2016, issue 4 при разработке и производстве микробиологических питательных сред актуальной задачей остается поиск сырья для белковых основ, не уступающих мясным по питательной ценности и удовлетворяющих требованиям стандартности и безопасности [1, 3]. основными аргументами использования альтернативных источников сырья являются экономические факторы, экологические соображения, а также то обстоятельство, что в ряде случаев получаемые основы по отдельным качественным показателям превосходят традиционно применяемые пептоны. в нашей стране было выполнено большое количество исследований по использованию различных видов белкового сырья для изготовления гидролизатов, применяемых в производстве питательных сред. показана целесообразность их использования для решения конкретных задач. однако, по нашему мнению, в доступной литературе недостаточно сравнительных данных о специфической активности пептонов, полученных из различных видов белкового сырья, в отношении микроорганизмов разных таксономических групп. для увеличения номенклатуры пептонов, применяемых при изготовлении микробиологических питательных сред, и определения путей их оптимального использования нами ранее была проведена БиотехнолоГия, иММунолоГия
Objectiveof the study was to test the nutrient medium based on the enzymatic hydrolysate of corn extract condensed for a scaled production of live plague vaccine and to check the quality of the obtained batches against the specified parameters.Materials and methods.A dense nutrient medium based on corn extract was used to grow biomass in the process of live plague vaccine production. The quality parameters of the vaccine preparation obtained were studied by the regulated methods set forth in the regulatory documentation.Results and conclusions. The vaccine was monitored at all stages of its manufacture, including control of the finished dosage form, in strict accordance with the approved regulatory documentation. All the experimental production series complied with the specified indices. Approbation of the production cycle environment for live plague vaccine manufacturing showed efficiency of the conditions and the possibility of environment’s application in the industrial production of the preparation.
5 experimental nutritious bases were prepared out of plant materials (soybeans, soymilk, sugar-beet molasses - sugar syrup), and their physical and chemical parameters and amino-acid composition were studied. Nutrient media developed on experimental basis met the nutritious demands of studied strains of plague, cholera, anthrax microbes and sporogenous saprophytes.
The aim was to carry out a comparative analysis of the immunogenic activity of the live plague vaccine obtained on various nutrient media.Materials and methods. The subject of the study was the blood of outbred white mice immunized with a series of live plague vaccine based on Yersinia pestis EV NIIEG strain, produced using experimental and regulated nutrient media. The immunogenic activity of vaccines was studied through flow cytometry. The intensity of antigen-reactivity of lymphocytes was determined in cell tests in vitro, analyzing the early activation marker CD25+ . For the specific activation of lymphocytes, a complex of water-soluble antigens of the plague microbe was used. To identify the interdependence between the presence of protective anti-plague immunity and the level of CD 25+ expression intensity, the ED50 of the series under study was determined by the standard method.Results and discussion. A comparative analysis of the immunogenic activity of the live plague vaccine obtained on the experimental nutrient medium with the vaccine produced on Hottinger’s agar has been performed. When animals were immunized with doses of 4·103 , 2·104 and 1·105 live microbial cells (regulated doses), the highest level of expression of CD25 marker by lymphocytes was on the day 14, with a subsequent decrease on the day 21 after vaccination. When determining immunogenicity using the conventional method, a high degree of direct correlation between the number of surviving animals and an increase in the level of lymphocytes expressing markers of early activation has been established. Comparison has revealed the general pattern: when the lowest immunizing dose (8·102 ) was administered, activation of early immunity markers was not observed. In case of immunization with higher doses on days 7, 14 and 21, a proportional increase in the number of CD25-positive lymphocytes after stimulation with a specific antigen under in vitro conditions is detected in the blood of biomodels.
СРАВНИТЕЛЬНАЯ ОЦЕНКА пОТЕНЦИАЛЬНыХ БЕЛКОВыХ ОСНОВ МИКРОБИОЛОГИЧЕСКИХ СРЕДФКУЗ «Ставропольский научно-исследовательский противочумный институт», Ставрополь, Российская ФедерацияЦелью работы является сравнительная оценка использования панкреатических гидролизатов белковых продуктов растительного и животного происхождения в качестве питательной основы микробиологических сред. В качестве исходного сырья использовали: желатин, сою, соевый концентрат, глютен кукурузный, рыбную кормовую муку, кильку каспийскую, кровь КРС. Гидролиз белкового сырья, очистку гидролизатов и их контроль по физико-химическим показателям проводили общепринятыми методами. Определение биологических показателей пептонов осуществляли на модели питательного агара с помощью тест-штаммов Shigella flexneri 1a 8516, Shigella sonnei «S form», Pseudomonas aeruginosa 27/99, Serratia plymuthica 1. Определены физико-химические показатели исследуемых гидролизатов. Выявлены различия в количестве, диаметре и частоте диссоциации колоний Shigella flexneri 1a 8516, Shigella sonnei «S form», пигментообразовании Pseudomonas aeruginosa 27/99 и Serratia plymuthica 1 на агаровых средах с изучаемыми гидролизатами. Получены сравнительные данные физикохимических и биологических показателей исследуемых гидролизатов, что позволит дифференцировать их применение в составе бактериологических питательных сред.Ключевые слова: гидролизат, питательная среда, физико-химические показатели, биологические показатели, пигментообразование.Objective of the work is to carry out comparative assessment of the pancreatic hydrolysates of protein-containing products, both phytogenous and zoogenous, as nutrient base for microbiological media. Gelatine, soy, soy concentrate, maize gluten, fish meal, common kilka, and bovine blood have been used as a feedstock. Protein stuff hydrolysis, hydrolysate purification, and validation of physical-chemical properties were performed in accordance with conventional techniques. Testing of peptone biological parameters has been carried out on the model of nutrient agar using Shigella flexneri 1a 8516, Shigella sonnei "S form", Pseudomonas aeruginosa 27/99, Serratia plymuthica 1 test strains. Identified have been physical-chemical parameters of the hydrolysates under study. Detected are the variations in quantity, diameter and frequency of dissociation among the colonies of Shigella flexneri 1a 8516, Shigella sonnei "S form", chromogenesis in Pseudomonas aeruginosa 27/99, Serratia plymuthica 1, cultivated on agar media with hydrolysates under study. Obtained are the comparative data on physical-chemical and biological parameters of all experimental hydrolysates, which offers an opportunity to differentiate their choice when adding them into bacteriological nutrient media.
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