scholarly journals Development of a probiotic for animals and aquaculture based on Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains

2021 ◽  
Vol 11 (3) ◽  
pp. 393-402
Author(s):  
A. V. Malkova ◽  
I. Yu. Evdokimov ◽  
M. V. Shirmanov ◽  
A. N. Irkitova ◽  
D. E. Dudnik
Keyword(s):  
No Value ◽  
Nutrient Medium ◽  
Bacillus Pumilus ◽  
Ph Value ◽  
Expiry Date ◽  
Maximum Value ◽  
Bacillus Genus ◽  
Number Of Bacteria ◽  
Bacillus Toyonensis ◽  
Selection Of

Abstract: This article aims to develop a probiotic for animals and aquaculture based on the Bacillus toyonensis B-13249 and Bacillus pumilus B-13250 strains. The selection of a nutrient medium was conducted for cultivating the inoculum of these microorganisms. Several bacteria fermentations of the Bacillus genus were performed in biological reactors with a capacity of 15 and 250 l. A technology for obtaining a finished probiotic for animals and aquaculture was developed. The results indicate that L-broth is the most optimal nutrient medium for cultivating the studied strains. The cultivation of B. toyonensis B-13249 and B. pumilus B-13250 strains in fermenters revealed that sporulation begins after 4–8 hours of fermentation. In contrast to the vegetative medium, the fermentative medium helped the bacilli develop a higher optical density (the maximum value in the B. pumilus strain – 2.400±0.149), pH value (maximum value in the B. toyonensis strain – 8.483±0.609) and titer (at least 1010 CFU/g). After 20–24 hours of incubation, both strains of bacilli in the fermenter, almost completely pass into endospores, which serve as a signal for the start of biomass centrifugation. This was indicated by the following: from a 15 l fermenter – 83.3±6.1 g of concentrate, from a 250 l fermenter – 499.8±51.4 g. The number of bacilli in a concentrated state was at least 1·1011 CFU/g for both strains. Obtaining a finished preparation required mixing bacterial concentrates with maltodextrin to a titer of at least 1·1010 CFU/g. The number of bacteria in the preparation checked every month during the year, recorded no value less than 1·1010 CFU/g. Thus, L-broth is most favorable for growing the mother culture of the B. toyonensis B-13249 and B. pumilus B-13250 strains, and fermentative nutrient medium – for the cultivation in fermenters. The expiry date of the bacilli-based biological preparation is at least 12 months, during which the drug’s polycomponence, color and consistency are preserved, in addition to the bacteria titer (at least 1·1010 CFU/g) and their viability.

scholarly journals Development of a microbiological preparation for crops based on Bacillus pumilus strains

2021 ◽  
Vol 36 ◽  
pp. 07012
Author(s):  
Angelina Malkova ◽  
Ivan Evdokimov ◽  
Maxim Shirmanov ◽  
Alena Irkitova ◽  
Dina Dudnik
Keyword(s):  
Russian Federation ◽  
Nutrient Medium ◽  
Bacillus Pumilus ◽  
Growth Stimulation ◽  
Active Components ◽  
Altai Region ◽  
Protective Medium ◽  
The Russian Federation ◽  
Microbial Preparation ◽  
Number Of Bacteria

Data of the microbial biopreparation development for protection and crop growth stimulation on the Bacillus bacteria basis are presented. Three B. pumilus strains isolated from the Altai region (the Russian Federation) plants rhizosphere were selected as active components of the bacterial preparation. L-bulone was chosen as the nutrient medium for flasks cultivation of the inoculum. A molasses-based nutrient medium was used to incubate the bacilli in a 15-liter fermenter. The finished microbial preparation was obtained in dry form. The biopreparation is a powder consisting of a lyophilically dried concentrates mixture of genus Bacillus spores. Bacilli biomass were pre-mixed with a protective medium based on gelatin and sucrose. The final number of bacteria in the microbial preparation is 1.29(±0.30) ×1012 CFU/g.

scholarly journals Investigation of the effect of hydrocinnamic (3-phenylpropanoic) acid on the microbiological corrosion of steel

2020 ◽  
Author(s):  
А.А. Грибанькова ◽  
М.А. Агиевич ◽  
О.А. Евтуховская ◽  
В.В. Гурченко ◽  
С. Чжай ◽  
...  
Keyword(s):  
Electrochemical Impedance ◽  
Ph Value ◽  
Inhibitory Effect ◽  
Bacterial Cells ◽  
Microbiological Corrosion ◽  
Sulfate Reducing ◽  
Corrosion Of Steel ◽  
Reducing Bacteria ◽  
Number Of Bacteria ◽  
Selection Of

В основе работы лежит исследование воздействия органического соединения (ОС) – гидрокоричной (3-фенилпропановой) кислоты на бактериальные клетки СРБ (сульфатредуцирующих бактерий) являющиеся стимуляторами коррозионных процессов. Подбор условий использования и количественная оценка эффективности ингибирующего (биоцидного) действия исследуемого ОС, которое может быть применено для защиты стали от коррозии в средах, содержащих сульфатредуцирующие бактерии. В ходе исследования было изучено влияние ОС на развитие сульфатредуцирующих бактерий и определены такие параметры, как изменение значения pH и окислительно-восстановительного потенциала образцов в среде, численности бактерий и содержания биогенного сероводорода в зависимости от концентрации используемого ОС. Сделаны основные выводы о ингибирующем коррозию и биоцидном действиях испытуемого органического соединения путем оценки скорости коррозии образцов стали и защитном эффекте в зависимости от выбранных концентраций органического соединения методом гравиметрии. Дополнительная оценка ингибирующего действия была проведена методом электрохимической импедансной спектроскопии и методом снятия поляризационных кривых испытуемых образцов. The study is based on the study of the effect of an organic compound (OC) – hydrocinnamic (3-phenylpropanoic) acid on the bacterial cells of SRB (sulfate-reducing bacteria), which are stimulants of corrosion processes. Selection of conditions of use and a quantitative assessment of the effectiveness of the inhibitory (biocidal) action of the studied OC, which can be used to protect steel from corrosion in environments containing SRB. In the course of the study, the effect of OC on the development of SRB was studied and such parameters as the change in the pH value and the redox potential of the samples in the medium, the number of bacteria and the content of biogenic hydrogen sulfide depending on the concentration of the used OC were determined. The main conclusions are drawn about the inhibitory corrosion and biocidal actions of the test OC by assessing the corrosion rate of steel samples and the protective effect depending on the selected concentrations of the OC by gravimetry. An additional assessment of the inhibitory effect was carried out by the method of electrochemical impedance spectroscopy and by the method of taking the polarization curves of the test samples.

scholarly journals Selection of Optimal Conditions for Cultivation of Bacillus megaterium UCM B-5710 – Producer of Keratinase

2021 ◽  
Vol 83 (6) ◽  
pp. 32-40
Author(s):  
K.V. Avdiyuk ◽  
◽  
A.O. Roy ◽  
Keyword(s):  
Soybean Meal ◽  
Nutrient Medium ◽  
Ph Value ◽  
Soluble Starch ◽  
Enzyme Synthesis ◽  
Cultivation Conditions ◽  
Additional Source ◽  
Carbon And Nitrogen ◽  
Initial Ph ◽  
Selection Of

Every year the volume of production of poultry products all over the world is growing steadily. This contributes to a constant increase in the amount of by-products of poultry processing in the form of down and feather waste, which are dangerous for the environment due to the hard-to-degrade keratin protein and a large number of microbial pathogens. Therefore, the use of environmentally friendly methods for the destruction of keratin substrates due to keratinases of microorganisms is an urgent area of research. The aim of this work was to select the optimal cultivation conditions for the Bacillus megaterium strain UCM B-5710 to increase the activity of the keratinase synthesized by it. Methods. The culture was grown at 28°C, 201 rpm for 7 days on a basic nutrient medium containing defatted chicken feathers as the only source of carbon and nitrogen. The selection of optimal cultivation conditions was carried out according to the following parameters: temperature (21°C, 28°C, 42°C), stirring speed (201 rpm, 212 rpm), amount of inoculum (5%, 10%, 15% , 20%, 25%), the initial pH value of the nutrient medium (4.0–11.0), concentration of keratin-containing substrate (0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2.0%), additional carbon source (glucose, galactose, lactose, maltose, sucrose, mannitol, potato and corn starch, soluble starch, soybean meal) and nitrogen (NH4Cl, NH4NO3, (NH4)2SO4, NaNO3, urea, peptone, tryptone, yeast extract and soybean meal) at a concentration of 1%. Keratinase activity was assessed by the UV absorption at 280 nm of the hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method. Results. The dynamics of the enzyme synthesis showed that the culture of B. megaterium UCM B-5710 exhibited the highest keratinase activity on the 3rd day, and complete splitting of feathers was observed on the 4–5th days. The selection of the concentration of the keratin-containing substrate showed that 0.5% is the optimal concentration. The study of the influence of the initial pH value of the nutrient medium indicates that the culture grew well at pH 6.0–7.0 and pH 9.0–11.0, but at pH 8.0 its growth was very weak. The culture exhibited the maximum keratinase activity at pH 10.0. In addition, at this pH value, complete splitting of feathers was visually observed. The influence of such a key factor as temperature on the growth and synthesis of the enzyme by B. megaterium UCM B-5710 culture demonstrated complete splitting of feathers already on the 2nd day of cultivation at 42°C, at 21°C the culture split feathers very poorly. The introduction of the inoculum into the composition of the nutrient medium in an amount of 15% of the volume of the medium and the mixing intensity of 212 rpm turned out to be optimal. Besides, it was shown that the introduction of an additional source of carbon or nitrogen had an ambiguous effect on the level of keratinase activity of B. megaterium UCM B-5710. Complete inhibition of enzyme synthesis was observed when ammonium sulfate was added to the nutrient medium, and partial inhibition was observed in the case of glucose, lactose, and maltose. Potato, corn, and soluble starch stimulated keratinase synthesis. The majority of inorganic nitrogen sources (ammonium chloride and nitrate) did not affect the synthesis of B. megaterium UCM B-5710 keratinase, while organic sources (urea, peptone, tryptone, yeast extract) increased the level of keratinase activity by 20–50%. However, the most effective result was obtained using soybean meal, the addition of which to the nutrient medium increased the keratinase activity by 2.5 times. Conclusions. As a result of the studies, the optimal conditions for cultivation of the B. megaterium UCM B-5710 strain were selected: the optimum temperature for the growth and development of the culture is 42°C, the initial pH value is 10.0, the stirring speed is 212 rpm and the amount of inoculum introduced is 15%, an additional source of carbon and nitrogen in the form of soybean meal at a concentration of 0.5%. This made it possible to increase the activity of keratinase by 4 times.

scholarly journals Development of polyvalent toxoid Clostridium perfringens against anaerobic enterotoxaemia in young cattle and small ruminants

2021 ◽  
Vol 282 ◽  
pp. 04009
Author(s):  
F.Kh. Pulotov ◽  
O.D. Nazarova ◽  
N.A. Akhmadov ◽  
A.I. Karimzoda
Keyword(s):  
Nutrient Medium ◽  
Small Ruminants ◽  
Farm Animals ◽  
Drug Preparation ◽  
Type D ◽  
Industrial Strains ◽  
Type C ◽  
Young Cattle ◽  
C 30 ◽  
Selection Of

The authors of the article presents the results of the development of a polyvalent toxoid from C.perfringens strains against anaerobic enterotoxaemia of young cattle and small ruminants and an assessment of its effectiveness. It is presented the data on the selection of the recipe composition of the nutrient medium, the cultivation of industrial strains, the process of drug preparation, the selection of the optimal ratio of components and the method of controlling the effectiveness. Sequential cultivation of industrial strains of C.perfringens was carried out by cultivating them in flasks, bottles and bioreactors in the nutrient medium developed by the authors, consisting of: 12.0 g/l - liver extract; 25.0 g/l - casein-peptone; 3.2g/l –K2HPO4; 1.8 g/l – KH2PO4; 0.5 g/l – MgSO4. The development of the drug included the selection of optimal and balanced ratios of C. perfringens type A toxoids - 15 IU/ml, C.perfringens type B - 20 IU/ml, C.perfringens type C - 30 IU/ml, C. perfringens type D - 30 IU/ml, which ensured 100% protection of the immunized animals. Studies on laboratory and farm animals revealed the harmlessness, areactogenicity of the drug, which stimulated the development of immunity and resistance to the toxic effects of all strains of C.perfringens.

The Use of Fluorogenic Substrates To Measure Fungal Presence and Activity in Soil

1998 ◽  
Vol 64 (2) ◽  
pp. 613-617 ◽  
Author(s):  
Morten Miller ◽  
Ansa Palojärvi ◽  
Andrea Rangger ◽  
Morten Reeslev ◽  
Annelise Kjøller
Keyword(s):  
Fungal Biomass ◽  
Phospholipid Fatty Acid ◽  
Soil Samples ◽  
Fluorogenic Substrates ◽  
Link Type ◽  
Solid Media ◽  
Enzyme Substrates ◽  
Agar Media ◽  
Number Of Bacteria ◽  
Selection Of

ABSTRACT Our objective was to determine if 4-methylumbelliferyl-labelled enzyme substrates could be used to detect and quantify specific components of chitinase and cellulase activities as specific indicators of the presence and activity of fungal biomass. The fluorogenic substrates 4-methylumbelliferyl (MUF)N-acetyl-β-d-glucosaminide and MUF β-d-lactoside were used for the detection and quantification of β-N-acetylglucosaminidase (EC 3.2.1.30 ) (NAGase) and endo 1,4-β-glucanase (EC 3.2.1.4 )/cellobiohydrolase (EC3.2.1.91 ) (CELase), respectively. Culture screenings on solid media showed a widespread ability to produce NAGase among a taxonomically diverse selection of fungi on media with and without added chitin. NAGase activity was expressed only in a limited number of bacteria and on media supplemented with chitin. The CELase activity was observed only in a limited number of fungi and bacteria. Bacterial CELase activity was expressed on agar media containing a cellulose-derived substrate. In soil samples, NAGase activity was significantly correlated with estimates of fungal biomass, based on the content of two fungus-specific indicator molecules, 18:2ω6 phospholipid fatty acid (PLFA) and ergosterol. CELase activity was significantly correlated with the PLFA-based estimate of fungal biomass in the soil, but no correlation was found with ergosterol-based estimates of fungal biomass.

scholarly journals Effect of Buckwheat and Potato as Forecrops on Soil Microbial Properties in Crop Rotation

2012 ◽  
Vol 66 (4-5) ◽  
pp. 185-191
Author(s):  
Vilhelmīne Šteinberga ◽  
Olga Mutere ◽  
Inga Jansone ◽  
Ina Alsiņa ◽  
Laila Dubova
Keyword(s):  
Enzymatic Activity ◽  
Crop Rotation ◽  
Microbial Respiration ◽  
Ph Value ◽  
Solanum Tuberosum L ◽  
Temperate Climate ◽  
Soil Microbial ◽  
Soil Microbial Respiration ◽  
Cereal Breeding ◽  
Number Of Bacteria

Field trials were carried out at the experimental cereal breeding fields in Stende (Latvia), an area characterized by a northern temperate climate. The soil had been under short-term organic or conventional crop management and was then subjected to crop rotation, using buckwheat (Fagopyrum esculentum Moench.) and potato (Solanum tuberosum L.) as the forecrops; wheat (Triticum aestivum L.) and oat (Avena sativa L.) as the following crops; and two fertilizer regimes under common organic and conventional practices. As the evaluation criteria of the soil quality, physico-chemical (pH value, organic matter, N, P, K) were tested. In addition, the following biological properties were estimated: plate counts of different physiological groups of microorganisms, soil microbial respiration, enzymatic activity (urease, dehydrogenase and fluoresceine diacetate hydrolysis) of soil microorganisms. Soil microbial respiration activity, FDA hydrolysis and urease activity showed a tendency to decrease in the soil after potato cultivation as the forecrop. The number of bacteria was higher in plots with wheat, and the actinomycete count was lower after oat cultivation. The number of bacteria, actinomycetes, fungi, as well as soil microbial respiration and enzymatic activity, fluctuated due to weather seasonality.

Concomitant Production, Partial Purification and Characterization of a Serine Protease and a Proteolysis-Resistant Metallolipase from Bacillus pumilus SG2

2010 ◽  
Vol 65 (1-2) ◽  
pp. 61-65 ◽  
Author(s):  
Ragupathy Sangeetha ◽  
Arumugam Geetha ◽  
Irulandi Arulpandi
Keyword(s):  
Bacterial Strain ◽  
Bacillus Pumilus ◽  
Genetic Relatedness ◽  
Ph Value ◽  
Temperature Response ◽  
Partial Purification ◽  
Rrna Gene ◽  
Molecular Weights ◽  
Fi Ltration

Our objective was to investigate the concomitant production of protease and lipase by a bacterial strain. A promising bacterial strain was isolated from a food-processing industrial effl uent, which can produce both protease and lipase. The isolate was characterized by sequencing the16S rRNA gene. The PCR amplifi ed gene was subjected to analysis by BLAST to ascertain the genetic relatedness of the isolate, Bacillus pumilus SG2. The enzymes were produced and subjected to purifi cation by ammonium sulfate precipitation and dialysis followed by gel fi ltration chromatography; twelve-fold purity was obtained. The lipase produced was found to be proteolysis-resistant. The partially purifi ed enzymes were characterized for their optimum pH value, temperature, response to inhibitors, surfactants and oxidants. The relative molecular weights of protease and lipase were determined as 28 kDa and 40 kDa, respectively, by zymogram studies.

Selection of a sterilization regime for the nutrient medium in lysine biosynthesis

1985 ◽  
Vol 19 (1) ◽  
pp. 71-74
Author(s):  
V. E. Matveev ◽  
V. E. Aksenovskaya ◽  
L. E. Petrosyan ◽  
A. �. Bulutyan ◽  
F. E. Nazaryan ◽  
...  
Keyword(s):  
Nutrient Medium ◽  
Lysine Biosynthesis ◽  
Selection Of ◽  
Sterilization Regime

scholarly journals Current state of bacterioplankton in Northern Caspian

2021 ◽  
Vol 0 (4) ◽  
pp. 31-38
Author(s):  
Svetlana Aleksandrovna Dyakova ◽  
Olga Borisovna Soprunova ◽  
Ecatherina Rafaelevna Galyautdinova ◽  
Anna Vitalievna Menkova ◽  
Dinara Gaidarovna Baubekova ◽  
...  
Keyword(s):  
Annual Runoff ◽  
Total Abundance ◽  
The North ◽  
Current State ◽  
Maximum Value ◽  
Simultaneous Decrease ◽  
Positive Correlations ◽  
Research Period ◽  
Number Of Bacteria

Microbiological monitoring of the Northern Caspian proved that for the research period in 2013-2018 the average annual value of the total number of bacteria (TBN) was 1.35 million cells/ml. The TBN maximum value (1.46 million cells/ml) was recorded in 2013, the minimum (1.19 million cells/ml) - in 2014. Concentration of saprotrophic bacteria in the waters of the Northern Caspian during the research period varied from 50.94 thousand CFU/ml in 2013 to 1.66 thousand CFU/ml in 2014. The concentration of saprotrophic bacteria remained practically unchanged within 2014-2018. The values of the ratio of the total abundance of microorganisms and saprotrophic bacteria show the eutrophication of the waters of the North Caspian in 2013 and in 2016, in other periods the water quality corresponded to the readings of an oligotrophic reservoir. The maximum number of oil-oxidizing bacteria (OOB) (8.28 thousand CFU/ml) in the waters of the Northern Caspian was recorded in 2013, the minimum (0.21 thousand CFU/ml) - in 2014. Starting from 2015 there was recorded an increasing number of OOBs and its stabilization until the end of the research period. The ratio of NOB and saprotrophs in water varied from 16.47 to 52.47%. Analysis of microbiological and hydrological-hydrochemical indicators revealed positive correlations of TBN and annual runoff (r = + 0.77), TBN and the content of mineral forms of nitrogen (r = + 0.60), the number of saprotrophic bacteria and nitrogen (r = + 0.83), the amount of NOB and mineral nitrogen and silicon (r = + 0.81). In the long-term aspect, an improvement in the microbiological situation was recorded against the background of a growing total number of bacterioplankton under a simultaneous decrease in the number of saprotrophic and oil-oxidizing bacteria.

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