CN1552234A - Feed additive for improving animal disease-resistant ability - Google Patents
Feed additive for improving animal disease-resistant ability Download PDFInfo
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- CN1552234A CN1552234A CNA031289576A CN03128957A CN1552234A CN 1552234 A CN1552234 A CN 1552234A CN A031289576 A CNA031289576 A CN A031289576A CN 03128957 A CN03128957 A CN 03128957A CN 1552234 A CN1552234 A CN 1552234A
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Abstract
A feed additive for increasing the resistance of animal to diseases is prepared from the fragments of the plants in pinaceae through pulverizing and extracting in hot water. Its contains polyphenol compound and polyose for regulating the balance between floras in animal's intestinal tract.
Description
One, technical field
The present invention relates to a kind of feed addictive, especially the feed addictive that contains polyphenol compound and glucose, mannose, wood sugar, Arabic galactolipin that extracts from Pinaceae abieteae pine genus plant fragment prepares the method for producing this feed addictive and the purposes of this feed addictive.
Two, background technology
People have realized that now the imbalance of flora in the animal intestinal is one of reason that causes animal diarrhoea.If the pathogenic microorganism in the enteron aisle, for example a large amount of breedings such as Escherichia coli, salmonella, perhaps the raised growth of corrupt mushroom can cause profitable strain, and for example decline of Bifidobacterium and Bacillus acidi lactici quantity is easy to cause animal diarrhoea like this.The conventional method of treatment animal diarrhoea is to feed to animal to use antibiotic, but long-term abuse of antibiotics, not only caused weakening of letting animals feed resistance against diseases, the decline of meat egg product quality and meat flavour, and improved the probability of carrying resistance factor in the pathogen significantly, it is that human health has been caused immeasurable infringement that animal is taken antibiotic consequence for a long time.Therefore not contain antibiotic feed addictive imperative in exploitation.Plant extracts contains the material of multiple physiologically active, does green feed additive with it, is subjected to people's attention.
People utilize the polyphenol compound in the plant extracts, existing history (Plantamedica 58 Supplemental 1 ppA691 for many years of treatment enteron aisle diarrhoea; Basic Life Science59:717-728).And there is the report Tea Polyphenols can reduce the content (Livestock production Science 68:217-229) of C.perfringens in human body and the animal intestinal effectively, and can improve the content of Bifidobacterium effectively, Tea Polyphenols has been widely used in human health care's food and the Animal nutrition as antioxidant and antiseptic, particularly be applied to immunopotentiator, as patent application 94107044.1, a kind of immunopotentiator that can treat disease caused by toxic reaction of excessive free radicals.
The noticeable compound of another one in the plant extracts just is the polysaccharide compound of non-starch based.Polysaccharide compound is the main source that constitutes string.String can be divided into two kinds of Soluble Fiber and insoluble fibers.Soluble Fiber is easier in animal intestinal is generated SCFA by the bacterium decomposing and fermenting, and some Soluble Fiber has the function of prebiotics, can optionally improve the activity and their growth and breeding of promotion of profitable strain in the intestines.Typical example is from the hemicellulose araboxylan that extracts in the wheat bran (Cereal Food World 38 (7): 490-492; Journal ofAgriculture Food Chemistry 48:1644-1652) and polyfructosan (Gastroenterology108:975-9821995).
Pine needle meal and extract from pine needles are widely used in the livestock culturing industry in China as feed addictive, a kind of pine needle agent and production method thereof of feeding pig of can be used for that number of patent application provides for 96118185.0 patent application, but in No. 96118185.0 patent applications active ingredient in the extract from pine needles is not measured, the influence of extract from pine needles to the animal intestinal bacterial structure in raising is not described simultaneously yet.Pine needle meal in the past and pine needle are applied in the feed addictive as a component of composition mostly, as number of patent application a kind of Chinese herbal medicine additives for compound pig fodder and the production method thereof of 95112493.5 patent application, and for example number of patent application is that patent application animal nutrient of 01118107.9 etc. is not always the case, their common ground is only as a component of feed addictive, and does not indicate the mechanism of action and concrete purposes.
Relate in the U.S. Pat 5614501 from European larch (genus Larix, Westernlartch) extract polyphenol compound and Arabic galactolipin can be regulated flora in the animal intestinal effectively as feed addictive balance in, reduce the quantity of harmful bacteria, thus prevention and treatment intestines problem.But because European larch belongs to Pinaceae larch subfamily larch, the distributed pole domestic in China is narrow, limited amount, and raw material can't satisfy mass-produced needs.China's pinaceae plant distributed more widely is the abieteae pine genus plant, as masson pine, Chinese pine, black pine, Korean pine, pinus yunnanensis, wet-land pine tree, Pinus taiwanesis, river deer pine etc.
Three, summary of the invention
The technical problem to be solved in the present invention provides a kind of green feed additive that can promote the fowl poultry kind healthy growth, produces the method for this green feed additive of preparation and the purposes of this green feed additive, this feed addictive can be regulated the balance of flora in the animal intestinal effectively, produces the shortcoming that green feed additive use Chinese herbal medicine causes cost high price costliness in the prior art to solve.For this reason, the present invention takes following technical scheme:
A kind of feed additive for improving animal disease-resistant ability, described feed addictive are the hot water extract after the powder process of Pinaceae abieteae pine genus plant fragment, contain polyphenol compound and polysaccharide among the described hot water extract.
Described Pinaceae abieteae pine genus plant can be one of following plant:
1. 2. 3. 4. 5. 6. Korean pine of pinus yunnanensis of wet-land pine tree of black pine of Chinese pine of masson pine
Extract from pine needles used in the present invention is meant the water-soluble extractive that extracts through pulverizing from the needle of Pinaceae abieteae pine genus plant and limb.Masson pine in the Pinaceae abieteae Pinus, Chinese pine, black pine, wet-land pine tree, pinus yunnanensis, Korean pine etc. all comprise in the present invention.Here the latin name of the Pinaceae abieteae pine genus plant of indication is as follows:
Masson pine: Pinus massoniana Lamb
Chinese pine: Pinus tabulaeformis Carr
Black pine: Pinus thunbergii Parl
Wet-land pine tree: Pinus elliottii Engelm
Pinus yunnanensis: Pinus yunnanensis Franch
Korean pine: Pinus Koraiensis Sieb.et Zucc
Huashan pine: Pinus armandi Franch
Pine tree fragment or pine needle that described Pinaceae abieteae pine genus plant fragment is pulverized for the pine branch xeromenia.
If adopt the limb (pine bark and xylem) of pine tree, then need through and pulverizing,
Described polyphenol compound is the mixture that comprises flavone compound, Lignanoids compounds, flavan-3-alcohol compounds.
Described polysaccharide is the mixture that contains glucose, mannose, wood sugar, Arabic galactolipin.
Ratio between described polyphenol compound and the polysaccharide can be between 1: 1 to 1: 50.
The present invention is to use the extract that extracts from pine needle and pine tree limb, wherein contain polyphenol compound and polysaccharide compound such as glucose, mannose, wood sugar, Arabic galactolipin as feed addictive, improve the quantity of beneficial bacterium in the animal intestinal and the growth of inhibition harmful bacteria.
A kind of method for preparing feed additive for improving animal disease-resistant ability, step is extracted in the following order:
1. fresh pine tree fragment flush away impurity, the segment that is cut into 1-5cm with chopper is made the pine tree meal, gets 1 part of pine tree meal, adds 2 parts-10 parts in water, and is pending;
2. pending liquid is smashed defibrination, and dope filtration is collected filtrate, and it is pending that filter residue adds 3 parts to 5 parts water;
3. the extraction of heating of pending liquid is extracted temperature and is controlled at 80-100 ℃, and extraction time is 30 minutes-2 hours, and extracting liquid filtering is collected filtrate;
4. filter residue with 3 parts-5 parts water extraction, extracts temperature and is controlled at 80-100 ℃ again, and extraction time is 30-60 minute, and extracting liquid filtering is collected filtrate for later use;
5. merge the filtrate of collecting previously, concentrate, obtain tan viscous liquid and promptly can be feed additive.
4. the preparation method of described feed addictive when described pine tree fragment is simple pine needle, can omit step.
2. the preparation method of described feed addictive when described pine tree fragment is simple pine branch dry crushing sheet, can omit step.
The preparation method of described feed addictive, described step amount of water preferred version 1. is 3 parts, and described step extraction time 3. is 60 minutes.
The preparation method of described feed addictive makes liquid form with the feed additive that 5. described step obtains.
The preparation method of described feed addictive makes solid form with the feed additive that 5. described step obtains.
Put it briefly, involved in the present invention to the extracting method of extract from pine needles be summarized as follows: fresh pine needle flush away impurity.Be twisted into the segment of 1-5cm with chopper, add the water of appropriate amount by a certain percentage, smash defibrination.The ratio of pine needle and water can be controlled in the bigger scope, from 1: 2 to 1: 10 all can, optimal proportion was controlled at 1: 3, homogenate filters, and collects filtrate.Filter residue is used 1: 3 to 1: 5 hot water extracting again, extracts temperature and is controlled at 80～100 ℃, and the time is 30 minutes to 2 hours, more after filtration, collects filtrate.Merge filtrate twice,, obtain tan viscous liquid through concentrating.
If adopt the limb (pine bark and xylem) of pine tree, then need add the water of appropriate amount by a certain percentage through pulverizing.Leaching process adopts high-temperature water to extract, and temperature is controlled at 80～100 ℃, and the time is 30 minutes to 2 hours, extracts the back and filters, and collects filtrate.Filter residue is used 1: 3 to 1: 5 80～100 ℃ of hot water extracting 30-60 minutes again.Twice filtrate merges the back and concentrates, and removes moisture.Concentration process can adopt atmospheric evaporation and two kinds of technologies of reduction vaporization, or adopts other effectively to concentrate means.
The feed addictive of making can be directly with the liquid form of above-mentioned extract, also can be processed into solid form, even extract directly can be added in the feed, being prepared into the animal feed that contains feed addictive of the present invention, can both attain the results expected with these mode feeding animals.
A kind of purposes of feed additive for improving animal disease-resistant ability, described feed addictive consumption can effectively reduce the quantity of pernicious bacteria such as Escherichia coli, salmonella and staphylococcus aureus greater than 0.1% dosage feeding animals of total forage volume; Also can stimulate the growth of profitable strain such as Bifidobacterium, Bacillus acidi lactici, bacillus subtilis and photosynthetic bacteria effectively.Described feed addictive preferable amount is the 0.1%-5% of total forage volume.
The purposes of described feed additive for improving animal disease-resistant ability can be suitable for one of following animal:
1. 2. 3. 4. 5. 6. 7. 8. 9. dove of cat of dog of horse of ox of goose of duck of chicken of pig
The present invention can effectively reduce pernicious bacteria quantity and stimulate the profitable strain growth effectively.Harmful bacteria among the present invention is meant the pathogen or the putrefactive fermentation bacterium that can cause intestines problem.This bacterioid can produce harmful substances such as ammonia, indoles by fermentation protein usually, and can improve the level of pH value in the enteron aisle, and the representative of the harmful bacterium of this class comprises Escherichia coli, salmonella and staphylococcus aureus.Beneficial bacteria among the present invention is meant that animal growth is had facilitation, especially to the helpful probiotics strain of animal intestinal digestive function.15 kinds of microbe additives mentioning in " allow use feed addictive kind catalogue " that comprises that the Ministry of Agriculture 2002 announces and the Bifidobacterium of food industry extensive use.
The content of Polyphenols accounts for more than 1.0% of total extract dry weight in the extract of the present invention.The position of the kind of pine tree, the season of collection and collection directly influences the content of Polyphenols in the product.Polyphenols is meant the poly phenols that contains two or more single phenol monomers.The polyphenol compound that occurring in nature has physiologically active is divided three classes by its chemical constitution: flavanol compound; Flavones and flavonols; The phenolic acid class comprises tannin, toxifolin, catechol, anthocyanin, catechin and amyrin.Contained polyphenol compound is the mixture of flavone compound, Lignanoids compounds and flavan-3-alcohol compounds in the pine extract.
Sugar contents accounts for more than 20% of total extract dry weight in the extract of the present invention.The position of the kind of pine tree, the season of collection and collection directly influences sugar contents in the product.Related total reducing sugar is the oligosaccharide and the polysaccharide of glucose, mannose, wood sugar, Arabic galactolipin among the present invention.Adopt alcohol precipitation can from extract, isolate polysaccharide, because glucose, mannose, wood sugar, Arabic galactolipin is a Bifidobacterium, Bacillus acidi lactici, the growth factor of bacillus subtilis and photosynthetic bacteria, so, among the present invention related from pine needle and limb, extract contain polyphenol compound and glucose, mannose, wood sugar, the feed addictive of Arabic galactolipin is not isolated polyphenol compound and glucose respectively, mannose, wood sugar, Arabic galactolipin, thereby as feed addictive with bidirectional modulation function, can suppress Escherichia coli effectively on the one hand, the growth of salmonella and staphylococcus aureus, and can promote the probio class on the other hand effectively, as Bifidobacterium, Bacillus acidi lactici, the growth of bacillus subtilis and photosynthetic bacteria.
Feed addictive of the present invention contains following characteristics:
1, the present invention is the feed addictive that contains polyphenol compound and polysaccharide.Contained polyphenol compound is the mixture of flavone compound, Lignanoids compounds and flavan-3-alcohol compounds in the pine extract.Related total reducing sugar is the oligosaccharide and the polysaccharide of glucose, mannose, wood sugar, Arabic galactolipin among the present invention.
2, the polysaccharide compound feed addictive that contains polyphenol compound among the present invention and contain glucose, mannose, wood sugar, Arabic galactolipin is to extract from Pinaceae abieteae pine genus plant five-leaved pine group, Chinese pine group pine needle and pine tree limb.
3, contain polysaccharide compounds such as polyphenol compound and glucose, mannose, wood sugar, Arabic galactolipin among the present invention and can reduce pernicious bacteria effectively as feed addictive, as Escherichia coli, the quantity of salmonella and staphylococcus aureus.
4, contain polysaccharide compounds such as polyphenol compound and glucose, mannose, wood sugar, Arabic galactolipin among the present invention as feed addictive, under certain concentration, can stimulate profitable strain effectively, as the growth of Bifidobacterium, Bacillus acidi lactici, bacillus subtilis and photosynthetic bacteria.
5, the ratio between polyphenol compound and the polysaccharide can be between 1: 1 to 1: 50 in the feed addictive that relates among the present invention.
6, the operable object of novel fodder additive comprises among the present invention: pig, bird (laying hen, fryer, duck, goose etc.), dove, ox, horse, dog and cat etc.Additive capacity can to 5% from accounting for 0.1% of total forage volume.
The present invention has the following advantages with respect to the existing feed addictive that contains pine needle:
1, the raw material wide material sources are cheap, are fit to mass industrialized production;
2, manufacture craft is simple, does not need to purify, and is refining;
3, preparation cost is low;
4, understand fully the mechanism of action of this invention substantially, can use this feed addictive pointedly;
5, improve the quantity of beneficial bacterium in the animal intestinal and the growth of inhibition harmful bacteria
Four, the specific embodiment
Embodiment 1
Relatively extraction time is determined most preferred extraction time to the influence of extract solid content:
With picking up from the fresh masson pine pine needle in mountain area, Deqing, Zhejiang is that raw material is determined (among the preparation method step 3. described extraction time) technology.
Get five parts of fresh masson pine pine needle equivalent amounts, remove other leaf, silt is cleaned.Earlier respectively pine needle is twisted into the segment of 1-5cm with chopper, the proportion control of pine needle and water was smashed defibrination at 1: 3.Obtain filtrate for the first time respectively, standby.Each filter residue uses 1: 3 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time was respectively 0,30 minute, 60 minutes, 90 minutes, 120 minutes, again through filtering, obtains filtrate for the second time.Merge filtrate respectively twice, concentrate, obtain the tan viscous liquid of equivalent through normal pressure.Recording containing of each viscous liquid is 25%, 31%, 40%, 42%, 42.5% admittedly.
So from producing actual consideration, 60 minutes 3. described extraction times of step are better among the preparation method.
Embodiment 2
With picking up from the fresh masson pine pine needle in mountain area, Deqing, Zhejiang is the feedstock production feed addictive
Get fresh masson pine pine needle, remove other leaf, silt is cleaned.Earlier respectively pine needle is twisted into the segment of 1-5cm with chopper, adds the water of appropriate amount by a certain percentage, smash defibrination.The ratio of pine needle and water can be controlled in 1: 3, obtained filtrate for the first time respectively, and is standby.Each filter residue uses 1: 3 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time is 60 minutes, again through filtering, obtains filtrate for the second time.Merge filtrate twice, concentrate, obtain tan viscous liquid through normal pressure.This pine needle water extract can be made feed addictive.
Experiment 1:
Get this pine needle water extract, measure polyphenol content by the vanillic aldehyde in Science Press's " plant polyphenol " (stone is green, and Di Ying writes, P19～20)-salt acid system; By phenolsulfuric acid colorimetric method for determining total sugar content (galactolipin is standard items); After measured, the polyphenol content in this pine needle water extract accounts for 12.5% of solid content, and total sugar content accounts for 38% of solid content.
Experiment 2:
Get this pine needle water extract, the dilution and with behind the dilute acid hydrolysis, by its monose composition of liquid chromatogram measuring, wherein chromatographic condition is:
Detector: differential post: the sugar-pak1 phase that flows: water
Column temperature: 90 ℃ of flow velocitys: 0.5ml/min sample size: 10ul
Instrument: 515-manual injector-2410
Draw from testing result, this pine needle water extract is after hydrolysis, have monose such as glucose, mannose, wood sugar, arabinose and galactolipin, wherein, glucose, mannose, wood sugar, arabinose and galactolipin respectively account for 0.18%, 0.10%, 0.21%, 0.15%, 0.06% in 1% total reducing sugar.
The feed addictive that contains polyphenol compound and glucose, mannose, wood sugar, Arabic galactolipin that extracts can join laying hen with liquid form, fryer, dove, duck, goose, pig, ox in the drinking-water of sheep, also can gavage or admix in the feed to animal.
Embodiment 3:
Observe extract from pine needles to the growth promotion effect of fryer and the influence of premunition:
The extract from pine needles test that makes with embodiment 2, fryer is with 1 age in days " Zhijiang River Huang ".For test strain is the salmonella of anti-30ppm kanamycins and 5ppm ampicillin.Feed does not add any antibiotic and coccidiostat with chick perfect compound feed (seeing Table 1).Test divides two groups with fryer: i.e. control group (not adding the extract from pine needles group) and add 2 ‰ extract from pine needles groups, every group is 40 chickens, and extract from pine needles is sneaked into feeding in the feed.2 ages in days are attacked salmonella 3 * 10
7The cfu/ plumage is weighed respectively and is detected salmonella at 9 ages in days and 15 ages in days, and the salmonella detection method is with reference to GB (GB13091-91).
Table 1: basic feed ration is formed
| Composition | Content (%) |
| Corn | ????64 |
| Dregs of beans | ????24.5 |
| Fish meal | ????2.5 |
| The dish dregs of rice | ????5 |
| Meat meal tankage | ????1 |
| Calcium monohydrogen phosphate | ????1 |
| Stone flour | ????0.7 |
| Salt | ????0.3 |
| Material is smart | ????1 |
| Add up to | ????100 |
Extract from pine needles is to the influence of broiler weight: when 9 ages in days the average weight of extract from pine needles group than control group many 10.2%, significant difference (p＜0.05), when 15 ages in days the average weight of extract from pine needles group than control group many 5.3% (seeing Table 2).
Table 2: extract from pine needles is to the influence of body weight
| Age in days | 9 ages in days | 15 ages in days | ||
| Group | The plumage number | Average weight | The plumage number | Average weight |
| Control group | ????20 | ????98 | ????16 | ????190 |
| The extract from pine needles group | ????20 | ????108 | ????17 | ????200 |
Extract from pine needles is to colibacillary influence in the fryer caecum: during 9 ages in days, extract from pine needles group cecal content sample is 10
6The drug resistance Escherichia coli recall rate of individual/gram level is 10%, and control group is 55%; During 15 ages in days, extract from pine needles group cecal content sample is 10
5The drug resistance Escherichia coli recall rate of individual/gram level is 47.1%, and control group is 62.5%.Be starkly lower than control group.
Table 3: the drug resistance Escherichia coli detect the result during different days
| Group | The plumage number | The cecal content sample is 10 during 9 ages in days 6The colibacillary result that detects of the horizontal drug resistance of individual/gram | The plumage number | The cecal content sample is 10 during 15 ages in days 5The colibacillary result that detects of the horizontal drug resistance of individual/gram |
| Control group | ????20 | ????55％(11/20) | ????16 | ????62.5％(10/16) |
| The extract from pine needles group | ????20 | ????10％(2/20) | ????17 | ????47.1％(8/17) |
Extract from pine needles is to the influence of drug resistance salmonella in the fryer caecum: the cecal content sample is 10 during 9 ages in days
6Individual/horizontal drug resistance salmonella recall rate of gram is identical, and during 15 ages in days 10
5Recall rate on individual/gram level is low nearly 13 percentage points.
Table 4: the drug resistance salmonella detects the result during different days
| Group | The plumage number | The cecal content sample is 10 during 9 ages in days 6The horizontal drug resistance salmonella of individual/gram detect the result | The plumage number | The cecal content sample is 10 during 15 ages in days 5The horizontal drug resistance salmonella of individual/gram detect the result |
| Control group | 20 | ????5％(1/20) | 16 | ????18.75％(3/16) |
| The extract from pine needles group | 20 | ????5％(1/20) | 17 | ????5.9％(1/17) |
Embodiment 4
With picking up from the fresh wet-land pine tree pine needle in mountain area, Jiangxi is the feedstock production feed addictive
Get fresh wet-land pine tree pine needle, remove other leaf, silt is cleaned.Earlier pine needle is twisted into the segment of 1-5cm with chopper, adds the water of appropriate amount by a certain percentage, smash defibrination.The ratio of pine needle and water can be controlled in 1: 5.Homogenate filters, and obtains filtrate for the first time.Filter residue uses 1: 4 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time is 90 minutes.Through filtering, obtain filtrate for the second time again.Merge filtrate twice, concentrate, obtain tan viscous liquid through normal pressure.This pine needle water extract can be made feed additive.
The method that this pine needle water extract is pressed among the embodiment 1 is surveyed polyphenol and sugar contents, and wherein polyphenol content accounts for 15.4% of solid content, and total sugar content accounts for 50% of solid content.
Embodiment 5
With picking up from the fresh Korean pine pine needle in northeast is the feedstock production feed addictive
Get fresh Korean pine pine needle, remove other leaf, silt is cleaned.Earlier pine needle is twisted into the segment of 1-5cm with chopper, adds the water of appropriate amount by a certain percentage, the proportion control of pine needle and water was smashed defibrination at 1: 3.Homogenate filters, and obtains filtrate for the first time.Filter residue uses 1: 3 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time is 60 minutes.Through filtering, obtain filtrate for the second time again.Merge filtrate twice, concentrate, obtain tan viscous liquid through normal pressure.This pine needle water extract can be made feed additive.
The method that this pine needle water extract is pressed among the embodiment 1 is surveyed polyphenol and sugar contents, and wherein polyphenol content accounts for 13.9% of solid content, and total sugar content accounts for 46% of solid content.
Embodiment 6
With picking up from the Moganshan Mountain, Deqing, Zhejiang green oil pine bark fragment is the feedstock production feed addictive
Get fresh lacebark pine pine bark fragment, remove other leaf, silt is cleaned.Earlier lacebark pine pine bark fragment is pulverized with pulverizer, extracted by 1: 7 water, controlled water temperature is built in 80-100 ℃, and extraction time is 90 minutes, obtains filtrate for the first time.Filter residue uses 1: 4 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time is 60 minutes, again through filtering, obtains filtrate for the second time.Merge filtrate twice, concentrate, obtain tan viscous liquid through normal pressure.This pine bark fragment extract can be made feed additive.
Get this pine bark fragment water extract, survey polyphenol and sugar contents (galactolipin is standard items) by the method among the embodiment 1; After measured, the polyphenol content in this pine needle water extract accounts for 25.1% of solid content, and total sugar content accounts for 45% of solid content.
Embodiment 7
With picking up from the fresh black pine pine tree of Zhejiang Siming Shan xylem is the feedstock production feed addictive
Get the xylem of fresh Chinese pine pine tree, through being ground into the pine tree fragment, in the water of 1: 8 ratio adding appropriate amount, leaching process adopts high-temperature water to extract, and temperature is controlled at 80～100 ℃, and the time is 1.5 hours, extracts the back filtration, deposits filtrate.Filter residue is used 80～100 ℃ of hot water extracting 60 minutes of 1: 5 again.Twice filtrate merges the back reduction vaporization and concentrates, and removes moisture, obtains tan viscous liquid.This extract that contains polyphenol compound and Arabic galactolipin is made particle or powder-form with spray-drying, add laying hen to, fryer, dove, duck, goose, pig, ox, the premixing feedstuff of sheep is also admixed in the feed.
This makes particle with spray-drying or powder-form is surveyed polyphenol and sugar contents by the method among the embodiment 1, and wherein polyphenol content accounts for 16.0% of solid content, and total sugar content accounts for 61% of solid content
Embodiment 8
With picking up from the fresh pinus yunnanensis pine needle in Anji, Zhejiang is the feedstock production feed addictive
Get fresh pinus yunnanensis pine needle, remove other leaf, silt is cleaned.Earlier pine needle is twisted into the segment of 1-5cm with chopper, adds the water of appropriate amount by a certain percentage, the proportion control of pine needle and water was smashed defibrination at 1: 7.Homogenate filters, and obtains filtrate for the first time.Filter residue uses 1: 5 water to extract again, extracts temperature and is controlled at 80～100 ℃, and extraction time is 60 minutes.Through filtering, obtain filtrate for the second time again.Merge filtrate twice, concentrate, obtain tan viscous liquid through normal pressure.This pine needle water extract is done following experiment.
The method that this pine needle water extract is pressed among the embodiment 1 is surveyed polyphenol and sugar contents, and wherein polyphenol content accounts for 14.2% of solid content, and total sugar content accounts for 56% of solid content.
Embodiment 9
Observe the pine tree water extract to pathogen, for example Escherichia coli, salmonella and aureus with inhibition.
Get the pine needle water extract test that embodiment 8 makes, get 20 milliliters test tube, 10 milliliters of broth bouillons of packing add above-mentioned extract from pine needles 0.0,0.1%, 0.3%, 0.5% respectively after the sterilization.Inoculate 2 * 10 respectively simultaneously
6Escherichia coli, salmonella or staphylococcus aureus.In temperature is 37 ℃, cultivates in the shaking table of 222r/min after 22 hours with the viable count in the decimal dilution method mensuration test tube.
Table 5: extract from pine needles is to colibacillary inhibition result of the test
| Extract from pine needles | ??0 | ??0.1％ | ??0.3％ | ??0.5％ |
| Escherichia coli (cfu/ml) | ??4.5×10 8 | ??3.9×10 8 | ??1.5×10 8 | ??2.0×10 4 |
Table 6: extract from pine needles is to the inhibition result of the test of salmonella
| Extract from pine needles | ??0 | ??0.1％ | ??0.3％ | ??0.5％ |
| Salmonella (cfu/ml) | ??4.8×10 8 | ??3.7×10 8 | ??1.3×10 8 | ??3.6×10 5 |
Table 7: extract from pine needles is to the inhibition result of the test of staphylococcus aureus
| Extract from pine needles | ??0 | ??0.1％ | ??0.3％ | ??0.5％ |
| Staphylococcus aureus (cfu/ml) | ??8.0×10 7 | ??5.0×10 7 | ??1.3×10 6 | ??2.8×10 4 |
From The above results, we can see, the extract from pine needles of variable concentrations all has in various degree inhibitory action to the growths of these pathogenic bacteria, especially extract from pine needles concentration is in 0.5%, its bacteriostasis is the strongest, and the quantity of Escherichia coli, salmonella and staphylococcus aureus obviously reduces.
Embodiment 10:
Observe the pine tree water extract to probio, for example the growth promoting function of Bifidobacterium, Bacillus acidi lactici, bacillus subtilis and photosynthetic bacteria.
Get the pine needle water extract test that embodiment 8 makes, get 20 milliliters test tube, 10 milliliters of broth bouillons of packing add extract from pine needles 0,0.05%, 0.1%, 0.3%, 0.5% respectively after the sterilization.Inoculating same quantity bacillus subtilis simultaneously respectively, is 37 ℃ in temperature, cultivates in the shaking table of 222r/min after 22 hours with the viable count in the decimal dilution method mensuration test tube.
Table 8: bacillus result of the test
| Extract from pine needles | ??0 | ??0.05％ | ??0.1％ | 0.3％ | 0.5％ |
| Bacillus (cfu/ml) | ??4.0×10 6 | ??4.0×10 7 | ??1.1×10 8 | 1.15×10 8 | 1.4×10 8 |
For anaerobic bacteria, as Bacillus acidi lactici and Bifidobacterium.Get the anaerobic bacteria culture test tube, adorn 10 milliliters lactobacillus culture medium or Bifidobacterium culture medium, after the sterilization, add extract from pine needles 0,0.1%, 0.2%, 0.3% respectively, inoculate 2 * 10 simultaneously
6Bacillus acidi lactici and Bifidobacterium.In the anaerobism incubator, cultivated 24 hours, take out, adopt the anaerobism decimal dilution method to measure viable count.
Table 9: lactobacillus result of the test
| Extract from pine needles | ????0 | ????0.1％ | ????0.2％ | ????0.3％ |
| Lactobacillus (cfu/ml) | ????2.3×10 8 | ????2.5×10 8 | ????2.8×10 8 | ????3.6×10 8 |
Table 10: bifidobacterium assays result
| Extract from pine needles | ????0 | ????0.1％ | ????0.2％ | ????0.3％ |
| Bifidobacterium (cfu/ml) | ????8.5×10 8 | ????9.5×10 8 | ????1.1×10 9 | ????7.8×10 8 |
See that from above result of the test the extract from pine needles of variable concentrations has the effect of different separately promotion growths to different probiotics strains.During low concentration, lactobacillus and Bifidobacterium had promote the growth effect preferably.
Table 11 pine tree water extract is to the growth promotion effect of photosynthetic bacteria
Get the reagent bottle of 5 1000ml, load onto 500 milliliters of photosynthetic bacteria culture mediums, add extract from pine needles 0,0.1%, 0.2% respectively, and inoculate 5 * 10
8Photosynthetic bacteria.Be positioned under the light and cultivate (cultivation temperature is controlled at 37 degree), cultivate after 24 hours that to observe bacterium liquid every day surperficial and with blood counting chamber microscopy total number of bacteria.
Table 11: photosynthetic bacteria result of the test
Result of the test shows: add extract from pine needles and accelerate clearly than the photosynthetic bacteria that does not add, and extract from pine needles is when 0.1% addition, effect is best.In addition, we find that also the bacterium liquid surface that adds extract from pine needles does not have varied bacteria growing, and the bacterium liquid surface microbiological contamination that does not add extract from pine needles is more.The extract from pine needles use except promoting photosynthetic bacterium growth, suppresses the effect of varied bacteria growing in addition in the production of photosynthetic bacteria.
Claims (15)
1, a kind of feed additive for improving animal disease-resistant ability is characterized in that described feed addictive is the hot water extract after the powder process of Pinaceae abieteae pine genus plant fragment, contains polyphenol compound and polysaccharide among the described hot water extract.
2, feed additive for improving animal disease-resistant ability as claimed in claim 1 is characterized in that described Pinaceae abieteae pine genus plant can be one of following plant:
1. 2. 3. 4. 5. 6. Korean pine of pinus yunnanensis of wet-land pine tree of black pine of Chinese pine of masson pine
3, feed additive for improving animal disease-resistant ability as claimed in claim 1 or 2 is characterized in that the pine tree fragment that described Pinaceae abieteae pine genus plant fragment is pulverized for the pine branch xeromenia.
4, feed additive for improving animal disease-resistant ability as claimed in claim 1 or 2 is characterized in that described Pinaceae abieteae pine genus plant fragment is a pine needle.
5, feed additive for improving animal disease-resistant ability as claimed in claim 1 is characterized in that described polyphenol compound is the mixture that comprises flavone compound, Lignanoids compounds, flavan-3-alcohol compounds.
6, feed additive for improving animal disease-resistant ability as claimed in claim 1 is characterized in that described polysaccharide is the mixture that contains glucose, mannose, wood sugar, Arabic galactolipin.
7, feed additive for improving animal disease-resistant ability as claimed in claim 1 is characterized in that the ratio between described polyphenol compound and the polysaccharide can be between 1: 1 to 1: 50.
8, a kind of method for preparing the described feed additive for improving animal disease-resistant ability of claim 1 is characterized in that preparation method's step extraction in the following order of described feed addictive:
1. fresh pine tree fragment flush away impurity, the segment that is cut into 1-5cm with chopper is made the pine tree meal, gets 1 part of pine tree meal, adds 2 parts-10 parts in water, and is pending;
2. pending liquid is smashed defibrination, and dope filtration is collected filtrate, and it is pending that filter residue adds 3 parts to 5 parts water;
3. the extraction of heating of pending liquid is extracted temperature and is controlled at 80-100 ℃, and extraction time is 30 minutes-2 hours, and extracting liquid filtering is collected filtrate;
4. filter residue with 3 parts-5 parts water extraction, extracts temperature and is controlled at 80-100 ℃ again, and extraction time is 30-60 minute, and extracting liquid filtering is collected filtrate for later use;
5. merge the filtrate of collecting previously, concentrate, obtain tan viscous liquid and promptly can be feed additive.
9,4. the preparation method of feed additive for improving animal disease-resistant ability as claimed in claim 8 when it is characterized in that the preparation method of described feed addictive, described pine tree fragment are simple pine needle, can omit step.
10,2. the preparation method of feed additive for improving animal disease-resistant ability as claimed in claim 8 when it is characterized in that the preparation method of described feed addictive, described pine tree fragment are simple pine branch dry crushing sheet, can omit step.
11, the preparation method of feed additive for improving animal disease-resistant ability as claimed in claim 8 is characterized in that the preparation method of described feed addictive, and described step amount of water 1. is 3 parts, and described step extraction time 3. is 60 minutes.
12, the preparation method of feed additive for improving animal disease-resistant ability as claimed in claim 8 is characterized in that the preparation method of described feed addictive the feed additive that 5. described step obtains being made solid form.
13, a kind of purposes of feed additive for improving animal disease-resistant ability according to claim 1, it is characterized in that described feed addictive is used for animal with 0.1% dosage greater than total forage volume, can effectively reduce the quantity of pernicious bacteria such as Escherichia coli, salmonella and staphylococcus aureus; Described feed addictive can stimulate the growth of profitable strain such as Bifidobacterium, Bacillus acidi lactici, bacillus subtilis and photosynthetic bacteria effectively.
14, the purposes of feed additive for improving animal disease-resistant ability as claimed in claim 13 is characterized in that described animal can be an one of the following:
1. 2. 3. 4. 5. 6. 7. 8. 9. dove of cat of dog of horse of ox of goose of duck of chicken of pig
15, the purposes of feed additive for improving animal disease-resistant ability as claimed in claim 13 is characterized in that described feed addictive dosage is the 0.1%-5% of total forage volume.
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| CNA031289576A CN1552234A (en) | 2003-05-27 | 2003-05-27 | Feed additive for improving animal disease-resistant ability |
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| CNA031289576A CN1552234A (en) | 2003-05-27 | 2003-05-27 | Feed additive for improving animal disease-resistant ability |
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| CN101828647A (en) * | 2010-06-03 | 2010-09-15 | 程永发 | Health care feedstuff additive for animals |
| CN102742550A (en) * | 2012-07-24 | 2012-10-24 | 师宗县雄壁蝇蛆微生态养殖基地 | Method for raising chickens with fresh pine needles |
| CN103283971A (en) * | 2012-02-24 | 2013-09-11 | Phylus公司 | Needle-leaved tree by-product fermentation liquid and preparation method thereof, forage containing fermentation liquid and drinking water |
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| CN101828647B (en) * | 2010-06-03 | 2013-09-04 | 程永发 | Health care feedstuff additive for animals |
| CN101828647A (en) * | 2010-06-03 | 2010-09-15 | 程永发 | Health care feedstuff additive for animals |
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| CN103283971B (en) * | 2012-02-24 | 2015-01-21 | Phylus公司 | Needle-leaved tree by-product fermentation liquid and preparation method thereof, forage containing fermentation liquid and drinking water |
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| CN104642735A (en) * | 2013-11-15 | 2015-05-27 | 中国科学院过程工程研究所 | Complex carbohydrate preparation for forage, forage containing complex carbohydrate preparation, and use of forage |
| CN104642732B (en) * | 2013-11-15 | 2018-10-02 | 中国科学院过程工程研究所 | A kind of fleshy duck fodder additive and its application |
| CN104171506A (en) * | 2014-07-03 | 2014-12-03 | 淮北嘉吉农牧科技有限公司 | Heat-clearing disease-resistant goose feed and preparation method thereof |
| CN105325738A (en) * | 2015-11-25 | 2016-02-17 | 广州聚注专利研发有限公司 | Feed additive special for pet dog |
| CN106047499A (en) * | 2016-05-16 | 2016-10-26 | 吉林森工集团泉阳泉饮品有限公司 | Extraction method for Pinus koraiensis volatile oil, forest air and application thereof |
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