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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
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
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 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
KHON KAEN UNIVERSITY
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
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 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
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 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
3.2.1 Experimental design 27
3.2.2 Diets formulation 27
3.2.3 Research methodology 27
3.2.4 Data record 28
3.3 Experiment III 28
3.3.1 Experimental design 28
3.3.2 Diets formulation 28
3.3.3 Research methodology 29
3.3.4 Data record 29
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
TABLE OF CONTENTS (Cont.)
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
4.2.2 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets on crude
protein digestibility (CPD) of broiler chickens 57
4.2.3 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets on ether
extract digestibility (EED) of broiler chickens 59
4.2.4 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets on gross
energy digestibility (GED) of broiler chickens 60
4.3 Experiment III 65
4.3.1 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets of broiler
chickens on the villi height 65
4.3.2 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets of broiler
chickens on the villi width 68
4.3.3 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets of broiler
chickens on crypt depth 70
xii
TABLE OF CONTENTS (Cont.)
Page
4.3.4 Effect of Andrographis paniculata and Zingiber
cassumunar mixture (Mu-Plus®) in the diets of broiler
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
Table 4.2 Analyzed nutrient composition of diets at 22-35d of age (proximate
analysis) 36
Table 4.3 Analyzed nutrient composition of diets at 36-42d of age (proximate
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
different age of broilers 45
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
different age of broilers 48
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 4.15 Effect of Mu-Plus® in the diets on nutrients digestibility of
broilers at 22-35 d of age 63
Table 4.16 Effect of Mu-Plus® in the diets on nutrients digestibility of
broilers at 36-42 d of age 64
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,
jejunum and ileum of broilers at different period of age 69
Table 4.19 Effect of Mu-Plus® in the diets on crypt depth of duodenum,
jejunum and ileum of broilers at different period of age 72
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.
1.3.2 To study the effects of different levels of Andrographis paniculata and
Zingiber cassumunar mixture (Mu-Plus®) in the diets on nutrients digestibility of
broiler chickens.
1.3.3 To study the effects of different levels of Andrographis paniculata and
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.
1.4.2 Results of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) supplemented diets will be improved carcass quality.
1.4.3 Results of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) supplemented diets will be improved nutrients digestibility.
1.4.4 Results of Andrographis paniculata and Zingiber cassumunar mixture
(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
148.33 Mathivanan and Edwin, 2012
53.86 Chumpawadee et al., 2008
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
125.00 Mathivanan and Edwin, 2012
104.48 Chumpawadee et al., 2008
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
Treatment 3. Basal diet + Mu-Plus® at 1.00 kg/ton diet
Treatment 4. Basal diet + Mu-Plus® at 1.50 kg/ton diet
Treatment 5. Basal diet + Mu-Plus® at 2.00 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.
3.2.1 Experimental design
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.
Treatment 1. Basal diet (Control group)
Treatment 2. Basal diet + Mu-Plus® at 0.50 kg/ton diet
Treatment 3. Basal diet + Mu-Plus® at 1.00 kg/ton diet
Treatment 4. Basal diet + Mu-Plus® at 1.50 kg/ton diet
Treatment 5. Basal diet + Mu-Plus® at 2.00 kg/ton diet
3.2.3 Research methodology
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.
3.3.1 Experimental design
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.
Treatment 1. Basal diet (Control group)
Treatment 2. Basal diet + Mu-Plus® at 0.50 kg/ton diet
Treatment 3. Basal diet + Mu-Plus® at 1.00 kg/ton diet
Treatment 4. Basal diet + Mu-Plus® at 1.50 kg/ton diet
Treatment 5. Basal diet + Mu-Plus® at 2.00 kg/ton diet
29
3.3.3 Research methodology
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)
Ingredients (%) Levels of Mu-Plus (kg/ton)
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
Full-fat soy bean (36 % CP) 20.00 20.00 20.00 20.00 20.00
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 0.22 0.22 0.22 0.22 0.22
L-lysine 0.21 0.21 0.21 0.21 0.21
Crude rice bran oil 1.50 1.50 1.50 1.50 1.50
Salt 0.40 0.40 0.40 0.40 0.40
Choline chloride 60% 0.08 0.08 0.08 0.08 0.08
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) 14.02 14.21 14.41 14.60 14.80
Chemical composition
CP, % 20.02 20.01 20.01 20.00 20.00
ME, kcal/kg 3,152 3,154 3,152 3,150 3,149 a presented in table 3.4, b presented in table 3.5
32
Table 3.3 Composition of the experimental diets for finisher period (36-42 d)
Ingredients (%) Levels of Mu-Plus (kg/ton)
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
Full-fat soy bean (36 % CP) 20.00 20.00 20.00 20.00 20.00
Monodicalcium phosphate, P21 2.38 2.38 2.38 2.38 2.38
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
Crude rice bran oil 2.00 2.00 2.00 2.00 2.00
Salt 0.40 0.40 0.40 0.40 0.40
Choline chloride 60% 0.08 0.08 0.08 0.08 0.08
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) 13.78 13.97 14.17 14.36 14.56
Chemical composition
CP, % 17.97 17.97 17.96 17.96 17.96
ME, kcal/kg 3,228 3,226 3,224 3,223 3,221 a presented in table 3.4, b presented in table 3.5
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
Table 4.2 Analyzed nutrient composition of diets at 22-35 d of age (proximate
analysis)
Nutrients Levels of Mu-Plus® (kg/ton)
0.00 0.50 1.00 1.50 2.00
Dry matter (%) 91.95 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
Total phosphorus (%) 0.77 0.96 1.01 0.92 0.84
Gross energy (kcal/kg) 4472 4481 4528 4641 4492
Table 4.3 Analyzed nutrient composition of diets at 36-42 d of age (proximate
analysis)
Nutrients Levels of Mu-Plus® (kg/ton)
0.00 0.50 1.00 1.50 2.00
Dry matter (%) 91.21 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
Total phosphorus (%) 0.83 0.80 0.82 0.88 0.89
Gross energy (kcal/kg) 4573 4569 4547 4564 4576
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)
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and
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
Treatments Mu-Plus®
(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)
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and
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
Treatments Mu-Plus®
(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)
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and
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
Treatments Mu-Plus®
(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
Treatments Mu-Plus®
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
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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
Treatments Mu-Plus®
(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
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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.
4.1.4 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(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
4.2.1 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets on dry matter digestibility (DMD) of broiler
chickens
This experiment investigated the effect of Andrographis paniculata and
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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.
4.2.2 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets on crude protein digestibility (CPD) of broiler
chickens
This experiment investigated the effect of Andrographis paniculata and
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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
4.2.3 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets on ether extract digestibility (EED) of broiler
chickens
This experiment investigated the effect of Andrographis paniculata and
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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
supplementation of Herbatob-Mix® (500 ppm) and Herbatob-Mix® + Mu-Plus® (500
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.
4.2.4 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets on gross energy digestibility (GED) of broiler
chickens
The experiment investigated the effect of Andrographis paniculata and
Zingiber cassumunar mixture (Mu-Plus®) at 0, 0.50, 1.00, 1.50 and 2.00 kg/ton diets
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
Treatments Mu-Plus®
(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
Table 4.15 Effects of Mu-Plus® in the diets on nutrients digestibility of broilers at
22-35 d of age
Treatments Mu-Plus®
(Kg/ton)
Nutrients digestibility (%)
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
Table 4.16 Effects of Mu-Plus® in the diets on nutrients digestibility of broilers at
36-42 d of age
Treatments Mu-Plus®
(Kg/ton)
Nutrients digestibility (%)
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
4.3.1 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(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
4.3.2 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets of broiler chickens on the villi width
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and 2.00
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 14 d of age), 102.96, 101.05, 98.64, 91.44 and 87.10 µm,
respectively (during 21 d of age), 79.39, 99.93, 98.40, 88.94 and 101.12 µm,
respectively (during 35 d of age), and 101.84, 76.61, 110.32, 103.58 and 106.21 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)
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 - 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.
* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant.
70
4.3.3 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets of broiler chickens on crypt depth
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and 2.00
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 14 d of age), 98.02, 95.81, 95.72, 104.95 and 74.79 µm,
respectively (during 21 d of age), 87.32, 97.13, 86.66, 94.57 and 93.67 µm,
respectively (during 35 d of age), and 85.32, 73.90, 78.92, 86.43 and 97.01 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
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 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)
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 - 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.
* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant.
73
4.3.4 Effect of Andrographis paniculata and Zingiber cassumunar mixture
(Mu-Plus®) in the diets of broiler chickens on the ratio of villi height:
crypt of liberkhun depth
The effect of herbal mixture Mu-Plus® at 0, 0.50, 1.00, 1.50 and 2.00
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
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 - 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.
* = P<0.05, ** = P<0.01, *** = P<0.001 and NS = non-significant.
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.
5.1.2 Supplement Andrographis paniculata and Zingiber cassumunar mixture
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
5.1.4 Supplement Andrographis paniculata and Zingiber cassumunar mixture
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.
REFERENCES
Ajaya Kumar, R., Sridevi, K., Vijaya, K.N., Nanduri, S. and Rajagopal, S. 2004.
Anticancer and immunostimulatory compounds from Andrographis paniculata.
Journal of Ethnopharmacology 92: 291–295.
AOAC. 2000. Association of Official Analytical Chemical. Official methods of
analysis of AOAC International (17th ed.). AOAC International, Gatithersburg,
MD.
Arkan, B.M., Al-Rubaee, M.A.M. and Jalil, A.Q. 2012. Effect of ginger (Zingiber
officinale) on performance and blood serum parameters of broiler.
International Journal of Poultry Science 11(2): 143–146.
Awad, W.A., Bohm, J., Razzazi-Fazeli, E., Ghareeb, K. and Zentek, J. 2006. Effect of
addition of a probiotic microorganism to broiler diets contaminated with
deoxynivalenol on performance and histological alterations of intestinal villi
of broiler chickens. Poultry Science 85: 974–979.
, Ghareeb, K., Abdel-Raheem, S. and Bohm, J. 2009. Effects of dietary
inclusion of probiotic and synbiotic on growth performance, organ weights,
and intestinal histomorphology of broiler chickens. Poultry Science 88: 49–55.
Balachandranm, P. and Govindarajan, R. 2005. Cancer an ayurvedic perspective.
Pharmacol Res, 51: 19–30.
Ballou, S.P. and Kushner, I. 2000. Laboratory evaluation of inflammation. In: Text
book of rheumatology. Kelley WN, Harris Jr ED, Ruddy S, Sledge CB,
editors. Philadelphia: WB Saunders Co.
Bhuiyan, M.N.I., Chowdhury, J.U. and Begum, J. 2008. Volatile constituents of
essential oils isolated from leaves and rhizome of Zingiber cassumunar.
Bangladesh Journal of Pharmacology 3: 69–73.
Bhusita, W., Pattra, M., Sirinan, T. and Gansuda, W. 2009. Antimicrobial activity of
essential oils extracted from Thai herbs and spices. Journal of Food Agro-
Industry 2(04): 677–689.
Bin, J.I., Mohd, Y.M.S., Chin, C.B. and Sim, N.L. 2003. Antifungal activity of the
essential oils of nine zingiberaceae species. Pharm Biol, 41: 392–397.
79
Bunyapraphatsara, N. 2000. Research strategies for the use of medicinal plants in
animal production. International Conference on Tropical Agriculture for
better health and environment at Kasetsart University, Kampaengsaen,
Nakornpathom, Thailand.
Chairul, P. and Sofnie, M.C. 2009. Phagocytosis effectivity test of phenylbutenoid
compounds isolated from bangle (Zingiber cassumunar Roxb. rhizome).
Biodiversity 10(1): 40–43.
Chaiwongkeart, D. 1997. Medicinal plant used in shrimp: Luk-Tai-Bai, Pha-Ya-
Yoe, and Fha-Talai-Jone. Department of Microbiology, Faculty of Science,
Kasetsart University, Bangkok. (Thai).
Chanda, R., Srivastava, V., Tandon, J.S. and Kapoor, N.K. 1995. Antihepatotoxic
activity of diterpenes of Andrographis paniculata (Kal-Megh) against
Plasmodium Berghei-induced hepatic damage in Mastomys nataalensis.
Pharm Boil, 33: 135–138.
Chang, H.M. and But, P.P.H. 1986. Pharmacology and applications of Chinese
materia medica Vol. 1. Singapore: World Scientific.
Chanjula, P., Chatreewong, D. and Waree, W. 2006. A study using Andrographis
paniculata in diet on growth performance of betong chicken: Starting and
growing period. Conference of Prince of Songkla University, Pattani
campus: 83–92.
, Chatreewong, D. and Waree, W. 2007. A study using Andrographis
paniculata in diet on growth performance of betong chicken: Starting period.
Songklanakarin J Sci Technol, 29(1): 197–206.
Chen, I.N., Chang, C.C., Ng, C.C., Wan, C.Y., Shyu, Y.T. and Chang, T.L. 2008.
Antioxidant and antimicrobial activity of zingiberaceae plants in Taiwan.
Plants Food Hun Nutr, 63: 15–21.
Chiou, W.F., Chen, C.F. and Lin, J.J. 1998. Andrographolide suppresses the
expression of inducible nitric oxide synthase in macrophage and restores the
vasoconstriction in rat aorta treated with lipopolysaccharide. British Journal
of Pharmacology 125: 327–334.
Chopra, R.N., Nayar, S.L. and Chopra, I.C. 1992. Glossary of Indian medicinal
plants. 3rd ed. CSIR, New Delhi, India.
80
Choudhury, B.R. and Poddar, M.K. 1983. Effect of kalmegh extract on rat liver and
serum enzymes. Clinical of Pharmacology 5(10): 727–730.
. and Poddar, M.K. 1984. Andrographolide and kalmegh (Andrographis
paniculata) extract: in vivo and in vitro effect on hepatic lipid peroxidation.
Methods find. Exp. Clinical of Pharmacology 6: 481–485.
. and Poddar, M.K. 1985. Andrographolide and kalmegh (Andrographis
paniculata) extract: effect on intestinal brush-border membrane-bound
hydrolases. Methods find. Exp. Clinical of Pharmacology 7(12): 617–621.
Chumpawadee, S., Chinrasri, O., Somchan, T., Ngamluan, S. and Soychuta, S. 2008.
Effect of dietary inclusion of cassava yeast as probiotic source on growth
performance, small intestine (ileum) morphology and carcass characteristic in
broilers. International Journal of Poultry Science 7(3): 246–250.
Cosentino, S., Barra, A., Pisano, B., Cabizza, M., Pirisi, F.M. and Palmas, F. 2003.
Composition and antimicrobial properties of sardinian juniperus essential oil
against foodborne pathogens and spoilage microorganisms. Journal of Food
Production Marketing 66: 1288–1291.
Dahanukar, S.A., Kulkarni, A. and Rege, N.N. 2000. Pharmacology of medicinal
plants and natural products. Indian Journal of Pharmacology 32: 81–118.
Deng, W.L. 1978. Preliminary studies on the pharmacology of the andrographis
product dihydroandrographolide sodium succinate. Newsletter Chinese
Herbal Medicine 8: 26–28.
, Nie, R.J. and Liu, J.Y. 1982. Comparison of pharmacological effect of four
andrographolides. Chin Pharm Bull, 17: 195–198.
Dhamma-Upakorn, P., Chaichantipyuth, C., Pongjunyakul, P., Sangalungkarn, V. and
Chaimongkol, S. 1992. Spasmolytic activity of some active substances from
Andrographis paniculata. J SPSNRCT Seminar: Pharmacological Active
Substance from natural source. Faculty of Pharmacy, Chulalongkorn University,
Bangkok, Thailand: 21.
Dhiman, A., Jugnu, G., Kavita, S., Arun, N. and Sandeep, D. 2012. A review on
medicinal prospective of Andrographis paniculata Nees. Journal of
pharmaceutical and scientific innovation: 1–4.
81
DMPRD. 1990. Division of Medical Plants Research and Development, department
of medical science, ministry of public health. Hand book of Medicinal Plant
for Primary Public Health . Text and Journal Corporation Co., Ltd. Press,
Bangkok: 53.
Duke, J.A. and Ayensu, E.S. 1985. Medicinal plants of China. Vol.1 Algonac, Mich,
Publications: 705.
Dutta, A. and Sukul, N.C. 1982. Filaricidal properties of a wild herb, Andrographis
paniculata. Journal of Helminthology 56(2): 81–84.
Fasina, Y.O., Hoerr, J., McKee, S.R. and Conner, D.E. 2010. Influence of salmonella
enterica serovar typhimurium infection on intestinal goblet cells and villous
morphology in broiler chicks. Avian Dis, 54: 841–847.
Golam, K., Nikkon, Farjana, Nikkon, F., Abdur Rashid, M. and Yeasmin, T. 2011.
Antimicrobial activities of the rhizome extract of Zingiber zerumbet Linn.
Asian Pacific Journal of Tropical Biomedicine: 409–412.
Gordana, U., Zikic, D., Peric, v., Nollet, L. and Vukic-Vranjes, M. 2004. Effect of
mannan oligosaccharides (Bio-Mos) supplementation on the small intestinum
morphology of broiler chickens. Faculty of Agriculture, University of Novi
Sad,: 362–364.
Han, A.-R., Kim, M.-S., Jeong, Y.H., Lee, S.K. and Seo, E.K. 2005. Cyclooxygenase-
2 inhibitory phenylbutenoids from the rhizomes of Zingiber cassumunar.
Chem Pharm Bull, 53(11): 1466–1468.
Handa, S.S. and Sharma, A. 1990. Hepatoprotective activity of andrographolide from
Andrographis paniculata against carbontetrachloride. Indian Journal of
Medicine Research 92: 276–283.
James, G.C. and Bradley, G.K. 2007. Digestion and absorption: the nonfermentative
process. Veterinary physiology.4th ed. Saunders, an imprint of Elsevier Inc.
Jeenapongsa, R., Yoovathaworn, K., Sriwatanakul, K.M., Pongprayoon, U. and
Sriwatanakul, K. 2003. Anti-inflammatory activity of (E)-1-(3, 4-dimeth-
oxyphenyl) butadiene from Zingiber cassumunar Roxb. Journal of
Ethnopharmacology 87: 143–148.
82
Jeurissen, S.H.M., Lewis, F., Van Der Klis, J.D., Mroz, Z., Rebel, J.M.J. and Ter
Huurne, A.A.H.M. 2002. Parameters and techniques to determine intestinal
health of poultry as constituted by immunity, integrity and functionality. Curr
Issues Intest Microbiol, 3: 1–14.
Kaewchoothong, A., Tewtrakul, S. and Panichayupakaranant, P. 2012. Inhibitory
effect of phenylbutanoid-rich Zingiber cassumunar extracts on nitric oxide
production by murine macrophage-like RAW264.7 cells. phytotherapy
Research.
Kanokwan, J. and Nobuo, N. 2008. Pharmacological aspect of Andrographis
paniculata on health and its major diterpenoid constituent andrographolide.
Journal of health Science 54(4): 370–381.
Kapil, A., Kiul, I.B., Banerjee, S.K. and Gupta, B.D. 1993. Antihepatotoxic effects of
major diterpenoid constituents of Andrographis paniculata. Biochemistry
Pharmacology 46: 182–185.
Kawalilak, L.T., Ulmer Franco, A.M. and Fasenko, G.M. 2010. Impaired intestinal
villi growth in broiler chicks with unhealed navels. Poultry Science 89: 82–87.
Khajarern, J. and Siriloaphaisan, S. 2007. Efficacy of herbal mixture (Herbatob-Mix®)
supplementation in layer diets on egg performance, egg quality and blood
analysis. In: Proceeding of the 3th Animal Science Conference, Department
of Animal Science, Faculty of Agriculture, Khon Kaen University: 50–57.
Kishore, N. and Zerumbone, D.R.S. 1992. A potential fungitoxic agent isolated from
Zingiber cassumunar Roxb. Mycopathologia 120: 155–59.
Kongkathip, N. 1995. Extraction and isolation of the most active substances of
Andrographis paniculata wall. Ex nees (National Research Report in 1995).
Department of Chemistry, Faculty of Science, Kasetsart University.
Koul, I.B. and Kapil, A. 1994. Effect of diterpenes from Andrographi paniculata on
antioxidant defense system and lipid peroxidation. Indian Journal of
Pharmacology 26: 296–300.
Kungsoksomboun, P. 2007. Effect of Zingiber cassumunar and Zingiber officinale
in broiler ration on carcass quality. Special problem, Department of Animal
Production Technology, Faculty of Agricultural Technology, King Mongkut’s
Institute of Technology, Chaokuntaharn Ladkrabang, Bangkok, Thailand.
83
Lala, S., Nandy, A.K., Mahato, S.B. and Basu, M.K. 2003. Delivery in vivo of 14-
deoxy-11-oxoandrographolide, antileshmanial agent by different carriers.
Indian Journal Biochemistry Biophysic 40: 169–174.
Liu, J., Wang, Z.T. and Ji, L.L. 2007. In vivo and in vitro anti-inflammatory activities
of neoandrographolide. Journal of Chinese Medicine 35(2): 317–328.
Madav, H.C., Tripathi, T. and Mishra, S.K. 1995. Analgesic and antiulcerogenic
effects of andrographolide. Indian Journal of Pharmaceutical Science 57:
121–125.
Manjunath, B.L., 1948. The wealth of India. A dictionary of India raw material and
industrial products. Council of Scientific and Industrial Research, New
Delhi.1: 77.
Marchini, C.F.P., Silva, P.L., Nascimento, M.R.B., Beletti, M.E., Silva, N.M. and
Guimaraes, E.C. 2011. Body weight, intestinal morphometry and cell
proliferation of broiler chickens submitted to cyclic heat stress. International
Journal of Poultry Science 10(6): 455–460.
Mathivanan, R. and Edwin, S.C. 2012. Effects of alternatives to antibiotic growth
promoters on intestinal content characteristics, intestinal morphology and gut
flora in broilers. Wudpecker Journal of Agricultural Research 1(7): 244–249.
, Edwin, S.C., Amutha, R. and Viswanathan, K. 2006. Panchagavya and
Andrographis paniculata as alternatives to antibiotic growth promoter on
broiler production and carcass characteristics. International Journal of
Poultry Science 5(12): 1144–1150.
, Edwin, S.C. and Viswanathan, K. 2008. Effect of Andrographis paniculata
supplementation on growth and feed conversion efficiency of broilers.
Abstract of Indian Journal of Poultry Science 43(2): 185–188.
. and Kalaiarasi, K. 2007. Panchagavya and andrographis paniculata as
alternatives to antibiotic growth promoters on haematological, serum
biochemical parameters and immune status of broilers. Journal of Poultry
Science 44: 198–204.
Mohamed Saleem, T. 2010. Hepatoprotective herbs: a review. Int J Res Pharm Sci,
1(1): 1–5.
84
MPRI. 1999. Medicinal Plant Research Institute, Department of Medical Science,
Ministry of Public Health. Standard of Thai Herbal Medicine: Andrographis
paniculata (Burm. f.) Nees. The War Veterans Organization Press, Bangkok.
Narkchamnarn, A. 2005. The effects of a combination of Andrographis paniculata,
Curcuma longa, Momordica charantia and Zingiber montanum on
immunity and growth performance in Broilers. M.Sc thesis, Department of
Animal Science, Faculty of Agriculture, Kasetsart University, Bangkok 10900
Thailand (Thai).
Narkchamnarn, A., Isariyodom, S., Tirawattanawanich, C., Kongkathip, N. and
Santisopasri, W. 2005. The effects of Herbatob-Mix® and Mu-Plus® on growth
performance in broilers. Seminar at Kasetsart University, Bangkok, Thailand
(Thai).
Nipon, P. 1985. The commercial broiler and swine industries in Thailand.
Nopparatmaitree, K. 2008. The effects of Andrographis paniculata and Zingiber
cassumunar mixture on productive performance, nutrient digestibility
and health status in weaning pigs. M.Sc thesis, Department of Animal
Science, Faculty of Agriculture, Khon Kaen University 40002, Khon Kaen,
Thailand (Thai).
NRC. 1994. Nutrient Requirement of Domestic Animals No1. Nutrient requirement
of poultry . National Academy Press, Washington D.C.
Nyamambi, B., Ndlovu, L.R., Naik, Y.S. and Kock, N.D. 2007. Intestinal growth and
function of broiler chicks fed sorghum based diets differing in condensed
tannin levels. South African Journal of Animal Science 37(3): 202–214.
Ojha, S.K., Nandave, M., Kumar, S. and Arya, D.S. 2009. Antioxidant activity of
Andrographis paniculata in ischemic mycocardium of rats. Global Journal of
Pharmacology 3(3): 154–157.
Ong-chai, S., Chotjumlong, P.M.S., Kongtawelert, P. and Krisanaprakornkit, S. 2008.
Zingiber cassumunar Roxb. inhibits hyaluronan production in human oral
fibroblasts. Chiang Mai Medical Journal 47(4): 177–187.
Ozaki, Y., Kawahara, N. and Harada, M. 1991. Anti-inflammatory effect of Zingiber
cassumunar Roxb., and it’s active principles. Chem Pharm Bull, (Tokyo) 39:
2353–2356.
85
Palawatvichai, N. 2007. Dietary supplement of Turmeric (Curcuma longa L.) on
growth performance, digestibility, carcass quality, meat quality and
cholesterol in broilers. M.Sc thesis, Department of Animal Science, Faculty
of Agriculture, Khon Kaen University, Khon Kaen, Thailand (Thai).
Pelicano, E.R.L., Souza, P.A., Souza, H.B.A., Figueiredo, D.F., Boiago, M.M.,
Carvalho, S.R. and Bordon, V.F. 2005. Intestinal mucosa development in
broiler chickens fed natural growth promoters. Brazilian Journal of Poultry
Science 7(4): 221–229.
Phatarapheecha, P., Jaikhan, W., Khajarern, J. and Porntrakulpipat, S. 2011. Efficacy
of herbal mixture (Herbatob-Mix®) supplementation in diets on productive
performance of sows and their piglets. Khon Kaen Agriculture journal 39:
31–37.
Pithayanukul, P., Tubprasert, J. and Wuthi-Udomlert, M. 2007. In vitro antimicrobial
activity of Zingiber cassumunar (Plai) oil and a 5% Plai oil gel. Phytother
Res, 21: 164–169.
Pluemjai, T. 1992. The antibacterial activity of Fah Talai Joan (Andrographis
Paniculata Nees.). J Dept Med Sci, 34: 9–15.
Potten, C.S. and Loeffler, M. 1990. Stem cells: Attributes, cycles, spirals, pitfalls and
uncertainties. Lessons from the crypt development: 110.
Pratoomtong, S. 2011. Effects of herbal mixture Andrographis paniculata, Curcuma
longa, Momardica charantia and Zingiber cassumunar supplementation in
diets on growth performance, nutrient digestibility and morphology of
small intestinal in weaning piglets. M.Sc thesis, Department of Animal
Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand
(Thai).
Puri, A., Saxena, R., Saxena, R.P., Saxena, K.C., Srivastava, V. and Tandon, J.S.
1993. Immunostimulant agents from Andrographis paniculata. J Nat Prod,
56(7): 995–999.
Rana, A.C. and Avadhoot, Y. 1991. Hepatoprotective effects of Andrographis
paniculata against carbontetrachloride-induced liver damage. Arch Pharm
Res, 14: 93–95.
86
Research Review. 1997. Andrographis vs the common cold. The journal of the
America Botanical Council and the Herb Research Foundation 40: 18.
Saengsophon, A., Sitthigripong, R., Srithaneadchai, P. and Srikitkasamewat, K. 2000.
Affect of Andrographis paniculata on the performance of broiler and native
crossbred chickens. Journal of King Mongkut’s Institute of Technology
Ladkrabang 8: 15–21.
SAS. 1996. User’s Guide: Statistic, Version 5. Edition. SAS. Inst Cary, NC., U.S.A.
Sawasdimongkol, K., Permpipat, U. and Kiatyingungsulee, N. 1990. Pharmacological
study of Andrographis paniculata Nees. National Institute of Health,
Bangkok, Thailand.
SCHRI. 1973. Sichuan Chinese Herb Research Institute. Review of chemical studies
on A. paniculata. Journal of New medicinal and new drugs: 23–30.
Semakanit, N. 2005. Effect of dietary substituting granulated creat (Andrographis
paniculata) extract for antibiotic on growth performance, digestibility,
immunity and health of weaning pigs. M.Sc thesis, Department of Animal
Science, Faculty of Agriculture, Khon Kaen University, Khon Kaen, Thailand
(Thai).
Sheeja, K., Guruvayoorappan, C. and Kuttan, G. 2007. Antiangiogenic activity of
Andrographis paniculata extract and andrographolide. International Immuno-
pharmacology 7: 211–221.
, Shihab, P.K. and Kuttan, G. 2006. Antioxidant and anti-inflammatory
activities of the plant Andrographis paniculata Nees. Immunopharmacol
Immunotoxicol 28: 129–40.
Shukla, B., Visen, P.K., Patnaik, G.K. and Dhawan, B.N. 1992. Choleretic effect of
andrographolide in rats and guinea pigs. Planta Med, 58(2): 146–148.
Sithisomwongse, N., Phengchata, J. and Cheewapatana, S. 1989. Acute and chronic
toxicity of Andrographis paniculata Nees. Journal of Thai Pharmacology
Science 14(2): 109– 117.
Sonaiya, E.B. and Swan, S.E.J. 2004. Small-scale poultry production. FAO Animal
Production and Health.
87
Suci, D.M., Nisa, Z., Wahdah, A.A. and Herman, W. 2012. Dietary supplementation
of Andrographis Paniculata Nees meal on performance and serum cholesterol
of laying hen. In: Proceeding of the 2nd International Seminar on Animal
Industry, Jakarta : 315–319.
Suebsasana, S., Pongnaratorn, P., Sattayasai, J., Arkaravichien, T., Tiamkao, S. and
Aromdee, C. 2009. Analgesic, antipyretic, anti-inflammatory and toxic effects
of andrographolide derivatives in experimental animals. Arch Pharm Res,
32(9): 1191–1200.
Surveswaran, S., Cai, Y.Z., Corke, H. and Sun, M. 2007. Systematic evaluation of
natural phenolic antioxidant from 133 Indian medicinal plants. Food
Chemistry 102(3): 938–953.
Tangmutthapattharakun, G. 2007. Effect of Zingiber cassumunar and Zingiber
officinale supplementation in broiler ration on performance during 0-3
weeks of age. Special problem, Department of Animal Production Technology,
Faculty of Agricultural Technology, King Mongkut’s Institute of Technology,
Chaokuntaharn Ladkrabang, Bangkok 10520 Thailand (Thai).
Tg Siti, A.T.K., Othman, A.S., Muhammad, T., Deny, S. and Haitham, Q. 2012.
Antimicrobial activity and essential oils of Curcuma aeruginosa, Curcuma
mangga, and Zingiber cassumunar from Malaysia. Asian Pacific Journal of
Tropical Medicine: 202–209.
Thamaree, S., Pachotikarn, C., Tankeyoon, M. and Itthipanichpong, C. 1985. Effect
on intestinal motility of thirty herbal medicines used in treatment of diarrhea
and dysentery. Chula Med J, 29: 9–51.
Tipakorn, N. 2002. Effects of Andrographis paniculata (Burm. f.) Nees on
performance, mortality and coccidiosis in broiler chickens. PhD thesis,
Institute of animal physiology and animal nutrition, Georg-August-University,
Göttingen, Germany.
Tongwiti, P. 2012. Effects of herbal mixture Androgrophis paniculata, Curcuma
longa and Momordica charantia (Herbatob-Mix ®) supplementation in
diets on productive performance and carcass quality of broilers. M.Sc
thesis, Department of Animal Science, Faculty of Agriculture, Khon Kaen
University, Khon Kaen, Thailand (Thai).
88
Tongwiti, P., Jaikhan, W., Khajarern, J. and Wongtangtintharn, S. 2012. Efficacy of
herbal mixture (Herbatob-Mix®) supplementation in diets on productive
performance in broilers. Khon Kaen Agriculture Journal 40(2): 289–294.
Tripathi, P., Dubey, N.K. and Shukla, A.K. 2008. Use of some essential oils as post-
harvest botanical fungicides in the management of grey mould of grapes
caused by Botrytis cinerea. World Journal of Microb Biotech, 24: 39–46.
Trivedi, N. and Rawal, U.M. 2000. Hepatoprotective and toxicological evaluation of
Andrographis paniculata on severe liver damage. Indian Journal of
Pharmacology 32: 288–293.
. and Rawal, U.M. 2001. Hepatoprotective and antioxidant property of
Andrographis paniculata (Nees) in BHC induced liver damage in mice.
Indian J Exp Biol, 39: 41–46.
Tummaruk, P. and Limtrajitt, V. 2010. The influence of Andrographis paniculata
compound on average daily feed intake of lactating sows and the litter weight
gain of piglets. In: Proceeding of the 48th Kasetsart University Annual
Conference, Thailand.
Udomlak, S., Prapassorn, R., Putthita, P., Sopida, C. and Vichien, K. 2009. Chemical
composition and physical properties of oil from Plai (Zingiber cassumunar
Roxb.) obtained by hydro distillation and hexane extraction. Kasetsart
Journal 43: 212–217.
Vaezi, G., Masoud, T., Shahab, B., Heyder, F. and Saeed, H. 2011. Effects of
different levels of lysine on small intestinal villous morphology in starter diet
of broiler chickens. Global Veterinaria 7(6): 523–526.
Wangboonskul, J., Daodee, S., Jarukamjorn, K. and Sripanidkulchai, B.O. 2006.
Pharmacokinrtic study of Andrographis paniculata tablets in healthy Thai
male volunteers. Thai Pharmacology Health Science Journal 1(3): 209–218.
Wasuwat, S., Wanisorn, P., Mahintorntep, B., Kuwaono, K. and Sanghirun, S. 1989.
Studies on antimicrobial and antifungal activities of terpinen-4-ol extracted
from Zingiber cassumunar Roxb. Thailand Institute of Scientific and
Technological Research, Research Project No. 30- 32/Rep. No.1.
89
Watanasit, S., Siriwathananukul, Y. and Itharat, A. 2005. Effect of Andrographis
paniculata and psidium guajava leaves on growth performance and carcass of
broiler chicken. Songklanakarin Journal of Science and Technology 27:
587–596.
William, O.R. 2005. Digestion and absorption.pp 312-368. In: Functional anatomy
and physiology of domestic animal. 3rd ed. Lippincott Williams & Wilkins.
Wungsintaweekul, J., Sitthithaworn, W., Putalun, W., Hartwig, W.P. and Brantner, A.
2010. Antimicrobial, antioxidant activities and chemical composition of
selected Thai spices. Songklanakarin Journal of Science Technology 32 (6):
589–598.
Xiaolun, S. 2004. Broiler performance and intestinal alterations when fed drug-
free diets. M.Sc thesis, Virginia Polytechnic Institute and State University.
Yang, M., Wang, J. and Kong, L. 2012. Quantitative analysis of four major
diterpenoids in Andrographis paniculata by 1H NMR and its application for
quality control of commercial preparations. Journal of Pharmaceutical and
Biomedical Analysis 70: 87–93.
Yanson, C.V., Summers, B.A. and Schat, K.A. 1987. Pathogenesis of not a virus
infection in various age groups of chickens and turkeys. Pathology. Am J Vet
Res, 6: 927–938.
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
2 80 % ethanol 1.30 Dehydration
3 95 % ethanol 1.50 Dehydration
4 95 % ethanol 1.30 Dehydration
5 Absolute ethanol 1.30 Dehydration
6 Absolute ethanol 1.30 Dehydration
7 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: laingdanet@yahoo.com 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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