CN105861399A - Lactobacillus plantarum for preventing necrotic enteritis of chickens and application thereof - Google Patents
Lactobacillus plantarum for preventing necrotic enteritis of chickens and application thereof Download PDFInfo
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Abstract
The invention discloses lactobacillus plantarum Lac16, the preservation name is Lactobacillus plantarum Lac16, and the preservation number is CCTCC NO: M 2016259. The lactobacillus plantarum can prevent occurrence of necrotic enteritis and inhibit growth of clostridium perfringens, so that the problem that production performance of meat chickens is lowered due to attacking toxin of the clostridium perfringens is relieved, and the structural damage of the intestinal mucosa is relieved; immunity of intestinal mucosa of chickens can also be improved, and growth of the meat chickens can be promoted.
Description
Technical field
The present invention relates to screening and the application thereof of a kind of Lactobacillus plantarum having suppression bacillus perfringens effect.
Background technology
Necrotic enteritis (Necrotic enteritis, NE) is mainly by A type or c-type gram-positive anaerobic bacterium perfringens shuttle
Bacterium is caused a disease the birds acute infectious intestinal disease caused, and is one of intestinal tract disease endangering maximum in poultry farming.Necrotic enteritis
Reported first, in Britain (Parish et al, 1961), is reported by world community subsequently in succession.The generation of chicken necrotizing enterocolitis mainly with
Intestinal severe haemorrhage, intestinal mucosa damage, ulcer, necrosis etc. are principal character (Guo S et al, 2015), and this disease is throughout the year
All can occur, especially with burning hot and rainy summer multiple, multiple be born in 2～6 week old broiler, 3～6 commodity egg (Cooper at monthly age
KK et al.,2013,2010).Acute chicken necrotizing enterocolitis may result in the unexpected mortality of chicken group, can after pathological anatomy
Find that intestinal mucosa has a large amount of diffusivity necrosis region；Chronic type chicken necrotizing enterocolitis is difficult to be authenticated, although be not result in that chicken group dashes forward
The most dead, but the reduction of chicken appetite, the decline of intestinal digestive and absorptive functions, feed efficiency reduction, weight loss can be caused,
Thus causing chicken group's growth retardation etc., the economic loss that this chronic type chicken necrotizing enterocolitis causes to poultry farming is bigger
(Savva CG et al.,2013).Recent decades, low dosage antibiotic was widely used as growth promoter (AGPs) always
In animal cultivation produces (Van et al., 2009；Pattison et al.,2002；Vigre et al.,2008；Der Sluis et al.,
2000) generation of multiple fowl bacterial disease, is controlled again simultaneously as antibacterials.But, along with a series of antibiotic
After negative effect is in the news, many countries have been gradually lowered or have prohibited use antibiotic and have been applied to animal as feed additive
In breeding production (Wade et al., 2015；Uzal FA et al.,2014；Shimizu et al.,2002；Myers et al., 2006), its
Result be cause many such as intestinal diseases such as necrotic enteritis on a large scale, epidemic generation (Si W et al., 2007；Nauerby
et al.,2003；Pedersen et al.,2003).It is reported, after antibiotic is prohibited to be applied to poultry farming production as AGPs,
The morbidity of chicken necrotizing enterocolitis steeply rises.At present, necrotic enteritis the economic loss of the global poultry farming caused by
2,000,000,000 dollars in 2000 rise to 2015 6,000,000,000 dollars (Naylor et al., 1998；Nagahama et al., 2002),
Necrotic enteritis has become one of most important economic disease in the poultry industry cultivation of the whole world, seriously hinders the sound development of domestic fowl farming,
Also endanger human health simultaneously.Therefore, prevent and treat chicken necrotizing enterocolitis by the method for non-antibiotic increasingly to be closed by people
Note.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of Lactobacillus plantarum preventing chicken necrotizing enterocolitis and application thereof；This plant
Lactobacillus can suppress bacillus perfringens growth, improve chicken macrophage immunity function, and after bacillus perfringens counteracting toxic substances can be alleviated
Meat chicken production performance is declined and the impact of mucosal structure damage, the generation of prevention chicken necrotizing enterocolitis.
In order to solve above-mentioned technical problem, one Lactobacillus plantarum Lac16 of the present invention, its preservation is entitled: Lactobacillus plantarum Lac16
Lactobacillus plantarum Lac16, its preserving number is: CCTCC NO:M 2016259.
The present invention provides the purposes of above-mentioned Lactobacillus plantarum Lac16 the most simultaneously: prevent the generation of chicken necrotizing enterocolitis.
The improvement of purposes as the Lactobacillus plantarum Lac16 of the present invention: Lactobacillus plantarum suppression bacillus perfringens growth, thus
After alleviating bacillus perfringens counteracting toxic substances, meat chicken production performance is declined and the impact of mucosal structure damage.
The further of purposes as the Lactobacillus plantarum Lac16 of the present invention is improved: Lactobacillus plantarum can improve chicken macrophage immunity
Function.
The further of purposes as the Lactobacillus plantarum Lac16 of the present invention is improved: promote growth of meat chicken.
The present invention with Lac16 bacterial strain STb gene as template, use universal primer:
Forward primer (P1): 5 '-AGAGTTTGATCCTGGTCAGAACGAACGCT-3 '
Downstream primer (P6): 5 '-TACGGCTACCTTGTTACGACTTCACCCC-3 '.
Carry out PCR amplification and obtain the 16s rDNA partial sequence of 1510bp.The antibacterial 16s included by Blast with GenBank
RDNA gene order comparison, finds that Lac16 bacterial strain is 99% with the homology of Lactobacillus plantarum.
The purposes of Lactobacillus plantarum Lac16 of the present invention: sending out of suppression bacillus perfringens growth, effectively prevention chicken necrotizing enterocolitis
Raw, and promote growth of meat chicken.
Accompanying drawing explanation
Below in conjunction with the accompanying drawings the detailed description of the invention of the present invention is described in further detail.
Fig. 1 is that lactobacillus to be screened is to Cp bacteriostatic test；
Fig. 2 is Lactobacillus plantarum Lac16 colonial morphology；
Fig. 3 is that Lactobacillus plantarum Lac16 is on chicken macrophage HD11 inflammation-related gene and the impact of iNOS gene expression；
Fig. 4 is that Lactobacillus plantarum Lac16 is to broiler ileum fine hair mucosa structure influence after Cp counteracting toxic substances；
Fig. 5 is that Lactobacillus plantarum Lac16 is to broiler ileum microvillus structure influence after Cp counteracting toxic substances；
Left figure is ileum microvillus regularity (5 μm)；Right figure is ileum gut barrier (TJ, AB, D)；TJ: compact siro spinning technology；
AB: adhesive tape；D: desmosome.
Detailed description of the invention
Embodiment 1, the acquisition of Lactobacillus plantarum Lac16
1, diluent is prepared:
Take Suburb Areas of Hangzhou vegetable field soil 1 gram to add in 90mL sterilized water, then be diluted to 10 with sterilized water0、10-1、10-2,
10-3、10-4、10-5、10-6Totally seven Concentraton gradient.
2, cultivate:
Take the diluent 100ul of each concentration be respectively coated in MRS solid medium (peptone 10g/L, yeast extract 5g/L,
Carnis Bovis seu Bubali cream 10g/L, sodium acetate 5g/L, dipotassium hydrogen phosphate 2g/L, ammonium citrate 2g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.2g/L,
Tween 80 1.0mL/L, glucose 20g/L, agar 20g/L, remaining is distilled water；PH value glacial acetic acid is adjusted to 6.2～6.6,
121 DEG C of sterilizings 20 minutes) flat board, it is inverted after the coating of sterilizing Glass rod in being placed on 35 DEG C of calorstats and cultivates.
3, the separation of bacterial strain, purification:
Cultivate at MRS and cultivate 48h based on 35 DEG C, i.e. have single bacterium colony to occur.With the single bacterium colony of Inoculating needle picking, train at MRS
Support base (formula is as above) planar surface the most streak culture, select the different bacterium colony of those forms, just can shape after being repeated several times by
Become the most isolated bacterium colony, thus obtain 6 strain pure culture lactic acid bacterias.
4, different time fungistatic effect measures:
By 6 strain bacterium colonies activation after be respectively placed in 50mL MRS fluid medium (peptone 10g/L, yeast extract 5g/L,
Carnis Bovis seu Bubali cream 10g/L, sodium acetate 5g/L, dipotassium hydrogen phosphate 2g/L, ammonium citrate 2g/L, magnesium sulfate 0.2g/L, manganese sulfate 0.2g/L,
Tween 80 1.0mL/L, glucose 20g/L, remaining is distilled water；PH value glacial acetic acid is adjusted to 6.2～6.6,121 DEG C of sterilizings 20
Minute) middle cultivation, initial inoculation concentration is 1.0 × 106Each bacterium solution is adjusted to 1.0 × 10 after CFU/mL, 24h9CFU/mL is standby.
LB culture medium (peptone 10g/L, yeast extract 5g/L, NaCl 10g/L, fine jade containing 1% (w/v) glucose
Fat 20g/L, remaining is distilled water, and pH value 5mol/L NaOH solution is adjusted to 7.2～7.4) flat board high temperature sterilize (121 DEG C
Sterilizing 20 minutes) after, after being cooled to 40～50 DEG C, add pathogenic A type perfringens shuttle according to the inoculum concentration of 1% (v/v)
Bacterium bacterium solution (cultivates the bacillus perfringens (Clostridium perfringens CVCC2030, Cp) of 15h), is down flat plate, puts
Put sterilizing room about 1h, be inverted and be put in 1h in 4 DEG C of refrigerators, Oxford cup (internal diameter 6mm, external diameter 8mm) is placed on flat board table
Face, then according to the sample-adding amount of every hole 200 μ l adds screened bacterial strain bacterium solution, cultivates at 35 DEG C of incubators, samples every 3h,
Measuring the antibacterial circle diameter change in 24h, each plate sets 3 repetitions.Pick out bacillus perfringens inhibition optimal
Bacterium colony, named Lac16.Lac16 is the most obvious (Fig. 1) to bacillus perfringens inhibition, 24h antibacterial ring average diameter
For 15.58mm.
5, strain identification:
(1) morphological characteristic
In uniformly muddiness and with microprecipitation in liquid medium within, it is shaken gently for precipitating and i.e. scatters.On MRS agar plate
Lac16 bacterium colony is the least, and for about 0.5mm, bacterium colony is that point-like is protruding, rounded, and smooth surface is fine and closely woven, milky, Fig. 2
Colonial morphology for Lactobacillus plantarum Lac16.
(2) 16S rRNA sequence analysis
To separation screening to bacterial strain Lac16 carry out further 16S rRNA sequence analysis and comparison.According to bacterial 16 S rDNA
Conserved sequence design pair of primers and by Shanghai raw work synthesis:
Forward primer (P1): 5 '-AGAGTTTGATCCTGGTCAGAACGAACGCT-3 '
Downstream primer (P6): 5 '-TACGGCTACCTTGTTACGACTTCACCCC-3 '.
PCR amplification is carried out for template with the bacterial strain that screening obtains.Reaction condition: 95 DEG C of denaturations 5min, 94 DEG C of degeneration 50s,
52 DEG C of annealing 1min, 72 DEG C extend 1min 30s, react 30 circulations.
Purpose segment conventionally (Sambrook, et al., 2001) carries out cloning and sequencing, measurement result with
The 16S rRNA sequence (accession number: NC_004567, NC_012984 and NC_020229 etc.) logged in GenBank
Comparing, genetic homology reaches more than 99%.According to " Bergey's Mannual of Determinative
Bacteriology " (Holt, et al., 1994) and " common bacteria system identification handbook " (the elegant pearl in east etc., 2001), pass through
Morphological characteristic and 16S rRNA sequence analysis, determine that bacterial strain Lac16 is Lactobacillus plantarum.
Being carried out preservation, preservation is entitled: Lactobacillus plantarum Lac16Lactobacillus plantarumLac16, depositary institution:
China typical culture collection center, preservation address: Wuhan, China Wuhan University；Preservation date: on May 11st, 2016, protects
Tibetan number: CCTCC NO:M 2016259.
In the ratio of 1% (v/v), preservation the Lactobacillus plantarum Lac16 that activates are inoculated in 50mL MRS fluid medium,
After 35 DEG C are cultivated 24 hours, it is thus achieved that containing 108The Lactobacillus plantarum Lac16 culture fluid of cfu/mL.
Above-mentioned Lactobacillus plantarum Lac16 culture fluid is processed 30min in 90 DEG C, Lactobacillus plantarum Lac16 must be inactivated.
Embodiment 2, the Lactobacillus plantarum Lac16 impact on chicken macrophage system HD11 immunologic function
Chicken macrophage system HD11 presses concentration 5 × 105Individual/mL is seeded to 12 porocyte culture plates, and every hole 1mL F12 cultivates completely
Base (88%F12 basal medium+10% chicken serum+1% mycillin+1% non essential amino acid (100x)), arranges comparison
Group, Lac16 group, Cp group (bacillus perfringens) and Lac16+Cp group；Particularly as follows:
Matched group: process without inactivation lactic acid bacteria, discard cultivation after only cultivating 3,6,12 hours respectively with F12 complete medium
Liquid, after washing 3 times with room temperature sterilizing PBS, adds after F12 complete medium continues to cultivate 3h and receives sample.
Lac16 group: add 1ml containing 108The F12 complete medium of cfu/mL inactivation Lactobacillus plantarum Lac16 processes 3h, 6h respectively
With receipts sample after 12h；
Cp group (bacillus perfringens): process without inactivation lactic acid bacteria, only cultivate 3,6,12 respectively with F12 complete medium
Discard culture fluid after hour, add 1ml after washing 3 times with room temperature sterilizing PBS containing 108The F12 of cfu/mL bacillus perfringens viable bacteria
Complete medium receives sample after processing 3h；
Lac16+Cp group: add 108After cfu/mL inactivation Lactobacillus plantarum Lac16 processes 3h, 6h and 12h respectively, abandon culture fluid,
Room temperature sterilizing PBS adds 1ml containing 10 after washing 3 times8The F12 complete medium of cfu/mL bacillus perfringens viable bacteria is received after processing 3h
Sample；
Often 3 repeating holes of group, 41 DEG C, 5%CO2Cultivate 6h to cell attachment.Discard F12 complete medium, sterilizing PBS afterwards
1mL F12 complete medium is respectively added: matched group and Cp group process without lactic acid bacteria, Lac16 group and Lac16+Cp after washing 3 times
Group interpolation 108Cfu/mL inactivates Lactobacillus plantarum Lac16, culture medium is removed, sterilizing PBS after respective pretreatment 3h, 6h, 12h
1mL F12 complete medium, Cp group and Lac16+Cp group interpolation 10 is respectively added after washing 3 times8At cfu/mL bacillus perfringens viable bacteria
Collect cell, (amendment) after reason 3h, carry out Total RNAs extraction, reverse transcription and qPCR, detect cytokine (IL-6, IFN-γ,
IL-10, TGF-β) and the expression of iNOS gene.
Compared with control treatment, as it is shown on figure 3, inactivation Lactobacillus plantarum Lac16 can raise cytokine IL-6, IFN-γ, IL-10,
TGF-β and iNOS gene expression (P < 0.05), bacillus perfringens can improve the gene expression (P < 0.05) of IFN-γ, suppression
The gene expression (P < 0.05) of iNOS.Compared with Cp group, after adding Cp again after lactobacillus Lac16 pretreatment, significantly reduce IFN-γ
Gene expression, improve IL-10 and the gene expression (P < 0.05) of TGF-β and iNOS, prompting Lactobacillus plantarum Lac16 can
Significantly improve the immunologic function of chicken macrophage system HD11, alleviate Cp and infect the proinflammatory reaction caused, and improve anti-inflammatory properties,
Strengthen the Scavenging activity to CP, protect cell.
Embodiment 3, the preparation method of Lactobacillus plantarum Lac16 microbial inoculum:
In the ratio of 1% (v/v), Lactobacillus plantarum is inoculated in MRS fluid medium, after 35 DEG C are cultivated 24 hours, 4 DEG C,
5000rpm/min is centrifuged, and takes bacterium precipitation, 50 DEG C of freeze-day with constant temperature 6～8 hours, mix by 4:1 (w:w) with corn starch and
Making dry bacterium powder, every gram of mycopowder viable count is 1.0 × 108cfu。
Embodiment 4, Lactobacillus plantarum Lac16 promote growth of meat chicken
Precious 500 broiler of section are purchased from Zhejiang University's honest broiler centre of development, and test site is laying hen test site, Zhejiang Hangzhou city.Adopt
Take single-factor design: 1 age in days health and body weight precious 500 broiler of healthy section without significant difference (P > 0.05) are chosen in test
315 plumages, are randomly divided into 3 test group, each test group 6 repetition, each repetition 35 plumage.Control group and Cp group are only
Feeding Basic drawing, Lac+Cp group adds Lactobacillus plantarum Lac16 preparation 1 000mg/kg on the basis of Basic drawing.Cp
Group and Lac+Cp group during 14～20 ages in days continuous 7 days oral A type bacillus perfringens (1.0ml/ pcs/day, 108cfu/mL)
Set up necrosis induced property colitis model.Experimental period is 21 days.Detect accordingly after test 21d, described in detail below.
Table 1, Lactobacillus plantarum Lac16 are on the impact of meat chicken production performance after Cp counteracting toxic substances
| Project | Average whole body weight | Average daily gain (g) | Average daily ingestion amount (g) | Feed-weight ratio |
| Items | Body weight/(g) | ADG/(g) | ADFI/(g) | F/G |
| Control | 637.88±15.00ab | 41.95±2.14 | 74.29±0.84 | 1.77±0.072 |
| Cp | 615.15±5.08b | 39.77±1.62 | 75.58±0.95 | 1.90±0.055 |
| Lac16+Cp | 648.48±15.96a | 41.79±5.31 | 76.67±2.28 | 1.86±0.243 |
The different letter person of same column shoulder note represents significant difference (P<0.05) (n=6), otherwise difference is not notable (P>0.05).Under
With.
As shown in Table 1, Cp group broiler average weight be less than Control group and Lac16+Cp (P < 0.05), average daily gain with
Feed efficiency is below Control group (P > 0.05).Lac16+Cp group broiler average weight, average daily gain, average day
Feed intake is similar to matched group with feed efficiency, is above Cp group (P > 0.05), the Lactobacillus plantarum Lac16 that prompting is added
The impact that meat chicken production performance is declined after contributing to alleviating Cp counteracting toxic substances by preparation.
Table 2, Lactobacillus plantarum Lac16 are on the impact of broiler shoot formation after Cp counteracting toxic substances
| Project | Index and spleen index | Bursal index |
| Items | Spleen index | IBD index |
| Control | 0.90±0.14 | 1.93±0.09b |
| Cp | 1.06±0.24 | 2.55±0.36a |
| Lac+Cp | 0.96±0.18 | 1.47±0.41b |
As seen in Table 2, Cp group broiler index and spleen index is higher than Control group and Lac+Cp group (P > 0.05), Lac+Cp group broiler spleen
Index is close with Control group；Cp group broiler bursal index is significantly higher than Control group and Lac+Cp group (P < 0.05), Lac+Cp
Group bursal index is less than Control group (P > 0.05).The above results is pointed out, and Lactobacillus plantarum Lac16 contributes to alleviating Cp to be infected
The enlargement of the broiler immune organ caused.
The generation of embodiment 5, Lactobacillus plantarum Lac16 prevention broiler necrotic enteritis
Control group, Cp group, Lac+Cp group set-up mode with embodiment 4.Detect accordingly after test 21d.
As Fig. 4 can find, Control group broiler ileum fine hair queueing discipline is orderly, surface normal, not damaged；Cp group broiler returns
Intestinal villus shortens, it is sparse to arrange, structural deterioration, surface damage serious (in figure, arrow indication represents ileum damage)；Lac16+Cp
Group broiler ileum fine hair queueing discipline, fine hair are thicker, surface is damaged without obvious.The above results is pointed out, Lactobacillus plantarum Lac16
Can effectively alleviate Cp and infect the damage of the broiler ileum fine hair caused.
As Fig. 5 can find, Control group and Lac16+Cp group broiler ileum microvillus marshalling, closely, Lac16+Cp group
Ileum microvillus is elongated, and Cp group broiler ileum microvillus is sparse, microvillus (left figure) different in size.With Control group phase
Ratio, Cp group broiler ileum compact siro spinning technology (TJ), adhesive tape (AB) shorten, thin out, and gap occurs in intercellular tight junction,
Adhesive tape gap broadens, and desmosome (D) is thin out, gap broadens；Compared with Control group and Cp group, Lac16+Cp group broiler
Ileum compact siro spinning technology (TJ), adhesive tape (AB) are elongated, thicker, and intercellular tight junction is seamless, and adhesive tape gap is full of
Material, desmosome (D) thicker (right figure).The above results is pointed out, and Lactobacillus plantarum Lac16 can be greatly reduced Cp infection and draw
The broiler ileum microvillus risen damages and strengthens Cell tracking (compact siro spinning technology, adhesive tape, desmosome etc.).
Contrast experiment, following Lactobacillus plantarum (table 3) is carried out according to method described in embodiment 1 experiment of fungistatic effect mensuration.
Acquired results is as described in Table 3.
Table 3, different lactic acid bacteria are to bacillus perfringens inhibition
Finally, in addition it is also necessary to be only several specific embodiments of the present invention it is noted that listed above.Obviously, the present invention is not
It is limited to above example, it is also possible to have many deformation.Those of ordinary skill in the art directly can lead from present disclosure
The all deformation gone out or associate, are all considered as protection scope of the present invention.
<110>Zhejiang University
<120>Lactobacillus plantarum and the application thereof of chicken necrotizing enterocolitis are prevented
<160> 2
<210> 1
<211> 29
<212> DNA
<213>artificial sequence
<220>
<223>forward primer (P1)
<400> 1
agagtttgat
cctggtcaga acgaacgct 29
<210> 2
<211> 28
<212> DNA
<213>artificial sequence
<220>
<223>downstream primer (P6)
<400> 2
tacggctacc
ttgttacgac ttcacccc 28
Claims (6)
1. Lactobacillus plantarum Lac16, is characterized in that, preservation is entitled: Lactobacillus plantarum Lac16Lactobacillus plantarum
Lac16, its preserving number is: CCTCC NO:M 2016259.
2. the purposes of Lactobacillus plantarum Lac16 as claimed in claim 1, is characterized in that: the generation of prevention chicken necrotizing enterocolitis.
3. the purposes of Lactobacillus plantarum Lac16 as claimed in claim 1, is characterized in that: suppression bacillus perfringens growth.
The purposes of Lactobacillus plantarum Lac16 the most according to claim 3, is characterized in that: after alleviating bacillus perfringens counteracting toxic substances
Meat chicken production performance declines, alleviates the impact of mucosal structure damage.
5. the purposes of Lactobacillus plantarum Lac16 as claimed in claim 1, is characterized in that: improve chicken macrophage immunity function.
6. the purposes of Lactobacillus plantarum Lac16 as claimed in claim 1, is characterized in that: promote growth of meat chicken.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109182220A (en) * | 2018-10-22 | 2019-01-11 | 东北农业大学 | One plant of inhibition C.perfringens proliferation growing plants lactobacillus and its application |
| CN112716942A (en) * | 2020-12-31 | 2021-04-30 | 江苏农牧科技职业学院 | Indole and application of duck feed containing indole in maintaining duck intestinal health and/or preventing necrotic enteritis of ducks |
| CN113170842A (en) * | 2021-04-26 | 2021-07-27 | 北京科为博生物科技有限公司 | Composite microecological preparation for preventing and treating necrotic enteritis of poultry and application thereof |
| CN113521113A (en) * | 2020-04-15 | 2021-10-22 | 丰华生物科技股份有限公司 | Use of liquid cultures of lactic acid bacterial strains for anti-inflammation and treatment of inflammatory disorders |
| FR3112557A1 (en) | 2020-07-20 | 2022-01-21 | Lesaffre Et Compagnie | COMBINATION OF LACTOBACILLI STRAINS AND ITS USE IN ANIMAL HEALTH |
| CN115886149A (en) * | 2023-01-06 | 2023-04-04 | 吉林农业科技学院 | Application of lactobacillus plantarum LPJZ-658 in improving growth performance, meat quality and disease resistance of broiler chickens |
| CN117448213A (en) * | 2023-10-24 | 2024-01-26 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
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| CN117448213A (en) * | 2023-10-24 | 2024-01-26 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
| CN117448213B (en) * | 2023-10-24 | 2024-06-04 | 山东宝来利来生物工程股份有限公司 | Lactobacillus plantarum for inhibiting clostridium perfringens and its progeny and application |
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