation

of Mu-Plus® at 0.1 and 0.2% of weaning pigs diets compared to the control group

found that both levels of Mu-Plus® were improved to decrease (P>0.05) FCR when

compared to the control group. Moreover, Pratoomtong (2011) also reported that

supplementation of Herbatob-Mix® (500 ppm) and Herbatob-Mix® + Mu-Plus® (500

ppm + 1,000 ppm) in the diets of weaning pigs indicated that both the Herbatob-Mix®

+ Mu-Plus® and Herbatob-Mix® showed lower (P>0.05) in FCR when compared with

positive control. Tongwiti et al. (2012) reported that the herbal mixture of Herbatob-

Mix ® had no effect (P>0.05) on FCR, but it shows greater FCR in all groups fed with

Herbatob-Mix® supplementation. Mathivanan et al. (2006) also reported that feeding

Andrographis paniculata at 2g/kg diet to broilers improved FCR. On the other hand,

Tangmutthapattharakul (2007) studied the effect of Zingiber cassumunar in 0.2, 0.4

and 0.8% diets of broilers at 0-3 weeks of age on FCR showed no significantly

different (P>0.05) among treatment groups. Nevertheless, the group that received

Zingiber cassumunar at 0.2% diet had a FCR higher than 0.4 and 0.8% diets, but it is

lower than the control group. Arkan et al. (2012) reported that broilers fed with

Zingiber officinale at 0.1 and 0.2% diets showed the best FCR in overall period, 1.98

and 1.90, respectively, while that of control was 2.25. Suci et al. (2012) reported that

Andrographis paniculata added 3 g/kg was given a positive response to decrease feed

consumption of laying hens. Tipakarn (2002) reported that supplementation of

Andrographis paniculata leaf powder at 0.1, 0.2, 0.3 and 0.4% diets of broilers had no

effect (P>0.05) on FCR, but it improved to decrease the FCR with increasing the

levels of Andrographis paniculata leaf powder in the diets. Narkchamnarn (2005) also

reported that supplement Herbatob-Mix® 1,000 ppm + Mu-Plus® 2,000 ppm in broiler

diets have no effect (P>0.05) on FCR between groups, but herbal-fed broilers trend to

43

have a FCR higher than control and antibiotic group. Semakanit (2005) also reported

that supplement different levels of andrographolide in the diets of weaning pigs,

indicated that weaning pigs that received andrographolide higher than 6 ppm shown to

increase (P<0.05) FCR when compared to control.

Table 4.6 Effect of Mu-Plus® in the diets on the feed conversion ratio (FCR) at

different age of broilers


(Kg/ton)

Feed conversion ratio

1-21 d 22-35 d 36-42 d 1-42 d

T1 - 1.352 1.780 3.120 1.817

T2 0.50 1.322 1.755 2.984 1.767

T3 1.00 1.318 1.797 2.835 1.775

T4 1.50 1.331 1.762 3.032 1.780

T5 2.00 1.321 1.774 2.854 1.784

SEM 0.038 0.097 0.403 0.054

--------------------------- Probability --------------------------

Contrast

Lin NS NS NS NS

Quad NS NS NS NS

Cubic NS NS NS NS

Quar NS NS NS NS

4.1.2.4 Survival rate (SR)


2.00 kg/ton diets on SR during 1-21 d, 22-35 d, and 36-42 d of broiler chickens are

presented in Table 4.7. During 1-21 d of age, the averages SR of broilers are 100,

98.96, 100, 97.92 and 98.96%, respectively. The results showed that the SR was not

significantly different (P>0.05) among treatment groups. During 22-35 d of age, the

averages SR of broilers are 96.88, 100, 97.92, 98.86 and 100%, respectively. The

results indicated that the SR was not significantly different (P>0.05) among treatment

groups. During 35-42 d of age, the averages SR of broilers are 94.51, 100, 100, 98.96

and 97.87%, respectively. The results revealed that it was significantly increased

44

(P<0.05) SR in four graded levels of Mu-Plus® supplement groups when compared to

the control group. In addition, the group fed with Mu-Plus® at 0.50 kg/ton diet

showed the highest SR. During the overall period (1-42 d of age), the averages SR of

broilers are 91.67, 98.96, 97.92, 95.83 and 96.88%, respectively. Supplementation of

Mu-Plus® for all of the four graded levels in the diets on SR of broilers showed no

significantly different (P>0.05) among treatment groups. Even though there was not

significant different, there was trended to improve SR for all groups that received

Mu-Plus® diets showed higher than the control group. Normally, mortality standard

not more than 5%, but Mu-Plus®-free group had a mortality rate higher than standard

and Mu-Plus® groups. This result of this study is in agreement with Chopra et al.

(1992); Chaiwongkeart (1997); Tipakorn (2002) and Mathivanan et al. (2006) who

reported Andrographis paniculata leaf supplementation reduced mortality among

broiler treatments. Otherwise, both Andrographis paniculata and Zingiber

cassumunar are well-known and widely used as medicinal plants to prevent or/and

treat diarrhea, bacteria and fungi-caused diseases that resulted in inflame gut wall and

damage digestive cells and suddenly increased mortality by its anti-inflammation,

antibacterial, antifungal activity and diarrhea prevention (Chopra et al., 1992; Tripathi

et al., 2008; Bhusita et al., 2009; Suebsasana et al., 2009 and Kaewchoothong et al.,

2012).

45

Table 4.7 Effect of Mu-Plus® in the diets on the survival rate (SR) at different age of

broilers


(Kg/ton)

Survival rate (%)

1-21 d 22-35 d 36-42 d 1-42 d

T1 - 100.00 96.88 94.51bb

91.67

T2 0.50 98.96 100.00 100.00aa 98.96

T3 1.00 100.00 97.92 100.00aa 97.92

T4 1.50 97.92 98.86 98.96ab

95.83

T5 2.00 98.96 100.00 97.87ab

96.88

SEM 1.61 2.32 2.96a 5.10

--------------------------- Probability --------------------------

Contrast

Lin NS NS NS NS

Quad NS NS * NS

Cubic NS NS NS NS

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

4.1.2.5 Summary of productive performance

Table 4.8 describes the effects of Andrographis paniculata and

Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets

of broilers on growth performance, the SR and PI at 42 d of age. Supplementation of

Mu-Plus® at all of the four graded levels in the diets on the SR of broilers indicated

that it was no significantly different (P>0.05) among treatment groups. Although there

was not significant different, there was trended to improve SR for all groups received

Mu-Plus® diets showed higher than the control group. The averages SR of broilers are

91.67, 98.96, 97.92, 95.83 and 96.88%, respectively, within the SR improvement are

ranging from +4.54 to +7.95%. Mortality standard usually not more than 5%, but

Mu-Plus®-free group had a mortality rate higher than standard and Mu-Plus® groups.

This result is in agreement with Chopra et al. (1992); Tipakorn (2002) and

Mathivanan et al. (2006) reported Andrographis paniculata leaf supplementation

reduced mortality among broiler treatments. Supplementation of Mu-Plus® at all of

46

the four graded levels in diets showed no effects (P>0.05) on BW, BWG, FI, FCR and

PI. The averages BW of broilers are 2,373, 2,428, 2,395, 2,397 and 2,478 g/bird,

respectively, within BW improvement are ranging from +0.93 to +4.43%. The

averages BWG of broilers are 2,333, 2,388, 2,355, 2,357 and 2,438 g/bird,

respectively, within BWG improvement are ranging from +0.94 to +4.37%. The

averages FI of broilers are 4,259, 4,221, 4,170, 4,190 and 4,349 g/bird, respectively,

within FI improvement are ranging from -2.11 to +2.09%. The averages FCR of

broilers are 1.817, 1.767, 1.775, 1.780 and 1.784, respectively, within the FCR

improvement are ranging from +1.66 to +2.76%. The averages PI of broilers are 282,

319, 309, 302 and 316, respectively, within the PI improvement are ranging from

+7.09 to 13.12%. Although it was no significant difference, but it trended to show

BW, BWG, FI and FCR better than the control group because of andrographolide

constituent in Andrographis paniculata play an important role to stimulate the

digestive system and enzyme lactase, maltase and sucrase to digest and absorb

carbohydrate in all part of the small intestine including duodenum, jejunum and ileum

(Choudhury and Poddar, 1983). PI in all groups that received Mu-Plus® diets showed

the highest when compare to the control group. We can then conclude that

supplementation of Mu-Plus® for all of the four graded levels in the diets showed

improved to increase growth performance of broilers by increase BW, BWG, feed

efficiency and PI of broilers when compared to the control group. Similar to the

results made by Tongwiti (2012) who reported that supplementation of Herbatob-

Mix ® at 500 and 1,000 ppm in the diets of broilers shown improvement PI higher than

the control group.

47

Table 4.8 Effects of Mu-Plus® in the diets of broilers on growth performance,

survival rate (SR) and productive index (PI) at 42 d of age


Kg/ton

BW

g/bird

BWG

g/bird

SR

%

FI

g/bird

FCR

PI*

T1 - 2,373 2,333 91.67 4,259 1.817 282

T2 0.50 2,428 2,388 98.96 4,221 1.767 319

Improvement** (%) +2.32 +2.36 +7.95 +0.89 +2.760 +13.12

T3 1.00 2,395 2,355 97.92 4,170 1.775 309

Improvement (%) +0.93 +0.94 +6.82 +2.09 +2.210 +9.57

T4 1.50 2,397 2,357 95.83 4,190 1.780 302

Improvement (%) +1.01 +1.03 +4.54 +1.62 +1.660 +7.09

T5 2.00 2,478 2,438 96.88 4,349 1.784 316

Improvement (%) +4.43 +4.37 +5.68 -2.11 +1.660 +12.05

SEM 211.63 241.82 5.10 396.41 0.054 39.27

----------------------------- Probability ------------------------

Contrast

Lin NS NS NS NS NS NS

Quad NS NS NS NS NS NS

Cubic NS NS NS NS NS NS

Quar NS NS NS NS NS NS * Productive index (PI) = [BWG (kg) x Survival rate] ÷ [Age (day) x FCR] x 100 ** Improvement = [(Mu-Plus® group – Control group) ÷ Control group] x 100

4.1.3 Effects of Andrographis paniculata and Zingiber cassumunar mixture

(Mu-Plus®) in the diets on feed cost and economic benefit returns

4.1.3.1 Feed cost per gain, (FCG)

Table 4.9 describes the effect of Andrographis paniculata and


on feed costs per gain (FCG) during 1-21 d, 22-35 d, 36-42 d and 1-42 d of age. It

indicated that all groups of all periods were not significantly different (P>0.05).

During 1-21 d and 22-35 d of age, the averages FCG of broilers are 19.22, 19.04,

19.24, 19.69, and 19.80 Baht/kgBW and 24.95, 24.96, 25.90, 25.74, and 26.26

Baht/kgBW, respectively. In both periods, broilers received dietary Mu-Plus® at 1.00,

48

1.50 and 2.00 kg/ton diets shown to increase FCG, exceptionally, broilers fed dietary

Mu-Plus® at 0.50 kg/ton diet shown to decrease FCG when compared with the group

without added Mu-Plus®. Otherwise, during 36-42 d and 1-42 d of age, the averages

FCG of broilers are 43.00, 41.72, 40.17, 43.57, and 41.56 Baht/kgBW and 27.52,

24.98, 25.66, 26.74, and 26.82 Baht/kgBW, respectively. In both of the periods,

groups that fed dietary Mu-Plus® had shown less FCG than the control group. Similar

the previous study reported by Tongwiti et al. (2012) who supplementation of

Herbatob-Mix® at 500 and 1,000 ppm in the diets of broilers at termination of 1-42

days of age had improved to decrease (P>0.05) FCG lower than the control group. On

one hand, this finding is in agreement with Nopparatmaitree (2008) who reported that

supplementation of Mu-Plus® at 0.1 and 0.2% diets of weaning piglets had trended to

decrease (P>0.05) FCG when compared with the control group. Pratoomtong (2011)

also investigated the effect of supplementation of Herbatob-Mix® (500 ppm) and

Table 4.9 Effect of Mu-Plus® in the diets on feed cost per gain (FCG) at different

age of broilers


(kg/ton)

Feed cost

(Baht/kg)

Feed cost per gain* (Baht/kgBW)

1-21 d 22-35 d 36-42 d 1-42 d

T1 - 14.00 19.22 24.95 43.00 27.52

T2 0.50 14.20 19.04 24.96 41.72 24.98

T3 1.00 14.39 19.24 25.90 40.17 25.66

T4 1.50 14.59 19.69 25.74 43.57 26.74

T5 2.00 14.78 19.80 26.26 41.56 26.82

SEM 0.56 1.40 5.72 1.84

---------------------------- Probability ---------------------------

Contrast

Lin NS NS NS NS NS

Quad NS NS NS NS NS

Cubic NS NS NS NS NS

Quar NS NS NS NS NS * Feed cost per gain (FCG) = [Feed cost (Baht/bird) ÷ (Survival x BW)] x 100

49

Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets on FCG of weaning

pigs shown that both of the Herbatob-Mix® and Herbatob-Mix® + Mu-Plus® groups

had trended to decrease in FCG when compared with the control group, but there was

not significant different (P>0.05). This finding is in agreement with Semakanit (2005)

who reported that supplement different level of andrographolide in the diets of

weaning pigs did not show any effect (P>0.05) on FCG. Therefore, all of

andrographolide-fed groups trend to have FCG higher than the control group.

4.1.3.2 Economic benefit returns

Table 4.10 describes the effect of Andrographis paniculata and


on salable bird returns and economic benefit returns at termination 42 d of age.

Supplementation of Mu-Plus® for all of the four graded levels in the diets showed no

effect (P>0.05) on salable bird returns (SBR), its averages are 90.19, 92.27, 91.01,

91.09 and 94.17 Baht/bird, respectively. However, it was not significantly different

among treatment groups, but all groups received Mu-Plus® diets showed SBR higher

than the control group. Supplementation of Mu-Plus® for all of the four levels in the

diets showed no effects (P>0.05) on net profit returns per bird (NPR1) and returns of

investment by comparing with the control group (ROI1), but all groups that received

Mu-Plus® diets showed NPR1 and ROI1 higher than the control group. The averages

NPR1 of broilers are 25.19, 31.69, 29.54, 26.84 and 27.80 Baht/bird, respectively

within the averages ROI1 of broilers are ranging from +1.65 to +6.50 Baht/bird.

Moreover, supplementation of Mu-Plus® for all of the four graded levels in the diets

also showed no effects (P>0.05) on net profit returns per kilogram body weight

(NPR2) and returns of investment by comparing with the control group (ROI2), but

all groups of broilers fed Mu-Plus® diets shown NPR2 and ROI2 higher than the

group that without added Mu-Plus®. The averages NPR2 of broilers are 10.48, 13.02,

12.34, 11.26 and 11.18 Baht/kgBW, respectively within the averages ROI2 of broilers

are ranging from +0.70 to +2.54 Baht/kgBW. This finding is in agreement with

Tongwiti et al. (2012) who reported that supplementation of Herbatob-Mix at 500 and

1,000 ppm in the diets of broilers during 1-42 d of age showed to improve NPR1 and

NPR2 higher than the group that without added Herbatob-Mix®, however, it was not

50

significantly different (P>0.05) among treatment groups. Moreover, Pratoomtong

(2011) investigated the effect of supplementation of Herbatob-Mix® (500 ppm) and

Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets on the economic

benefit returns of weaning pigs showed that Herbatob-Mix® and Herbatob-Mix® +

Mu-Plus® groups had improved to increase in economic benefit returns when

compared to the control group. Nevertheless, it was not significantly different

(P>0.05). Similar report made by Nopparatmaitree (2008) who reported that

Table 4.10 Effects of Mu-Plus® in the diets of broilers on economic benefit returns

for overall 42 d of testing

Mu-Plus®

Kg/ton

FCG*

Baht/bird

SBR**

Baht/bird

NPR1***

Baht/bird

ROI1****

Baht/bird

NPR2***

Baht/kgBW

ROI2****

Baht/kgBW

T1 - 65.00 90.19 25.19 - 10.48 -

T2 0.50 60.58 92.27 31.69 +6.50 13.02 +2.54

T3 1.00 61.47 91.01 29.54 +4.35 12.34 +1.86

T4 1.50 64.24 91.09 26.84 +1.65 11.26 +0.78

T5 2.00 66.36 94.17 27.80 +2.61 11.18 +0.70

SEM 6.94 9.182 5.55 - 1.84 -

---------------------------- Probability ------------------------------------

Contrast

Lin NS NS NS NS NS NS

Quad NS NS NS NS NS NS

Cubic NS NS NS NS NS NS

Quar NS NS NS NS NS NS * Feed cost per gain (FCG) = [Feed cost (Baht/bird) ÷ Survival] x 100 ** Salable bird returns (SBR) = Price of live chicken (38 Baht) x BW *** Net profit returns per bird (NPR)

NPR1= SBR – FCG

NRP2 = NPR1 ÷ BW **** Returns of investment by comparing with the control group (ROI)

ROI1 = NPR1 (Added Mu-Plus®) – NPR1 (Control)

ROI2 = NPR2 (Added Mu-Plus®) – NPR2 (Control)

51

supplementation of Mu-Plus® at 0.1 and 0.2% diets of weaning pigs on economic

benefit returns indicated that it was not significantly different (P>0.05) among groups.

This finding is contrast from Semakanit (2005) reported on supplement different

levels of andrographolide in the diets of weaning pigs had an effect (P<0.05) to

decrease economic benefit returns within increasing the level of andrographolide.


(Mu-Plus®) in the diets on carcass quality of broilers

4.1.4.1 Effects of Mu-Plus® in the diets on dressing percentage and

carcass characteristics of broilers

Table 4.11 describes the effects of Mu-Plus® supplementation at

0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets full period (1-42 d of age) on carcass

characteristics, it's shown that dressing percentage, percentage of breast, drumstick,

thigh, wings, feet and edible meat were not significantly different (P>0.05) among

treatment groups. The average dressing percentages of broilers are 85.41, 86.62,

87.02, 86.56 and 85.57%, respectively. The average percentages of the breast are

23.28, 23.73, 23.54, 23.15 and 23.55%, respectively. The average percentages of the

thigh are 12.96, 13.34, 13.05, 12.77 and 12.97%, respectively. The average

percentages of the drumstick are 10.25, 10.46, 10.32, 10.68 and 10.63%, respectively.

The average percentages of the wings are 7.45, 7.49, 7.70, 7.66 and 7.50%,

respectively. The average percentages of the feet are 3.36, 3.46, 3.49, 3.56 and 3.46%,

respectively. The average percentages of the edible meat of broilers are 53.94, 54.73,

54.06, 52.97 and 54.65%, respectively. The average percentage of the carcass of

broilers showed no significantly different (P>0.05) among treatment groups. It may be

caused by experimental diets contained a similar percentage of protein and energy

among treatment groups that why it did not influence on carcass quality. On the other

hand, the bioactive ingredient of both herbs prevented isoproterenol induced elevated

lipid peroxidation and elevated the activities of antioxidant enzymes, superoxide

dismutase, calatase, glutathione peroxidase and the level of reduced glutathione level

in hearts (Ojha et al., 2009 and Dhiman et al., 2012). There was trend to have higher

in percentage of carcass characteristics for all groups that received dietary Mu-Plus®.

Similar to reporting made by Tongwiti (2012) who had studied the effect of Herbatob-

52

Mix ® at 250, 500 and 1,000 ppm in the diets of broilers compared with a control group

on carcass characteristics indicated that dressing percentage, percentage of breast,

thigh, drumstick, wings and total edible meat of all groups were not significantly

different (P>0.05). This finding is in agreement with Kungsoksomboun (2007)

reported on supplementation of Zingiber cassumunar at 0.2, 0.4 and 0.8% diets of

broilers showed no significantly differ (P>0.05) on dressing percentage, breast, thigh,

drumstick, wings and total edible meat when compared to the control group.

Table 4.11 Effects of Mu-Plus® in the diets on carcass quality of broilers at

termination (42 d of age)

Mu-Plus®

(Kg/ton)

Dressing

Percentage

Carcass quality (%)

Breast

Thigh

Drumstick

Wings

Feet

Total edible

meat*

T1 - 85.41 23.28 12.96 10.25 7.45 3.36 53.94

T2 0.50 86.62 23.73 13.34 10.46 7.49 3.46 54.73

T3 1.00 87.02 23.54 13.05 10.32 7.70 3.49 54.06

T4 1.50 86.56 23.15 12.77 10.68 7.66 3.56 52.97

T5 2.00 85.57 23.55 12.97 10.63 7.50 3.46 54.65

SEM 2.02 1.45 0.68 0.79 0.41 0.29 2.17

---------------------------- Probability ------------------------------------

Contrast

Lin NS NS NS NS NS NS NS

Quad NS NS NS NS NS NS NS

Cubic NS NS NS NS NS NS NS

Quar NS NS NS NS NS NS NS * Total edible meat = Breast + Drumstick + Thigh + Wings

Nevertheless, all groups of broilers that received Zingiber cassumunar trend to have

carcass characteristics higher than the control group. A similar previous study report

by Tipakorn (2002) also mentioned that it was not significantly different (P>0.05) in

carcass characteristics between each group while broilers was fed at varying levels of

Andrographis paniculata leaf powder in the diets (0.1, 0.2, 0.3 and 0.4% diets).

53

Narkchamnarn (2005) also reported that supplement Herbatob-Mix® 1,000 ppm +

Mu-Plus® 2,000 ppm in broiler diets showed no effect (P>0.05) on dressing

percentage, wings, breast and drumstick. However, herbal-fed broilers trended to have

dressing percentage and drumstick better than control and antibiotic group.

4.1.4.2 Effects of Mu-Plus® in the diets on the organ weights and

abdominal fat of broilers

Table 4.12 describes the effects of Mu-Plus® at 0, 0.50, 1.00,

1.50 and 2.00 kg/ton diets on the percentage of organ weights and percentage of

abdominal fat of broilers at termination 42 d of age. The results indicated that the

average percentage of organ weights and abdominal fat were not significantly

different (P>0.05) among treatment groups. The average percentages of abdominal fat

of broilers are 1.20, 1.14, 1.16, 1.17 and 1.17%, respectively. The average percentages

of liver of broilers are 1.70, 1.83, 1.90, 1.80 and 1.72%, respectively. The average

percentages of spleen of broilers are 0.10, 0.08, 0.11, 0.09 and 0.08%, respectively.

The average percentages of heart of broilers are 0.41, 0.41, 0.40, 0.39 and 0.40%,

respectively. The average percentages of the gizzard of broilers are 1.34, 1.34, 1.49,

1.39 and 1.37%, respectively. The average percentages of the thymus of broilers are

0.40, 0.36, 0.37, 0.38 and 0.35%, respectively. The average percentages of the bursa

of broilers are 0.09, 0.07, 0.09, 0.08 and 0.08%, respectively. The average percentage

of organ weights and abdominal fat of broilers were not significantly different

(P>0.05) among treatment groups, it may be caused by the diets of all groups contain

similar values of protein and energy and then it showed no effects on organ weights

and abdominal fat. However, supplement 0.50 kg/ton diet showed a good level that

provides best results when compared with the control group. Andrographolide and

phenylbutenoid compounds (Zingiber cassumunar) function as an immune-

stimulatory activity to stimulate antibody (Puri et al., 1993 and Chairul et al., 2009).

On the other hand, Andrographolide also play role as a potent stimulator of

gallbladder function by producing a significant increase in bile flow, bile salts and

improved liver function (Dhiman et al., 2012), hepatoprotective by reducing a lipid

peroxidation (Kapil et al., 1993). This finding is in agreement with Tongwiti (2012)

who studied the effects of Herbatob-Mix® at 250, 500 and 1,000 ppm in the diets of

54

broilers when compared with a control group on carcass characteristics indicated that

percentage of abdominal fat, liver, spleen, heart, gizzard in all groups were not

significantly different (P>0.05). Similar previous studies was reported by

Kungsoksomboun (2007) who supplementation of Zingiber cassumunar at 0.2, 0.4

and 0.8% diets of broilers showed no significantly differ (P>0.05) on abdominal fat,

liver, heart and gizzard when compared with the control group. Nevertheless, all

groups of broilers that received Zingiber cassumunar trend to have organ weights and

abdominal fat higher than the control group. Narkchamnarn (2005) also reported that

supplement Herbatob-Mix® 1,000 ppm + Mu-Plus® 2,000 ppm in broiler diets showed

no effect (P>0.05) on abdominal fat, heart, liver and gizzard.

Table 4.12 Effects of Mu-Plus® in the diets on organ weights and abdominal fat of

broilers at termination (42 d of age)

Mu-Plus®

(Kg/ton)

Organ weights (%BW)

Abdominal

fat

Liver Spleen Heart Gizzard Thymus Bursa of

fabriciums

T1 - 1.20 1.78 0.10 0.41 1.34 0.40 0.09

T2 0.50 1.14 1.83 0.08 0.41 1.34 0.36 0.07

T3 1.00 1.16 1.90 0.11 0.40 1.49 0.37 0.09

T4 1.50 1.17 1.80 0.09 0.39 1.39 0.38 0.08

T5 2.00 1.17 1.72 0.08 0.40 1.37 0.35 0.08

SEM 0.30 0.25 0.04 0.06 0.16 0.09 0.04

---------------------------- Probability ------------------------------------

Contrast

Lin NS NS NS NS NS NS NS

Quad NS NS NS NS NS NS NS

Cubic NS NS NS NS NS NS NS

Quar NS NS NS NS NS NS NS

55

4.1.4.3 Effects of Mu-Plus® in the diets on composition of breast meat

and liver of broilers at termination

The composition of breast meat and liver of broiler chickens at

termination, including the percentage of crude protein and fat are presented in Table

4.13. It shown that broilers fed with Mu-Plus® at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton

diets were significantly different (P<0.05) with linear decrease in the percentage of

crude protein and highly significantly different (P<0.01) with linear decrease in

percentage of fat of breast meat while increasing the level of Mu-Plus® in basal diet.

The averages CP of breast meat of broilers are 26.41, 25.60, 26.54, 26.08 and 24.97%,

respectively. The average fats by ether extract of breast meat of broilers are 3.37,

3.11, 3.02, 2.92 and 2.82%, respectively. In contrast, in the liver, it found that broilers

Table 4.13 Effects of Mu-Plus® in the diets on composition of breast meat and liver

of broilers at termination (42 d of age)

Mu-Plus®

(Kg/ton)

Composition of breast meat (%) Composition of Liver (%)

CP1 IM 3 Fat2 IM 3 CP1 IM 3 Fat2 IM 3

T1 - 26.41aa - 3.37

aa - 18.16bb - 3.95

a -

T2 0.50 25.60ab -3.06 3.11

ab -7.71 18.84ab +3.74 3.68

a -6.83

T3 1.00 26.54aa +0.49 3.02

ab -10.38 18.96aa +4.40 3.97

a +0.51

T4 1.50 26.08aa -1.25 2.92

bb -13.35 19.02aa +4.73 3.02

b -23.54

T5 2.00 24.97bb -5.45 2.82

bb -16.32 18.88ab +3.96 2.81

b -28.86

SEM 0.69aa

0.25aa

0.46ab 0.30

a

---------------------------- Probability ------------------------------------

Contrast

Lin * ** * **

Quad NS NS NS NS

Cubic * NS NS NS

Quar NS NS NS ** a, b column means with different superscripts differ significantly at P <0.05.

* = P<0.05, ** = P<0.01 and NS = non-significant, 1 CP = crude protein, 2 Fat by ether extract 3 Improvement (IM, %) = [(Mu-Plus® group – Control group) ÷ Control group] x 100

56

fed Mu-Plus® at different levels in the diets were highly significantly different

(P<0.01) with linear decrease in percentage of fat and significant different (P<0.05)

with linear increase in CP of liver when increasing the levels of Mu-Plus® in basal

diet. The averages CP of liver of broilers are 18.16, 18.84, 18.96, 19.02 and 18.88%,

respectively. The average fats by ether extract of liver of broilers are 3.95, 3.68, 3.97,

3.02 and 2.81%, respectively. However, Mu-Plus® supplementation at 1.50 kg/ton diet

was a good level to improve the composition of liver and breast meat including CP

and fat when compared with the control group. From these results, It may be caused

by andrographolide have capable to improve liver function (Dhiman et al., 2012) and

hepatoprotective activity by reducing a lipid peroxidation (Kapil et al., 1993).

Moreover, active ingredients of both herbs play an important role to elevate the

activities of antioxidant enzymes, superoxide dismutase, calatase, glutathione

peroxidase (Ojha et al., 2009). This finding is in contrast from Tongwiti (2012) who

investigated the effect of Herbatob-Mix® at 250, 500 and 1,000 ppm in the diets of

broilers compared with the control group on the composition of breast meat, the

results indicated that percentage of CP and fat in all groups were not significantly

different (P>0.05) among treatment groups.

4.2 Experiment II. Effects of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on nutrients digestibility of broiler chickens


(Mu-Plus®) in the diets on dry matter digestibility (DMD) of broiler

chickens

This experiment investigated the effect of Andrographis paniculata and


during 1-21 d, 22-35 d and 36-42 d of age of broiler chickens on dry matter

digestibility (DMD). During 1-21 d of age, the averages DMD of broilers are 60.56,

62.30, 62.07, 59.84 and 60.60%, respectively. Broilers fed with Mu-Plus® diet at 0.50,

1.00 and 2.00 kg/ton diets shown to have DMD higher than the control group, but

there was not significant different (P>0.05) (Table 4.14). During 22-35 d of age, the

averages DMD of broilers are 72.99, 75.68, 73.06, 68.95 and 69.10%. Broilers that

received Mu-Plus® diet at 0.50 kg/ton diet shown to have DMD higher (P<0.01) than

57

the control group and other groups (Table 4.15). During 35-42 d of age, DMD of

broilers was highly significantly different (P<0.01) among treatment groups. The

averages DMD of broilers are 74.63, 77.10, 74.11, 63.78 and 71.76%, respectively.

Broilers fed with Mu-Plus® diet at 0.50 kg/ton diet showed the highest in DMD when

compared to the other group and the control group (Table 4.16). Similar results also

reported by Pratoomtong (2011) showed that supplementation of Herbatob-Mix® (500

ppm) and Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets show effect

(P<0.01) on DMD of weaning pigs. Then, both of the Herbatob-Mix® and Herbatob-

Mix ® + Mu-Plus® showed improving increase DMD of weaning pigs when compared

to the control group. On the other hand, Semakanit (2005) also reported that

supplement andrographolide in the diets of weaning pigs had an effect (P<0.05) to

decrease DMD when increasing the level of andrographolide higher than 9 ppm in the

diets. This result is in contrast from Nopparatmaitree (2008) who reported that

supplement Mu-Plus® at 0.1 and 0.2% diets showed no effect (P>0.05) on DMD of

weaning pigs, but Mu-Plus® trend to have DMD higher than the control group.

Palawatvichai (2007) also reported that supplement Thai herbal (Turmeric) had no

any effect (P>0.05) on DMD in broilers.


(Mu-Plus®) in the diets on crude protein digestibility (CPD) of broiler

chickens



during 1-21 d, 22-35 d and 36-42 d of age of broiler chickens on crude protein

digestibility (CPD). During 1-21 d of age, the averages CPD of broilers are 64.29,

65.89, 62.97, 63.54 and 63.73%, respectively. Broilers fed with Mu-Plus® at 0.50

kg/ton diet shown to have CPD higher (P>0.05) than the control group and another

group (Table 4.14). During 22-35 d of age, CPD of broilers showed significant linear

decreased (P<0.01) by increasing the levels of Mu-Plus® in the diets when compared

to the control group. The averages CPD of broilers are 76.25, 76.98, 72.34, 72.31 and

69.54%, respectively (Table 4.15). During 36-42 d of age, CPD of broilers was

significant decrease (P<0.05) in linear, quadratic and cubic when increasing the levels

58

of Mu-Plus® in the diets when compared to the control group. The averages CPD of

broilers are 70.74, 73.00, 66.54, 58.11 and 68.42%, respectively (Table 4.16). These

results showed decreasing the CPD when increasing the levels of Mu-Plus® in the

diets. It may be caused by andrographolide constituents in Andrographis paniculata

act functions to stimulate the digestive system and enzyme lactase, maltase and

sucrase to digest and absorbed carbohydrate in the small intestine (Choudhury and

Poddar, 1983). Furthermore, Andrographolide and phenylbutenoid compounds

(Zingiber cassumunar) act function immunostimulatory activity to stimulate antibody

(Puri et al., 1993 and Chairul et al., 2009). On the other hand, andrographolide also

play role as a potent stimulator of gallbladder function by producing a significant

increase in bile flow, bile salts and improved liver function (Shukla et al., 1992). This

finding is in agreement with Semakanit (2005) who reported that supplement different

level of andrographolide in the diets of weaning pigs indicated that weaning pigs that

received andrographolide showed decrease (P<0.05) protein utilization when

compared to the control group. Consequently, It will decrease in crude protein

digestibility if the supplement andrographolide more than 9 ppm in the diets. It is

contrasted from Nopparatmaitree (2008) who reported that supplement Mu-Plus® at

0.1 and 0.2% diets had no effect (P>0.05) on CPD of weaning pigs, but Mu-Plus®-fed

weaning pigs shown to increase CPD when compared to the control group.

Pratoomtong (2011) also reported that supplementation Herbatob-Mix® (500 ppm)

and Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets had no effect

(P>0.05) on CPD of weaning pigs. Nonetheless, both of the Herbatob-Mix® and

Herbatob-Mix® + Mu-Plus® groups showed to increase CPD of weaning pigs when

compared to the control group. Tangmutthapattharakul (2007) studied the effect of

Zingiber cassumunar at 0.2, 0.4 and 0.8% diets of broilers at 0-3 weeks of age on

protein utilization shown no significantly different (P>0.05) among treatment groups,

but groups that received Zingiber cassumunar diets showed FI higher than the control

group. Palawatvichai (2007) also reported that supplement Thai herbal (Turmeric)

showed no any effect (P>0.05) on CPD of broilers when compared to the control

group.

59


(Mu-Plus®) in the diets on ether extract digestibility (EED) of broiler

chickens



during 1-21 d, 22-35 d and 36-42 d of age of broiler chickens on ether extract

digestibility (EED). During 1-21 d of age, the averages EED of broilers are 70.37,

72.33, 73.89, 71.51 and 73.35%, respectively (Table 4.14). All of the groups of

broilers fed with Mu-Plus® diet shown increase EED when compared with the control

group, but no significantly different (P>0.05) among treatment groups. During 22-35

d of age, EED of broilers was significant increased (P<0.01) in quartic and cubic by

increasing the levels of Mu-Plus® in the diets when compared to the control group.

The averages EED of broilers are 79.69, 83.83, 84.63, 80.58 and 82.45%, respectively

(Table 4.15). During 36-42 d of age, EED of broilers was significant increased

(P<0.01) in linear, cubic, quartic and significantly increased (P<0.05) in quadratic

when increasing the levels of Mu-Plus® in the diets when compared to the control

group. The averages EED of broilers are 82.33, 87.06, 83.35, 75.22 and 84.31%,

respectively (Table 4.16). The results shown that Mu-Plus®-supplemented diet

improved fat digestion in the small intestine because of Andrographis paniculata and

Zingiber cassumunar play role as antibacterial, antimicrobial and hepatoprotective

that it helps to protect liver and stimulate the gallbladder to increase bile flow, bile

acid and bile salt (Shukla et al., 1992). Oral administration of andrographolide to adult

male rats produced a dose-related and time-dependent characteristic activation of

brush-border membranes-bound hydrolases and stimulates lactases, maltase and

sucrase in all parts of the small intestine (Choudhury and Poddar, 1985). It also

showed good response to the enterocytes (intestinal absorptive cell) developed.

Enterocytes also act function as secretary and contain digestive enzymes including

peptidase, sucrase, maltase, lactase and lipase. On the other hand, andrographolide act

function role as an emulsifier, to increase the surface area of fat that making available

to digest by lipase. These results similar to reports made by Nopparatmaitree (2008)

who reported that supplement Mu-Plus® at 0.1 and 0.2% diets had no effect (P>0.05)

on EED of weaning pigs, but Mu-Plus® shown to increase EED when compared to the

60

control group. This result is in agreement with Pratoomtong (2011) who reported that


ppm + 1,000 ppm) showed no effect (P>0.05) on EED of weaning pigs. Nonetheless,

both of the Herbatob-Mix® and Herbatob-Mix® + Mu-Plus® groups improved EED of

weaning pigs when compared to the control group. Semakanit (2005) also reported

that supplement andrographolide in the diets of weaning pigs showed no effect

(P>0.05) on EED when compared to the control group. There was a trend to increase

EED when increasing the levels of andrographolide in the diets even though there was

no significant between groups. Palawatvichai (2007) also reported that supplement

Thai herbal (Turmeric) increased (P>0.05) EED of broilers when compared to the

control group.


(Mu-Plus®) in the diets on gross energy digestibility (GED) of broiler

chickens

The experiment investigated the effect of Andrographis paniculata and


during 1-21 d, 22-35 d and 36-42 d of age of broiler chickens on gross energy

digestibility (GED). During 1-21 d of age, the averages GED of broilers are 69.93,

72.20, 70.79, 71.27 and 70.76%, respectively (Table 4.14). All of the broilers fed with

Mu-Plus® diets showed increase GED when compared with the control group, but no

significantly different (P>0.05) among treatment groups. During 22-35 d of age, the

averages GED of broilers are 82.48, 82.97, 81.36, 79.87 and 79.47%, respectively

(Table 4.15). The level of Mu-Plus® at 0.50 kg/ton diet showed the highest GED

when compared with the control group. During 36-42 d of age, GED of broilers was

significantly different (P<0.01) in linear, cubic and quartic among groups when

compared with control without added Mu-Plus® in the diets. The averages GED of

broilers are 81.90, 83.63, 81.45, 72.90 and 79.63%, respectively (Table 4.16).

Andrographis paniculata and Zingiber cassumunar play role as antibacterial,

antimicrobial and hepatoprotective which helps to protect liver and stimulate the

gallbladder to increase bile flow, bile acid and bile salt (Shukla et al., 1992). It also

has a good response to the enterocytes (intestinal absorptive cell) developed.

61

Enterocytes also act function as secretary and contain digestive enzymes including

sucrase, maltase, lactase and lipase. On the other hand, andrographolide act function

as emulsifier, to increase the surface area of fat that making available to digest by

lipase. Nopparatmaitree (2008) reported that supplement Mu-Plus® at 0.1 and 0.2%

diets had no effect (P>0.05) on GED of weaning pigs, but Mu-Plus® showed to

increase GED when compared to the control group. These results are in agreement

with Pratoomtong (2011) who reported that supplementation Herbatob-Mix® (500

ppm) and Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets had an

effect (P<0.05) on GED of weaning pigs. In addition, both of the Herbatob-Mix® and

Herbatob-Mix® + Mu-Plus® groups improved GED of weaning pigs when compared

with the control group. Moreover, Semakanit (2005) also reported that supplement

andrographolide in the diets of weaning pigs had an effect (P<0.05) to increase GED

within increasing the levels of andrographolide in the diets. Palawatvichai (2007) also

reported that supplement Thai herbal (Turmeric) had no any effect (P>0.05) on GED

of broilers when compared to the control group.

62

Table 4.14 Effects of Mu-Plus® in the diets on nutrients digestibility of broilers at

1-21 d of age


(Kg/ton)

Nutrients digestibility (%)

DMD1 CPD2 EED3 GED4

T1 - 60.56 64.29 70.37 69.93

T2 0.50 62.30 65.89 72.33 72.20

T3 1.00 62.07 62.97 73.89 70.79

T4 1.50 59.84 63.54 71.51 71.27

T5 2.00 60.60 63.73 73.35 70.76

SEM 2.62 2.61 2.64 2.64

--------------------------------- Probability -------------------------------------

Contrast

Lin NS NS NS NS

Quad NS NS NS NS

Cubic NS NS NS NS

Quar NS NS NS NS 1 DMD = Dry matter digestibility, 2 CPD = Crude protein digestibility, 3 EED = Ether extract digestibility, 4 GED = Gross energy digestibility

63


22-35 d of age


(Kg/ton)


DMD1 CPD2 EED3 GED4

T1 - 72.99b 76.25

a 79.69cc 82.48

ab

T2 0.50 75.68a 76.98

a 83.83aa 82.97

aa

T3 1.00 73.06b 72.34

b 84.63aa 81.36

bb

T4 1.50 68.95c 72.31

b 80.58bc 79.87

cc

T5 2.00 69.10c 69.54

c 82.45ab 79.47

cc

SEM 0.85 c 0.89

c 0.87

bc 0.49

aa

--------------------------------- Probability -------------------------------------

Contrast

Lin *** *** NS ***

Quad * NS ** NS

Cubic ** NS ** *

Quar NS * * NS a, b, c column means with different superscripts differ significantly at P <0.05.

* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant. 1 DMD = dry matter digestibility, 2 CPD = crude protein digestibility, 3 EED = ether extract digestibility, 4 GED = gross energy digestibility

64


36-42 d of age


(Kg/ton)


DMD1 CPD2 EED3 GED4

T1 - 74.63b 70.74

ab 82.33cc 81.90

b

T2 0.50 77.10a 73.00

aa 87.06aa 83.63

a

T3 1.00 74.11b 66.54

cc 83.35bc 81.45

b

T4 1.50 63.78d 58.11

dd 75.22dd 72.90

d

T5 2.00 71.76c 68.42

bc 84.31bb 79.63

c

SEM 0.76c 1.06

cc 0.52bb 0.56

c

--------------------------------- Probability -------------------------------------

Contrast

Lin *** *** ** ***

Quad NS ** * NS

Cubic *** *** *** **

Quar ** NS ** *** a, b, c, d column means with different superscripts differ significantly at P <0.05.

* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant. 1 DMD = dry matter digestibility, 2 CPD = crude protein digestibility, 3 EED = ether extract digestibility, 4 GED = gross energy digestibility

65

4.3 Experiment III. Effect of Andrographis paniculata and Zingiber cassumunar

mixture (Mu-Plus®) in the diets on small intestinal histomorphology of

broiler chickens


(Mu-Plus®) in the diets of broiler chickens on the villi height

The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and 2.00

kg/ton diets on the villi height of duodenum, jejunum and ileum at four stages of age

of broiler chickens are presented in Table 4.17. The average duodenal villi heights of

broilers are 718.88, 837.25, 767.38, 999.64 and 968.33 µm, respectively (during 14 d

of age), 831.36, 745.39, 782.19, 1,037.57 and 930.64 µm, respectively (during 21 d of

age), 919.00, 945.68, 1,180.36, 1,033.96 and 1,184.21 µm, respectively (during 35 d

of age), and 1,012.53, 925.22, 1,037.77, 1,222.43 and 1,371.41 µm, respectively

(during 42 d of age). The average jejunal villi heights of broilers are 698.47, 685.20,

794.12, 575.42 and 649.57 µm, respectively (during 14 d of age), 834.87, 742.27,

1,039.73, 1,019.83 and 894.18 µm, respectively (during 21 d of age), 884.70, 919.57,

983.51, 1,076.51 and 1,397.76 µm, respectively (during 35 d of age), and 810.57,

543.17, 594.23, 1,314.61 and 1,049.48 µm, respectively (during 42 d of age). The

average ileal villi heights of broilers are 474.30, 475.16, 461.43, 537.42 and 571.61

µm, respectively (during 14 d of age), 510.15, 601.46, 680.72, 633.76 and 725.54 µm,

respectively (during 21 d of age), 589.98, 652.68, 599.77, 639.43 and 799.97 µm,

respectively (during 35 d of age), and 491.56, 408.29, 673.37, 590.56 and 499.04 µm,

respectively (during 42 d of age). The results showed that there was significant

increase (P<0.01) in villi height of duodenum in all periods with increasing the levels

of Mu-Plus® in dietary treatments when compared to the control group. Thus,

supplementation of Mu-Plus® at 1.50 and 2.00 kg/ton diets provided the highest villi

height of duodenum in starter, grower and finisher period, respectively when

compared with the control group. In the jejunum, supplementation of Mu-Plus® at

1.00 and 2.00 kg/ton diets provided the highest (P<0.01) villi height in starter, grower

and finisher period, respectively when compared with the control group. In the ileum,

supplementation of Mu-Plus® at 2.00 kg/ton diet provided the highest (P<0.01) villi

height in all periods of testing when compared with the control group. All graded

levels of Mu-Plus® supplemented in the diets was linear significant increased (P<0.01)

66

villi height of duodenum, jejunum and ileum when increasing the level of herbal

mixture Mu-Plus® in the diets. Therefore, we can suppose thinking that all the three

parts of small intestine of broilers fed Mu-Plus® diets show a good response to

enterocytes (intestinal absorptive cell) developed. Enterocytes are simple columnar

epithelial cell located in muscularis mucosa layer of the small intestine and microvilli

on the apical surface increase surface area for the digestion and transport of molecules

from the intestinal lumen. Enterocytes also play role as secretor and contain digestive

enzymes including peptidase, sucrase, maltase, lactase and lipase. These results are in

agreement with Nopparatmaitree (2008) who investigated the effect of Mu-Plus® at

0.1 and 0.2% diets in weaning pigs showed that both levels of Mu-Plus® in the diets

have an effect (P<0.01) to increase the villi height of duodenum, jejunum and ileum

when compared to the control group. Similar previous results reported by

Pratoomtong (2011) who investigated the effect of supplementation of Herbatob-

Mix ® (500 ppm) and Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets

of weaning pigs showed that both Herbatob-Mix® and Herbatob-Mix® + Mu-Plus®

trended to have a villi height of duodenum, jejunum and ileum higher than the control

group, but showed no significant different (P>0.05) among treatment groups. Similar

reports made by Mathivanan and Edwin (2012) reported that broilers fed

Andrographis paniculata and probiotic diet shown higher significantly increased

(P<0.05) in ileal villi than the control group. This finding is in agreement with Awad

(2009) reported that the probiotics supplemented groups had taller villi in the

duodenum and jejunum in broilers. Narkchamnarn (2005) also reported that

supplement Herbatob-Mix® 1,000 ppm + Mu-Plus® 2,000 ppm in broiler diets showed

increase (P<0.05) the villi height of duodenum and jejunum when compared to the

control group.

67

Table 4.17 Effects of Mu-Plus® in the diets on the villi height of duodenum, jejunum and ileum of broilers at different period of age

Treatments

Mu-Plus®

(Kg/ton)

Villi height (µm)

Duodenum Jejunum Ileum

14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d

T1 - 718.88cc 831.36

bc 919.00

bb 1,012.53

bc 698.47abb 834.87

b 884.70cc 810.57

c 474.30 510.15dd

589.98b 491.56

c

T2 - 0.50 837.25bc 745.39

cc 945.68

bb 925.22cc 685.20

abc 742.27c 919.57

cc 543.17

d 475.16 601.46

cc 652.68

b 408.29

d

T3 - 1.00 767.38cc 782.19

bc 1,180.36aa 1,037.77

bc 794.12

aaa 1,039.73a 983.51

bc 594.23d 461.43 680.72

ab 599.77

b 673.37

a

T4 - 1.50 999.64aa 1,037.54

aa 1,033.96ab 1,222.43

ab 575.42

ccc 1,019.83a 1,076.51

bb 1,314.61

a 537.42 633.76

bc 639.43

b 590.56

b

T5 - 2.00 968.33ab 930.64

ab 1,184.21aa 1,371.41

aa 649.57

bcc 894.18

b 1,397.76

aa 1,049.48b 571.61 725.54

aa 799.97

a 499.04

c

SEM 126.74aa 121.30

aa 174.09

a 210.23

a 112.56

bcc 69.50

b 104.52

aa 149.39

b 93.99 61.54

aa 85.44

a 72.09

a

--------------------------------------------------------------- Probability ---------------------------------------------------------------------

Contrast

Lin *** ** *** *** NS *** *** *** NS *** *** *

Quad NS NS NS NS NS *** *** ** NS NS * ***

Cubic NS * NS NS NS *** NS *** NS * * ***

Quar NS NS ** NS ** *** NS *** NS * NS *** a, b, c, d column means with different superscripts differ significantly at P <0.05.

* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant.

68


(Mu-Plus®) in the diets of broiler chickens on the villi width


kg/ton diets on the villi width of duodenum, jejunum and ileum at four stages of age

of broiler chickens are presented in Table 4.18. The average duodenal villi widths of

broilers are 81.07, 92.65, 91.68, 102.83 and 92.60 µm, respectively (during 14 d of

age), 118.19, 86.74, 115.71, 116.24 and 106.27 µm, respectively (during 21 d of age),

93.08, 89.20, 96.89, 89.94 and 113.59 µm, respectively (during 35 d of age), and

130.46, 99.38, 113.08, 113.64 and 106.78 µm, respectively (during 42 d of age). The

average jejunal villi widths of broilers are 88.59, 105.88, 91.72, 99.53 and 94.46 µm,




respectively (during 42 d of age). The average ileal villi widths of broilers are 106.36,

94.30, 93.98, 98.24 and 90.93 µm, respectively (during 14 d of age), 104.51, 98.25,

118.03, 107.05 and 100.63 µm, respectively (during 21 d of age), 89.41, 116.41,

84.18, 106.46 and 108.28 µm, respectively (during 35 d of age), and 98.16, 88.36,

93.10, 91.58 and 99.39 µm, respectively (during 42 d of age). All groups fed

Mu-Plus® diets showed relatively higher in villi width of duodenum, jejunum and

ileum when compared with the control group. These results are similar with

Nopparatmaitree (2008) who investigated the effect of Mu-Plus® at 0.1 and 0.2% diets

in weaning pigs showed both levels of Mu-Plus® in the diets have an effect (P<0.01)

to increase the villi width of duodenum, jejunum and ileum when compared with the

control group. Mathivanan and Edwin (2012) reported that broilers received

Andrographis paniculata and probiotic diets showed increase the ileal villi width

higher than the control group, but it was not significantly different (P>0.05) among

treatment groups. Narkchamnarn (2005) also reported that supplement Herbatob-Mix®

1,000 ppm + Mu-Plus® 2,000 ppm in the broiler diets have no effect (P>0.05) on the

villi width of duodenum, jejunum and ileum when compared to the control group.

69

Table 4.18 Effects of Mu-Plus® in the diets on villi width of duodenum, jejunum and ileum of broilers at different period of age

Treatments

Mu-Plus®

(Kg/ton)

Villi width (µm)


14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d

T1 - 81.07bb

118.19a 93.08

b 130.46 88.59

bb 102.96aa 79.39

c 101.84a 106.36

aa 104.51

bb 89.41

b 98.16

T2 - 0.50 92.65ab 86.74

b 89.20

b 99.38 105.88aa 101.05

ab 99.93

a 76.61

b 94.30

bb 98.25

bb 116.41

a 88.36

T3 - 1.00 91.68ab 115.71

a 96.89b 113.08 91.72

bb 98.64ab 98.40

a 110.32a 93.98

bb 118.03aa 84.18

b 93.10

T4 - 1.50 102.83aa 116.24

a 89.94

b 113.64 99.53

ab 91.44bc 88.94

b 103.58

a 98.24

ab 107.25

ab 106.46

a 91.58

T5 - 2.00 92.60ab 106.27

a 113.59a 106.78 94.46

ab 87.10

cc 101.12

a 106.21a 90.93

bb 100.63

bb 108.28

a 99.39

SEM 12.22ab 11.59

a 11.72

a 22.91 12.25

ab 10.86

aa 7.78

a 19.88

a 7.17

bb 12.71

bb 14.41

a 18.41

--------------------------------------------------------------- Probability ---------------------------------------------------------------------

Contrast

Lin * NS *** NS NS *** *** NS ** NS * NS

Quad NS NS ** NS NS NS * NS NS NS NS NS

Cubic NS *** NS NS NS NS *** NS * NS * NS

Quar NS * * NS * NS NS * NS * *** NS a, b, c column means with different superscripts differ significantly at P <0.05.


70


(Mu-Plus®) in the diets of broiler chickens on crypt depth


kg/ton diets on crypt depth of duodenum, jejunum and ileum during 14 d, 21 d, 35 d

and 42 d of age of broiler chickens are presented in Table 4.19. The average crypt

depths of duodenum are 122.43, 111.53, 95.54, 115.48 and 114.00 µm, respectively

(during 14 d of age), 89.71, 96.46, 117.19, 95.12 and 98.75 µm, respectively (during

21 d of age), 92.82, 90.10, 85.25, 88.91 and 81.45 µm, respectively (during 35 d of

age), and 90.70, 94.10, 86.83, 94.48 and 89.99 µm, respectively (during 42 d of age).

The average crypt depths of jejunum are 95.86, 80.35, 89.82, 76.06 and 80.13 µm,




respectively (during 42 d of age). The average crypt depths of ileum are 103.21,

74.72, 98.63, 95.10 and 87.35 µm, respectively (during 14 d of age), 90.73, 88.07,

100.47, 88.04 and 85.58 µm, respectively (during 21 d of age), 86.40, 76.54, 75.18,

86.65 and 96.21 µm, respectively (during 35 d of age), and 79.75, 65.79, 76.05, 76.86

and 70.56 µm, respectively (during 42 d of age). Both of the Andrographis paniculata

and Zingiber cassumunar have anti-inflammatory activity that it is important to

prevent crypt in the small intestine from inflammation caused cryptitis lead to crypt

abscess. It had mentioned that a crypt depth is indicative of a faster tissue turnover

and a higher demand for new tissue. Crypts are covered by epithelium which contains

two types of cells, goblet cells (secreting mucus) and enterocytes (absorbing water and

electrolytes). The enterocytes in the mucosa contain digestive enzymes that digest

specific food while they are being absorbed through the epithelium. These enzymes

include peptidase, sucrase, maltase, lactase and intestinal lipase. Renewal of cells for

the villi is provided by the migration of new cells from the crypt toward the tips of the

villi. Moderate physical or functional loss of villus cells, either through attrition or

disease, can be replaced by the dividing cells at the crypt (William, 2005 and James

and Bradley, 2007). The response of intestinal microstructure to dietary or lumen

factors may be affected by battery cages stress, dietary Mu-Plus® level inclusion and

age of birds when taking tissue samples for histomorphology. Therefore, it is

71

necessary to determine the optimum dietary inclusion level of Mu-Plus® in poultry

diets. These results are similar to Nopparatmaitree (2008) who investigated the effect

of Mu-Plus® at 0.1 and 0.2% diets in weaning pigs showed that both levels of

Mu-Plus® in the diets have an effect (P<0.01) to increase crypt depth of duodenum,

jejunum and ileum when compared with the control group. This finding is in

agreement with Mathivanan and Edwin (2012) reported that broilers received

Andrographis paniculata and probiotic diets shown higher significantly different

(P<0.05) in ileal crypt depth when compared to the control group. Narkchamnarn

(2005) reported that supplement Herbatob-Mix® 1,000 ppm + Mu-Plus® 2,000 ppm in

the broiler diets have no effect (P>0.05) on the crypt depth of duodenum, jejunum and

ileum when compared to the control group. However, crypt depth of control was

deeper than herbal-fed group. Pratoomtong (2011) investigated the effect of


ppm + 1,000 ppm) in the diets of weaning pigs showed that both Herbatob-Mix® and

Herbatob-Mix® + Mu-Plus® trended to have a crypt depth of duodenum, jejunum and

ileum lower than the control group but there was not significant different (P>0.05)

among treatment groups.

72

Table 4.19 Effects of Mu-Plus® in the diets on crypt depth of duodenum, jejunum and ileum of broilers at different period of age

Treatments

Mu-Plus®

(Kg/ton)

Crypt of liberkhun depth (µm)


14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d

T1 - 122.43aa 89.71

b 92.82

aa 90.70 95.86

aa 98.02a 87.32

85.32bb 103.21

aa 90.73

b 86.40

b 79.75

aa

T2 - 0.50 111.53ab 96.46

b 90.10

ab 94.10 80.35bc 95.81

a 97.13 73.90

cc 74.72

cc 88.07

b 76.54

c 65.79

cc

T3 - 1.00 95.54bb

117.19a 85.25

ab 86.83 89.82

ab 95.72a 86.66 78.92

bc 98.63ab 100.47

a 75.18

c 76.05

ab

T4 - 1.50 115.48aa 95.12b

88.91ab 94.48 76.06

cc 104.95a 94.57 86.43

ab 95.10

ab 88.04

b 86.65

b 76.86

ab

T5 - 2.00 114.00aa 98.75

b 81.45bb

89.99 80.13bc 74.79

b 93.67 97.01

aa 87.35

bc 85.58

b 96.21

a 70.56bc

SEM 14.58aa 10.27

a 11.37

aa 16.34 9.09

aa 9.19

a 11.32 8.98

aa 11.04

aa 9.97

a 9.12

a 5.86

aa

--------------------------------------------------------------- Probability ---------------------------------------------------------------------

Contrast

Lin NS NS * NS ** *** NS ** NS NS ** NS

Quad * ** NS NS NS *** NS ** NS NS *** NS

Cubic NS NS NS NS NS *** NS NS *** NS NS ***

Quar NS ** NS NS ** * NS NS * * NS NS a, b, c column means with different superscripts differ significantly at P <0.05.


73


(Mu-Plus®) in the diets of broiler chickens on the ratio of villi height:

crypt of liberkhun depth


kg/ton diets on the ratio of villi height: crypt depth of duodenum, jejunum and ileum

during 14 d, 21 d, 35 d and 42 d of age of broiler chickens are presented in Table

4.20. The average villi height: crypt depth ratio of duodenum are 5.91, 7.56, 8.16,

8.79 and 8.80, respectively (during 14 d of age), 9.33, 7.73, 6.79, 10.86 and 9.47,

respectively (during 21 d of age), 9.88, 10.71, 13.86, 11.82 and 14.68, respectively

(during 35 d of age), and 11.20, 10.65, 12.08, 13.08 and 15.47, respectively (during

42 d of age). The average villi height: crypt depth ratio of jejunum are 7.63, 8.73,

8.86, 7.59 and 8.32, respectively (during 14 d of age), 8.64, 7.85, 11.06, 9.80 and

12.15, respectively (during 21 d of age), 10.06, 9.42, 11.32, 11.59 and 15.03,

respectively (during 35 d of age), and 9.54, 7.41, 7.68, 15.33 and 10.99, respectively

(during 42 d of age). The average villi height: crypt depth ratio of ileum are 4.70,

6.32, 4.77, 5.70 and 6.55, respectively (during 14 d of age), 5.62, 6.82, 6.83, 7.27 and

8.58, respectively (during 21 d of age), 6.93, 8.60, 7.99, 7.44 and 8.34, respectively

(during 35 d of age), and 6.14, 6.39, 8.87, 7.96 and 7.23, respectively (during 42 d of

age). These results revealed that supplementation of herbal mixture Mu-Plus® at

different levels in the diets showed to increase the ratio of villi height: crypt depth of

duodenum, jejunum and ileum with linear significant (P<0.01) when increasing the

levels of herbal mixture among treatment groups. However, herbal mixture Mu-Plus®

supplementation at 2.00 kg/ton diet showed the best response to villi height: crypt

depth ratio of duodenum, jejunum and ileum at different stages of age. Villi increase

the internal surface area of the intestinal walls. Increased surface area allows for an

increased intestinal wall area that it is available for absorption. Thus, increasing the

ratio of villi height: crypt depth showed a good result to increase surface digestive

area. The increased digestive area is useful for nutrients absorption because digested

nutrients (monosaccharide and amino acid) pass into the semipermeable villi.

Furthermore, it also considered that a villi height: crypt depth ratio is associated with

well-developed intestinal mucosa with high digestion and absorption (Yanson et al.,

1987; Potten and Loeffler, 1990; Jeurissen et al., 2002 and Fasina et al., 2010). These

74

results are similar to Nopparatmaitree (2008) who investigated the effect of Mu-Plus®

at 0.1 and 0.2% diets in weaning pigs showed that Mu-Plus® supplementation at 0.2%

diet showed to increase (P<0.01) the ratio of villi height: crypt depth of duodenum,

jejunum and ileum when compared with the control group. This data is in agreement

with Narkchamnarn (2005) who reported that supplement Herbatob-Mix® 1,000 ppm

+ Mu-Plus® 2,000 ppm in the broiler diets increase (P<0.05) the ratio of villi height:

crypt depth of duodenum and jejunum higher than the control group. Pratoomtong

(2011) also investigated the effect of supplementation of Herbatob-Mix® (500 ppm)

and Herbatob-Mix® + Mu-Plus® (500 ppm + 1,000 ppm) in the diets of weaning pigs

showed both Herbatob-Mix® and Herbatob-Mix® + Mu-Plus® trended (P>0.05) to

have the ratio of villi height: crypt depth of duodenum, jejunum and ileum higher than

the control group.

75

Table 4.20 Effects of Mu-Plus® in the diets on villi height: crypt depth ratio of duodenum, jejunum and ileum of broilers at different

period of age

Treatments

Mu-Plus®

(Kg/ton)

Ratio of villi height: crypt of liberkhun depth


14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d 14 d 21 d 35 d 42 d

T1 - 5.91bb

9.33ab 9.88

b 11.20

bb 7.63 8.64bc 10.06

bc 9.54bc 4.70

bb 5.62

c 6.93

bb 6.14

cc

T2 - 0.50 7.56ab 7.73

bc 10.71

b 10.65bb

8.73 7.85cc 9.42

cc 7.41

cc 6.32

aa 6.82

b 8.60

aa 6.39

cc

T3 - 1.00 8.16ab 6.79

cc 13.86a 12.08

bb 8.86 11.06

aa 11.32

bb 7.68cc 4.77

bb 6.83

b 7.99

ab 8.87

aa

T4 - 1.50 8.79aa 10.86

aa 11.82

b 13.08

ab 7.59

9.80bb

11.59bb

15.33aa 5.70

ab 7.27

b 7.44

ab 7.96

ab

T5 - 2.00 8.80aa 9.47

ab 14.68a 15.47

aa 8.32 12.15

aa 15.03

aa 10.99bb

6.55aa 8.58

a 8.34

aa 7.23

bc

SEM 1.80aa 1.36

aa 2.12

a 2.84

aa 1.30 1.21

aa 1.48

aa 2.11

aa 1.19

aa 0.83

a 1.28

aa 1.11

aa

--------------------------------------------------------------- Probability ---------------------------------------------------------------------

Contrast

Lin ** NS *** ** NS *** *** *** 0.05 *** NS **

Quad NS * NS NS NS NS ** NS NS NS NS **

Cubic NS ** NS NS NS NS NS *** 0.05 NS ** NS

Quar NS ** ** NS NS *** NS ** NS *** NS * a, b, c column means with different superscripts differ significantly at P <0.05.


CHAPTER V

CONCLUSION AND RECOMMENDATION

This experiment was conducted to investigate the effects of Andrographis

paniculata and Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and

2.00 kg/ton diets on growth performance, carcass quality, nutrients digestibility and

small intestinal histomorphology of broiler chickens compare to the control group.

5.1 Conclusion

Based on the results and discussions, it could be concluded as follows:

5.1.1 Supplement Andrographis paniculata and Zingiber cassumunar mixture

show no effects (P>0.05) on the growth performance, FCG and PI. However, it was

trended to enhance by the level of concentrate at 0.50 kg/ton diet leaded to the higher

in FBW, BWG, FI, PI, feed efficiency and economic benefit returns when compared

with the control group. Moreover, it has enhanced to decrease (P>0.05) FCG when

compared to the control group in the overall period. However, the SR was

significantly increased (P<0.05) by the four graded levels of Andrographis paniculata

and Zingiber cassumunar mixture than the control group.


showed no effect (P>0.05) on the weight of carcass characteristics and organ weights

of broilers at termination. The level at 0.50 kg/ton diet showed her in thigh, breast,

drumstick, wings, total edible meat, and gizzard but showed lower in abdominal fat,

liver, and heart.

5.1.3 Carcass quality was significantly (P<0.05) with the level of Andrographis

paniculata and Zingiber cassumunar mixture at 0.50 kg/ton diet showed the lowest fat

composition of breast meat and liver of broilers. In addition, the level at 0.50 kg/ton

diet also showed the higher in CP of breast meat and liver of broilers.

77


showed effect (P<0.05) on nutrients digestibility including CP, fat, GE and DM in

grower and finisher period. In addition, nutrients digestibility was significantly

enhanced (P<0.05) by the level of concentrate at 0.50 kg/ton diet showed the highest

DM, CP, GE and fat digestibility in three cycle periods (starter, grower and finisher).

5.1.5 Intestinal villi height and crypt of liberkhun depth were significant

enhanced (P<0.01) by the levels of Andrographis paniculata and Zingiber

cassumunar mixture when compared to the control group. In addition, supplement at

2.00 kg/ton diet leaded to the higher in villi height, villi width and villi height: crypt

depth ratio of duodenum, jejunum and ileum than the control group.

5.2 Recommendation and suggestion

5.2.1 Andrographis paniculata and Zingiber cassumunar mixture could be used

as an animal for feed additives.

5.2.2 Should be study more on immune activity, plasma concentration and fatty

acid profile.

5.2.3 Should be considering on the bioactive ingredient of each herb for the

efficacy of the activities.

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APPENDIX

91

CHROMIUM OXIDE (Cr 2O3) ANALYSIS

1. Materials

1.1 Crucible

1.2 Desiccators

1.3 Muffle furnace

1.4 Vulometric flask 25 ml

1.5 Beaker 250 ml

1.6 Pipet 5 ml

1.7 Filter paper Whatman No. 40

1.8 Funnel

1.9 Spectrophotometer

2. Chemical substance

Reagent mixture Tri-potassium phosphate (K3PO4) 250g and Potassium

hydroxide pellets (KOH) 125g soluble in 500ml of distill water.

3. Analyzed method

3.1 Weigh Cr2O3- contained sample 0.5g put into the crucible (feed and fecal

should be conducted at the same time)

3.2 Added reagent mixture 2ml into the crucible (dry samples by oven at 100 ˚C

overnight if samples are wet)

3.3 Bring samples to burn in 800 ˚C during 90 minutes

3.4 Soluble warm ash and transfer to beaker 250 ml and then adjust solution till

100 ml by distill water after that keep overnight.

3.5 Filter solution with filter paper Whatman No. 40 into a volumetric flask

250ml then adjust by distill water.

3.6 Pipet solution in point 3.5 amount 5 ml into a volumetric flask 25ml and

then adjust by distill water

92

3.7 Prepare Cr2O3 standard by weighing Cr2O3 at mass 2, 4, 6, 8, 10, 12, 14 and

16 mg put into crucible and take procedure same feed samples. Thus, 2 ml of the

solution will contain the amount of Cr2O3

2 ml of 2 mg Cr2O3 = (2x2)/250 = 0.016 mg

2 ml of 4 mg Cr2O3 = (2x4)/250 = 0.032 mg

2 ml of 6 mg Cr2O3 = (2x6)/250 = 0.048 mg

2 ml of 8 mg Cr2O3 = (2x8)/250 = 0.064 mg

2 ml of 10 mg Cr2O3 = (2x10)/250 = 0.080 mg

2 ml of 12 mg Cr2O3 = (2x12)/250 = 0.096 mg

2 ml of 14mg Cr2O3 = (2x14)/250 = 0.112 mg

2 ml of 16 mg Cr2O3 = (2x16)/250 = 0.128 mg

mg = 0.072

3.8 Bring ready samples solution and standard to measure by using a

spectrophotometer with infrared wavelength 370 nm in the blank so that compare to

standard

Formulation

mg Cr2O3 = b(A) – b(A) + (mg)

% Cr2O3 = (mg Cr2O3 x 250 x 100) ÷ (ml, used x g, sample x 1000)

Where :

b(A) = Absorbance of sample solution

b(A) = Absorbance of standard solution

mg = Average of Cr2O3 in the standard solution

mg = Average of Cr2O3 in sample solution

93

VILLI HEIGHT AND CRYPT DEPTH OF SMALL INTESTINE

Small intestinal segment collections are duodenum, jejunum and ileum at

midpoint about 3 cm immediately fixed into 10 % buffer neutral formalin until

analysis process.

1. Materials

1.1 Automatic tissue processor 1.14 Surgical instruments

1.2 Paraffin dispenser 1.15 Cylinder

1.3 Rotary microtome 1.16 Staining jar

1.4 Refrigerator 1.17 Beaker

1.5 Hood 1.18 Cassette

1.6 Oven 1.19 Mold

1.7 Slide warmer 1.20 Embedded ring

1.8 Magnetic stirrer 1.21 Microtome

1.9 Hot plate 1.22 funnel

1.10 Compound light microscope 1.23 Filter paper

1.11 pH meter 1.24 Glass slide

1.12 Weights 1.25 Cover slide

1.13 Sample container box 1.26 Slide box

2. Methods

2.1Tissue washing

Cleaning fix active of 10% buffer neutral formalin with running water during 10

mn (fixed 24 hours) more than 30 mn (fixed longer than 24 hours).

2.2 Tissue preparing

2.2.1 Dehydration is the excessive loss of water by ethanol and ethanol start

from low concentrate ethanol to high concentrate ethanol. Ethanol concentration

depends on a tissue slice including 30-40 % (large tissue slice) 15-20 % (small tissue

slice) such as 70 % ethanol, 80 % ethanol, 95 % ethanol and absolute ethanol.

94

2.2.2 Clearing or dealcoholization is the process for free ethanol by xyline,

play a role as a paraffin transporter into the tissue. If tissue still contains water, xyline

will thick so need to turn to clean with new absolute ethanol and then put into new

xyline again.

2.2.3 Infiltration is the process that allows paraffin to absorb to inside of

tissue. Put paraffin wax into paraplast 56-60˚C so that to cut in 5-7 µm by rotary

microtome. Paraffin machine will be working step by step automatically last 22 hours

and 30 mn.

Table 1 Automatic tissue processing

Step Solution Duration (hour) Process

1 70 % ethanol 3.00 Dehydration




5 Absolute ethanol 1.30 Dehydration



8 Xyline I 1.00 Clearing

9 Xyline II 1.30 Clearing

10 Xyline III 1.30 Clearing

11 Paraffin I 3.00 Infiltration

12 Paraffin II 3.00 Infiltration

2.3 Embedding

A paraffin-infiltrated sample was embedded in liquid paraffin into a mould

connect with embedded ring by paraffin dispenser and freezes it till tissue cutting

process. Consequently, Samples must stand in the middle side of paraffin block.

95

2.4 Rotary microtome

Rotary microtome is a paraffin block cutting instrument with thick about 5-7

µm. The tissue-contained ribbon will be floated in water bath with gelatin 0.5 % in

43–45 ˚C. This technique has helped tissue ribbon good stick to a glass slide. Glass

slide was dry by oven in 60 ˚C during 30 minutes and warm slide before staining.

2.5 Staining

There are many steps in heamatoxyline and eosin slide staining as bellow

2.5.1 Deparaffinnization in xyline twice, 2 minutes per each.

2.5.2 Dehydrate and xyline cleaning by absolute ethanol

2.5.3 95 % ethanol, 80 % ethanol and washing with running water last 2

minutes per each step.

2.5.4 Heamatoxyline staining during 6 minutes then washes by running

water during 2 minutes.

2.5.5 Heamatoxyline clearing by 1% acid alcohol, fast doubles staining if

color not clear should one more time stain and then wash by running water last 2

minutes.

2.5.6 Eosin staining (working solution) last 2-5minutes, tissue color will be

turning red or pink.

2.5.7 Dehydration by 70 % ethanol during the 1 minute (if take a long time,

the color will turn slightly red or pink) and stain by 95 % ethanol and absolute ethanol

two steps (2 minutes per each).

2.5.8 Tissue clearing by xyline three times, 3 minutes per each.

2.6 Villi heigh and crypt dept measurement by light microscope

Ready tissue slide was measured 20 villi heights and 20 crypt depth per bird

with AxioCam ERc 5s equipped microscope. The unit of villi height and crypt depth

is micrometer (µm).

96

APPENDICES FIGURE

Experiment I. Productive performance and carcass quality trial

Mu-Plus® floor-pen open house Broiler at 21 day of age

Figure 1 Abdominal fats of broilers with different treatments

97

Liver T1 Liver T2 Liver T3

Liver T4 Liver T5

Experiment II. Nutrients digestibility trial

Battery-raised broilers Cr2O3 mixture Cr2O3 -fed broilers

98

Fecal sampling Fecal-collected sample

Experiment III. Small intestinal histomorphology trial

Battery cage Sample collection Sample cutting

Tissue block preparing Rotary microtome Staining

99

Tissue slide Microscopy

Duodenum T1 Duodenum T2 Duodenum T3

Duodenum T4 Duodenum T5 Jejunum T1

100

Jejunum T2 Jejunum T3 Jejunum T4

Jejunum T5 Ileum T1 Ileum T2

Ileum T3 Ileum T4 Ileum T5

101

Table 2 Temperature and humidity throughout the experiment

dd/mm/yy

Temperature and Humidity

Temperature Humidity

Maxi Mini Average Maxi Mini Average

09/01/2013 28.00 20.00 24.00 90.00 74.00 82.00

10/01/2013 28.00 20.00 24.00 91.00 89.00 90.00

11/01/2013 28.50 20.50 24.500 90.00 74.00 82.00

12/01/2013 27.00 20.00 23.50 91.00 89.00 90.00

13/01/2013 23.00 19.00 21.00 90.00 89.00 89.50

14/01/2013 30.00 21.50 25.75 91.00 90.00 90.50

15/01/2013 30.00 21.00 25.50 89.00 75.00 82.00

16/01/2013 29.50 22.00 25.75 91.00 80.00 85.50

17/01/2013 30.00 20.00 25.00 91.00 89.00 90.00

18/01/2013 27.50 18.50 23.00 91.00 89.00 90.00

19/01/2013 28.50 18.00 23.25 91.00 89.00 90.00

20/01/2013 30.00 20.00 25.00 91.00 89.00 90.00

21/01/2013 32.00 18.00 25.00 89.00 69.00 79.00

22/01/2013 32.50 20.50 26.50 80.00 76.00 78.00

23/01/2013 31.50 21.00 26.25 89.00 75.00 82.00

24/01/2013 32.50 21.00 26.75 89.00 69.00 79.00

25/01/2013 31.00 23.00 27.00 90.00 75.00 82.50

26/01/2013 31.00 24.00 27.50 90.00 75.00 82.50

27/01/2013 27.00 24.50 25.75 90.00 82.00 86.00

28/01/2013 22.50 20.00 21.25 90.00 89.00 89.50

29/01/2013 28.50 20.50 24.50 89.00 82.00 85.50

30/01/2013 28.00 22.00 25.00 91.00 90.00 90.50

31/01/2013 26.50 23.00 24.75 91.00 90.00 90.50

01/02/2013 28.00 21.00 24.50 89.00 67.00 78.00

02/02/2013 28.50 21.00 24.75 89.00 74.00 81.50

03/02/2013 29.00 22.00 25.50 90.00 67.00 78.50

04/02/2013 33.00 23.00 28.00 80.00 63.00 71.50

102

Table 2 Temperature and humidity throughout the experiment (Cont.)

dd/mm/yy

Temperature and Humidity

Temperature Humidity

Maxi Mini Average Maxi Mini Average

05/02/2013 33.00 23.00 28.00 80.00 63.00 71.50

06/02/2013 36.00 23.00 29.50 90.00 57.00 73.50

07/02/2013 37.00 23.50 30.25 90.00 58.00 74.00

08/02/2013 37.00 23.00 30.00 90.00 57.00 73.50

09/02/2013 32.00 24.50 27.25 90.00 69.00 79.50

10/02/2013 31.50 22.00 26.75 90.00 62.00 76.00

11/02/2013 33.00 22.00 27.50 90.00 69.00 79.50

12/02/2013 31.00 21.50 26.25 89.00 68.00 78.50

13/02/2013 31.00 20.00 25.50 89.00 75.00 82.00

14/02/2013 32.00 22.00 27.00 90.00 69.00 79.50

15/02/2013 34.00 22.00 28.00 90.00 63.00 76.50

16/02/2013 33.00 22.00 27.50 80.00 69.00 74.50

17/02/2013 33.00 22.00 27.50 90.00 69.00 79.50

18/02/2013 35.00 26.00 30.50 90.00 64.00 77.00

CURRICULUM VITAE

Full name : LAING DANET Gender : Male Date of Birth : June 24, 1987 Place of Birth : Potaoun Village, Srayov Commune, Kampong Svay District, Kampong Thom Province, Cambodia. Nationality : Cambodian Marital status : Single Contact : (+855)92 810 745, E-mail: [email protected] ACADEMIC BACKGROUND Level of study Institution/Place and

Country Year

Attended Year of study

(i.e. 1st, 2nd)

Field of Study

Graduated 2006-2010

Royal University of Agriculture / Phnom Penh, Cambodia.

4

1-4

Bachelor of Science in Animal Science and Veterinary Medicine

High School 2000-2006

Hun Sen Balaing High School / Kampong Thom Province, Cambodia.

6

7-12

General Education

Primary School 1994-2000

Hun Sen Trapaing Veng Primary School, Kampong Thom Province, Cambodia.

6

1-6

General Education

LANGUAGE AND DEGREE PROFICIENCY Khmer : Excellent speaking and writing (Mother tongue) English : Good Comprehension Thai : Good Comprehension WORK EXPERIENCE 1 year : Veterinary Medicine Internship at the private Animal Clinic, Phnim Penh, Cambodia. Dec04, 09-Jan14, 10 : Research Volunteer, Chicken raising Project of Dr. Chhum Phith Loan, Dean of Faculty of Animal Science and Veterinary medicine, RUA, Cambodia. Cambodia. Corporate with CP Cambodia Co, Ltd 2010- 2012 : Secretary and Administrative Affairs Officer and Personal Assistant to Dean of faculty of Agriculture and Food processing at the University of Battambang, Battambang province, Kingdom of Cambodia. 2010-2012 : Lecturer at University of Battambang.

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