CN107164294B - Inducing microbial inoculum and method for relieving ruminant rumen acidosis by using same - Google Patents
Inducing microbial inoculum and method for relieving ruminant rumen acidosis by using same Download PDFInfo
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Abstract
The invention discloses an induction microbial inoculum and a method for relieving rumen acidosis of ruminants by using the same. The anaerobic culture method has the advantages that the anaerobic culture is kept in the whole process from the seed culture to the induction of the strain, the steps are simple, the operation is convenient, the supplementation of special gas is not needed in the induction process, the lactic acid is used as the only carbon source, the production cost is saved, and the high adaptability and the lactic acid decomposition capability can be obtainedM.elsdeniiThe LC1 strain can relieve rumen acidosis by perfusing rumen with acidosis ruminant, and has great production, practice and application value and economic benefit.
Description
Technical Field
The invention belongs to the technical field of applied microorganisms, and particularly relates to a microbial fertilizerM. elsdeniiLC1 inducing microbial inoculum and method for relieving ruminant rumen acidosis by using the same.
Background
In modern intensive ruminant breeding, grain feed is often used in excess as a feed for ruminants in pursuit of high productivity. The feed rich in a large amount of grains is used for feeding the ruminant to obviously improve the production performance to a certain extent, but due to the physiological particularity of the rumen of the ruminant, the excess grain feed easily causes the rumen microbial fermentation unbalance, and the specific expression is as follows: the mass production of organic acids such as lactic acid is unbalanced with the absorption of rumen microorganisms and rumen epithelium, and the organic acids are accumulated in rumen to cause the decrease of rumen pH value and induce acidosis. Ruminant rumen acidosis can induce clinical symptoms such as laminitis, rumen epithelial abscission, diarrhea and the like, and if the clinical symptoms cannot be found in time and treated in a targeted manner, the production, reproduction and use cycles of the ruminant can be directly influenced, so that direct economic loss is caused.
The existing methods for relieving rumen acidosis are mainly divided into a buffer method and a microbial agent method. The buffered salt method mainly aims to achieve the purpose of improving the pH of rumen by feeding weak alkaline buffered salt such as baking soda and the like, has obvious effect in a short period, but cannot essentially relieve acidosis because the method does not play a role in reducing the amount of organic acid or changing the composition mode of organic acid in rumen; the microbial agent method mainly refers to feeding giant coccus aegypti (E) ((E))Megasphaera elsdenii) And the lactic acid utilization bacteria are used, so that the aims of regulating a rumen fermentation mode, reducing rumen lactic acid accumulation, improving rumen pH and finally relieving rumen acidosis are fulfilled. However, the problem is that the use effect of the microbial inoculum in the current market is poor due to poor adaptability and other reasons. To this end, high adaptability was developedM. elsdeniiMicrobial agents are essential to relieve rumen acidosis.
Disclosure of Invention
The invention aims to improve the adaptability and the lactic acid decomposition capacity of a strain M.elsdenii LC1 through low-nutrition enrichment induction, and achieve the purpose of relieving rumen acidosis induced by high-starch diet, in particular to a high-adaptability strain M.elsdeniiInducing and using the method for relieving rumen acidosis.
The related strains of the invention are ATCC standard strains, and the strain numbers are as follows: ATCC25940, web site obtained: https:// www.atcc.org/products/all/25940.aspx
The technical scheme for realizing the purpose of the invention is as follows:
a kind ofM. elsdeniiLC1 induces a microbial inoculum, and the preparation process of the microbial inoculum comprises the following steps: after seed culture is carried out in an anaerobic workstation, a seed culture solution is taken out, the seed culture solution is directly inoculated into a high-nutrition induction culture medium for high-nutrition enrichment induction, then the seed culture solution is inoculated into a medium-nutrition induction culture medium for medium-nutrition enrichment induction, and finally the seed culture solution is inoculated into a low-nutrition induction culture medium for low-nutrition enrichment induction to obtain the seed culture solutionM. elsdeniiLC1 induction microbial inoculum; in the induction process, the induction tank is closed without introducing other gases, lactic acid is used as a fermentation substrate to perform gradient induction of different lactic acid concentrations of the substrate, and the whole induction process is kept anaerobic.
Further, the seed culture process is as follows: taken out for freezingM. elsdeniiInoculating the strain seed bacterial liquid of the LC1 strain into a seed culture medium by using an inoculation amount of 1%, carrying out anaerobic culture at 36-39 ℃, wherein the culture time is 8-10 hours, when the OD value of the turbidity of the seed culture medium is 0.38-0.42, taking out the seed culture liquid, inoculating the seed culture liquid into a new seed culture medium, continuing the culture, wherein the culture time is 8-10 hours, and when the OD value of the turbidity of the culture liquid is 0.38-0.42, taking out the seed culture liquid, and carrying out high-nutrition enrichment induction.
Further, the seed culture medium is water as a solute, and 1.0% of tryptone, 0.5% of yeast extract, 0.5% of beef extract, 0.5% of glucose, 0.05% of cysteine hydrochloride, 0.4% of Na2HPO4, 5.0% of sterile defibrinated horse blood, 0.002% of D, L-lactic acid, and 0.0005% of resazurin are added thereto in mass ratio. Sterilizing the prepared seed culture medium liquid at a high temperature of 105-110 ℃, removing oxygen, sealing, cooling, and adjusting the pH value to 6.8-7.0.
Further, the high-nutrition induction culture medium takes water as solute, and 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine and 0.09% of K are added into the high-nutrition induction culture medium according to the mass ratio2HPO4，0.09% KH2PO4，0.18% NaCl，0.024% CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO43.0% fetal bovine serum, 0.3% glucose and 0.36% D, L-lactic acid. Sterilizing the prepared fermentation acidogenic culture medium liquid at the high temperature of 105-110 ℃ for 10 minutes, removing oxygen, sealing, cooling, and adjusting the pH to 6.7-7.0.
Further, the medium nutrient induction culture medium takes water as solute, and 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine and 0.09% of K are added into the medium nutrient induction culture medium according to the mass ratio2HPO4，0.09% KH2PO4，0.18% NaCl，0.024% CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO4And 0.18% D, L-lactic acid. Sterilizing the prepared fermentation acidogenic culture medium liquid at the high temperature of 105-110 ℃ for 10 minutes, removing oxygen, sealing, cooling, and adjusting the pH to 6.7-7.0.
Further, the low-nutrition induction culture medium takes water as solute, and 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine and 0.09% of K are added into the low-nutrition induction culture medium according to the mass ratio2HPO4，0.09% KH2PO4，0.18% NaCl，0.024% CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO4And 0.09% D, L-lactic acid. Sterilizing the prepared fermentation acidogenic culture medium liquid at the high temperature of 105-110 ℃ for 10 minutes, removing oxygen, sealing, cooling, and adjusting the pH to 6.7-7.0.
Further, the high-nutrition enrichment induction is to inoculate a seed culture solution to a high-nutrition induction culture medium in an amount of 0.3 to 1 percent, the temperature of an induction water bath is controlled to be 37 to 39 ℃, no special gas is required to be introduced, the stirring speed of the culture solution is set to be 100 to 200rpm, the culture solution is inoculated to a new high-nutrition induction culture medium after induction growth is carried out for 18 to 24 hours, and after the steps are repeated for 3 to 4 times, the induction culture solution is taken out for medium-nutrition enrichment induction.
Further, the medium nutrition enrichment induction is to inoculate high nutrition induction bacteria liquid into a medium nutrition induction culture medium in an amount of 0.5-1%, the temperature of induction water bath is controlled to be 37-39 ℃, no special gas is needed to be introduced, the stirring speed of the culture liquid is set to be 100-200 rpm, after the induction growth is carried out for 20-26 hours, the bacteria liquid is taken and inoculated into a new medium nutrition induction culture medium, and after the steps are repeated for 3-4 times, the induction bacteria liquid is taken out to carry out low nutrition enrichment induction.
Further, the low-nutrition enrichment induction is to inoculate the medium-nutrition inducing bacteria liquid to the low-nutrition inducing culture medium in the amount of 1-2%, the temperature of the inducing water bath is controlled to be 37-39 ℃, no special gas is needed to be introduced, the stirring speed of the culture liquid is set to be 100-200 rpm, the bacteria liquid is taken to inoculate a new medium-nutrition inducing culture medium after the induction growth is carried out for 24-30 hours, and the steps are repeated for 5-7 times to obtain the low-nutrition enrichment inductionM. elsdeniiLC1 induced microbial inoculum.
The invention also provides a method for applying the aboveM. elsdeniiLC1 inducing microbial inoculum for relieving rumen acidosisA method.
The method specifically comprises the following steps:
taking outM. elsdeniiLC1 inducing microbial inoculum for rumen perfusion, selecting 8 adult Saanen milk goats in dry period, adopting self-control test design, setting control period, acidosis induction period andM. elsdeniiand (3) testing the bacteria liquid in a perfusion period. Fed at 08:00 and 20:00 pairs per day. The control period was fed TMR diet containing 20% cracked corn and 80% oat grass. TMR diet containing 50% of crushed corn and 50% of oat grass was fed during induction period. The feeding period is the same as the induction period, and 200ml of low-nutrition enriched bacterial liquid is perfused through the rumen fistula before feeding (the perfusion process is completed within 15 s). Samples were collected on day 10 after 9d continuous feeding for the control period. The induction period was set to 10 days. Wherein the proportion of broken corn in the diet is increased from 20% to 50% in the first 3 days, then the feed is continuously fed for 7 days, and samples are collected on the 7 th day. The perfusion period was set to 10 days and samples were taken on day 10. The sampling time is set as 08:00 (the first sampling is before feeding), 5 mL of rumen and abdominal sac rumen juice is collected through the rumen fistula every 1.5 h to determine the pH, and 20 mL of rumen and abdominal sac rumen juice is collected through the rumen fistula every 3.0h to determine the related index of rumen juice.
The invention has the beneficial effect of providing a utilizationM. elsdeniiThe LC1 strain induces high adaptability and lactic acid decomposition capacity, thereby relieving rumen acidosis. The anaerobic culture method has the advantages that the anaerobic culture is kept in the whole process from the seed culture to the induction of the strain, the steps are simple, the operation is convenient, the supplementation of special gas is not needed in the induction process, the lactic acid is used as the only carbon source, the production cost is saved, and the high adaptability and the lactic acid decomposition capability can be obtainedM. elsdeniiThe LC1 strain can relieve rumen acidosis by perfusing rumen with acidosis ruminant, and has great production, practice and application value and economic benefit.
Drawings
FIG. 1 rumen perfusion oligotrophic enrichmentM. elsdeniiInfluence of bacterial liquid on pH change of rumen fluid (● control phase, ■ SARA phase, tangle-solidup-perfusion phase).
FIG. 2 rumen perfusion oligotrophic enrichmentM. elsdeniiInfluence of bacterial liquid on the concentration of lactic acid in rumen fluid (● control period)■ SARA stage, tangle-solidup-shrubby stage).
FIG. 3 rumen perfusion oligotrophic enrichmentM. elsdeniiInfluence of bacterial liquid on LPS concentration of rumen fluid (● control phase, ■ SARA phase, tangle-solidup end bacteria perfusion phase).
Detailed Description
This example illustrates the use ofM. elsdeniiLC1 strain is subjected to low nutrition induction enrichment, and rumen fistula is perfused with acidosis goat fed with high starch diet.
Example 1: preparation of seed Medium
Seed culture medium: water is used as a solute, and 1.0% of tryptone, 0.5% of yeast extract, 0.5% of beef extract, 0.5% of glucose, 0.05% of cysteine hydrochloride, 0.4% of Na2HPO4, 5.0% of sterile defibrinated horse blood, 0.002% of D, L-lactic acid and 0.0005% of resazurin are added into the water. Sterilizing the prepared seed culture medium liquid at the high temperature of 110 ℃, removing oxygen, sealing, cooling and adjusting the pH to 7.0.
Example 2: preparing high-nutrient enrichment induction culture medium
High nutrient enrichment induction medium: adding water as solute into the mixture, and adding tryptone 0.1%, yeast extract 0.1%, cysteine 0.06%, and K0.09%2HPO4，0.09% KH2PO4，0.18% NaCl，0.024%CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO43.0% fetal bovine serum, 0.3% glucose and 0.36% D, L-lactic acid. And sterilizing the prepared fermentation acid-producing culture medium liquid at the high temperature of 110 ℃ for 10 minutes, removing oxygen, sealing, cooling and adjusting the pH to 7.0.
Example 3: nutrient enrichment induction culture medium in preparation
Medium nutrient enrichment induction medium: adding water as solute into the mixture, and adding tryptone 0.1%, yeast extract 0.1%, cysteine 0.06%, and K0.09%2HPO4，0.09% KH2PO4，0.18% NaCl，0.024%CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO4And 0.18% D, L-lactic acid. Sterilizing the prepared fermentation acid-producing culture medium liquid at 110 ℃ for 10 minutes at high temperature, removing oxygen, and sealingCooled and the pH was adjusted to 7.0.
Example 4: preparation of Low nutrient enrichment Induction Medium
Low nutrient enrichment induction medium: adding water as solute into the mixture, and adding tryptone 0.1%, yeast extract 0.1%, cysteine 0.06%, and K0.09%2HPO4，0.09% KH2PO4，0.18% NaCl，0.024%CaCl·2H2O，0.083% MgSO4·7H2O，0.18% (NH3)2SO4And 0.09% D, L-lactic acid. And sterilizing the prepared fermentation acid-producing culture medium at the high temperature of 110 ℃ for 10 minutes, removing oxygen, sealing, cooling and adjusting the pH to 7.0.
Example 5:
a kind ofM. elsdeniiLC1 induces a microbial inoculum, and the preparation process of the microbial inoculum comprises the following steps:
1) seed culture: the culture process is to be frozenM. elsdeniiInoculating the LC1 strain into a seed culture medium, culturing at 36 ℃ for 8 hours, transferring 1mL of the strain into 10mL of a new seed culture medium when the OD of the turbidity of the seed culture medium reaches 0.38, continuously culturing for 10 hours, and taking out a seed culture solution for induced enrichment when the OD of the turbidity of the seed culture medium reaches 0.42.
2) The inducing and enriching steps are as follows: first, 1% of the above seed culture (OD 0.42 in logarithmic growth phase) was inoculated into a high nutrient medium. After 15 h of culture, 2 mL of culture solution is sucked and inoculated in a new high nutrient medium (the culture medium of example 2), and the steps are repeated for 3 times; inoculating 2 mL of the last high-nutrient enriched bacterial liquid into a medium nutrient medium (the medium in example 3), culturing for 20 h, then absorbing 2 mL of the culture liquid and inoculating the culture liquid into a new medium nutrient medium, and repeating the steps for 3 times; inoculating 2 mL of the final medium-enriched bacterial liquid into a low-nutrient medium (the medium in example 4), after culturing for 24 h, sucking 2 mL of the culture liquid and inoculating into a new low-nutrient medium, repeating the process for 5 times and retaining the final bacterial liquid to obtain the final bacterial liquidM. elsdeniiLC1 induced microbial inoculum for rumen perfusion. The whole enrichment culture process is carried out in a constant-temperature shaking table, the temperature is set to be 37 ℃, and the shaking rotating speed is 160 rpm/min.
Example 6
And (3) designing a perfusion test: selecting 8 adult dry-period Saaner milk goats, adopting self-control experimental design, setting a control period, an acidosis induction period andM. elsdeniiand (3) testing the bacteria liquid in a perfusion period. Fed at 08:00 and 20:00 pairs per day. The control period was fed TMR diet containing 20% cracked corn and 80% oat grass. TMR diet containing 50% of crushed corn and 50% of oat grass was fed during induction period. The feeding period is the same as the induction period, and 200ml of low-nutrition enriched bacterial liquid is perfused through the rumen fistula before feeding (the perfusion process is completed within 15 s). Samples were collected on day 10 after 9d continuous feeding for the control period. The induction period was set to 10 days. Wherein the proportion of broken corn in the diet is increased from 20% to 50% in the first 3 days, then the feed is continuously fed for 7 days, and samples are collected on the 7 th day. The perfusion period was set to 10 days and samples were taken on day 10. The sampling time is set as 08:00 (the first sampling is before feeding), 5 mL of rumen and abdominal sac rumen juice is collected through the rumen fistula every 1.5 h to determine pH, and 20 mL of rumen and abdominal sac rumen juice is collected through the rumen fistula every 3.0h to determine rumen juice related indexes (the collection process is completed within 30 s).
Index measurement method and results: the pH value of rumen fluid was measured with a pH meter (Seven Excellence, InLab route Pro-ISMMeter, Mettler-Toledo, Switzerland), and a pH change curve was plotted according to the change in pH at each time point. The results show that compared with the acidosis group induced by high starch diet, the perfusion of the low nutrient enrichment inductionM. elsdeniiThe bacterial liquid significantly increased the pH of rumen fluid (fig. 1).
Centrifuging 1mL rumen fluid at 10000 rpm/min for 5 min, collecting supernatant, titrating pH to 2.5 with concentrated hydrochloric acid, filtering with 0.22 um water phase filter membrane, and retaining filtrate for determination of organic acid. The organic acid content of rumen fluid was determined by high performance liquid chromatography (HPLC, LC-15C, SHIMADZU, Japan). An organic acid chromatographic column (PN. 062902, Thermo, USA) is selected, an ultraviolet detector (UV 210 nm) is selected for detection, the temperature of a column box is set to be 30 ℃, and a mobile phase (100 mM/L Na) is adopted2SO4Methanesulfonic acid corrected pH to 2.65) flow rate was 0.6 ml/min. The results show that compared with the acidosis group induced by high starch diet, the perfusion of the low nutrient enrichment inductionM. elsdeniiObvious bacterial liquidThe rumen fluid lactate concentration was reduced (figure 2).
Taking 1mL rumen fluid, centrifuging at room temperature 10000 rpm/min for 5 min, collecting supernatant, and diluting the sample by 10000 times with endotoxin-free water gradient for measuring the content of endotoxin (LPS) in rumen fluid. The measurement was carried out using a chromogenic substrate limulus kit (PN. CE80545, Limulus tridentatus reagent) and the measurement procedure was carried out according to the instruction manual. The results show that compared with the acidosis group induced by high starch diet, the perfusion of the low nutrient enrichment inductionM. elsdeniiThe bacterial solution significantly reduced the free LPS concentration in rumen fluid (fig. 3).
Claims (6)
1. A kind ofM. elsdeniiThe LC1 induced microbial inoculum is characterized in that the preparation process of the microbial inoculum is as follows: after the seed culture is carried out in an anaerobic workstation, the seed culture medium is prepared by taking water as a solute and adding 1.0% of tryptone, 0.5% of yeast extract, 0.5% of beef extract, 0.5% of glucose, 0.05% of cysteine hydrochloride, 0.4% of Na2HPO4, 5.0% of sterile defibrinated horse blood, 0.002% of D, L-lactic acid and 0.0005% of resazurin into the seed culture medium by mass ratio, sterilizing the prepared seed culture medium at a high temperature of 105-110 ℃, removing oxygen, sealing, cooling and adjusting the pH to 6.8-7.0; taking out a seed culture solution, directly inoculating the seed culture solution into a high-nutrition induction culture medium to perform high-nutrition enrichment induction, wherein the high-nutrition induction culture medium takes water as a solute, 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine hydrochloride, 0.09% of K2HPO4, 0.09% of KH2PO4, 0.18% of NaCl, 0.024% of CaCl.2H2O, 0.083% of MgSO 4.7H2O, 0.18% (NH3)2SO4, 3.0% of fetal calf serum, 0.3% of glucose and 0.36% of D, L-lactic acid are added into the high-nutrition induction culture medium in a mass ratio, the prepared fermentation acid-production culture medium is sterilized at a high temperature of 105 to 110 ℃ for 10 minutes, sealed and cooled after oxygen removal, and the pH is adjusted to be 6.7 to 7.0; then inoculating the strain into a medium nutrition induction culture medium to perform medium nutrition enrichment induction, wherein the medium nutrition induction culture medium takes water as a solute, and 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine hydrochloride, 0.09% of K2HPO4, 0.09% of KH2PO4, 0.18% of NaCl, 0.024% of CaCl 2H2O, 0.083% of MgSO4 & 7H2O, 0.18% (NH3)2SO4 and 0.18% of D, L-lactic acid are added into the medium nutrition induction culture medium in a mass ratio; finally inoculating to low-nutrition induction culture medium for low-nutrition enrichment induction, whereinThe low-nutrition induction culture medium takes water as a solute, and 0.1% of tryptone, 0.1% of yeast extract, 0.06% of cysteine hydrochloride, 0.09% of K2HPO4, 0.09% of KH2PO4, 0.18% of NaCl, 0.024% of CaCl.2H2O, 0.083% of MgSO 4.7H2O, 0.18% (NH3)2SO4 and 0.09% of D, L-lactic acid are added into the low-nutrition induction culture medium in a mass ratio; to obtainM. elsdeniiLC1 induction microbial inoculum; in the induction process, the induction tank is closed without introducing other gases, lactic acid is used as a fermentation substrate to perform gradient induction of different lactic acid concentrations of the substrate, and the whole induction process is kept anaerobic.
2. The method of claim 1M. elsdeniiThe LC1 induced microbial inoculum is characterized in that the seed culture process comprises the following steps: taken out for freezingM. elsdeniiInoculating the strain seed bacterial liquid of the LC1 strain into a seed culture medium by using an inoculation amount of 1%, carrying out anaerobic culture at 36-39 ℃, wherein the culture time is 8-10 hours, when the OD value of the turbidity of the seed culture medium is 0.38-0.42, taking out the seed culture liquid, inoculating the seed culture liquid into a new seed culture medium, continuing the culture, wherein the culture time is 8-10 hours, and when the OD value of the turbidity of the culture liquid is 0.38-0.42, taking out the seed culture liquid, and carrying out high-nutrition enrichment induction.
3. The method of claim 1M. elsdeniiThe LC1 induced microbial inoculum is characterized in that the high-nutrition enrichment induction is to inoculate a seed culture solution to a high-nutrition induced culture medium in an amount of 0.3-1%, the temperature of an induced water bath is controlled to be 37-39 ℃, no special gas is required to be introduced, the stirring speed of the culture solution is set to be 100-200 rpm, after 18-24 hours of induced growth, a bacterial solution is taken and inoculated to a new high-nutrition induced culture medium, and after the steps are repeated for 3-4 times, the induced bacterial solution is taken out for medium-nutrition enrichment induction.
4. The method of claim 1M. elsdeniiThe LC1 induced microbial inoculum is characterized in that the medium nutrition enrichment induction is to inoculate high nutrition induced bacterial liquid into medium nutrition induced culture medium in an amount of 0.5-1%, the temperature of induced water bath is controlled to be 37-39 ℃, no special gas is needed to be introduced, the stirring speed of the culture solution is set to be 100-200 rpm, and after the induced growth is carried out for 20-26 hoursInoculating the bacterial liquid to a new medium nutrition inducing culture medium, repeating the steps for 3 to 4 times, and taking out the inducing bacterial liquid to perform low nutrition enrichment induction.
5. The method of claim 1M. elsdeniiThe LC1 induced microbial inoculum is characterized in that the low-nutrition enrichment induction is to inoculate medium-nutrition induced microbial liquid to a low-nutrition induced culture medium in an amount of 1-2%, the temperature of an induced water bath is controlled to be 37-39 ℃, no special gas is required to be introduced, the stirring speed of the culture medium is set to be 100-200 rpm, after 24-30 hours of induced growth, the microbial liquid is taken to inoculate a new medium-nutrition induced culture medium, and after the steps are repeated for 5-7 times, the low-nutrition enrichment induction is obtainedM. elsdeniiLC1 induced microbial inoculum.
6. The method according to claim 1 to 5M. elsdeniiApplication of LC1 induced microbial inoculum in preparing medicine for relieving ruminal acidosis of ruminant is provided.
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| US4138498A (en) * | 1976-12-07 | 1979-02-06 | W. R. Grace & Co. | Ruminant feed additive |
| CN100582215C (en) * | 2002-07-18 | 2010-01-20 | 农业调查委员会 | Megasphaera elsdenii strain and its uses |
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