biotechnologies for improving animal growthunapcaem.org/activities files/a16/biotechnology...

2005-11-1 Intern.Seminar on Anim.Feed Biotechnology,Beijing

1

Biotechnologies for Biotechnologies for Improving Animal GrowthImproving Animal Growth

Daiwen Chen

Institute of Animal NutritionSichuan Agricultural University

Ya’an Sichuan, 625014China


2

Asian-Australasian J.Anim.Sci.14(12):1794-1802,2001


3

Why is biotechnology needed for animal production?


4

1. Great challenges faced2. increase of animal protein


5

Great challenges faced


6

Flow of nutrients through animals

Cows

Input 100% Output 15 -20%

Excretion to envi. 80-85%


7

Animal production

Low efficiency

HighPollution

Use of growth-promotingingredients

MoreResources

Consumption

High Riskon Human

Health


8

Pollution

Solidmanure, dustwasted feed

Liquidurine,wasted

water

GasCH4,CO2,NH3NO,NO2,SO2

Noise

Odour

Water PollutionGreenhouse

EffectDirect harms

to life


9

Food Safety

• Bovine Spongiform Encephalopathy • Dioxin contamination of animal feeds • Drug residues• Mycotoxins • E. coli O157:H7 • Salmonella


10

challenges

• Ban of growth promotants (antibiotics)• Ban of animal feeds(bone and meat meal)

How to maintain productivity ?


11

1. Great challenges faced2. increase of animal protein


12

Increasing World Population

10popul.(bil.) 8

6

4

2

1920 1940 1960 1980 2000 2020 2040


13

Per capita protein consumption (g/d) 120

total protein100

80

60

40 animal protein

20

500 1000 1500 2000 annual income ($)Pond,2000


14

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


15

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


16

Where to go for animal agriculture?


17

Answer: Biotechnology

biotechnology


18

biotechnology biotechnology

biotechnology biotechnology


19

1. Administration of recombinant somatotropin,IGF,and growth hormone-releasing peptides(GHRPs)


20

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)


21

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 %


22

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


23

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


24

Administration of ST, IGF and GHRPs

• Advantage: high efficient • disadvantage:

---intensive labor---high stress on animals---unacceptable by the public


25

2. Transgenesis


26


27


28

29g

44g

GH-transgenic mouse


29

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%


30

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.


31

• GH-transgenic fish (Chen and Lu,1998) :

growth rate 30-50% G/F: 6-19%


32

Other transgenes

• GRF、 IGF-1、 cSki• Lys 、Thr synthases• cellulase


33

Problems of gene transfer

low efficiencyhealth troubles


34


35

• GH-transgenic pigs had severe health and reproductive problems


36

3. Gene knockout


37

GENE KNOCKOUT

Target gene: general growth-inhibiting gene:

Type Ⅱ IGF receptor gene (Igf2r)local (tissue-specific) growth-inhibiting gene

myostatin


38

Gene Knockout

Igf2r knockout mice (Ludwig et al.,1996).

--- IGF-2 levels--- birth weight being 1.4 times higher


39

Gene Knockout

myostatin -- knockout mice:

muscle mass : doubled


40


41


42


43

Gene Knockout

• in vitro study(Taylor et al.,2001):recombinant MS protein

inhibited in a dose-dependent mannercell proliferationDNA synthesis protein synthesis


44

Gene Knockout

• Double-muscled cattle：Belgian Blue and Piedmontese

20-25% more muscle growth


45


46

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.


47


48

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


49

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


50

4. Immunomodulation


51

---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）


52

--- 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


53

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


54

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



55

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



56

----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



57


-----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


58

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


59

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


60

5 Gene Therapy


61

Skeletal muscle

DNA

Active proteins

artificial endocrine tissue


62

• 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。


63

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）


64


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）


65


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


尿排

泄N（

gN/d）


66


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


N沉

积率

（%）


67


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


68


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)


69


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



70


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



71

Conclusion

• Biotechnology is greatest potential,perhaps the key approach to improve animal growth potential and solve the problems of animal agriculture


72

Thanks and Goodbye !

biotechnologies for improving animal growthunapcaem.org/activities files/a16/biotechnology...

Documents