CN108041330A - A kind of additive of substitute antibiotics, preparation method and applications - Google Patents
A kind of additive of substitute antibiotics, preparation method and applications Download PDFInfo
- Publication number
- CN108041330A CN108041330A CN201711458324.XA CN201711458324A CN108041330A CN 108041330 A CN108041330 A CN 108041330A CN 201711458324 A CN201711458324 A CN 201711458324A CN 108041330 A CN108041330 A CN 108041330A
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- Prior art keywords
- bacillus
- chicken
- preparation
- bacteria suspension
- additive
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K50/00—Feeding-stuffs specially adapted for particular animals
- A23K50/70—Feeding-stuffs specially adapted for particular animals for birds
- A23K50/75—Feeding-stuffs specially adapted for particular animals for birds for poultry
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/02—Breeding vertebrates
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
Abstract
The present invention discloses a kind of additive of substitute antibiotics, preparation method and applications, including powder and bacteria suspension, specific steps：1) preparation of powder：By weight, 3 parts of umbellate pore furgus, 3 parts of Poria cocos, 1 part of malt are selected, powder is beaten in three's mixing, and packing preserves, spare；2) preparation of bacteria suspension：By the bacillus pumilus activated and Bacillus megatherium, beef extract-peptone fluid nutrient medium is inoculated in respectively, and 150r/min cultivates 72h under 37 DEG C of environment, and bacterium solution 100mL is taken to centrifuge 5min with 6000r/min, abandoning supernatant, by lower part precipitation normal saline dilution into 1 × 105The bacteria suspension of cfu/mL, it is spare.The present invention has only selected 3 taste Chinese herbal medicines, and the in vivo bacillus of compatibility chicken can improve chicken body immunity, can effectively control the generation of the common diseases such as salmonellosis, colibacillosis in hen cultivation, achieve the effect of significant substitute antibiotics.
Description
Technical field
The present invention relates to a kind of additive of substitute antibiotics, preparation method and applications, belong to poultry breeding technique neck
Domain.
Background technology
In China's livestock and poultry cultivation industry because antibiotic it is universal in violation of rules and regulations, be excessively used, caused by bacterial drug resistance and
Medicament residue phenomenon has seriously affected the sustainable development of the safety and poultry husbandry of chicken product, thus related antibiotic substitutes
The research of product is very burning hot.But current research focuses primarily upon the single of probiotics, Chinese herbal medicine or plant extracts
Aspect, it is seldom as the research of Substitutes For Antibiotic with compound Chinese herb compatibility especially with respect to prebiotic bacillus, apply to
Hen cultivate reality and obtain good antibiotic substitution effect report it is even more rare.
The content of the invention
In view of the above-mentioned problems of the prior art, the present invention provides a kind of additive of substitute antibiotics, preparation method
And its apply, achieve the effect of significant substitute antibiotics.
To achieve these goals, the preparation method of the additive for a kind of substitute antibiotics that the present invention uses, the addition
Agent includes herb powder and bacteria suspension, specifically includes following steps：
1) preparation of herb powder：By weight, 3 parts of umbellate pore furgus, 3 parts of Poria cocos, 1 part of malt are selected, powder is beaten in three's mixing,
Packing preserves, spare；
2) preparation of bacteria suspension：Choose healthy chicken from free-ranging Huainan Spotted-brown Chickens, sterile dissection after fasting for 24 hours, take liver,
Spleen, lung tissue organ, routinely sterile working are inoculated on blood agar plate, and for 24 hours, picking individual colonies are respectively in wheat for 37 DEG C of cultures
Purifying culture is carried out on Kang Kai culture mediums and sheep blood medium, then selects single suspicious bacterium colony Gram's staining, microscope
Lower observation ne ar, through the identification of MALDIbiotyper analyzers, 16S rRNA, sequence results compare after PCR amplification, determine
Two bacillus are respectively bacillus pumilus and Bacillus megatherium, are then inoculated in activated two kinds of bacterial strains respectively
Beef extract-peptone fluid nutrient medium, 150r/min cultivates 72h under 37 DEG C of environment, respectively takes bacterium solution 100mL respectively with 6000r/
Min centrifuges 5min, abandoning supernatant, by lower part precipitation normal saline dilution into 1 × 105The bacteria suspension of cfu/mL is to get short
Microspore bacillus bacteria suspension and Bacillus megatherium bacteria suspension, it is spare.
The present invention also provides a kind of additives of the substitute antibiotics according to made from above-mentioned preparation method.
In addition, the present invention also provides the application that additive is made in a kind of above-mentioned preparation method, by the additive for grass
Chicken cultivates.
As an improvement, specifically include following steps：Huainan Spotted-brown Chickens is taken to search for food day in chicken since 1 age in days of brood time
4 grams of powder are added in grain per kg body weight by chicken fully to mix thoroughly, then by material, water than 1:1 mix wet feed feeding, with mix with feed；
Since 1 age in days of brood time, 400mL bacillus pumiluses bacteria suspension or huge is added by every liter of water in drinking-water
Bacillus bacteria suspension, chicken group carry out chicken Newcastle disease live vaccine one in 7d, 28d and exempt to exempt from two.
Compared to more existing Substitutes For Antibiotic, the present invention has only selected 3 taste Chinese herbal medicines, the in vivo bacillus of compatibility chicken
(bacillus pumilus, Bacillus megatherium) can improve chicken body immunity, can effectively control salmonella in hen cultivation
The generation of the common diseases such as disease, colibacillosis achieves the effect of significant substitute antibiotics；The spice feeding selected simultaneously
Method is greatly reduced traditional Chinese herbal medicine and decocts the time and efforts that juice gavages, be suitable for the operation of currently common raiser with
With.
Description of the drawings
Fig. 1 is the bacillus pumilus in the embodiment of the present invention 1；
Fig. 2 is the Bacillus megatherium in the embodiment of the present invention 1.
Specific embodiment
Understand to make the object, technical solutions and advantages of the present invention clearer, below by accompanying drawings and embodiments, to this
Invention is further elaborated.However, it should be understood that the specific embodiments described herein are merely illustrative of the present invention,
The scope being not intended to limit the invention.
Unless otherwise defined, all technical terms and scientific terminology used herein are led with belonging to the technology of the present invention
The normally understood meaning of technical staff in domain is identical, and used term is intended merely to retouch in the description of the invention herein
State the purpose of specific embodiment, it is not intended that in the limitation present invention.
A kind of preparation method of the additive of substitute antibiotics, which includes herb powder and bacteria suspension, specific to wrap
Include following steps：
1) preparation of herb powder：By weight, 3 parts of umbellate pore furgus, 3 parts of Poria cocos, 1 part of malt are selected, powder is beaten in three's mixing,
Packing preserves, spare；
2) preparation of bacteria suspension：Choose healthy chicken from free-ranging Huainan Spotted-brown Chickens, sterile dissection after fasting for 24 hours, take liver,
The histoorgans such as spleen, lung, routinely sterile working are inoculated on blood agar plate, and for 24 hours, picking individual colonies exist respectively for 37 DEG C of cultures
Mai Kangkai culture mediums separate the upgrowth situation and colony morphology characteristic of bacterium with carrying out purifying culture, observation on sheep blood medium,
Single suspicious bacterium colony Gram's staining is selected again, and micro- Microscopic observation ne ar is reflected through MALDI biotyper analyzers
Fixed, 16S rRNA sequence results after PCR amplification compare, and it is respectively bacillus pumilus and huge bud to determine two bacillus
Then activated two kinds of bacterial strains are inoculated in beef extract-peptone fluid nutrient medium, under 37 DEG C of environment by born of the same parents bacillus respectively
150r/min cultivates 72h, and bacterium solution 100mL is respectively taken to centrifuge 5min with 6000r/min respectively, and lower part is precipitated and used by abandoning supernatant
Normal saline dilution is into 1 × 105The bacteria suspension of cfu/mL hangs to get bacillus pumilus bacteria suspension and Bacillus megatherium bacterium
Liquid, it is spare.
The present invention also provides a kind of additives of the substitute antibiotics according to made from above-mentioned preparation method.
In addition, the present invention also provides the application that additive is made in a kind of above-mentioned preparation method, by the additive for grass
Chicken cultivates.
As an improvement, specifically include following steps：Huainan Spotted-brown Chickens is taken to search for food day in chicken since 1 age in days of brood time
In grain by chicken per kg body weight add 4 grams of powder fully mix thoroughly, with mix with feed；
Since 1 age in days of brood time, 400mL bacillus pumiluses bacteria suspension or huge is added by every liter of water in drinking-water
Bacillus bacteria suspension, chicken group carry out chicken Newcastle disease live vaccine one in 7d, 28d and exempt to exempt from two.
Embodiment 1
A kind of additive of substitute antibiotics, specifically includes following preparation process：
1) 3 parts of Polyporus, 3 parts of Poria cocos, 1 part of malt are selected, powder is beaten in three's mixing, is sub-packed in polybag after tying and is done
Dry shady place preserves；
2) separated out of free-ranging, healthy Huainan Spotted-brown Chickens body and (such as attached drawing, Fig. 1 are inoculated in spleen tissue through microscopy
Occur bacterium colony-bacillus pumilus of complete hemolysis on blood plate, Fig. 2 be liver inoculation blood plate occur it is anhemolytic
Larger rough bacterium colony-Bacillus megatherium), biochemical test and etc. be accredited as bacillus pumilus and huge brood cell
Bacillus, respectively with normal saline dilution, be prepared into 1 × 105The bacteria suspension of cfu/mL is preserved in dry shady place.
The performance study of the obtained additive of the present invention：
Healthy 1 age in days Huainan Spotted-brown Chickens are divided into 6 groups immediately, are respectively common diet group (I), Chinese medicine group (II), short and small bud
Born of the same parents bacillus group (III), Bacillus megatherium group (IV), bacillus pumilus+Chinese medicine group (V), Bacillus megatherium+Chinese medicine group
(Ⅵ)；
I group of chicken group only feeds perfect compound feed, normal water；II group of chicken group presses every kilogram of body of chicken in searched for food daily ration
4 grams of herb powders are re-added fully to mix thoroughly, then by material, water than 1:1 mix wet feed feeding, with mix with feed, if having in feeder every
Night clout adds virgin material again after need to emptying；III group of chicken group's drinking-water is physiological saline of the 400mL containing bacillus pumilus, after having drunk
Water is added again；IV group of chicken group's drinking-water is physiological saline of the 400mL containing Bacillus megatherium, adds water after having drunk again, changes water daily
Shi Qingxi water fountains, after ensureing water appliance health；V group of chicken group's drinking-water is physiological saline of the 400mL containing bacillus pumilus,
Water is added after having drunk again, VI group of chicken group's drinking-water is physiological saline of the 400mL containing Bacillus megatherium, water is added after having drunk again.Separately
Outside, V group, VI group is added 4 grams of herb powder spice feedings in daily ration all by chicken per kg body weight respectively, and method is same as above.
By the feeding of entire brood time, the detection data of the index and spleen index of 6 groups of chicken groups, bursal index and IgG content
1, table 2 and table 3 are shown in Table respectively.
The influence (g/kg) of 1 bacillus pumilus of table, Bacillus megatherium and Chinese herbal medicine compatibility to chicken spleen index
Note：Colleague's shoulder mark contains the identical lowercase letter not notable (P of difference between the two>0.05) it is on the contrary, difference
Significantly, similarly hereinafter.
The influence (g/kg) of 2 bacillus pumilus of table, Bacillus megatherium and Chinese herbal medicine compatibility to chicken bursa index
The influence (mg/mL) of 3 bacillus pumilus of table, Bacillus megatherium and Chinese herbal medicine compatibility to chicken IgG content
Spleen and the bursa of farbricius are immune organs important in chicken body, and the development condition of the two directly decides the immune water of chicken
Gentle disease resistance, spleen and bursa of farbricius relative weight increase namely index and spleen index, bursal index improve, then reflects exempting from for chicken
Epidemic disease function enhances.IgG (immunoglobulin G) also plays an important role in terms of immunity of organism, the content in serum
Increase then reflects that immune level is promoted.
By testing Huainan Chicken immune performance, the results show：Bacillus pumilus, Bacillus megatherium and the two
Respectively can promote in various degree with Chinese herbal medicine compatibility spleen, bursa of farbricius immune organ growth and development, improve IgG in chicken body
Content, and bacillus pumilus compatibility Chinese herbal medicine is the most apparent to improving index and spleen index, bursal index and IgG content effect.
Wherein, bacillus pumilus group (III), bacillus pumilus+Chinese medicine group (V) to promote chicken brood the middle and later periods spleen grow hair
It is the most apparent to educate effect；Bacillus pumilus group (III), bacillus pumilus+Chinese medicine group (V), Bacillus megatherium+Chinese medicine group
(VI) it is particularly evident to Cu Jinfashi capsule growth and development effects in 35 and 42 age in days of chicken；Compare blank control group, in only adding in
The test group of medicine or bacillus (II, III, IV) is slightly promoted to chicken IgG content, but after drug matching bacillus, i.e.,
Bacillus pumilus+Chinese medicine group (V), Bacillus megatherium+Chinese medicine group (VI) are with obvious effects.
The present invention provides a kind of drug combination method of probiotics and Chinese herbal medicine, and probiotics supports hen from this area
It is internal, while abandoned traditional Chinese herbal medicine and decoct juice and removed slag the cumbersome methods for gavaging or drinking water, accomplished that Chinese medicine preparation is succinct
Efficiently, probiotics separation is simple and feasible, is suitble to prevent common disease in practice for substitute antibiotics in local hen, obtain
Good using effect.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modification, equivalent substitution or improvement made within refreshing and principle etc., should all be included in the protection scope of the present invention.
Claims (4)
1. a kind of preparation method of the additive of substitute antibiotics, which is characterized in that the additive includes herb powder and bacterium is hanged
Liquid specifically includes following steps：
1) preparation of herb powder：By weight, 3 parts of umbellate pore furgus, 3 parts of Poria cocos, 1 part of malt are selected, three's mixing is beaten powder, dispensed
It preserves, it is spare；
2) preparation of bacteria suspension：Choose healthy chicken from free-ranging Huainan Spotted-brown Chickens, sterile dissection after fasting for 24 hours, take liver, spleen,
Lung tissue organ, routinely sterile working are inoculated on blood agar plate, and for 24 hours, picking individual colonies are respectively in Mai Kang for 37 DEG C of cultures
Purifying culture is carried out on triumphant culture medium and sheep blood medium, then selects single suspicious bacterium colony Gram's staining, under microscope
Ne ar is observed, sequence results compare after PCR amplification through the identification of MALDI biotyper analyzers, 16S rRNA, determine
Two bacillus are respectively bacillus pumilus and Bacillus megatherium, are then inoculated in activated two kinds of bacterial strains respectively
Beef extract-peptone fluid nutrient medium, 150r/min cultivates 72h under 37 DEG C of environment, respectively takes bacterium solution 100m L respectively with 6000r/
Min centrifuges 5min, abandoning supernatant, by lower part precipitation normal saline dilution into 1 × 105The bacteria suspension of cfu/mL is to get short
Microspore bacillus bacteria suspension and Bacillus megatherium bacteria suspension, it is spare.
2. a kind of additive of substitute antibiotics made from preparation method according to claim 1.
3. the application of additive is made in a kind of preparation method according to claim 1, which is characterized in that is used for the additive
Hen cultivates.
4. the application of additive according to claim 3, which is characterized in that specifically include following steps：Take Huainan Spotted-yellow
Chicken is added per kg body weight by chicken in chicken searches for food daily ration and powder is made in 4 grams of claims 1 since 1 age in days of brood time
Fully mix thoroughly, then by material, water than 1:1 mix wet feed feeding, with mix with feed；
It is daily to add in bacillus pumilus bacteria suspension or huge obtained in 400m L claims 1 since 1 age in days of brood time
Bacillus bacteria suspension, adds water again after having drunk, chicken group carries out chicken Newcastle disease live vaccine one in 7d, 28d and exempts to exempt from two.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711458324.XA CN108041330A (en) | 2017-12-28 | 2017-12-28 | A kind of additive of substitute antibiotics, preparation method and applications |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711458324.XA CN108041330A (en) | 2017-12-28 | 2017-12-28 | A kind of additive of substitute antibiotics, preparation method and applications |
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| CN108041330A true CN108041330A (en) | 2018-05-18 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109077018A (en) * | 2018-08-02 | 2018-12-25 | 紫云自治县水塘镇小文生态养殖场 | A kind of purple cloud flower swine rearing method |
Citations (2)
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| CN103349165A (en) * | 2013-07-31 | 2013-10-16 | 刘范涛 | Chinese herbal medicine and biological bacterium compound feed additive, and preparation method and application of compound feed additive |
| CN106318887A (en) * | 2016-08-22 | 2017-01-11 | 山东省农业科学院畜牧兽医研究所 | SPF (specific pathogen free) chicken intestinal micro-ecological preparation, method for preparing same and application of SPF chicken intestinal micro-ecological preparation |
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2017
- 2017-12-28 CN CN201711458324.XA patent/CN108041330A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103349165A (en) * | 2013-07-31 | 2013-10-16 | 刘范涛 | Chinese herbal medicine and biological bacterium compound feed additive, and preparation method and application of compound feed additive |
| CN106318887A (en) * | 2016-08-22 | 2017-01-11 | 山东省农业科学院畜牧兽医研究所 | SPF (specific pathogen free) chicken intestinal micro-ecological preparation, method for preparing same and application of SPF chicken intestinal micro-ecological preparation |
Non-Patent Citations (3)
| Title |
|---|
| 杨若岚: "乌鸡源芽孢杆菌与中草药配伍对乌鸡生长性能与免疫性能的影响", 《中国优秀硕士学位论文全文数据库（电子期刊），农业科技辑》 * |
| 白涛，等: "中草药免疫增强剂在鸡疫病防治中的应用", 《中国畜禽种业》 * |
| 陈江，等: "香猪肺源性枯草芽孢杆菌的分离鉴定及体外抑菌作用", 《贵州农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109077018A (en) * | 2018-08-02 | 2018-12-25 | 紫云自治县水塘镇小文生态养殖场 | A kind of purple cloud flower swine rearing method |
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Application publication date: 20180518 |