approach to authorization of novel technologies on ... · approach to authorization of novel...
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Approach to Authorization of
novel technologies on
alternatives to antibiotic in China
Hui yi Cai, Yuan Wang
Deputy President National Feed Industry Association
President National Feed Industry Economy Committee
General Director Feed Research Institute of the CAAS
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1. Animal husbandry and feed
industry of china
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0
40
80
120
160
1990 1995 1998 1999 2000 2001 2002 2003 2004 2005 2006 2007 2008 2009 2010 2011
Feed production (MT) in China (1990-2011)
Fe
ed
pro
du
ctio
n (M
T)
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Compound feed product pass rate are steadily increasing
Feed quality improved
89.38 88.97 88.63
90.93
95.5193.9
80
85
90
95
100
2006 2007 2008 2009 2010 2011合格率(%)
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Feed Row Material
Feed Safety
Excreta Pollution
Animal Product Safety Animals
Today’s Challenges
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61
The balance of protein resources
0
0.1
0.2
0.3
0.4
0.5
0.6
0.7
0.8
2000 2010 2020
Requirment
Avalable
2010 imported
540 million T.
100 million T
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71
0
1
2
3
4
5
6
2000 2010 2020
Requirment
Avalable
1
0
0
M
T
The balance of energy resources
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New technology adoption
Government now encourage the feed industry enterprises adopt new high technology to go along with a professional, standard and the industrial model, the animal farm scale will grow bigger and bigger. Integrator farm play an important role in our country and will keep growing.
New technology such as producing biofeed, nonpolluting feed, no residue feed, and functional feed will be developed to meet overcoming the environment pollution and the animal product quality improving.
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2.General situation on
alternatives to antibiotic
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Fear of the antibiotics use
Over 90 percent of antibiotics consumption are used for feed all over world.
50 percent of the world feed antibiotics were consumed in China.
The increased use of antibiotics has given rise to a fear of the development of resistant pathogenic bacterial strains and residual contamination of the food chain with antibiotics.
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Growth promoting effect of antibiotics
Animal Daily gain FCR
piglet +4.2~136%
(15%)
+1.7~42.7%
(6.5%)
Growing pig
+0~8.9%
(3.6%)
-1.8~3.8%
(2.4%)
Source:Thaler & Miller,2000
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Animals Daily gain FCR
4-20kg piglet(62trials) 11.12±1.19% 8.15±0.82%
20-60kg growing pig(73trials)
9.80±0.80% 8.02±0.70%
60-90kg finish pig(39trials)
6.24±1.68% 8.15±1.36%
chicken(73trials) 4.36±0.44% 3.88±0.47%
Broiler medium(61trials) 3.81±0.44% 3.93±0.44%
Broiler large(35trials) 4.21±1.01% 4.12±0.87%
Duck(41trials) 5.31±0.84% 3.88±0.61%
(1994-2008 trials)
Growth promoting effect of antibiotics in China
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Development of alternative strategies in China
Many countries have a comprehensive ban of antibiotics in feed.
The feeding environment in China is different.
Feed antibiotics is still a meaningful option to prevention of animal diseases.
China will have to face the antibiotic disabling problem.
Intensive research has focused on the development of alternative strategies with the aim of maintenance of animal health and performance.
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3. Management on
alternatives to antibiotic
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The organization of feed additives and animal medicine in china
The management for feed ingredient and feed
additive are under the animal husbandry
department, MOA.
Any new feed additives have to be approved by the
new feed evaluation committee , the ministry of
agriculture of china (MOA).
Presently china has published a< feed additive list>,
<the use guide for feed additive> and <feed
ingredient list >.
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New feed
evaluation committee
Department of
veterinary
Department of
animal husbandry
Feed additive list
Feed resources list guide
for feed additive
Alternatives
to antibiotic
Animal new medicine
evaluation committee
The ministry of agriculture
of China (MOA).
Producers
Organization of feed additives and animal medicine in china
Antimicrobial peptides(AMP).
Lysozyme (溶菌酶).
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Feed quality supervision & detecting system
National Center (1)
MOA Centers (7)
Provincial Agencies (30)
In order to control the feed quality, China has established the
Feed quality supervision & detecting system.
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Feed quality supervision & detecting items by MOA
General feed safety.
Detecting of nutrients and hygiene criteria.
Detecting of clenbuterol and ractopamine.
Detecting of illegal additives:e.g. melamine etc.
Detecting of cattle, sheep & goat derived
ingredients in ruminant feed.
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Feed Additives Supervision items by MOA
Routine supervision and management
Administrative law enforcement
Reporting complaints
Information feedback and the Urgent events.
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4.Novel technologies on alternatives
to antibiotic
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Antibiotics抗生素
Residue
残留 Resistance
耐药性
Harmful to animal and human health
影响动物和人类的健康
Reduce the use of antibiotics
减少抗生素的使用 EU
欧盟 China
中国
Current situation and future development
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Alternatives to antibiotics in China
Various natural materials have been
investigated as efficient alternatives to
antibiotic growth promoters. Many of which
are commercially available.
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Prebiotics
Probiotics
Enzymes
Acidifying agents
Antimicrobial Peptides
Herbal extracts
Alternatives to Antibiotics in China
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1. Probiotics
Probiotics are one of the most important
alternatives to antibiotic in China., which was
widely used in feed mills and animal farms.
Its output is about 50000 tons a year.
Focused on three categories:
Bacillus, Lactic acid bacteria & Yeasts.
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Direct-fed microorganisms by MOA
1. Lactobacillus casei (干酪乳杆菌)
2. Streptococcus faecalis (粪链球菌)
3. Pediococcus acidilacticii (乳酸片球菌)
4. Bacillus natto (纳豆芽孢杆菌)
5. Streptococcus lactis (乳链球菌 )
6. Candida utilis (产朊假丝酵母 )
7. Lactobacillus Plantarum (植物乳杆菌)
8. Streptococcus faeciun (屎链球菌)
9. Bacillus subtilis (枯草芽孢杆菌)
10. Lactobacillus acidophilus (嗜酸乳杆菌 )
11. Sacchaeomyces cerevisiae (啤酒酵母)
12. Rhodop seudanonas palustris (沼泽红假单胞菌 )
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Functional synbiotics
Synbiotics
Bacillus subtilis
枯草芽孢杆菌
Astragalus, aloe
polysaccharide, etc.
黄芪多糖,芦荟多糖等
Immunity
免疫力 Antibacterial substance
抑菌物质 Organic acid
有机酸
Animal Health
动物健康
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Challenge of the probiotics use
Unstable effects
Zn, Cu in the feed Stomach acid
and salt High temp. pressure
In feed processing
Microbial strains
Gene stability
Bad environment of
trans. and store
low live microbes
lower resistance
Higher cost
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② Higher Micro encapsulation of efficiency
③ Higer stability
④ Lower cost
① High live microbes
New processing: pre-fermentation- encapsulation
Pre-fermentation- encapsulation
Realized the integration of fermentation
and micro-capsules
Make the probiotics industrialization possible
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Microscopy photos for preparation of micro-capsule
1 h
8 h
10 h
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Encapsulation technique Processing characteristic
Pre-fermentation
encapsulation
1. High efficiency of Micro encapsulation
2. Effective living bacteria content in the
product of up to 1011cfu/g.
3. Simplify post-processing and easier to
industrialization
4. Low cost and versatile device
5. Well product uniformity
Post-fermentation-coating (fluidized laver coating and spray
coating)
1. Easier to industrialization
2. Low rate of embedding
3. Effective living bacteria content in the
product of up to 109~1010cfu/g
4. Large energy-consuming, high cost
Comparison of Encapsulation techniques
Under the same conditions, consumes the same fermentation raw materials, but 10-100 times live bacteria available
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2. Prebiotics
In China, most of the studied on prebiotic
oligosaccharides are focused:
Fructo oligosaccharides (FOS)
Mannan oligosaccharides (MOS)
Arabinoxylo- oligosaccharides
Xylo-oligosaccharide.
Oligosaccharides can be available on the
market. Some of the feed mills start to use.
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Study on preparative technology of pectic oligosaccharides(果胶寡糖)
and chito-oligosaccharide(壳寡糖)
On the basis of structure-activity relationships
study to pectin and oligosaccharides, Endo-
pectate lyase and commercialization were
selected respectively for Apple pectin and
Chitosan papain degradation, and through
targeted monitoring of polymerization degree
of degradation products, access to the desired
target saccharides.
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Fructo oligosaccharides
(FOS)
FOS can be found naturally in some cereal crops and
onions.
Widely used in swine, poultry and aquatic feed in
China.
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Effects of FOS
Increase
Bifidobacterium,
Lactobacillus
Increase the content
of Lactic acid &
Short chain fatty acids
Intestinal fermentation
FOS
Increase
digestibility
Inhibit pathogenic
bacterial growth Reduce burden
on the kidneys
Promote absorption
of isoflavones
Increased excretion of cholesterol
Decreased cholesterol synthesis Improve
immune function
Promote
Ca, Mg, Fe
absorption
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Mannan oligosaccharides(MOS)
MOS is obtained from the cell wall of yeast (Saccharomyces
cerevisiae).
In recent years, with the development of hemicellulose
resources in feed industry and aquaculture, the application
and research of β – mannanase enter a new stage.
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1. Regulation of immune defense
a. Activated macrophages
b. Improve the antibody response ability, strengthening the protection
of vaccine efficacy
c. Stimulation of hepatic secretion of mannose binding protein, affects
the immune system.
2. Regulation of non-specific immune defense
Interference with intestinal pathogen colonization, prompted the
bacteria become intestinal predominant flora.
3. Adsorption of mycotoxin
Through physical adsorption or direct binding of mycotoxin, eliminate
toxin on the body of the harmful effects
Effects of MOS
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Arabinoxylo-oligosaccharides
A new oligosaccharides, has been highlighted
oligosaccharide on the basis of current state at home
and domestic.
Considered as food ingredients, arabinoxylo-
oligosaccharides have favorable technological
properties and cause prebiotic effects derived from
their ability to modulate the intestinal function.
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Xylo-oligosaccharide
Xylo-oligosaccharide, a kind of prebiological
substance, which can' t be digested but can
selectively activate bacterial reproduction within
intestines.
It can obviously improve intestinal micro
ecological balance, proliferate bifid bacteria and
gastric function.
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3.Enzyme preparations 酶制剂
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3. Enzyme preparation
The feed enzymes developed in recent 10 years
in china, there are a lot of reports about their
application .
Enzymes can degrade the anti-nutrient factors
in feedstuff, increase nutrient digestibility, and
reduce pollution to environment, and have
been widely supplemented in animal feed.
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12 kinds feed enzyme preparations
Presently, 12 kinds feed enzyme preparations are used in China:
amylase, cellulase, ß- glucan, glucose oxidase, lipase, maltase, mannanase, pectinase, protease, amylase, amylopectin, phytase xylanase.
Although the enzyme preparation on the microbial itself does not have significant impact, but it can improve the feed digestibility, reduced nutrient residues in digestive tract weight, shorten the residual time, indirect reduction of pathogen growth opportunities, and realize the effect of disease prevention.
The use of feed enzymes as antimicrobial growth promoters will need to be studied further before this strategy finds widespread use within the feed industry.
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Phytase
Phytase is now most important enzyme
because of its relatively easy manufactured
and be used in all kind feed of animals.
There are 4 companies have set up a
international business with 40 countries.
Granular, powder, Instant Soluble.
5,000u/g - 200,000u/g are avaiable.
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Instant Soluble Phytase
SPECIFICATIONS 99.99%
soluble. (see the left solubility
test picture) Colour: Buff or
white powder Odour: Normal
fermentation odour Enzymatic
Activity: 100,000u/g Activity
Temperature:20-
70℃,favorable at 50℃ Carrier
Material: Instant soluble
Mineral.
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Heat-resistant acid Phytase
Temperature, ℃
Activ
ity, %
Temperature, ℃
Activ
ity,%
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Heat-resistant acid Phytase
High temperature
resistant acid Phytase
pepsin and trypsin-
resistant features,
pH3.5~pH7.0 range
optimum pH4.0, have
a higher stability.
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Existing questions and the methods to increase the yield of -mannanase存在问题及提高-甘露聚糖酶产量的方法
Low yield and enzymatic activity, difficult for massive
application of -mannanase
产量低,酶活力不高,使-甘露聚糖酶不能大量推广使用
Screening high yielding A. sulphureus mannanase strains by a
mutant treatment
诱变选育高产硫色曲霉MAFIC001 -甘露聚糖酶菌种
Modifying -mannanase gene sequence according to codon bias
根据密码子偏好性,优化-甘露聚糖酶基因序列
-mannanase
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Pichia pastoris expression vector
毕赤酵母表达载体
A
Fermentation device (10 L)
发酵装置(10升)
Highest protein expression level (最高蛋白浓度):3 g/L Highest enzymatic activity (最高酶活力):1100 U/mL
Purity (纯度):96%
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Enzyme Microb. Tech., P. Deng, Defa Li, 2006
Xylanase木聚糖酶
Aspergillus niger Xylanase gene
Realization of expression in Escherichia coli and pichia pastoris
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+ Modified xynA 改造xynA
pGAPZαA
载体
expression in Pichia pastoris
毕赤酵母中表达
Aspergillus sulphureus硫色曲霉
Thermostability of the modified recombinant xylanase
改造的重组木聚糖酶热稳定性研究
0
20
40
60
80
100
10 30 50 70 90
Time (min)
Relative activity (%)
Enzymatic activity: 120 U/mL
最高酶活力达120 U/mL
Protein concentration: 3.78 mg/mL
蛋白浓度为3.78 mg/mL
Good stability: 80ºC for 90 min
热稳定性好,80ºC保持90
min
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Alpha-Galactose glucoside enzyme
Alpha-Galactose glucoside enzyme could resistant the degradation of neutral and alkaline protease. it has high specific activity under the condition of PH value 2-8, and can withstand 80 ℃ high temperature.
Experimental results show that addition in the broiler diet with different levels of Galactose glucoside enzyme (150 U/kg, 300 U/kg, 1500 U/kg group), it improves FCR, increase energy, protein apparent metabolic rate as compared with the control group.
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High cell density fermentation of recombinant pichia pastoris yeast production technology
Established
Established the high level expression of
recombinant strain of Alpha-Galactose
glucoside enzyme, Cellulase and Pectinase.
The expression of 3 kinds of enzyme in the
fermentation tank is up to 10000U/ml, 30000
U/ml and 10000 U/ml respectively.
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4.Antimicrobial peptides (AMPs)抗菌肽
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4. Antimicrobial peptides (AMPs)
AMPs are another major group of promising novel alternatives to antibiotics based on their effectiveness, safety, and enormous diversity.
AMPS :Important components of the innate immune
system 先天免疫系统的重要组成成分
Abundant and diverse groups of molecules that are encoded by definite genes
特定编码基因产生的一类小分子多肽
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Current situation and existing problems 现状及存在问题
Research of higher structure and antibacterial
mechanism of AMPs
研究抗菌肽高级结构和抗菌机理
Short half-life, easy to lose activity
半衰期短,易失活
Toxicity toward prokaryotic cell
抗菌肽对原核细胞的毒性
Low yield, difficult to purify
天然抗菌肽纯化难度大,产量低
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Modify the molecule of AMPs
优化抗菌肽分子
Select definite strain, and fusion expression
原核系统中选育特定抗性菌株,真核系统中采用融合
表达以提高其表达率
Special induction mode, reduce product cost
以特殊的诱导模式,降低生产成本
Key points for product development
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Antimicrobial peptides-activity feed products development
Through the Bacillus subtilis modified
system, antibacterial peptide expression
volume reached 1.8mg/ml above.
The key technology to the low expression
volume and low stability solved.
① Two Antimicrobial Peptide expression platform
establishment of Bacillus subtilis and Yeast .
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② Clear features molecular structure is available.
The mass spectrometric analysis for Antibacterial
peptide :
cecropin consists of 37 amino acid composition,
molecular weight of 3.866KD.
Bacillus subtilis consists of 37 amino acid, molecular
weight: 3.879KD, its amino acid sequence is:
GLGKAQCAALWLQCASGGTIGCGGGAVACQNYRQFCR.
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Figure 1. Antibacterial peptide
cecropin mass spectra
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Figure 2. Bacillus subtilis bacteria
antimicrobial peptide mass spectra
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Three-dimensional structure of the two antibacterial peptides
Figure 3 cecropin three-
dimensional structure
Figure 4 Bacillus subtilis three-
dimensional structure
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③ Complete the antibacterial peptide cecropin and subtilin part antimicrobial spectrum and stability test
Division bacteria Minimum inhibitory
concentration(μg/ml)
E.coli K12D31 (大肠杆菌K12D31 ) 1.8
E.coli K88 (大肠杆菌K88) 2.0
E. coli K99 (大肠杆菌K99) 2.0
Salmonella typhimurium (鼠伤寒沙门氏菌) 8.0
Salmonella enteritidis (肠炎沙门氏菌) 16.0
Staphylococcus aureus (金黄色葡萄球菌) 0.2
Streptococcus faecalis (粪链球菌) 24.0
Table 1, cecropin antibacterial peptides against pathogenic bacteria
of the minimum inhibitory concentration
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Table 2. Minimum inhibitory concentration of the
Bacillus subtilis against pathogenic bacteria
Division bacteria Gram attributes
Minimum inhibitory
concentration(ug/ml)
Staphyloccocus aureus CMCC26003 G+ 1.3
Staphyloccocus aureus CVCC1882 G+ 4.69
Staphyloccocus aureus ATCC29213 G+ 23.43
Staphyloccocus aureus ATCC25923 G+ 11.7
Staphyloccocus aureus ATCC43300 G+ 187.5
Staphyloccocus aureus野生株(吉林) G+ 23.43
Staphyloccocus aureus野生株(华中) G+ 11.7
A.hydrophila (嗜水气单胞菌 ) G- 46.9
B.cereus (蜡样芽孢杆菌) G+ 0.2
Vibrio anguillarum (鳗弧菌 ) G- 12.8
C.perfringens (魏氏梭菌) G+ 1.4
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pH value effect on the activity of two antibacterial peptide
0.0
1.0
2.0
3.0
4.0
5.0
6.0
抑菌
圈(
mm)
Figure 9 pH value effect on the activity
of antibacterial peptide cecropin Figure 10 pH value on effect
of Bacillus subtilis bacteria activity
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Antimicrobial peptides-activity feed products development
④ Obtained pure purity greater than 99.5%
of antibacterial peptide, established two
methods for detection of antimicrobial
peptides, complete isolation and purification
of antibacterial peptide cecropin pilot
⑤ Completed the feed safety evaluation of
antibacterial peptide cecropin, established
product quality standards.
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1: control with no insert; 2-6: positive recombinants induced 72 h
1:对照;2-6:重组菌诱导72 h的正对照组(抗菌肽)
Process biochemistry, J. Zhang, 2006
Antibacterial assay
抑菌分析
E. coli K12D31
AMPs-sensitive strain
抗菌肽敏感菌株
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Studies on other bioactive peptides and defense peptides
① Recombinant expression of host defense peptides of Palustrin-OG2
Improved OG2 peptide expression in Escherichia coli , purification and activity determination.
② Recombinant expression of Apidaecin(蜜蜂肽) and Spheniscin(企鹅防御肽).
③ Recombinant expression of BuforinII(牛乳铁蛋白素) in the photorhabdus luminescens .
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5.Herbal extracts (Astragalus polysaccharide, APS)
天然提取物(黄芪多糖)
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5. Chinese herbal extracts
Practice has proved it can promote appetite, enhance immunity, prevent and cure diseases etc..
Traditional Chinese medicine additives is over 50 kinds.
Slow effect ,Much required dose and antibiotics-like effects.
the main problems : bad palatability and high cost.
two important representives : oregano oil and Astragalus polysaccharide.
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APS Hypothesis
Inflammation reaction
of LPS
LPS炎性反应
Disorder of GH/IGF-I axis
GH / IGF-I轴异常 Immune suppress (cyclophosphamide)
免疫抑制(环磷酰胺,CY )
Immune enhancement
免疫增强
Immune type
免疫类型
APS 黄芪多糖
Inflammation factor secretion
炎性因子分泌
-?
+
- -?
-?
+
+ ?
-
Cell culture
细胞培养
?
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1
1.2
1.4
CD
4+
/CD
8+
0 100 250 500 750 1000
APS (mg/kg)
(Linear P < 0.05)
1
1.1
1.2
1.3
1.4
1.5
T-ly
mp
ho
cy
te
pro
lifera
tio
n(O
D)
0 100 250 500 750 1000
APS (mg/kg)
(Linear P < 0.05)
Effects of APS on ratio of CD4+/CD8+ and
Lymphocyte proliferation in piglets
Anim. Sci., S. L. Yuan, X. S. Piao, Defa Li, 2006
Improve cell immune function, the optimal APS level was
500 mg/kg
APS在断奶仔猪日粮中的最适添加量为500 mg/kg
CD4+/CD8+ 淋巴细胞增殖
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Effects of dietary supplement of APS on serum cytokines in weaned pigs received cyclophosphamide (CY)
APS(注射环磷酰胺, CY)对断奶仔猪血清细胞因子的影响 (pg/mL)
Anim. Sci., S. L. Yuan, X. S. Piao, Defa Li, 2006
-CY +CY
-APS +APS -APS +APS
d 14
IL-2 55.4 79.8 50.6 77.8
IFN-γ 133.3 160.1 111.7 138.9
Reverse CY-induced cell immune suppress and humour
immune suppress 环磷酰胺免疫抑制模型研究发现,APS可促进猪的正常细胞免 疫,也可逆转CY造成的免疫抑制
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Effects of APS on serum index in piglets APS对仔猪血清指标的影响
Recover the normal function of GH/IGF-I axis
APS通过降低炎性细胞因子、GH和皮质醇的含量,增加抗炎性
因子的合成,恢复GH/IGF-I轴的正常而发挥其免疫抗炎作用
Anim. Sci., S. L. Yuan, X. S. Piao, Defa Li, 2006
-LPS +LPS
-APS +APS -APS +APS
d 14
Cortisol 115.3 113.0 155.5 136.0
GH 3.82 3.83 4.56 3.94
IGF-I 235.6 240.0 215.9 224.9
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b
Increase NO and iNOS activity of the peripheral blood
lymphocytes in piglets
APS显著增加仔猪外周血淋巴细胞NO和iNOS的活性,并呈剂量依赖性
Effects of APS on NO and iNOS activity in piglets APS对仔猪NO和iNOS的影响
J. Anim. Sci., X. F. Mao, Defa Li, 2005
0.5
0.6
0.7
0.8
0.9
1
1.1
0 40 80 120 160 200 240 280 320 360多糖浓度 Polysaccharide concentration
(ug/mL)
iNOS 活
性 iNOS bioactivity
(U/mg.pro)
1
1.2
1.4
1.6
1.8
2
2.2
0 40 80 120 160 200 240 280 320 360多糖浓度 Polysaccharide concentration(ug/mL)
NO产
生量
No production
(pmol/L)
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0
20
40
60
80
100
120
140
160
180
200
6 12 24 48 72
Time (h)
IF
N-γ
(p
g/m
L)
0 μg/mL APS
40 μg/mL APS
80 μg/mL APS
160 μg/mL APS
* *
* *
*
(P < 0.05)
0
10
20
30
40
50
60
70
80
90
6 12 24 48 72Time (h)
IL-2
(p
g/m
L)
0 μg/mL APS
40 μg/mL APS
80 μg/mL APS
160 μg/mL APS
*
* *
*
* *
*
(P < 0.05)
Effects of APS on different cytokines APS对不同细胞因子的影响
Increase the IL-2 and IFN-γ secretion
APS可在体外促进细胞因子IL-2和IFN-γ的分泌,这 可能是APS的免疫调节作用之一
Anim. Sci., S. L. Yuan, X. S. Piao, Defa Li, 2006
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J. Anim. Sci., X. F. Mao, Defa Li, 2005
Mechanism of immune function of APS
APS的免疫作用机理
APS
iNOS NO
细胞因子表达
淋巴细胞增殖
+
改善动物免疫
IL-2 etc. IFN-γ
+ +
Receptor
Gene knockout
基因敲除
Signal transduction pathway
信号传导通路
TLRsToll 样受体
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6. Acidifying agent
Research shows, lactic acid, citric acid or acetic acid and other organic acids can promote the animal appetite, improve weight gain and feed conversion. However, there are some problems of feed supply organic acids must be considered, such as corrosion of equipment and higher cost.
Acidification agent application in China feed industry is still in the initial stage, usage and methods have not yet be standardized.
Organic acid is added in feed and feed formula to consider the compatibility, high protein and salt minerals will be of the acid buffer action; moreover some special acidification agent will cause the animal reduction of feed intake
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5. Conclusions
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1. Wide variety of antibiotic replacements
available.
2.Each product or class has distinct
characteristics, responses, advantages, and
disadvantages.
3.Scientific literature increasing worldwide,
and effectiveness of products is continually
improving.
Conclusions
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Probiotics, Antimicrobial peptides, Prebiotics, Herbal
extracts, Enzyme preparations, Acidifying agents
酶制剂,天然提取物,益生素,抗菌肽, 寡糖, 酸化剂
Recombinant technique
重组DNA技术 Microbiology
微生物学
Antibiotics
抗生素
Extraction
提取
Conclusions
Safety Animal products
绿色畜产品
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