WO2004080200A1 - Micro-organismes probiotiques et leurs utilisations - Google Patents

Micro-organismes probiotiques et leurs utilisations Download PDF

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Publication number
WO2004080200A1
WO2004080200A1 PCT/CA2004/000368 CA2004000368W WO2004080200A1 WO 2004080200 A1 WO2004080200 A1 WO 2004080200A1 CA 2004000368 W CA2004000368 W CA 2004000368W WO 2004080200 A1 WO2004080200 A1 WO 2004080200A1
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WIPO (PCT)
Prior art keywords
probiotic
composition
spores
animal
food
Prior art date
Application number
PCT/CA2004/000368
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English (en)
Inventor
Ben Saïda Ali HALIDI
Blaise Ouattara
Original Assignee
Inatech International Inc.
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Inatech International Inc. filed Critical Inatech International Inc.
Priority to CA002559183A priority Critical patent/CA2559183A1/fr
Publication of WO2004080200A1 publication Critical patent/WO2004080200A1/fr

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Classifications

    • 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
    • A23K50/30—Feeding-stuffs specially adapted for particular animals for swines
    • 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
    • A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/189—Enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K40/00—Shaping or working-up of animal feeding-stuffs
    • A23K40/30—Shaping or working-up of animal feeding-stuffs by encapsulating; by coating

Definitions

  • This invention relates to a new natural food additive for animal such as swine, poultry and the like. More particularly, the invention relates to a probiotic composition including a carrier fraction and microbial spores, a method for modulating digestion in a human or an animal and the use of microbial spores in the manufacture of the probiotic composition.
  • Probiotic micro-organisms are micro-organisms which beneficially affect a host by improving its intestinal microbial balance (Fuller, 1989; Drago et al., 1997). Many probiotic strains are able to significantly reduce disorders caused by infectious agents including Escherichia coli, Listeria monocytogenes, Salmonella, Shigella, Yersinia, and Campylobacter (Con et al, 2001; Drago et al., 1997). Extensive literature is also available on the beneficial effect of probiotic micro-organisms intestinal metabolic activities (hypocholesterolemic effect, improvement of lactose digestion), inhibition of carcinogenesis, and stimulation of immune response (Klaver and ven der Meer, 1993).
  • Probiotics used for human and animal nutrition include Lactobacillus, bifidobacterium, Bacillus, Treptococcus, Pediococcus, Enterococcus and yeast such as Saccharomyces cerevisiae and S. boulardii .
  • probiotic bacteria achieves antipathogenic activity involves active colonisation of attachment sites on the mucosa of the gut wall and a resulting out competition of potential pathogens.
  • Probiotics also produce i) organic acids (lactic and acetic acids) which show antimicrobial activity towards many microorganisms, decrease the pH and the redox potential in the gut, creating suboptimal conditions for pathogens, ii) metabolites such as volatile fatty acids and hydrogen peroxide which is antagonistic to some pathogens through inhibition of their ability to take up nutrients and ions , and iii) bacteriocins and bacteriocin-like products which exert their effects by disruption of energy production systems, macromolecul ⁇ synthesis and membrane permeability.
  • probiotic micro-organisms into foodstuffs.
  • many fermented milk products which contain probiotic micro-organisms are commercially available. Usually these products are in the form of yogurts and an example is the LC1.RTM. yogurt (Societe des Produits Nestle SA).
  • LC1.RTM. yogurt Societe des Produits Nestle SA.
  • infant and follow-up formulas which contain probiotic micro-organisms are also commercially available.
  • probiotic micro-organisms into animal feeds.
  • Canadian patents 2,275,507, 2,222,758 and US patents 5,688,502 and 6,241,983 Bl disclose processes and food additives formulations based on probiotic bacteria and various other ingredients including prebiotics and enzymes.
  • microorganisms incorporated into foodstuffs must fulfill a number of criteria including ability to survive in the gastrointestinal tract and during prolonged periods of storage. Therefore, the stability of commercial probiotic strains is important to assure that stated levels of viable cells are delivered in probiotic products.
  • the survival of ingested probiotics at different levels of the gastrointestinal tract differs between strains. Although some strains such as Bifidobacteria or Lactobacillus can pass through the entire gut at very high concentrations, it is well known that vegetative forms of probiotics bacteria are highly sensitive to environmental conditions.
  • the formulation should be a food composition.
  • One aspect of the present invention is to provide a probiotic composition comprising a carrier fraction and microbial spores.
  • Another aspect of the present invention is to provide a method for modulating digestion in a human or an animal comprising orally administrating to said human or animal a portion of a probiotic food composition as defined above in a quantity sufficient to obtain a desired level of modulation.
  • the modulation may improve intake of nutrients, or reduce or improve the growth of microorganisms.
  • microbial spore in the manufacture of a composition to be orally administered to a human or an animal.
  • the probiotic composition of the present invention comprises at least one digestive enzyme.
  • the carrier fraction can be a food component or composition.
  • the spores can originate from bacteria or from fungi.
  • the spores can also be spores of lactic acid bacteria and can be thermoresistant.
  • the spores can be at least one of endospores or conidia, and can be from microorganisms selected from the group consisting of Lactobacillus, Bifidobacterium, Lactobacillus, Saccharomyses, and Bacillus.
  • the composition of the invention contains the spores at a concentration between 3 to 8 log CFU/g.
  • the spores and digestive enzymes can be encapsulated or coated into a biodegradable or bioresorbable polymer or a biopolymer in order to protect them against digestive enzymes and conditions into the digestive tube.
  • the enzyme contained in the probiotic composition of the present invention can be selected from but are not limited to, the group of amylase, phytase, xylanase, glucanase, and galactosidase.
  • the probiotic food composition of the present invention may comprise in addition, for example, a biological or biochemical active agent, such as a food additive or a supplement, a vitamin or cofactors, an antibiotic, an antifungal compound, an antibody, an antimicrobial product, a pH neutralizing agent.
  • a biological or biochemical active agent such as a food additive or a supplement, a vitamin or cofactors, an antibiotic, an antifungal compound, an antibody, an antimicrobial product, a pH neutralizing agent.
  • probiotic as used herein is intended to mean live microbial feed supplements which beneficially affect the host animal by improving its intestinal microbial balance. It will be recognized by someone skilled in the art that the probiotic microorganisms of the present invention are found in the form of spores, or accompanied by microbial spores.
  • a spore is generally defined in the art as being a type of reproductive cell produced by some plants, fungi and protozoa. Bacterial spores are thick-walled, dormant forms which are capable of surviving unfavorable conditions.
  • endospore as used herein is intended to mean a spore intracellularly formed. It will be understood that spore includes endospores and exospores.
  • spore-forming lactic acid producing bacteria is an interesting alternative in the development of resistant probiotic food formulations.
  • a number of species allocated to the genus Bacillus are known to produce lactic acid. Most of them have been isolated from soil, spoiled food or milk, and from intestine of crayfish. Due to their ability of forming endospores resistant to air drying and other stresses, and which enable them to survive long terms under adverse conditions, spore-forming lactic acid bacteria are good candidates for the development of heat and pressure resistant probiotic foods. Strains described so far belong to the genus Bacillus (B. coagulans, B. licheniformis, B. subtilis, B. stearotherrnophilus, B. smithii) and Sporolactobacillus (Spl. inulinus).
  • One embodiment of the present invention is to provide a food, probiotic composition comprising thermoresistant probiotic bacteria that can survive during the extrusion process and be used as supplements for extrudable food (swine, chicken, turkey).
  • probiotic-organisms examples include spore-forming lactic acid bacteria including Bacillus subtilis, B. licheniformis, Bacillus cereus var toyoi, Lactobacillus sporogenes or a mixture of them.
  • the probiotic micro-organisms are preferably in powdered dried form. It is also an object of the invention to encapsulate or microencapsulate vegetative forms of probiotic micro-organisms to increase their probability of survival in gastro-intestinal conditions (low pH, bile) and under adverse processing and storage conditions (high temperature and pressure, relative humidity).
  • the micro-organisms may be incorporated in a sugar matrix, a fat matrix or a polysaccharide matrix.
  • Suitable vegetative probiotic micro-organisms for encapsulation or microencapsulation processes are Lactobacillus acidophilus, Lb rhamnosus, Lb salivarius, Lb reuteri, Bifidobacterium bifidum, B. animalis, B. longum, B. infantis, Enterococcus faecium, E. faecalis, Pediococcus acidilactis, Sacchaomyces cerevisiae, S. boulardii.
  • thermoprotectant compounds are glycine, betaine, choline, sorbitol, mannosylglycerate, or a mixture of these compounds.
  • Another embodiment of the invention is to provide a new formulation of animal food supplement based on a synergistic and/or additional beneficial effect between probiotic, prebiotic, and other ingredients such as enzymes, vitamins, and minerals.
  • the different constituents of the supplement may be used directly or may be treated following the thermoprotection procedure described previously.
  • nutritional supplements in powdered form comprising prebiotic, enzymes and a probiotic system comprising Lactobacillus acidophilus, Pediococcus acidilactici, Enterococcus faecium, Bacillus licheniforniis, Bacillus subtilis, and Bacillus toyoi.
  • the powdered nutritional supplements contain at least 10 8 viable organism grams.
  • the bacteria in the supplement are stable and can withstand room temperature storage and extreme environmental conditions such as heat, moisture and pH.
  • the probiotic bacteria in the supplement are also stable during the preparation of premixes and complete pelleted feed.
  • Probiotic bacteria were evaluated for their inhibitory effects against pathogenic bacteria.
  • Probiotic bacteria under study included one or several strains of the following species: Lactobacillus acidophilus, Lactobacillus rhamnosus, Lactobacillus salivarius, Pediococcus acidilactici, Enterococcus faecium, Bifidobacterium bifidum, and Bifidobacterium longum.
  • Target pathogenic bacteria included Escherichia coli O157:H7, Salmonella typhimurium, Salmonella enteritidis, Yersinia enterocolitica, Listeria monocytogenes, Aeromonas hydrophila, Bacillus cereus, Staphylococcus aureus, and Shigella sonei.
  • Agar spot test was used to determine the inhibitory efficacy of probiotic bacteria. Antagonistic effects between probiotic strains were also evaluated. The results indicated that most of probiotic bacteria had consistent inhibitory effects against the target pathogenic bacteria. The most efficient of the strains appeared to be Lb acidophilus, Lb rhamnosus, P. acidilactici, and. Ec.
  • YE Yersinia enerocolitica
  • LM Listeria monocytogenes.
  • a feed mixture is supplemented with powdered probiotic preparations, moistened and submitted to different temperatures (70 to 100°C) and times of exposure (0 to 1 min). Viable counts of probiotic bacteria were determined in control samples and samples were submitted to temperature and pressure treatment in order to evaluate the residual percentage of viable cells.
  • the test for high temperature tolerance was performed at Comptoir Agricole de St-Hyacintlie (Quebec, Canada).
  • the probiotic formulation containing Bacillus subtilis and Bacillus licheniformis was incorporated in poultry food and pelleted at a temperature of 78°C for 45s.
  • Samples before and after heat treatment were collected for viable probiotic numeration.
  • the microbiological analysis were done by Bodycote Essais de Materiaux Canada Inc. (Pointe-Claire, Quebec, Canada). The results obtained are presented in Table 2. No significant reduction of viability was observed between samples submitted to heat treatment and control samples.
  • Probiotic counts before heat treatment was 5.59 ⁇ 0.16 lg CFU/g compared to 5.02 ⁇ 0.19 lg CFU/g after heat treatment.
  • Probiotic supplement and complete animal feed containing the probiotic supplement are evaluated for stability during storage.
  • the test preparations consisted of both pelleted and non pelleted feed. Samples are stored at 2 different temperatures (25°C and 45°C) and relative humidity (56 % and 100 %). Residual viable counts were performed monthly or every two months during a total storage period of one year.
  • formulation 1 contained a mixture of vegetative probiotic bacteria Lactobacillus acidophilus, Enterococcus faecium and Pediococcus acidilactici;
  • formulation 2 contained sporulated probiotic bacteria Bacillus subtilis and Bacillus licheniformis. The initial bacterial concentration was determined and other microbial tests were performed after 2 weeks 4 weeks and 6 months. The results of the stability tests are presented in Table 3.
  • Results are expressed in log CFU/g of viable probiotic bacteria in the formulations
  • Trials are performed using 1 day old chickens and young piglets until weaning.
  • the animals were separated in 3 groups. One group is fed with complete chicken or piglets feed incorporated with probiotic supplements at the concentration of 1 kg/ton of animal feed. The second group is fed with animal feed supplemented with an antibiotic growth promoter. The third group is fed with the animal feed without any additive. The animals were weighted periodically and the amount of feed consumed was recorded is order to calculate the growth performance and the food conversion rate in each treatment group.
  • ANOVA Analysis of variance
  • Experiment 2 was performed only with the most interesting concentration of Nutraflore-FTM obtained from experiment 1 i.e 5 kg / ton.
  • Two treatment groups comprising 25 chicken each were tested: treatment 1 (control, 11 ppm virginiamycin) and treatment 2 (Nutraflore-FTM, 5 kg/ton).
  • the experiment was performed in four separate replications.
  • the diets were given ad libitum (day 1-21 starter diet; day 22-35 grower diet, and day 36-42 finisher diet). At day 1, 7, 18, 28 and 42, five (5) chickens were randomly selected in each group and weighted.
  • the global weights of all the chicken was also taken and divided by the number of chickens to obtain the average body weight in each experimental group.
  • treatment 3 (5 kg of Nutraflore-F TM /ton) resulted in significant improvement of growth performance at day 18 Table 2). After day 18, no significant difference was observed between control and treatment 3.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Polymers & Plastics (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Food Science & Technology (AREA)
  • Microbiology (AREA)
  • Animal Husbandry (AREA)
  • Birds (AREA)
  • Health & Medical Sciences (AREA)
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  • Biomedical Technology (AREA)
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Abstract

La présente invention se rapporte à des compositions alimentaires probiotiques, En particulier, la composition probiotique selon l'invention contient des spores microbiennes. Les spores peuvent être également mélangées à des enzymes digestives ou d'autres agents actifs biologiques ou biochimiques utiles pour moduler la digestion chez un être humain ou un animal.
PCT/CA2004/000368 2003-03-11 2004-03-11 Micro-organismes probiotiques et leurs utilisations WO2004080200A1 (fr)

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CA002559183A CA2559183A1 (fr) 2003-03-11 2004-03-11 Micro-organismes probiotiques et leurs utilisations

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US45321703P 2003-03-11 2003-03-11
US60/453,217 2003-03-11

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Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100391356C (zh) * 2005-11-21 2008-06-04 武汉烁森生物科技有限公司 啤酒糟生物改性生产饲料的方法
WO2008087173A1 (fr) 2007-01-17 2008-07-24 Chr, Hansen A/S Procédé de production de poulets
EP1987726A1 (fr) * 2007-05-01 2008-11-05 Friesland Brands B.V. Produit alimentaire de goût agréable contenant un agent de neutralisation des composés indésirables
EP2011858A1 (fr) * 2007-07-06 2009-01-07 Chr. Hansen A/S Composition de bacille résistant à la bile secrétant des concentrations élevées de phytase
EP2103226A1 (fr) 2008-03-18 2009-09-23 Friesland Brands B.V. Produit alimentaire probiotique de longue durée
WO2010070005A1 (fr) * 2008-12-19 2010-06-24 Chr. Hansen A/S Composition de bacille résistante à la bile
CN102771627A (zh) * 2011-05-09 2012-11-14 北京奕农顺丰生物技术有限公司 一种含有复合酶的饲料添加剂
EP2545930A1 (fr) * 2010-03-12 2013-01-16 Calpis Co., Ltd. Agent de régulation de l'augmentation et de la diminution de lactobacillus bifidus dans le côlon
EP2593187A2 (fr) * 2010-07-16 2013-05-22 The Board of Trustees of The University of Arkansas Procédés et compositions comprenant des bactéries formant des spores destinées à améliorer la santé des animaux
RU2564127C2 (ru) * 2008-12-19 2015-09-27 Кр. Хансен А/С Композиция палочковидных бактерий, устойчивых к желчи и секретирующих высокие уровни незаменимых аминокислот
US9393275B2 (en) 2012-08-01 2016-07-19 Novozymes A/S Probiotic for amelioration of coccidiosis vaccine reaction
WO2017011489A1 (fr) * 2015-07-14 2017-01-19 Novozymes A/S Compositions, formulations probiotiques et méthodes pour favoriser la digestion et améliorer la nutrition chez la volaille
US10596209B2 (en) 2017-12-15 2020-03-24 Solarea Bio, Inc. Microbial compositions and methods for treating type 2 diabetes, obesity, and metabolic syndrome
CN114423442A (zh) * 2019-09-16 2022-04-29 诺维信公司 在狗中的基于孢子的益生菌补充和内毒素血症的控制
CN114766613A (zh) * 2022-04-18 2022-07-22 佛山播恩生物科技有限公司 一种适用于泥鳅高密度养殖的生物制剂及其制备方法
WO2023191736A1 (fr) * 2022-03-28 2023-10-05 Sayar Organi̇k Bi̇yoloji̇k Ürünler İmalat Sanayi̇ Ve Ti̇caret Li̇mi̇ted Şi̇rketi̇ Produit biotechnologique contenant une famille de bactéries bacillus subtilis
US11819524B2 (en) 2018-09-05 2023-11-21 Solarea Bio, Inc. Methods and compositions for treating musculoskeletal diseases
US11938158B2 (en) 2021-11-22 2024-03-26 Solarea Bio, Inc. Methods and compositions for treating musculoskeletal diseases, treating inflammation, and managing symptoms of menopause
US11980647B2 (en) 2018-09-05 2024-05-14 Solarea Bio, Inc. Methods and compositions for treating musculoskeletal diseases, treating inflammation, and managing symptoms of menopause

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Cited By (35)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN100391356C (zh) * 2005-11-21 2008-06-04 武汉烁森生物科技有限公司 啤酒糟生物改性生产饲料的方法
WO2008087173A1 (fr) 2007-01-17 2008-07-24 Chr, Hansen A/S Procédé de production de poulets
US8420138B2 (en) 2007-01-17 2013-04-16 Chr. Hansen A/S Method to produce chickens
EP1987726A1 (fr) * 2007-05-01 2008-11-05 Friesland Brands B.V. Produit alimentaire de goût agréable contenant un agent de neutralisation des composés indésirables
WO2008133501A1 (fr) * 2007-05-01 2008-11-06 Friesland Brands B.V. Produit alimentaire agréable au goût contenant un agent destiné à limiter la fixation des glucides et compositions contenant ledit agent
JP2010532161A (ja) * 2007-07-06 2010-10-07 セーホーエル．ハンセン アクティーゼルスカブ フィターゼを高レベルで分泌する胆汁耐性バチルス組成物
US8642305B2 (en) 2007-07-06 2014-02-04 Chr. Hansen A/S Bile resistant bacillus composition secreting high levels of phytase
RU2506307C2 (ru) * 2007-07-06 2014-02-10 Кр. Хансен А/С ШТАММ БАКТЕРИЙ Bacillus subtilis С ВЫСОКИМ УРОВНЕМ ПРОДУЦИРОВАНИЯ ФИТАЗЫ (ВАРИАНТЫ), КОМПОЗИЦИЯ ДЛЯ КОРМЛЕНИЯ ЖИВОТНЫХ И СПОСОБ КОРМЛЕНИЯ ЖИВОТНЫХ
US8334123B2 (en) 2007-07-06 2012-12-18 Chr. Hansen A/S Bile resistant Bacillus composition secreting high levels of phytase
WO2009007192A1 (fr) * 2007-07-06 2009-01-15 Chr. Hansen A/S Composition de bacille résistant à la bile sécrétant des taux élevés de phytase
CN101688173B (zh) * 2007-07-06 2013-04-03 科.汉森有限公司 分泌高水平植酸酶的耐胆汁芽孢杆菌组合物
EP2011858A1 (fr) * 2007-07-06 2009-01-07 Chr. Hansen A/S Composition de bacille résistant à la bile secrétant des concentrations élevées de phytase
EP2103226A1 (fr) 2008-03-18 2009-09-23 Friesland Brands B.V. Produit alimentaire probiotique de longue durée
WO2009116864A1 (fr) * 2008-03-18 2009-09-24 Friesland Brands B.V. Produit alimentaire probiotique de longue conservation
RU2494645C2 (ru) * 2008-03-18 2013-10-10 Фризлэнд Брэндз Б.В. Пробиотический пищевой продукт с длительным сроком хранения
WO2010070005A1 (fr) * 2008-12-19 2010-06-24 Chr. Hansen A/S Composition de bacille résistante à la bile
US8802079B2 (en) 2008-12-19 2014-08-12 Chr. Hansen A/S Bile resistant Bacillus composition
RU2564127C2 (ru) * 2008-12-19 2015-09-27 Кр. Хансен А/С Композиция палочковидных бактерий, устойчивых к желчи и секретирующих высокие уровни незаменимых аминокислот
EP2545930A4 (fr) * 2010-03-12 2013-10-02 Calpis Co Ltd Agent de régulation de l'augmentation et de la diminution de lactobacillus bifidus dans le côlon
EP2545930A1 (fr) * 2010-03-12 2013-01-16 Calpis Co., Ltd. Agent de régulation de l'augmentation et de la diminution de lactobacillus bifidus dans le côlon
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