2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Biotechnologies for Biotechnologies for Improving Animal GrowthImproving Animal Growth
Daiwen Chen
Institute of Animal NutritionSichuan Agricultural University
Ya’an Sichuan, 625014China
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Asian-Australasian J.Anim.Sci.14(12):1794-1802,2001
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Why is biotechnology needed for animal production?
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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1. Great challenges faced2. increase of animal protein
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Flow of nutrients through animals
Cows
Input 100% Output 15 -20%
Excretion to envi. 80-85%
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Animal production
Low efficiency
HighPollution
Use of growth-promotingingredients
MoreResources
Consumption
High Riskon Human
Health
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Pollution
Solidmanure, dustwasted feed
Liquidurine,wasted
water
GasCH4,CO2,NH3NO,NO2,SO2
Noise
Odour
Water PollutionGreenhouse
EffectDirect harms
to life
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Food Safety
• Bovine Spongiform Encephalopathy • Dioxin contamination of animal feeds • Drug residues• Mycotoxins • E. coli O157:H7 • Salmonella
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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challenges
• Ban of growth promotants (antibiotics)• Ban of animal feeds(bone and meat meal)
How to maintain productivity ?
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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1. Great challenges faced2. increase of animal protein
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Increasing World Population
10popul.(bil.) 8
6
4
2
1920 1940 1960 1980 2000 2020 2040
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Per capita protein consumption (g/d) 120
total protein100
80
60
40 animal protein
20
500 1000 1500 2000 annual income ($)Pond,2000
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Sustainable Animal Agriculture
• Reduce input/ cost. • Increase output/ productivity. • Reduce environmental burden/cost. • Maintain safety. • Improve quality. • Increase awareness and confidence • Maintain competitiveness• Compatible with other industry
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Goal Shift of Animal Agriculture
One goal in the past: high yield of productsMulti-goals from now on:
1. high yield of safe animal products, 2. high utilization of natural resources 3. minimum environmental pollution
sustainability
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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biotechnology biotechnology
biotechnology biotechnology
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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1. Administration of recombinant somatotropin,IGF,and growth hormone-releasing peptides(GHRPs)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Effects of pST in pigs
(Etherton and Bauman,1998)
daily retention (g) stage dose( ug/kg) ADG (g) G / F Protein Fat
10-25 kg 0 680 0.61 96 89120 680 (0) 0.61 (0) 113 ( +17) 61 ( -31)
20-50 kg 0 900 0.43 120 207150 990 ( +10) 0.49 ( +13) 150 ( +25) 122 ( - 41)
50-100 kg 0 1,140 0.33 135 340150 1,334 ( +17) 0.44 ( +33) 235 ( +74) 61 ( -82)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Injection of exogenous IGF-1
Klindt (1998):
Injection of IGF-1 into Meishan pigs for 28 days
growth rate 22 % carcass protein accretion 33 % trimmed lean cuts 5 %
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Growth hormone-releasing peptides(GHRPs)
GHRP-6: His-D-Trp-Ala-Trp-D-Phe-Lys-NH2
GHRP-1: D-Ala-His-D-βNal-Ala-Trp-D-Phe-Lys-NH2
GHRP-2: D-Ala-D-βNal-Ala-Trp-D-Phe-Lys-NH2
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Phung et al.(2000): pigs of 69 kg 30μg GHRP-2 /kg BW s.c. once daily for 30 days
ADG: 22.35% (P<0.05) G/F: 20.64% (p<0.01) Plasma GH peak concentration :12-15 folds
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Administration of ST, IGF and GHRPs
• Advantage: high efficient • disadvantage:
---intensive labor---high stress on animals---unacceptable by the public
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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GH-transgenic pigs (Pursel and Solomon,1993) :
ADG:13%G/F: 18%At 92kg, total fat in carcass 85%
s- fatty acids 85% ms-fatty acids 91%PUFA 66% IMF in ham 43%
in loin 66%in shoulder 64% in belly 69%
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Total fat in carcass(%)
BW kg 14 26 48 88
Transgenic 6.19 7.62 7.54 3.27
Control 10.03 12.32 15.50 19.55Solomon,1992.
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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• GH-transgenic fish (Chen and Lu,1998) :
growth rate 30-50% G/F: 6-19%
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Other transgenes
• GRF、 IGF-1、 cSki• Lys 、Thr synthases• cellulase
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Problems of gene transfer
low efficiencyhealth troubles
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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• GH-transgenic pigs had severe health and reproductive problems
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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GENE KNOCKOUT
Target gene: general growth-inhibiting gene:
Type Ⅱ IGF receptor gene (Igf2r)local (tissue-specific) growth-inhibiting gene
myostatin
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Gene Knockout
Igf2r knockout mice (Ludwig et al.,1996).
--- IGF-2 levels--- birth weight being 1.4 times higher
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Gene Knockout
myostatin -- knockout mice:
muscle mass : doubled
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Gene Knockout
• in vitro study(Taylor et al.,2001):recombinant MS protein
inhibited in a dose-dependent mannercell proliferationDNA synthesis protein synthesis
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Gene Knockout
• Double-muscled cattle:Belgian Blue and Piedmontese
20-25% more muscle growth
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Mechanism for double-muscling
• Belgian Blue:11 nucleotides in exon 3 deleted
null activity of MS protein. • Piedmontese:
guanosine in exon 3 adenosine
tyrosine cysteine.
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Gene Knockout
In Pigs (Ji et al., 1998):low birth weight (0.57 kg )
vshigh birth weight(1.37kg).
MS expression in LD: 65% higher (P<0.04) for low weight
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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High expression of MS is associated with low birth weight in pigs
MS 165% mRNA
in LM 100% Ji et al.1998
Am.J.Physiol.
1.37kg 0.57kg pig birth weight
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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---Immunomodulation of GH
• Active immunization against GH,GH receptor、GH binding protein、GHRIH
higher GH and anabolic activity higher growth and protein accretion
• pST antibodyantibody to pST antibody better growthsynthetic peptide (pGH54-95)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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--- immunization against• IGF-1、IGF-2: • faster growth ,less fat deposition• adrenocorticotropin: better growth• LH: removing boar taint
avoiding surgical castration maintaining high leanness and G/F
• adipose plasma membrane: • more lean gain, less fat gain
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Active immunization against CCK in pigs Active immunization against CCK in pigs 0f 270f 27--90 kg90 kg
0.00
500.00
1000.00
1500.00
2000.00
2500.00
1-2周 3-4周 5-6周 7-8周 9-10周
AD
FI(
g)
实验组 对照组
---- feed intake
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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0.00100.00200.00300.00400.00500.00600.00700.00800.00900.00
1-2周 3-4周 5-6周 7-8周 9-10周
ADG(
g)
实验组 对照组
----ADG
Active immunization against CCK in pigs Active immunization against CCK in pigs 0f 270f 27--90 kg90 kg
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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0.001.002.003.004.005.00
1-2周 3-4周 5-6周 7-8周 9-10周
F/G
实验组 对照组
--- F/G
Active immunization against CCK in pigs Active immunization against CCK in pigs 0f 270f 27--90 kg90 kg
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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----CCK antibody titer in serum抗体滴度变化曲线
0.000.200.400.60
0.801.001.20
1 15 29 43 58 71天
OD值
处理组 对照组
Active immunization against CCK in pigs Active immunization against CCK in pigs 0f 270f 27--90 kg90 kg
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Active immunization against CCK in pigs Active immunization against CCK in pigs 0f 270f 27--90 kg90 kg
-----CCK gene expression
.空肠中CCK mRNA的体积
3000
4000
5000
6000
7000
8000
9000
control对照组 CCK-8实验组
垂体中CCK mRNA的体积
30003500
40004500
50005500
60006500
7000
control对照组 CCK-8实验组
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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ADFI,g
70
80
90
control cck yolk
final BW,g
1950
2000
2050
2100
2150
control cck yolk
F/G
1.55
1.6
1.65
1.7
1.75
control cck yolk
Effect of 100 ppm CCK yolk in the diet on performance of broiler of d 1-42
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Immunomodulation
• advantage:-- no side effects-- more acceptable than GH injection
• disadvantage:-- active immunization: poor control of level
and duration of immune response
-- passive immunization: more expensive
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Skeletal muscle
DNA
Active proteins
artificial endocrine tissue
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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• plasmid DNA-encoding function genes into muscle tissue
• In mice(DraghiaAkli et al.,1997)injection of 100 ug GHRH-pDNA
serum GH: 3-4 fold for 2 wks;body gain: 10% higher。
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
pGRF基因质粒注射剂量对猪生产性能的影响
0.246c
0.194ab
0.179a
0.256c
0.243c
0.314a
0.366b
0.400c
0.386bc
0.349ab
0.1
0.2
0.3
0.4
0 0.25 0.5 1.0 2.0
注射剂量(mg)日增重(Kg) 日采食量(Kg)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
料肉比
1.76a
1.89a
1.62ab
1.51b
1.44b
1.4
1.5
1.6
1.7
1.8
1.9
2
0 0.25 0.5 1.0 2.0
注射剂量(mg)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
pGRF基因质粒注射剂量对猪氮代谢平衡的影响
4.8154a
4.6250a4.7700a
3.8900c
4.2200b
3.5
3.9
4.3
4.7
5.1
0 0.25 0.5 1.0 2.0
注射剂量(mg)
尿排
泄N(
gN/d)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
46.9a 48.4a
54.3b
61.4c
56.8b
40
45
50
55
60
65
0 0.25 0.5 1.0 2.0
注射剂量(mg)
N沉
积率
(%)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
注射pGRF基因质粒对猪蛋白质代谢的影响
7.8956a 8.5307ab10.2487c 10.7963c
10.1034c
18.8034b
14.5107a15.7056a
19.8363b
18.6628b
5
7
9
11
13
15
17
19
21
0 0.25 0.5 1.0 2.0注射剂量(mg)
速率(gN/kgW0.75.d)
蛋白合成速率 蛋白周转速率
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
蛋白沉积速率(gN/kgW0.75.d)
1.2805a 1.3359
b
1.694c 1.7563
c
1.5439bc
1
1.2
1.4
1.6
1.8
0 0.25 0.5 1.0 2.0
注射剂量(mg)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
内源尿N排泄速率(gN/kgW0.75
.d)
0.6582b
0.5925c
0.7198b0.7421ab
0.7917a
0.55
0.6
0.65
0.7
0.75
0.8
0.85
0 0.25 0.5 1.0 2.0
注射剂量(mg)
2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing
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Researches in Sichuan.Agri.Univ.
氨基酸重复利用速率(gN/kgW0.75.d)
5.8234a
6.4328a
7.8349ab
8.4475b
7.9012b
5
6
7
8
9
0 0.25 0.5 1.0 2.0
注射剂量(mg)
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Conclusion
• Biotechnology is greatest potential,perhaps the key approach to improve animal growth potential and solve the problems of animal agriculture
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