RU2217493C2 - Biopreparation- probiotic, method for its preparing and strain streptococcus faecium te-17 for preparing biopreparation-probiotic for animals and poultry - Google Patents
Biopreparation- probiotic, method for its preparing and strain streptococcus faecium te-17 for preparing biopreparation-probiotic for animals and poultry Download PDFInfo
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- RU2217493C2 RU2217493C2 RU2001104862/13A RU2001104862A RU2217493C2 RU 2217493 C2 RU2217493 C2 RU 2217493C2 RU 2001104862/13 A RU2001104862/13 A RU 2001104862/13A RU 2001104862 A RU2001104862 A RU 2001104862A RU 2217493 C2 RU2217493 C2 RU 2217493C2
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Abstract
Description
Изобретение относится к животноводству и ветеринарии, может быть использовано в биотехнологии, промышленном и фермерском способе выращивания с/х животных для оздоровления молодняка, получения дополнительного привеса животных. The invention relates to animal husbandry and veterinary medicine, can be used in biotechnology, an industrial and farm method of growing agricultural animals for the recovery of young animals, obtaining additional weight gain of animals.
Известно, что некоторые молочнокислые бактерии подавляют рост и развитие патогенной и условно патогенной микрофлоры кишечника животных и птицы. Так, штамм бактерий Streptococcus faecium M-3185 используют для борьбы с дисбактериозом с/х животных [1]. It is known that some lactic acid bacteria inhibit the growth and development of pathogenic and conditionally pathogenic intestinal microflora of animals and birds. So, the bacterial strain Streptococcus faecium M-3185 is used to combat dysbacteriosis of agricultural animals [1].
Известны биопрепараты на основе молочнокислых бактерий, используемые в животноводстве. Так, например, известен препарат на основе Lactobacillus acidophilus для профилактики кишечных заболеваний молодняка с/х животных [2] . Known biological products based on lactic acid bacteria used in animal husbandry. For example, a drug based on Lactobacillus acidophilus is known for the prevention of intestinal diseases of young animals of agricultural animals [2].
Известен биопрепарат на основе молочнокислых бактерий Streptococcus и Lactobacillus-галлиферм, подавляющий развитие условно-патогенной и гнилостной микрофлоры кишечника животных [3]. Known biological product based on lactic acid bacteria Streptococcus and Lactobacillus galliferm, inhibiting the development of conditionally pathogenic and putrefactive intestinal microflora of animals [3].
Известна также питательная среда для выращивания штамма Streptococcus faecium СТФ-1/56 при получении пробиотика, содержащая молочную сыворотку, морскую воду как источник микроэлементов Zn, Mg, Fe, Cu, Mn, Со, глюкозу, однозамещенный фосфорнокислый калий, хлористый аммоний и воду водопроводную [4]. A nutrient medium is also known for growing a strain of Streptococcus faecium STF-1/56 upon receipt of a probiotic containing milk whey, sea water as a source of trace elements Zn, Mg, Fe, Cu, Mn, Co, glucose, monosubstituted potassium phosphate, ammonium chloride and tap water [4].
Однако препараты на основе вышеуказанных штаммов не получили широкого применения; во-первых, используемые штаммы имеют невысокую продуктивность, во-вторых, предлагаемая технология требует исключительно высоких энергозатрат, так как форма препаратов сухая, с использованием сублимационной или распылительной сушки. However, preparations based on the above strains are not widely used; firstly, the strains used have low productivity, and secondly, the proposed technology requires exceptionally high energy consumption, since the form of the preparations is dry, using freeze-drying or spray drying.
Из кишечника цыпленка были выделены изоляты, содержащие молочнокислые бактерии, которые после ступенчатой селекции продуктивных вариантов были идентифицированы как Streptococcus faecium [5]. Isolates containing lactic acid bacteria were isolated from the chicken intestine, which, after a stepwise selection of productive variants, were identified as Streptococcus faecium [5].
Для выделения молочнокислых бактерий применяли прямой высев из образцов, накопительные культуры на жидких средах со спиртом, последующий высев на агаризованные среды с мелом. To isolate lactic acid bacteria, direct seeding from samples, accumulative cultures on liquid media with alcohol, and subsequent seeding on agar media with chalk were used.
1. Культурально-морфологические особенности. 1. Cultural and morphological features.
Streptococcus faecium относится к роду Streptococcus Rosenbach. Представляет собой кокки размером 0.8-1.0 мкм, расположенные короткими цепочками, группами или попарно, спор не образуют, неподвижны, окраска по Грамму - положительная. На агаризованных питательных средах образуют блестящие плотные диски правильной формы с ровными краями диаметром 1-2 мм. При выращивании на жидкой питательной среде через 12-24 часа образуют однородную муть, оседающую на 2-3 сутки. Streptococcus faecium belongs to the genus Streptococcus Rosenbach. It is a cocci with a size of 0.8-1.0 μm, arranged in short chains, groups or in pairs, does not form a spore, is motionless, Gram stain is positive. On agarized nutrient media form shiny dense discs of regular shape with smooth edges with a diameter of 1-2 mm. When grown on a liquid nutrient medium after 12-24 hours, a uniform turbidity forms, settling for 2-3 days.
2. Физиологические свойства. 2. Physiological properties.
Факультативный анаэроб, грамположительный, сбраживает маннозу, галактозу, арабинозу, трегалозу, мальтозу, глюкозу, лактозу, салицин, рибозу, целлобиозу, амигдалин, фруктозу, декстрин, маннит; не сбраживает сахарозу, раффинозу, дульцит, инулин, инозит, сорбит, сорбозу, ксилозу. Не содержит цитохром дыхательного пути, каталазоотрицательный, не восстанавливает нитраты, образует аммиак из аргинина, не разжижает желатин, сквашивает и слабо пептонизирует молоко, восстанавливает молоко с лакмусом и растет на среде, содержащей 40% желчи, 6,5% хлористого натрия, 4% таурохолата. Образует молочную кислоту 0,1-0,15% на 7-9 сутки ферментации. Optional anaerobic, gram-positive, ferments mannose, galactose, arabinose, trehalose, maltose, glucose, lactose, salicin, ribose, cellobiose, amygdalin, fructose, dextrin, mannitol; does not ferment sucrose, raffinose, dulcite, inulin, inositol, sorbitol, sorbose, xylose. Does not contain cytochrome of the respiratory tract, catalase-negative, does not reduce nitrates, forms ammonia from arginine, does not thin the gelatin, ferments and weakly peptones milk, restores milk with litmus and grows on an environment containing 40% bile, 6.5% sodium chloride, 4% taurocholate. It forms lactic acid 0.1-0.15% at 7-9 days of fermentation.
Особенностью штамма Streptococcus faecium ТЕ-17 является высокая удельная скорость роста, гомоферментативное молочнокислое брожение, антагонизм по отношению к некоторым патогенным и условно патогенным микроорганизмам: Ps aeruginosa, Str. aureus, Вас. mesentericus, Salmonella typhimurium, E. coli, Mycobacterium. smegmatis, Klebsiella pneumonia. A feature of the Streptococcus faecium TE-17 strain is a high specific growth rate, homoenzymatic lactic acid fermentation, antagonism against certain pathogenic and conditionally pathogenic microorganisms: Ps aeruginosa, Str. aureus, you. mesentericus, Salmonella typhimurium, E. coli, Mycobacterium. smegmatis, Klebsiella pneumonia.
Устойчив к антибиотикам: тетрациклину, ампициллину, левомицину, пенициллину, эритромицину. Resistant to antibiotics: tetracycline, ampicillin, levomycin, penicillin, erythromycin.
Условия хранения штамма: лиофизированная культура, выращенная на среде, содержащей 20% дрожжевого автолизата, 3,0% глюкозы, сухой обрат (или молочная-подтворожная) сыворотка - 10%, вода - до 1 литра. Хранение в ампулах или флаконах при температуре не выше минус 2oС.Strain conditions for the strain: lyophilized culture grown on a medium containing 20% yeast autolysate, 3.0% glucose, dry return (or milk-under-cure) serum - 10%, water - up to 1 liter. Storage in ampoules or vials at a temperature not exceeding minus 2 o C.
Сравнительная оценка продуктивности штамма Streptococcus faecium ТЕ-17 представлена в таблице 1. A comparative assessment of the productivity of the strain Streptococcus faecium TE-17 is presented in table 1.
Штамм хранится в коллекции микроорганизмов ЗАО "БИОФЛОРА", коллекционный В-07. The strain is stored in the collection of microorganisms of CJSC "BIOFLORA", collection B-07.
Выявленные свойства штамма дают основание использовать его для получения биопрепарата для животноводства в качестве пробиотика с лечебно-профилактическими свойствами. The revealed properties of the strain give reason to use it to obtain a biological product for animal husbandry as a probiotic with therapeutic and prophylactic properties.
Пример 1. Для получения 1 л жидкой культуры штамма Streptococcus faecium ТЕ-17 (коллекция ЗАО "Биофлора В-07") с целью получения биопрепарата готовили жидкую питательную среду: В 1 л молочной сыворотки вносили 200 г дрожжевого автолизата, 3,0 г глюкозы, среду стерилизовали 20 минут при 0,5 кгс/см2. В охлажденную стерильную воду вносили витаминные добавки в виде стерильного раствора витаминов: тиамина, парааминобензойной кислоты, рибофлавина, аскорбиновой кислоты в равных количествах суммарно 0,1 г/л среды, рН среды 6,5. Среду инокулировали приготовленной посевной культурой Streptococcus faecium ТЕ-17 (коллекция ЗАО "Биофлора В-07") из расчета 10% инокулюма, т. е. 100 мл. Ферментацию проводили при 41oС. На 6 часу ферментации стерильно, при слабом перемешивании вносили 2 л спиртового раствора непредельных жирных кислот концентрацией 0,005 М. Выращивание проводили в течение 18 часов, рН полученной жидкой культуры 3,8, число КОЕ 2•1010мл-1. Полученную жидкую культуру использовали для получения биопрепарата.Example 1. To obtain 1 l of a liquid culture of a strain of Streptococcus faecium TE-17 (collection of Bioflora V-07 CJSC) in order to obtain a biological product, a liquid nutrient medium was prepared: 200 g of yeast autolysate, 3.0 g of glucose were added to 1 l of whey , the medium was sterilized for 20 minutes at 0.5 kgf / cm 2 . Vitamin supplements were added to chilled sterile water in the form of a sterile solution of vitamins: thiamine, para-aminobenzoic acid, riboflavin, ascorbic acid in equal amounts for a total of 0.1 g / l of medium, pH of 6.5. The medium was inoculated with the prepared seed culture of Streptococcus faecium TE-17 (collection of Bioflora V-07 CJSC) at the rate of 10% inoculum, i.e. 100 ml. Fermentation was carried out at 41 o C. At 6 hours the fermentation was sterile, with gentle stirring, 2 l of an alcohol solution of unsaturated fatty acids with a concentration of 0.005 M was introduced. The cultivation was carried out for 18 hours, the pH of the obtained liquid culture was 3.8, the number of CFU was 2 • 10 10 ml -1 . The resulting liquid culture was used to obtain a biological product.
Пример 2. Жидкую культуру, полученную, как описано в примере 1, разбавляли водопроводной водой в соотношении 1:2 и вносили, при перемешивании, 20 г альгинатов фукуса и 20 мл жидкой культуры Bacillus mucilaginosus (6) c вязкостью 43•103 пуаз, карбамид из расчета 10 г/л и микроэлементы в виде глицинатов Zn, Mg, Mn, Fe, Cu, В, Se, из расчета 10 мг/л, стабилизаторы пропионат кальция, бензоат натрия, аскорбиновую кислоту в количестве 100 мг/л суммарно. После перемешивания доводили рН препарата до значения 3,5 добавлением молочной кислоты. Полученный препарат использовали для дальнейших опытов.Example 2. The liquid culture obtained as described in example 1 was diluted with tap water in a ratio of 1: 2 and 20 g of fucus alginates and 20 ml of a liquid culture of Bacillus mucilaginosus (6) with a viscosity of 43 • 10 3 poise were added with stirring. carbamide at a rate of 10 g / l and trace elements in the form of glycinates Zn, Mg, Mn, Fe, Cu, B, Se, at a rate of 10 mg / l, stabilizers calcium propionate, sodium benzoate, ascorbic acid in an amount of 100 mg / l in total. After stirring, the pH of the preparation was adjusted to a value of 3.5 by the addition of lactic acid. The resulting preparation was used for further experiments.
Пример 3. Исследовали сохранность жизнеспособных клеток в препарате в процессе длительного хранения при температуре 7±2oС. Полученные результаты представлены в таблице. 2
Пример 4. Биопрепарат, полученный, как описано в примере 2, вносили в корм бройлерам в количестве 3,0 и 1,5 мл на 1 кг корма в течение всего периода откорма цыплят. Результаты опыта представлены в таблице 3.Example 3. Investigated the safety of viable cells in the preparation during long-term storage at a temperature of 7 ± 2 o C. The results are presented in the table. 2
Example 4. The biological product obtained as described in example 2, was introduced into the broiler feed in the amount of 3.0 and 1.5 ml per 1 kg of feed during the entire period of feeding of chickens. The results of the experiment are presented in table 3.
Пример 5. Биопрепарат, полученный, как описано в примере 2, вносили в корм цыплятам с признаками диареи в дозе 3 и 1,5 мл на 1 кг корма в течение всего периода откорма цыплят. Вели наблюдения в течение 7 дней. Результаты опыта представлены в таблице 4. Example 5. The biological product obtained as described in example 2 was introduced into the feed for chickens with signs of diarrhea at a dose of 3 and 1.5 ml per 1 kg of feed during the entire period of feeding the chickens. Observed for 7 days. The results of the experiment are presented in table 4.
Пример 6. Биопрепарат, полученный, как описано в примере 2, вносили в корм поросят с признаками диареи в дозе 3 и 1,5 мл на 1 кг корма в течение всего периода откорма поросят. Вели наблюдения в течение 14 дней. Результаты опыта представлены в таблице 5. Example 6. The biological product obtained as described in example 2, was introduced into the feed of piglets with signs of diarrhea at a dose of 3 and 1.5 ml per 1 kg of feed during the entire period of feeding piglets. Observed for 14 days. The results of the experiment are presented in table 5.
Таким образом, применение заявляемого биопрепарата позволяет получить прибавку в весе (массе) птицы 11-15%, при этом заявляемый препарат подавляет рост патогенной микрофлоры, оказывая оздоравливющий эффект. Thus, the use of the claimed biological product allows to obtain an increase in weight (weight) of the bird 11-15%, while the claimed drug inhibits the growth of pathogenic microflora, providing a healing effect.
Пример 7. Биопрепарат, полученный, как описано в примере 2, вносили в корм поросятам в количестве 1,5 мл на 1 кг корма в течение 1 месяца. Результаты опыта представлены в таблице 6. Example 7. The biological product obtained as described in example 2 was introduced into the feed to piglets in an amount of 1.5 ml per 1 kg of feed for 1 month. The results of the experiment are presented in table 6.
Источники информации
1. Патент РФ 2152433, опубл. 10.07.2000 г.Sources of information
1. RF patent 2152433, publ. 07/10/2000
2. Л.Ф. Абызова, М.С. Полонская, В.В. Леонович. Сборник трудов ВНИИБактпрепарат, вып. 1. М., 1973 г., с. 97-105. 2. L.F. Abyzova, M.S. Polonskaya, V.V. Leonovich. Proceedings of VNIIBaktpreparat, issue. 1. M., 1973, with. 97-105.
3. Наставление по применению галлиферма в ветеринарии. М., 1990 г. 3. Manual on the use of galliferm in veterinary medicine. M., 1990
4. Патент РФ 2086647, опубл. 10.08.97 г. 4. RF patent 2086647, publ. 08/10/97
5. Определитель бактерий Берджи. М., 1987 г. 5. The determinant of bacteria Bergey. M., 1987
6. Патент РФ 2081867, опубл. 20.06.97 г. 6. RF patent 2081867, publ. 06/20/97
Claims (3)
Priority Applications (1)
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| RU2001104862/13A RU2217493C2 (en) | 2001-02-22 | 2001-02-22 | Biopreparation- probiotic, method for its preparing and strain streptococcus faecium te-17 for preparing biopreparation-probiotic for animals and poultry |
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| RU2001104862/13A RU2217493C2 (en) | 2001-02-22 | 2001-02-22 | Biopreparation- probiotic, method for its preparing and strain streptococcus faecium te-17 for preparing biopreparation-probiotic for animals and poultry |
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| RU2217493C2 true RU2217493C2 (en) | 2003-11-27 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2317328C1 (en) * | 2006-05-11 | 2008-02-20 | Государственное научное учреждение Всероссийский научно-исследовательский институт ветеринарной санитарии, гигиены и экологии Российской академии сельскохозяйственных наук (ГНУ ВНИИВСГЭ РОССЕЛЬХОЗАКАДЕМИИ) | Strain streptococcus vestibularis slk-92 used in preparing probiotic preparation designated for prophylaxis and treatment of bacterial intestine infection in agricultural young stock animals |
| RU2352136C2 (en) * | 2006-01-11 | 2009-04-20 | Елизавета Васильевна Чекасина | Feed additive based on beet pulp and method of producing feed additive through fermentation of beet pulp |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2383354C1 (en) * | 2008-09-22 | 2010-03-10 | Институт биологии моря им. А.В.Жирмунского Дальневосточного отделения Российской академии наук | Method of obtaining prostaglandins from alga gracilaria verrucoza |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2086647C1 (en) * | 1994-11-01 | 1997-08-10 | Прикаспийский зональный научно-исследовательский ветеринарный институт | Nutrient medium for growing the strain of streptococcus faecium stf-1/56 |
| RU2091075C1 (en) * | 1995-06-28 | 1997-09-27 | Государственный научно-исследовательский институт генетики и селекции промышленных микроорганизмов | Complex bacterial preparation for treatment and prophylaxis of gastroenteric disease in animals |
| RU2093571C1 (en) * | 1991-09-20 | 1997-10-20 | Пайонир Хай-Бред Интернэшнл, Инк. | Method of stimulation of poultry growth and the probiotic-base preparation |
| WO1997045530A1 (en) * | 1996-05-27 | 1997-12-04 | UZILOVA, Irina Semenovna, Heiress of UZILOV | Use of streptococcus faecium strains and composition containing the same |
| RU2152433C1 (en) * | 1993-03-30 | 2000-07-10 | Товарищество с ограниченной ответственностью "Алэф" | Strain of bacterium streptococcus faecium used for control of dysbacteriosis in agriculture animals |
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2001
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2093571C1 (en) * | 1991-09-20 | 1997-10-20 | Пайонир Хай-Бред Интернэшнл, Инк. | Method of stimulation of poultry growth and the probiotic-base preparation |
| RU2152433C1 (en) * | 1993-03-30 | 2000-07-10 | Товарищество с ограниченной ответственностью "Алэф" | Strain of bacterium streptococcus faecium used for control of dysbacteriosis in agriculture animals |
| RU2086647C1 (en) * | 1994-11-01 | 1997-08-10 | Прикаспийский зональный научно-исследовательский ветеринарный институт | Nutrient medium for growing the strain of streptococcus faecium stf-1/56 |
| RU2091075C1 (en) * | 1995-06-28 | 1997-09-27 | Государственный научно-исследовательский институт генетики и селекции промышленных микроорганизмов | Complex bacterial preparation for treatment and prophylaxis of gastroenteric disease in animals |
| WO1997045530A1 (en) * | 1996-05-27 | 1997-12-04 | UZILOVA, Irina Semenovna, Heiress of UZILOV | Use of streptococcus faecium strains and composition containing the same |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| RU2352136C2 (en) * | 2006-01-11 | 2009-04-20 | Елизавета Васильевна Чекасина | Feed additive based on beet pulp and method of producing feed additive through fermentation of beet pulp |
| RU2317328C1 (en) * | 2006-05-11 | 2008-02-20 | Государственное научное учреждение Всероссийский научно-исследовательский институт ветеринарной санитарии, гигиены и экологии Российской академии сельскохозяйственных наук (ГНУ ВНИИВСГЭ РОССЕЛЬХОЗАКАДЕМИИ) | Strain streptococcus vestibularis slk-92 used in preparing probiotic preparation designated for prophylaxis and treatment of bacterial intestine infection in agricultural young stock animals |
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