CN102093967A - Mink source enterococcus faecium and application thereof - Google Patents
Mink source enterococcus faecium and application thereof Download PDFInfo
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Abstract
The invention relates to an enterococcus faecium and application thereof, in particular to an enterococcus faecium MDF1104 with the patent collection number of CGMCC (China General Microbiological Culture Collection) No.4237. The invention also relates to the application of the mink source enterococcus faecium MDF1104 on fermenting carbohydrate and degrading cholesterol, in particular to the single application or combination with the common carrier of the enterococcus faecium MDF1104 to increase mink feed intake and daily gain for the mink, enhance organism immunity performance, and prevent and cure mink seasonal diarrhea.
Description
Technical field
The present invention relates to a strain and in the mink body, separate the faecium that obtains, and be particularly related to its application in the furbearer daily ration.
Technical background
Probiotics is to bring into play the probiotics preparation of the work of beneficial effect by improving host's intestinal microflora balance, with its green safety, have no side effect, the advantage of noresidue progressively becomes the main force that substitutes the antibiotics additive.The enterococcus spp bacterium distributes very wide at occurring in nature, common in some vegetables and plant, also is a kind of microorganism of normal presence in the humans and animals digestive tube, and main field planting is at ileum and colon position.Faecalis mainly concentrates on enterococcus faecalis and faecium as the application of probiotic bacterium, and faecium belongs to milk-acid bacteria, because its fast growth has the ability of sticking preferably, can produce lactic acid and some antimicrobial substances, is used as probiotics preparation widely.Studies show that the probiotic bacterium that faecium is formed can improve the body weight gains of livestock and poultry childhood, improve the nutrient digestion rate, improve the price of deed, reduce the diarrhoea ratio, alleviation stress situation, reduces mortality ratio.Faecalis also can be participated in the metabolism of cholesterol directly except that possessing above function, and abnormal metabolism is adjusted, and makes it to recover normal.Faecalis is widely used in fryer, growing and fattening pigs, finished cattle, piglet and the milk cow production as probiotics.In addition, probiotics has tangible immunologic enhancement to body, can promote the growth of newborn infant's intestinal tract immune system, also can promote intestinal mucosa part and systemic immune function, specificity and nonspecific immune response all there are regulating effect, keep the normal level of body immunologic function when state of health.Although probiotic bacterium has above numerous immunoregulation effect, concerning bacterial classification, not only immune effect is variant between the kind, and also has different between bacterial strain and the bacterial strain.In addition, aspects such as absorption time, supply mode and addition are also influential to the immunomodulatory effect.The host is also because of the difference of kind, age, physiological situation, living environment, and produces different effects.
Faecium extensively exists in environment, and belongs to the enteron aisle fungal component, is in the ascendance in 2～3d of the new birth of a lot of animals, and is a kind of amphimicrobian milk-acid bacteria, is very suitable for producing and using.Comprise manure enterococcin strain in existing in the market a lot of products, as " Medilac-Vita " of Korea S, probiotic composition Ventrux, Bioflorin that Germany and Sweden produce and Fiormil etc., its viable count is approximately 10
8-9Individual/the g product, the goods that contain faecium at present also have been widely used in during animal produces, the fodder additives of producing as Korea S " ratio of greater inequality he " etc.Perinatal period, the milk cow direct microorganism fodder that contains faecium of feeding can promote that forage digests in cud, increased milk yield.The beef cattle direct microorganism fodder that contains faecium of feeding can promote beef cattle to adapt to high fine fodder daily ration, prevents oxypathy.Using proper faecium not only can promote nutrient digestion to absorb, reduce serum cholesterol content, keep the intestinal microflora balance, can also regulate body's immunity, stimulate the growth of immune organs such as thymus gland, spleen and the fabricius bursa, strengthen the activate the phagocytic capacity of scavenger cell, activation gut-associated lymphoid tissue, promote the generation of immunoglobulin A, regulate body's immunological function.
About lactic bacteria additive, research and application in furbearer produces are studied less both at home and abroad, the present invention is based on the prebiotic effect effect of probiotic bacterium additive, proposed a strain and derived from the intravital safety of furbearer, stable, highly active milk-acid bacteria, and its advantage function detected.
Summary of the invention
Separating lactic acid bacterium in the healthy adult mink enteron aisle, get small intestine contents 20g, add in the 100ml sterilized water, shaking table concussion 30min, particulate matter is fully dissolved, gradient dilution, utilize pour plate method to carry out separation and Culture, cultivate the bacterium colony that 72h selects to have molten calcium circle for 37 ℃, adopt the mode purifying of plate streaking, purifying 3-4 generation, microscopy inserts the milk-acid bacteria behind the purifying in the MRS liquid nutrient medium, detects its Microbiological Characteristics, as morphology, the ability of physiology characteristic (table 1) and sugar decomposition metabolism (table 2) detects the degradation function (table 3) of its vitro culture to cholesterol.Carry out that mouse security (table 4) detects and to the influence of mink production performance (table 5).
This faecium MDF1104 on October 21st, 2010 in China Committee for Culture Collection of Microorganisms common micro-organisms center (address: No. 3, No. 1 institute in North Star West Road, Chaoyang District, BeiJing, China city, Institute of Microorganism, Academia Sinica) preservation, preserving number CGMCC No.4237.Its called after of classifying: faecium, latin name: Enterococcusfaecium.
Description of drawings
To the degraded of cholesterol, X-coordinate is incubation time (unit: hour) to Fig. 1 faecium MDF1104 under condition of in vitro culture, and ordinate zou is degradation rate (%).
Further specify the present invention below in conjunction with embodiment and Tu:
The embodiment I
Faecium MDF1104 morphological observation, microscope inspection and physiological and biochemical index detect
Aseptic condition is cultivated the faecium MDF1104 of 18-24h down with the transfering loop picking, is applied on the clean slide glass, and film-making, gramstaining carries out morphology and coloration result is observed.Under the aseptic condition faecium is inoculated under corresponding substratum or the culture condition, carries out the detection of physiological and biochemical index and observe, detected result sees Table 1
Table 1 faecium MDF1104 morphology and physics and chemistry test-results
| Detect index | Characteristic |
| Morphology | Sphere, two chainings or be dispersed in |
| Gramstaining | Positive |
| Form gemma | Not |
| Catalase | - |
| Oxydase | - |
| Grow in the air | + |
| 65 ℃ of growths | + |
| 45 ℃ of growths | + |
| 15 ℃ of growths | + |
| PH value (cultivating 24h) | 4.29 |
| The growth of 6.5% sodium-chlor | + |
| The pH9.6 growth | + |
| The pH4.5 growth | + |
The embodiment II
Faecium MDF1104 detects different carbohydrate fermentation activities
Under the aseptic condition, a little is eugonic with the transfering loop picking, cultivate the faecium MDF1104 of 18-24h, inoculation is gone in the different sugar fermentation tube of Beijing Luqiao Technology Co., Ltd.'s production, 37 ℃ of constant temperature culture 18-24h, observe the fermented liquid colour-change, to judge fermentation situation to different carbohydrate.
The different carbohydrate fermentation activities of table 2 faecium MDF1104
| Detect index | Characteristic | Detect index | Characteristic |
| Glucose | + | Raffinose | + |
| Seminose | + | Sorbyl alcohol | + |
| Melizitose | - | Melibiose | + |
| Fructose | + | Semi-lactosi | + |
| Saligenin | + | N.F,USP MANNITOL | + |
| Amygdaloside | + | Pectinose | + |
| Ribose | + | Sunmorl N 60S | - |
| Trehalose | + | Sucrose | + |
| Wood sugar | + | Cellobiose | + |
| Rhamnosyl | + | Polychrom | + |
| Maltose | + | The glucose fermentation aerogenesis | - |
| Lactose | + | ? | ? |
By table 1,2 as can be known, faecium MDF1104, and Gram-positive can be grown in than wide temperature range, and it is strong that 24h cultivates acid producing ability, can extensively utilize the carbohydrate generate energy, is the good milk-acid bacteria of a strain proterties.
The embodiment III
Faecium MDF1104 in the vitro culture process to the Degradation of cholesterol
With the faecium MDF1104 after the purifying cultivation, inoculation is gone in the MRS liquid nutrient medium of new preparation and sterilization, cultivates 18-24 hour for 37 ℃, every sampling in four hours, the centrifuging and taking supernatant utilizes o-phthalaldehyde method to measure the degradation capability of the interior milk-acid bacteria of 24h to cholesterol.
Table 3 faecium MDF1104 is to the degradation rate of cholesterol
| Sample time | Cholesterol degradation rate % |
| 4 | 38.45 |
| 8 | 32.05 |
| 12 | 32.05 |
| 16 | 32.55 |
| 24 | 31.07 |
By table 3 and accompanying drawing 1 as can be known, faecium MDF1104, the cholesterol of under condition of in vitro culture, can degrading, but and in the equal efficient degradation of whole incubation period, average degradation rate reaches 33.23%, reaches in the logarithm division stage to be up to 38.45%, illustrates that thus mink source faecium MDF1104 has degradation property preferably to cholesterol, and vitro culture in the phase its degradation efficiency descend not quite, illustrate also that thus MDF1104 has its unique advantage to the degraded of hypercholesterolemia food or feed.
The embodiment IV
Faecium MDF1104 is to the influence of small white mouse security and production performance index
Faecium MDF1104 is inserted in the 10ml liquid nutrient medium, and inoculum size is cultivated 18h for 1%, 37 ℃, and centrifugal 10 minutes of 5000rmp/min is 10 with the physiological saline dilution
7Cfu/ml and 10
9Cfu/ml.Choose healthy adult Kunming white mouse, 15 every group, the MDF1104 diluent 0.3ml of experimental group every per os perfusion every day different concns, control group pours into the physiological saline of equivalent every day, and continous pouring 20 days was observed 10 days, observe every day, writes down the situation of searching for food, the body weight situation.Routine weighing, monitor the situation of searching for food every day, butchers experiment during 30d, weighing thymus gland, spleen weight calculates the immune organ index.
Table 4. faecium MDF1104 is to small white mouse security and the influence of production performance index
| Group | Dead number of elements | Average daily gain g | Average daily ingestion amount g | The immune organ index |
| Control group | 2 | 9.90 | 12.98 | 0.57％ |
| MDF1104-10 7 | 0 | 10.80 | 18.19 | 0.56％ |
| MDF1104-10 9 | 0 | 11.63 | 18.27 | 0.59％ |
As shown in Table 4, the faecium MDF1104 in mink source, with various dose perfusion mouse, dead mouse is not arranged, and daily ingestion amount and the average daily gain of adding MDF1104 group mouse all are significantly higher than control group, immune organ index low dose group is lower than control group, and high dose group is a little more than control group.Hence one can see that, and faecium MDF1104 does not have lethality to small white mouse.And day weight gain and average daily ingestion amount to small white mouse have promoter action, and high dose group has the effect that promotes immune function of mice.
The embodiment V
Faecium MDF1104 is to the influence of mink production performance
Faecium by 1% inoculum size, is inserted in the liquid nutrient medium, and 37 ℃ of fermentation 24h choose healthy finishing period mink, and 20 every group, every of test group every day is raised 10 with feed
9The faecium MDF1104 of cfu/g dry-matter, control group does not add, routine weighing, monitor the situation of searching for food every day, follows the tracks of every group of mink diarrhoea situation, fed continuously 30 days, butchers, and the weighing immune organ weight is calculated the immune organ index.
Table 5 faecium MDF1104 is to the influence of mink production performance
| Group | The diarrhoea number of elements | Dead number of elements | Average daily gain g | Average daily ingestion amount g | Immune organ index g |
| Control group | 11 | 4 | 22.28 | 368.81 | 0.36 |
| MDF1104-10 9cfu | 4 | 1 | 23.33 | 384.38 | 0.40 |
As shown in Table 5, in finishing period mink daily ration, add faecium MDF1104, the sickness rate of the seasonal diarrhoea of mink during can being reduced at the beginning of 8 the end of month to 9 month, compare with control group, diarrhea rate descends 40%, and mortality ratio descends 10%, can obviously improve the mink daily ingestion amount more than 4%, it is about 5% that average daily gain speed increases, and the immune organ index slightly increases.
Conclusion
The above results shows, faecium MDF1104 involved in the present invention, can increase daily ingestion amount and the day weight gain of mouse and mink, strengthen the immunity function of animal, faecium MDF1104 use separately or with conventional carrier combinations, can prevent the generation of the seasonal diarrhoea of mink, improve the production performance of animal.
Claims (8)
1. a faecium (Enterococcus faecium) MDF1104 preserves numbering CGMCC No.4237.
2. according to claim 1, described milk-acid bacteria separates in the mink body, and the vitro culture growth can extensively utilize carbohydrate rapidly.
3. according to claim 1 and 2 described faecium MDF1104, has the cholesterol degradation function.
4. according to claim 1 and 2 described faecium MDF1104, detect, can improve the mouse food consumption, increase day weight gain and improve the immune organ index by the mouse security.
5. according to claim 1 and 2 described faecium MDF1104, can improve the mink daily ingestion amount, body weight gains and day weight gain increase immune organ weight.
6. according to claim 3,4 described faecium MDF1104, can with other conventional carrier combinations, have the function that reduces the animal serum cholesterol.
7. according to claim 4,5,6 described, faecium mixes the back and adds in the furbearer daily ration with other conventional carriers, has the function that improves breeding performonce fo animals.
8. according to claim 7, a kind of feed pre-mixing material, feed pre-mixing material contain the described faecium MDF1104 of claim 1.
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