Pub Date : 2023-02-28DOI: 10.15407/microbiolj84.06.062
T. Matviykiv, M. Rozhko, R. Kutsyk, V. Gerelyuk
The oral cavity, like the lungs, is often referred to as the «ecological niche of commensal, symbiotic, and pathogenic organisms», and the emigration and elimination of microbes between them are constant, ensuring a healthy distribution of saprophytic microorganisms that maintains organ, tissue, and immune homeostasis. The prolonged hospital stays due to COVID-19 complications, cross-infection, oxygenation therapy through the mask or incubation, and long-term intravenous infusions limit the patient’s ability to care about the oral cavity, regularly clean teeth, floss interdental, etc., which creates extremely favorable conditions for colonization by aerobic and anaerobic pathogens of the oral cavity and periodontal pockets and leads to the rapid progression of chronic generalized periodontitis in this category of patients in the future. The goal of the study was to assess the state of the microbiome of the periodontal pockets of dental patients in the post-covid period. Methods. The object of the study was 140 patients with generalized periodontitis of the I and II stages of development in the chronic course (GP), among which 80 patients had coronavirus disease in the closest past. The patients were randomized by age, sex, and stage of GP development. The diagnosis of periodontal disease was established according to the classifi cation by Danilevskyi. The bacteriological material for aerobic and facultative anaerobic microflora and yeast-like fungi was collected from periodontal pockets with a calibrated bacteriological loop and immediately seeded on blood agar. Results. Significant qualitative and quantitative changes in the nature of the oral microbiocenosis were observed in patients with GP after the recent coronavirus disease, compared with similar patients who did not suffer from COVID-19. We have noticed almost complete disappearance of bacteria that belong to the transient representatives of the oral microflora such as Neisseria, corynebacteria (diphtheria), micrococci, and lactobacilli. The main resident representatives of the oral microflora, i.e., α-hemolytic Streptococci of the mitis group, were found in all healthy individuals and patients of groups A and C, but in 30.0±4.58% of patients in group B, α-hemolytic streptococci in the contents of periodontal pockets are present in quantities not available for detection by the applied method (<2.7 lg CCU/mL). In terms of species, Streptococcus oralis and Streptococcus salivarius are more characteristic in gingival crevicular fluid in healthy individuals (93.8% of selected strains). In 68.4±3.32% of patients in group A, 64.0±3.43% of patients in group B, and 67.5±3.76% of patients in group C, the dominant species were Streptococcus gordonii and Streptococcus sanguinis (p<0.01), which increased pathogenic potential as they produce streptolysin-O, inhibit complement activation, bind to fibronectine, actively form biofilms on the surface of tooth enamel and gum epithelial surface, and can act
{"title":"Peculiarities of Periodontal Pocket Microbiome in Patients with Generalized Periodontitis in the Post-COVID Period","authors":"T. Matviykiv, M. Rozhko, R. Kutsyk, V. Gerelyuk","doi":"10.15407/microbiolj84.06.062","DOIUrl":"https://doi.org/10.15407/microbiolj84.06.062","url":null,"abstract":"The oral cavity, like the lungs, is often referred to as the «ecological niche of commensal, symbiotic, and pathogenic organisms», and the emigration and elimination of microbes between them are constant, ensuring a healthy distribution of saprophytic microorganisms that maintains organ, tissue, and immune homeostasis. The prolonged hospital stays due to COVID-19 complications, cross-infection, oxygenation therapy through the mask or incubation, and long-term intravenous infusions limit the patient’s ability to care about the oral cavity, regularly clean teeth, floss interdental, etc., which creates extremely favorable conditions for colonization by aerobic and anaerobic pathogens of the oral cavity and periodontal pockets and leads to the rapid progression of chronic generalized periodontitis in this category of patients in the future. The goal of the study was to assess the state of the microbiome of the periodontal pockets of dental patients in the post-covid period. Methods. The object of the study was 140 patients with generalized periodontitis of the I and II stages of development in the chronic course (GP), among which 80 patients had coronavirus disease in the closest past. The patients were randomized by age, sex, and stage of GP development. The diagnosis of periodontal disease was established according to the classifi cation by Danilevskyi. The bacteriological material for aerobic and facultative anaerobic microflora and yeast-like fungi was collected from periodontal pockets with a calibrated bacteriological loop and immediately seeded on blood agar. Results. Significant qualitative and quantitative changes in the nature of the oral microbiocenosis were observed in patients with GP after the recent coronavirus disease, compared with similar patients who did not suffer from COVID-19. We have noticed almost complete disappearance of bacteria that belong to the transient representatives of the oral microflora such as Neisseria, corynebacteria (diphtheria), micrococci, and lactobacilli. The main resident representatives of the oral microflora, i.e., α-hemolytic Streptococci of the mitis group, were found in all healthy individuals and patients of groups A and C, but in 30.0±4.58% of patients in group B, α-hemolytic streptococci in the contents of periodontal pockets are present in quantities not available for detection by the applied method (<2.7 lg CCU/mL). In terms of species, Streptococcus oralis and Streptococcus salivarius are more characteristic in gingival crevicular fluid in healthy individuals (93.8% of selected strains). In 68.4±3.32% of patients in group A, 64.0±3.43% of patients in group B, and 67.5±3.76% of patients in group C, the dominant species were Streptococcus gordonii and Streptococcus sanguinis (p<0.01), which increased pathogenic potential as they produce streptolysin-O, inhibit complement activation, bind to fibronectine, actively form biofilms on the surface of tooth enamel and gum epithelial surface, and can act ","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"42 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83388526","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-28DOI: 10.15407/microbiolj84.06.003
N. Chuiko, A. Chobotarov, I. Kurdish
Bacteria of the Bacillus genus can synthesize specific phytase enzymes. This property is especially important for soil bacteria. It helps to mineralize phytin and phytates and to provide these bacteria and plants (in the root zone of which they live) with the available phosphorus. Our previous studies have demonstrated that the Bacillus subtilis IMV B-7023 strain exhibits a phytase activity and can use phytate as a nutrition source. It is a component of the «Azogran» complex bacterial preparation for crop production. As known, abiotic environmental factors can influence the phytase activity of bacteria. In particular, the phytase activity changes significantly under different pH and temperatures. Solid soil particles, including nanosized minerals, can also influence bacteria’s enzymatic activity. The influence of abiotic factors on Bacillus subtilis IMV B-7023 phytase activity has not previously been studied, so this was the aim of our research. Methods. The phytase activity of bacteria was studied by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction, and the nanomaterials’ influence on growth — by cultivation methods. Results. The highest B. subtilis IMV B-7023 phytase activity was observed at 28°C. Also, there was no B. subtilis IMV B-7023 phytase activity at pH 4—6. However, this activity increased at pH 7 and did not change significantly with increasing the buffer system pH to 12. Silicon dioxide influence on the B. subtilis IMV B-7023 growth activity during cultivation in a media with phytate as a phosphorus source depended on the nanomaterial concentration. Thus, at 0.05 and 0.5 g/L of silicon dioxide in the medium, this strain growth activity increased by 8—18%, and at 5.0 g/L of these nanoparticles, bacteria growth inhibition by 19% was observed. At the same time, clay mineral bentonite did not affect the B. subtilis IMV B-7023 growth under the studied cultivation conditions. In addition, silicon dioxide and bentonite stimulated B. subtilis IMV B-7023 phytase activity at all studied concentrations. So, phytase activity increased by 1.82—3.34 times upon adding silicon dioxide and by 2.54—5.83 times upon adding bentonite into the medium. Since the optimal values for phytase activity of most genus Bacillus bacteria are within neutral pH values and temperatures within 50—55°C, a property of B. subtilis IMV B-7023 to show maximum phytase activity at alkaline pH and lower temperatures (28°C) and also stimulation of this activity by soil minerals increases competitiveness of this strain as a component of a bacterial preparation for crop production. Conclusions. Abiotic environmental factors influence the B. subtilis IMV B-7023 supernagrowth and phytase activity. Optimal physicochemical factors for the phytase activity of these bacteria are temperature 28°C and pH 7—12. The concentrations 0.05, 0.5, and 5.0 g/L of silicon dioxide and bentonite increase B. subtilis IMV B-7023 phytase activity. The effect
{"title":"Abiotic Factors Influence on Bacillus subtilis IMV B-7023 Phytase Activity","authors":"N. Chuiko, A. Chobotarov, I. Kurdish","doi":"10.15407/microbiolj84.06.003","DOIUrl":"https://doi.org/10.15407/microbiolj84.06.003","url":null,"abstract":"Bacteria of the Bacillus genus can synthesize specific phytase enzymes. This property is especially important for soil bacteria. It helps to mineralize phytin and phytates and to provide these bacteria and plants (in the root zone of which they live) with the available phosphorus. Our previous studies have demonstrated that the Bacillus subtilis IMV B-7023 strain exhibits a phytase activity and can use phytate as a nutrition source. It is a component of the «Azogran» complex bacterial preparation for crop production. As known, abiotic environmental factors can influence the phytase activity of bacteria. In particular, the phytase activity changes significantly under different pH and temperatures. Solid soil particles, including nanosized minerals, can also influence bacteria’s enzymatic activity. The influence of abiotic factors on Bacillus subtilis IMV B-7023 phytase activity has not previously been studied, so this was the aim of our research. Methods. The phytase activity of bacteria was studied by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction, and the nanomaterials’ influence on growth — by cultivation methods. Results. The highest B. subtilis IMV B-7023 phytase activity was observed at 28°C. Also, there was no B. subtilis IMV B-7023 phytase activity at pH 4—6. However, this activity increased at pH 7 and did not change significantly with increasing the buffer system pH to 12. Silicon dioxide influence on the B. subtilis IMV B-7023 growth activity during cultivation in a media with phytate as a phosphorus source depended on the nanomaterial concentration. Thus, at 0.05 and 0.5 g/L of silicon dioxide in the medium, this strain growth activity increased by 8—18%, and at 5.0 g/L of these nanoparticles, bacteria growth inhibition by 19% was observed. At the same time, clay mineral bentonite did not affect the B. subtilis IMV B-7023 growth under the studied cultivation conditions. In addition, silicon dioxide and bentonite stimulated B. subtilis IMV B-7023 phytase activity at all studied concentrations. So, phytase activity increased by 1.82—3.34 times upon adding silicon dioxide and by 2.54—5.83 times upon adding bentonite into the medium. Since the optimal values for phytase activity of most genus Bacillus bacteria are within neutral pH values and temperatures within 50—55°C, a property of B. subtilis IMV B-7023 to show maximum phytase activity at alkaline pH and lower temperatures (28°C) and also stimulation of this activity by soil minerals increases competitiveness of this strain as a component of a bacterial preparation for crop production. Conclusions. Abiotic environmental factors influence the B. subtilis IMV B-7023 supernagrowth and phytase activity. Optimal physicochemical factors for the phytase activity of these bacteria are temperature 28°C and pH 7—12. The concentrations 0.05, 0.5, and 5.0 g/L of silicon dioxide and bentonite increase B. subtilis IMV B-7023 phytase activity. The effect ","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"5 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88509158","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-23DOI: 10.15407/microbiolj85.01.036
M. Gulich, N. L. Yemchenko, V.G. Kaplinenko, O.O. Kharchenko
Within the conditions of the ongoing COVID-19 pandemic, when many questions regarding prevention and treatment strategies remain unsolved and the search for the best antiviral agents is underway, attention should be paid to the role of trace elements zinc and selenium in increasing the body’s resistance to viral infections and their direct antiviral activity against SARS-CoV-2. Experimental data show that trace elements zinc and selenium not only act through regulating the immune response at all levels of humoral and cellular immunity, but also can play a significant role in adjuvant therapy for viral diseases. This is especially relevant in the case of COVID-19. Studies of the direct antiviral effect of these microelements testify to its 3 main ways to SARS-CoV-2: I — counteraction to virus replication and its transcription through: (i) their covalent binding to the SH-group of the cysteine of the main protease M(Pro) of the virus; (ii) inhibition of its RNA polymerase activity by zinc; II — preventing the penetration of the virus into cells due to blocking SH-groups of protein disulfide isomerase (RDI) of the protein of its spikes (peplomers); III — decreasing the adsorption capacity of the virus due to the blocking of the electrostatic interaction of SARS-CoV-2 peplomers and angiotensin-converting enzyme (ACE-2) in ultra-low, uncharacteristic oxidation states (Zn+1 and Se-2). The intensity of the antiviral action of these trace elements may depend on their chemical form. It was found that zinc citrate (a five-membered complex of zinc with citric acid) and monoselenium citric acid obtained with the help of nanotechnology have a greater intensity of action and higher chemical purity. Taking into account the immunostimulating and direct antiviral effect of zinc and selenium, their use in the form of pharmaceuticals and dietary supplements should be considered as adjunctive therapy for SARS-CoV-2 in patients, or as a preventive strategy for uninfected people from risk groups during the spread of COVID-19.
{"title":"Trace Elements Zinc and Selenium: Their Significance in the Conditions of the COVID-19 Pandemic","authors":"M. Gulich, N. L. Yemchenko, V.G. Kaplinenko, O.O. Kharchenko","doi":"10.15407/microbiolj85.01.036","DOIUrl":"https://doi.org/10.15407/microbiolj85.01.036","url":null,"abstract":"Within the conditions of the ongoing COVID-19 pandemic, when many questions regarding prevention and treatment strategies remain unsolved and the search for the best antiviral agents is underway, attention should be paid to the role of trace elements zinc and selenium in increasing the body’s resistance to viral infections and their direct antiviral activity against SARS-CoV-2. Experimental data show that trace elements zinc and selenium not only act through regulating the immune response at all levels of humoral and cellular immunity, but also can play a significant role in adjuvant therapy for viral diseases. This is especially relevant in the case of COVID-19. Studies of the direct antiviral effect of these microelements testify to its 3 main ways to SARS-CoV-2: I — counteraction to virus replication and its transcription through: (i) their covalent binding to the SH-group of the cysteine of the main protease M(Pro) of the virus; (ii) inhibition of its RNA polymerase activity by zinc; II — preventing the penetration of the virus into cells due to blocking SH-groups of protein disulfide isomerase (RDI) of the protein of its spikes (peplomers); III — decreasing the adsorption capacity of the virus due to the blocking of the electrostatic interaction of SARS-CoV-2 peplomers and angiotensin-converting enzyme (ACE-2) in ultra-low, uncharacteristic oxidation states (Zn+1 and Se-2). The intensity of the antiviral action of these trace elements may depend on their chemical form. It was found that zinc citrate (a five-membered complex of zinc with citric acid) and monoselenium citric acid obtained with the help of nanotechnology have a greater intensity of action and higher chemical purity. Taking into account the immunostimulating and direct antiviral effect of zinc and selenium, their use in the form of pharmaceuticals and dietary supplements should be considered as adjunctive therapy for SARS-CoV-2 in patients, or as a preventive strategy for uninfected people from risk groups during the spread of COVID-19.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"23 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"81380146","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-23DOI: 10.15407/microbiolj85.01.012
S. O. Syrchin, O. Yurieva, A. K. Pavlychenko, I. M. Kurchenko
Lignocellulosic biomass can be utilized as a low-cost, renewable, and sustainable feedstock for obtaining non-fossil energy sources with low CO2 emission. One of the most promising technologies for producing 2G biofuels is the saccharification of agricultural waste materials with the help of cellulolytic enzymes, followed by yeast fermentation of sugars into cellulosic ethanol. Cellulases are multi-component enzymes involved in the degradation of cellulose, which can synergistically degrade cellulose and includes three major categories: endoglucanase (EC 3.2.1.4), exoglucanase or cellobiohydrolase (EC 3.2.1.91), and β-glucosidase (EC 3.2.1.21). The core enzyme used for the degradation of the xylan skeleton of hemicellulose is endo-β-1,4-xylanase (EC 3.2.1.8). The high cost of enzymes synthesized by fungi is a bottleneck for the production of cellulosic ethanol. Optimization of the nutrient medium composition is an important factor in increasing the production of enzymes and the efficiency of lignocellulosic biomass hydrolysis. The aim of the current study was to optimize the production of cellulolytic and xylanolytic enzymes through cultivation of filamentous fungus Talaromyces funiculosus on low-cost nutrient media with non-pretreated agricultural waste materials. Methods. Filamentous fungus Talaromyces funiculosus was grown on potato-dextrose agar for 10—14 days at 26±2 °С. To obtain the culture filtrate, the fungus was cultivated under submerged conditions in an Erlenmeyer flask for 4 days. The nutrient medium composition was varied according to the factor experiment design. A two-step optimization of the nutrient medium composition was used. A screening experiment with the Plackett-Burman fractional factorial design and response surface methodology with the Box-Behnken design were used to optimize cellulase production. The enzymatic activity was determined by measuring the reduced sugar production after the enzymes hydrolysis with specific substrates: exoglucanase with filter paper, endoglucanase with carboxymethylcellulose, and xylanase with beech wood xylan, using the colorimetric DNS method with glucose or xylose as a standard. The activity of β-glucosidase was determined by the hydrolysis reaction of p-nitrophenyl-β-D-glucopyranoside, which results in the formation of p-nitrophenol, quantified at 410 nm. Results. As a result of experiments with using agricultural waste, including wheat straw, corn stalk, and corn cob as carbon sources of the culture medium, it was shown that T. funiculosus is able to grow and produce cellulase and xylanase on all non-pretreated substrates studied. The two-step sequential optimization of the nutrient medium composition for T. funiculosus cultivation according to the Plackett-Berman and Box-Behnken designs made it possible to increase the activity of cellulolytic and xylanolytic enzymes by 2.4—2.6 times. The optimized cultivation medium does not contain such expensive components as Avicel, peptone, and yeas
{"title":"Statistics-Based Optimization of Cellulase and Xylanase Production by the Endophytic Fungus Talaromyces Funiculosus using Agricultural Waste Materials","authors":"S. O. Syrchin, O. Yurieva, A. K. Pavlychenko, I. M. Kurchenko","doi":"10.15407/microbiolj85.01.012","DOIUrl":"https://doi.org/10.15407/microbiolj85.01.012","url":null,"abstract":"Lignocellulosic biomass can be utilized as a low-cost, renewable, and sustainable feedstock for obtaining non-fossil energy sources with low CO2 emission. One of the most promising technologies for producing 2G biofuels is the saccharification of agricultural waste materials with the help of cellulolytic enzymes, followed by yeast fermentation of sugars into cellulosic ethanol. Cellulases are multi-component enzymes involved in the degradation of cellulose, which can synergistically degrade cellulose and includes three major categories: endoglucanase (EC 3.2.1.4), exoglucanase or cellobiohydrolase (EC 3.2.1.91), and β-glucosidase (EC 3.2.1.21). The core enzyme used for the degradation of the xylan skeleton of hemicellulose is endo-β-1,4-xylanase (EC 3.2.1.8). The high cost of enzymes synthesized by fungi is a bottleneck for the production of cellulosic ethanol. Optimization of the nutrient medium composition is an important factor in increasing the production of enzymes and the efficiency of lignocellulosic biomass hydrolysis. The aim of the current study was to optimize the production of cellulolytic and xylanolytic enzymes through cultivation of filamentous fungus Talaromyces funiculosus on low-cost nutrient media with non-pretreated agricultural waste materials. Methods. Filamentous fungus Talaromyces funiculosus was grown on potato-dextrose agar for 10—14 days at 26±2 °С. To obtain the culture filtrate, the fungus was cultivated under submerged conditions in an Erlenmeyer flask for 4 days. The nutrient medium composition was varied according to the factor experiment design. A two-step optimization of the nutrient medium composition was used. A screening experiment with the Plackett-Burman fractional factorial design and response surface methodology with the Box-Behnken design were used to optimize cellulase production. The enzymatic activity was determined by measuring the reduced sugar production after the enzymes hydrolysis with specific substrates: exoglucanase with filter paper, endoglucanase with carboxymethylcellulose, and xylanase with beech wood xylan, using the colorimetric DNS method with glucose or xylose as a standard. The activity of β-glucosidase was determined by the hydrolysis reaction of p-nitrophenyl-β-D-glucopyranoside, which results in the formation of p-nitrophenol, quantified at 410 nm. Results. As a result of experiments with using agricultural waste, including wheat straw, corn stalk, and corn cob as carbon sources of the culture medium, it was shown that T. funiculosus is able to grow and produce cellulase and xylanase on all non-pretreated substrates studied. The two-step sequential optimization of the nutrient medium composition for T. funiculosus cultivation according to the Plackett-Berman and Box-Behnken designs made it possible to increase the activity of cellulolytic and xylanolytic enzymes by 2.4—2.6 times. The optimized cultivation medium does not contain such expensive components as Avicel, peptone, and yeas","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"65 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89071923","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-23DOI: 10.15407/microbiolj85.01.003
O. Lohosha, Yu. О. Vorobei, N. Leonova
The efficiency of the introduction of nodule bacteria, microsymbionts of legumes in agrocenoses, largely depends on the activity of biologically active substances’ biosynthesis by diazotrophs. Seed bacterization with effective rhizobia strains capable of synthesizing exometabolites for phytostimulating activity not only promotes the formation and functioning of symbiosis but also creates the conditions for increasing plant resistance to adverse environmental conditions. The aim of the work was to research the symbiotic activity, efficiency and ability of chickpea rhizobia new strains to biosynthesize phytohormonal exometabolites of cytokinin nature. Methods. Microbiological, physiological, cytological, biochemical, and physicochemical. Results. New strains of Mesorhizobium ciceri ND-101 and Mesorhizobium ciceri ND-64 were shown to have different symbiotic activity. The efficiency of inoculation of Skarb chickpea seeds with bacterial suspension of Mesorhizobiu mciceri ND-101 was at the same level with the industrial strain of Mesorhizobium ciceri H-12. Bacterization of Mesorhizobium ciceri ND-64 increased the chickpea roots nodules by 69%, their weight by 74%, and nitrogenase activity by 73% relative to the positive control (inoculation with Mesorhizobium ciceri H-12), as well as increased chickpeas yield by 22%. It was established that Mesorhizobium ciceri ND-64 strain exhibits the highest cytokinin activity in the bioassay. Cytokinins in the total amount of 174.94 μg/g of completely dry biomass were detected in the culture medium of Mesorhizobium ciceri ND-64, which is 53% higher than that of Mesorhizobium ciceri ND-101 strain and 99% higher than that of Mesorhizobium ciceri H-12 strain. Conclusions. Mesorhizobium ciceri ND-64 strain with high nitrogen-fixing activity and symbiotic efficiency is capable to synthesize a relatively high amount of extracellular cytokinins. The high concentration of cytokinins indicates their important role in the formation and functioning of nodules, as they stimulate the proliferation of root tissues and, in this way, have a positive effect on the chickpea productivity.
{"title":"Symbiotic Efficiency and Cytokinin Activity of New Mesorhizobium cicerі Strains","authors":"O. Lohosha, Yu. О. Vorobei, N. Leonova","doi":"10.15407/microbiolj85.01.003","DOIUrl":"https://doi.org/10.15407/microbiolj85.01.003","url":null,"abstract":"The efficiency of the introduction of nodule bacteria, microsymbionts of legumes in agrocenoses, largely depends on the activity of biologically active substances’ biosynthesis by diazotrophs. Seed bacterization with effective rhizobia strains capable of synthesizing exometabolites for phytostimulating activity not only promotes the formation and functioning of symbiosis but also creates the conditions for increasing plant resistance to adverse environmental conditions. The aim of the work was to research the symbiotic activity, efficiency and ability of chickpea rhizobia new strains to biosynthesize phytohormonal exometabolites of cytokinin nature. Methods. Microbiological, physiological, cytological, biochemical, and physicochemical. Results. New strains of Mesorhizobium ciceri ND-101 and Mesorhizobium ciceri ND-64 were shown to have different symbiotic activity. The efficiency of inoculation of Skarb chickpea seeds with bacterial suspension of Mesorhizobiu mciceri ND-101 was at the same level with the industrial strain of Mesorhizobium ciceri H-12. Bacterization of Mesorhizobium ciceri ND-64 increased the chickpea roots nodules by 69%, their weight by 74%, and nitrogenase activity by 73% relative to the positive control (inoculation with Mesorhizobium ciceri H-12), as well as increased chickpeas yield by 22%. It was established that Mesorhizobium ciceri ND-64 strain exhibits the highest cytokinin activity in the bioassay. Cytokinins in the total amount of 174.94 μg/g of completely dry biomass were detected in the culture medium of Mesorhizobium ciceri ND-64, which is 53% higher than that of Mesorhizobium ciceri ND-101 strain and 99% higher than that of Mesorhizobium ciceri H-12 strain. Conclusions. Mesorhizobium ciceri ND-64 strain with high nitrogen-fixing activity and symbiotic efficiency is capable to synthesize a relatively high amount of extracellular cytokinins. The high concentration of cytokinins indicates their important role in the formation and functioning of nodules, as they stimulate the proliferation of root tissues and, in this way, have a positive effect on the chickpea productivity.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85939006","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-23DOI: 10.15407/microbiolj85.01.026
E. Janbakhsh, M. Mehrabi
Background and objective. Escherichia coli (E. coli) O157: H7 as an enterohemorrhagic pathogen causes severe damage to the gastrointestinal tract and dangerous diseases in humans such as hemolytic uremic syndrome (HUS) and acute renal failure, which is associated with increased blood creatinine levels. This study aimed to evaluate antibiotic resistance of E. coli O157: H7 pathotypes to detect the virulence of gene rfbE and to study variations in its expression. Methods. The isolates were first inoculated on eosin methylene blue (EMB) agar and then identified using the Microgen kit and the presence of rfbE gene. Antibiotic susceptibility of the identified strains was tested by the disk diffusion technique, followed by inoculating E. coli O157: H7 strains at concentrations of 1, 3, and 6 mg dl–1 in BHI broth. DNA and RNA were then extracted from the bacteria, and cDNA was prepared from purified RNA. Then, the rfbE gene expression was evaluated using a real-time PCR approach, and the data were analysed with Rest software. Results. The research results revealed high resistance of isolated strains against some of the studied antibiotics, and variations in the expression of the rfbE gene were found to be different at different creatinine concentrations and at different time points. A significant decrease in variations in the rfbE gene expression was observed at low concentrations (1 mg dl-1), but, on the contrary, a significant increase in variations in the rfbE gene expression was found at higher concentrations (3 and 6 mg dl-1) (p<0.05). Conclusions. The rfbE gene is one of the factors affecting the bacterial virulence. We believe that a secondary increase in creatinine for any reason can exacerbate kidney disease and failure by affecting the rfbE gene expression while producing O antigen or bacterial endotoxin.
{"title":"Molecular Evaluation of rfbE Gene Expression Changes under Different Creatinine Concentrations in Escherichia coli Strains Via Real-Time PCR","authors":"E. Janbakhsh, M. Mehrabi","doi":"10.15407/microbiolj85.01.026","DOIUrl":"https://doi.org/10.15407/microbiolj85.01.026","url":null,"abstract":"Background and objective. Escherichia coli (E. coli) O157: H7 as an enterohemorrhagic pathogen causes severe damage to the gastrointestinal tract and dangerous diseases in humans such as hemolytic uremic syndrome (HUS) and acute renal failure, which is associated with increased blood creatinine levels. This study aimed to evaluate antibiotic resistance of E. coli O157: H7 pathotypes to detect the virulence of gene rfbE and to study variations in its expression. Methods. The isolates were first inoculated on eosin methylene blue (EMB) agar and then identified using the Microgen kit and the presence of rfbE gene. Antibiotic susceptibility of the identified strains was tested by the disk diffusion technique, followed by inoculating E. coli O157: H7 strains at concentrations of 1, 3, and 6 mg dl–1 in BHI broth. DNA and RNA were then extracted from the bacteria, and cDNA was prepared from purified RNA. Then, the rfbE gene expression was evaluated using a real-time PCR approach, and the data were analysed with Rest software. Results. The research results revealed high resistance of isolated strains against some of the studied antibiotics, and variations in the expression of the rfbE gene were found to be different at different creatinine concentrations and at different time points. A significant decrease in variations in the rfbE gene expression was observed at low concentrations (1 mg dl-1), but, on the contrary, a significant increase in variations in the rfbE gene expression was found at higher concentrations (3 and 6 mg dl-1) (p<0.05). Conclusions. The rfbE gene is one of the factors affecting the bacterial virulence. We believe that a secondary increase in creatinine for any reason can exacerbate kidney disease and failure by affecting the rfbE gene expression while producing O antigen or bacterial endotoxin.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"24 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"85881430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-17DOI: 10.15407/microbiolj84.05.038
M.P. Taranukho, Yu.M. Kovalyshyna, Yevhen Zaika
One of the significant reserves for further increasing the yield of berry crops is to protect them from pests and diseases. Among the latter, viral ones are especially dangerous. Therefore, methods of virus diagnostics and especially electron microscopy are of great importance, which makes it possible to see viral particles, determine their shape, size, localization in tissues, and identify anomalies in affected plant cells. Objective. To conduct a comparative study of healthy and diseased blackcurrant leaves in order to determine the degree of influence of the two viruses on the anatomical structure of organelles and inclusions, which can be used in the diagnosis and identifi cation of viruses affecting plants. Methods. The material was blackcurrant plants with symptoms of reversion and green speckles, which are detected visually when examining the plantings of this crop. The morphology of viral particles, the anatomical structure of organelles, and inclusions were studied using the method of electron microscopy of ultrathin sections. Detected ultrastructural changes in cells can be used as diagnostic signs when identifying viruses. Also, viruses were identified by external signs and biological testing. Results. The study of ultrathin sections of leaf tissue and abnormal petals of the blackcurrant flower with symptoms of reversion revealed a bacillus-visible virus (Blackcurrant reversion virus) from the Rabdoviridae family, which is easy to identify due to its large size and appearance, in which it differs from similar features in ordinary cellular components. Typical locations of virus particles are the cytoplasm, nucleus, and perinuclear zone. The size of viral particles on ultrathin sections was 271±7.19 nm long and 78±2.31 nm in diameter. According to electron microscopic methods of studying artificially infected plants of Chenopodium quinoa, an inoculum of affected blackcurrant leaves, virions of Cucumis virus 1 Smith were observed in cells, which were freely located in the cytoplasm of the cell interspersed with ribosomes. Individual areas of the cytoplasm with a high virus concentration were also found in the affected parenchymal cells. Zones surrounded by a double membrane differ in the size and degree of virus saturation. When studying the pathogen morphology in the native preparations, the viral particles had a spherical shape with smparticles showed that they had an average size of 29.6±0.59 nm. During the study of the ultrastructure of blackcurrant plant cells affected by Cucumis virus 1. Smith, myelin-like bodies were found not only in the cytoplasm of affected cells but also in the extra-plasma space. Analysis of the morphology of chloroplasts of blackcurrant plants affected by green speckles and reversion shows that chloroplasts with outgrowths and cup-shaped formations are present in many cells. Under various viral infections, there is a wide variety of mitochondria’s shapes: they are elongated, cup- or club-like, etc. At the sam
{"title":"Effect of Viral Infection on the Ultrastructural Organization of Black Currant Leaf Tissue Cells","authors":"M.P. Taranukho, Yu.M. Kovalyshyna, Yevhen Zaika","doi":"10.15407/microbiolj84.05.038","DOIUrl":"https://doi.org/10.15407/microbiolj84.05.038","url":null,"abstract":"One of the significant reserves for further increasing the yield of berry crops is to protect them from pests and diseases. Among the latter, viral ones are especially dangerous. Therefore, methods of virus diagnostics and especially electron microscopy are of great importance, which makes it possible to see viral particles, determine their shape, size, localization in tissues, and identify anomalies in affected plant cells. Objective. To conduct a comparative study of healthy and diseased blackcurrant leaves in order to determine the degree of influence of the two viruses on the anatomical structure of organelles and inclusions, which can be used in the diagnosis and identifi cation of viruses affecting plants. Methods. The material was blackcurrant plants with symptoms of reversion and green speckles, which are detected visually when examining the plantings of this crop. The morphology of viral particles, the anatomical structure of organelles, and inclusions were studied using the method of electron microscopy of ultrathin sections. Detected ultrastructural changes in cells can be used as diagnostic signs when identifying viruses. Also, viruses were identified by external signs and biological testing. Results. The study of ultrathin sections of leaf tissue and abnormal petals of the blackcurrant flower with symptoms of reversion revealed a bacillus-visible virus (Blackcurrant reversion virus) from the Rabdoviridae family, which is easy to identify due to its large size and appearance, in which it differs from similar features in ordinary cellular components. Typical locations of virus particles are the cytoplasm, nucleus, and perinuclear zone. The size of viral particles on ultrathin sections was 271±7.19 nm long and 78±2.31 nm in diameter. According to electron microscopic methods of studying artificially infected plants of Chenopodium quinoa, an inoculum of affected blackcurrant leaves, virions of Cucumis virus 1 Smith were observed in cells, which were freely located in the cytoplasm of the cell interspersed with ribosomes. Individual areas of the cytoplasm with a high virus concentration were also found in the affected parenchymal cells. Zones surrounded by a double membrane differ in the size and degree of virus saturation. When studying the pathogen morphology in the native preparations, the viral particles had a spherical shape with smparticles showed that they had an average size of 29.6±0.59 nm. During the study of the ultrastructure of blackcurrant plant cells affected by Cucumis virus 1. Smith, myelin-like bodies were found not only in the cytoplasm of affected cells but also in the extra-plasma space. Analysis of the morphology of chloroplasts of blackcurrant plants affected by green speckles and reversion shows that chloroplasts with outgrowths and cup-shaped formations are present in many cells. Under various viral infections, there is a wide variety of mitochondria’s shapes: they are elongated, cup- or club-like, etc. At the sam","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"27 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"80218469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-17DOI: 10.15407/microbiolj84.05.010
M. Bilan, M. Lieshchova, V.E. Podliesnova, V. Brygadyrenko
Millions of tons of microplastics get into the environment, being eaten by many species of mammals and humans. One of the main types of plastic, polystyrene, and its monomer, bisphenol, have been fairly well studied in terms of their effects on metabolism, but changes in the intestinal microbiota under the influence of its addition to the diet remain insufficiently studied. The aim of this article is to describe the changes in the main components of the mice intestinal microbiota in the conditions of adding different concentrations of crushed polystyrene foam to their diet. Methods. Four groups of white laboratory mice ate crushed particles of polystyrene foam (10% of the polymer by weight of the feed, 1%, 0.1%, and the control group — without addition of plastic) as part of the compound feed for 42 days. At the end of the experiment, cultures of animal feces samples were analyzed. Results. Polystyrene foam particles in the main mice diet, especially at a higher concentration (10%), have changed the number.
{"title":"The Effect of Polystyrene Foam on the White Mice’s Intestinal Microbiota","authors":"M. Bilan, M. Lieshchova, V.E. Podliesnova, V. Brygadyrenko","doi":"10.15407/microbiolj84.05.010","DOIUrl":"https://doi.org/10.15407/microbiolj84.05.010","url":null,"abstract":"Millions of tons of microplastics get into the environment, being eaten by many species of mammals and humans. One of the main types of plastic, polystyrene, and its monomer, bisphenol, have been fairly well studied in terms of their effects on metabolism, but changes in the intestinal microbiota under the influence of its addition to the diet remain insufficiently studied. The aim of this article is to describe the changes in the main components of the mice intestinal microbiota in the conditions of adding different concentrations of crushed polystyrene foam to their diet. Methods. Four groups of white laboratory mice ate crushed particles of polystyrene foam (10% of the polymer by weight of the feed, 1%, 0.1%, and the control group — without addition of plastic) as part of the compound feed for 42 days. At the end of the experiment, cultures of animal feces samples were analyzed. Results. Polystyrene foam particles in the main mice diet, especially at a higher concentration (10%), have changed the number.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"24 3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83657333","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-17DOI: 10.15407/microbiolj84.05.003
O. Gudzenko, N. Borzova, L. Varbanets, I. Seifullina, E. Afanasenko, E.V. Martsinko
In recent years, the particular interest of researchers is focused on such enzymes as α-L-rhamnosidase and α-galactosidase. These enzymes are considered useful for various applications. α-L-rhamnosidases may be applied for debittering of citrus fruit juices, due to the less bitter taste of the derhamnosylated flavonones, for rhamnose production, and for the determination of the anomeric configuration in polysaccharides, glycosides and glycolipids. These enzymes may enhance wine aroma and flavonoid bioavailability, or assist in the synthesis of pharmaceuticals. α-Galactosidase finds application in many areas. It is widely used in the food industry to improve the quality of soy products by hydrolyzing indigestible galactosides such as raffinose and stachyose, in the processing of raw materials in order to increase the yield of sugar from molasses, and for the biotransformation of human blood erythrocytes of group B (III) in universal donor erythrocytes, as well as in enzyme therapy of some congenital disorders of sphingolipid metabolism. Earlier, as a result of screening microorganisms of different taxonomic groups, we has selected active α-L-rhamnosidase and α-galactosidase producers. One way to increase their activity is using various effector compounds capable of modifying the enzyme activity. The study of the influence of various effectors is one of the priority areas of modern research in biochemistry, biocoordination chemistry, and biotechnology. Recent advantages in the area of biocoordination chemistry revealed high activating properties of double heterometallic mixed-ligand coor dination compounds with germanium(IV)/tin(IV) tartaric complex anions and 1,10-phenanthroline/2,2`-bipyridine d-metallic cations. The aim is to estimate the enzyme-effector activity of five similar tartratostannates for the α-L-rhamnosidases of Cryptococcus albidus, Eupenicillium erubescens, and α-galactosidase of Penicillium restrictum. Methods. The activity of α-Galactosidase was determined using p-nitrophenyl-α-D-galactopyranoside («Sigma», USA) as a substrate. The activity of α-L-rhamnosidase was determined using the Davis method. As modifiers of enzyme activity, [Fe(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (1), [Co(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·8H2O (2), [Ni(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (3), [Cu(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (4), and [Zn(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·6H2O (5) were used. Results. The study of the effect of complexes 1—5, which are supramolecular salts consisting of the same tartrate stannate anion (electrophilic agent) and a 1,10-phenanthroline d-metal cation (nucleophilic agent), on the Cryptococcus albidus, Eupenicillium erubescens α-L-rhamnosidases, and Penicillium restrictum α-galactosidase showed that the compounds tested had a different influence on the enzymes’ activity. The catalytic activity of α-L-rhamnosidase is significantly influenced by all complexes. The effectiveness of compounds 1—5 for P. restrictum α-galactosid
近年来，研究人员特别关注α- l -鼠李糖酶和α-半乳糖苷酶等酶。这些酶被认为对各种应用都很有用。α- l -鼠李糖苷酶可用于柑桔果汁的脱臭，因为鼠李糖化黄酮的苦味较小，也可用于鼠李糖的生产，并可用于测定多糖、糖苷和糖脂中的头聚糖构型。这些酶可以提高葡萄酒的香气和类黄酮的生物利用度，或协助药物的合成。α-半乳糖苷酶在许多领域都有应用。它广泛应用于食品工业，通过水解棉子糖和水苏糖等难消化的半乳糖苷来提高豆制品的质量，用于原料加工以提高糖蜜的糖产量，用于人血液B (III)族红细胞在通用供体红细胞中的生物转化，以及用于一些先天性鞘脂代谢障碍的酶治疗。在此之前，通过对不同分类群微生物的筛选，我们筛选出了α- l -鼠李糖苷酶活性菌和α-半乳糖苷酶活性菌。增加其活性的一种方法是使用能够修饰酶活性的各种效应化合物。研究各种效应物的影响是生物化学、生物配位化学和生物技术现代研究的重点领域之一。锗(IV)/锡(IV)酒石酸配合物阴离子和1,10-菲罗啉/2,2′-联吡啶金属阳离子的双杂金属混合配体配合物具有较高的活化性能，是近年来生物配位化学领域的研究热点。目的是估计五种相似的酒石酸盐对隐球菌、绿绿青霉和限制青霉α- l -鼠李糖苷酶的酶效活性。方法。α-半乳糖苷酶的活性以对硝基苯-α- d -半乳糖苷(Sigma, USA)为底物测定。采用Davis法测定α- l -鼠李糖苷酶活性。作为酶活性的修饰符,[Fe(苯酚的)3]2 [Sn2(μ馅饼)2(Н2馅饼)2)·2水(1),[有限公司(苯酚的)3]2 [Sn2(μ馅饼)2(Н2馅饼)2)8·h2o (2), (Ni(苯酚的)3]2 [Sn2(μ馅饼)2(Н2馅饼)2)·2水(3),[铜(苯酚的)3]2 [Sn2(μ馅饼)2(Н2馅饼)2)·2水(4),和[锌(苯酚的)3]2 [Sn2(μ馅饼)2(Н2馅饼)2)6·h2o(5)。结果。由酒石酸盐、锡酸盐阴离子(亲电剂)和1,10-菲罗啉d-金属阳离子(亲核剂)组成的超分子盐类配合物1 - 5对隐球菌、绿绿青霉α- l -鼠李糖酶和限制性青霉α-半乳糖苷酶活性的影响研究表明，所测化合物对这些酶的活性有不同的影响。α- l -鼠李糖苷酶的催化活性受到各配合物的显著影响。化合物1 ~ 5对限制草α-半乳糖糖苷酶的抑制作用较差，而对杜鹃α- l -鼠李糖糖苷酶的抑制作用较差。它主要是在控制水平。配合物对隐球菌α- l -鼠李糖苷酶的影响有一定的规律。化合物2和5是最有效的酶，激活率为500-900%。结论。化合物2[Co(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·8H2O作为α- l -鼠李糖苷酶活性的影响因子有进一步的应用前景。
{"title":"Supramolecular 3-d Metal 1,10-Phenanthroline Tartratostannates(IV) as Modifiers of α-L-Rhamnosidase Activity of Cryptococcus albidus, Eupenicillium erubescens and α-Galactosidase Activity of Penicillium restrictum","authors":"O. Gudzenko, N. Borzova, L. Varbanets, I. Seifullina, E. Afanasenko, E.V. Martsinko","doi":"10.15407/microbiolj84.05.003","DOIUrl":"https://doi.org/10.15407/microbiolj84.05.003","url":null,"abstract":"In recent years, the particular interest of researchers is focused on such enzymes as α-L-rhamnosidase and α-galactosidase. These enzymes are considered useful for various applications. α-L-rhamnosidases may be applied for debittering of citrus fruit juices, due to the less bitter taste of the derhamnosylated flavonones, for rhamnose production, and for the determination of the anomeric configuration in polysaccharides, glycosides and glycolipids. These enzymes may enhance wine aroma and flavonoid bioavailability, or assist in the synthesis of pharmaceuticals. α-Galactosidase finds application in many areas. It is widely used in the food industry to improve the quality of soy products by hydrolyzing indigestible galactosides such as raffinose and stachyose, in the processing of raw materials in order to increase the yield of sugar from molasses, and for the biotransformation of human blood erythrocytes of group B (III) in universal donor erythrocytes, as well as in enzyme therapy of some congenital disorders of sphingolipid metabolism. Earlier, as a result of screening microorganisms of different taxonomic groups, we has selected active α-L-rhamnosidase and α-galactosidase producers. One way to increase their activity is using various effector compounds capable of modifying the enzyme activity. The study of the influence of various effectors is one of the priority areas of modern research in biochemistry, biocoordination chemistry, and biotechnology. Recent advantages in the area of biocoordination chemistry revealed high activating properties of double heterometallic mixed-ligand coor dination compounds with germanium(IV)/tin(IV) tartaric complex anions and 1,10-phenanthroline/2,2`-bipyridine d-metallic cations. The aim is to estimate the enzyme-effector activity of five similar tartratostannates for the α-L-rhamnosidases of Cryptococcus albidus, Eupenicillium erubescens, and α-galactosidase of Penicillium restrictum. Methods. The activity of α-Galactosidase was determined using p-nitrophenyl-α-D-galactopyranoside («Sigma», USA) as a substrate. The activity of α-L-rhamnosidase was determined using the Davis method. As modifiers of enzyme activity, [Fe(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (1), [Co(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·8H2O (2), [Ni(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (3), [Cu(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·2H2O (4), and [Zn(phen)3]2[Sn2(μ-Tart)2(Н2Tart)2]·6H2O (5) were used. Results. The study of the effect of complexes 1—5, which are supramolecular salts consisting of the same tartrate stannate anion (electrophilic agent) and a 1,10-phenanthroline d-metal cation (nucleophilic agent), on the Cryptococcus albidus, Eupenicillium erubescens α-L-rhamnosidases, and Penicillium restrictum α-galactosidase showed that the compounds tested had a different influence on the enzymes’ activity. The catalytic activity of α-L-rhamnosidase is significantly influenced by all complexes. The effectiveness of compounds 1—5 for P. restrictum α-galactosid","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"53 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72547935","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-02-17DOI: 10.15407/microbiolj84.05.030
S. Zahorodnia, K. Naumenko, O. Zaychenko, P. Zaremba, G. Baranova, A. V. Holovan'
Today, one of the topical areas of research is the search for antiviral drugs to fi ght against virus-associated oncological manifestations. One of the viruses for which a role in the transformation of cells is proved is Epstein-Barr virus (EBV), which is associated with a variety of lymphoproliferative diseases. The use of drugs that not only inhibit the replication of the virus but also stimulate the elimination of tumor cells is important for the treatment of tumors associated with the viruses. The purpose of this work was to investigate the ability of silver and gold nanoparticles to inhibit EBV replication under conditions of chronic infection. Methods. The objects of the study were 5 to 20 nm gold and silver nanoparticles stabilized with tryptophan, sodium dodecyl sulfate, and citrate. Th e investigations were performed in P3HR-1 (virus-productive) lymphoblastoid cells. MTT-assay, neutral red and trypan blue dyeing were used to study cell viability. Antiviral activity was estimated by the real-time polymerase chain reaction (RT-PCR). Transmissive electron microscopy was used to visualize nanoparticle-virus binding. Results. It was found that nanoparticles of silver and gold stabilized by tryptophan and citrate were low-toxic for the used cell cultures; the vitality of the cells was in the range of 65—100%. Silver nanoparticles in a citrate buffer were more effective against EBV because the used concentrations inhibited replication of the virus up to 70%. Gold nanoparticles reduced the amount of EBV DNA by a maximum of 16% at the lowest concentration of 0.00001 μg/mL, indicating a dose-dependent effect. The virucidal effect of gold nanoparticles against EBV was shown using transmissive electron microscopy. It was found that the interaction of the virus with 5 nm gold nanoparticles for 2 hr leads to damage of EBV virion, which indicates their virus-static effect. Conclusions. Thus, the cytotoxic and antiviral activity of silver and gold nanoparticles in different stabilizers was analyzed. Citrate buffer-stabilized silver and gold NPs were more effective against EBV.
{"title":"Effect of Metal Nanoparticles on EBV-Associated Cell Culture","authors":"S. Zahorodnia, K. Naumenko, O. Zaychenko, P. Zaremba, G. Baranova, A. V. Holovan'","doi":"10.15407/microbiolj84.05.030","DOIUrl":"https://doi.org/10.15407/microbiolj84.05.030","url":null,"abstract":"Today, one of the topical areas of research is the search for antiviral drugs to fi ght against virus-associated oncological manifestations. One of the viruses for which a role in the transformation of cells is proved is Epstein-Barr virus (EBV), which is associated with a variety of lymphoproliferative diseases. The use of drugs that not only inhibit the replication of the virus but also stimulate the elimination of tumor cells is important for the treatment of tumors associated with the viruses. The purpose of this work was to investigate the ability of silver and gold nanoparticles to inhibit EBV replication under conditions of chronic infection. Methods. The objects of the study were 5 to 20 nm gold and silver nanoparticles stabilized with tryptophan, sodium dodecyl sulfate, and citrate. Th e investigations were performed in P3HR-1 (virus-productive) lymphoblastoid cells. MTT-assay, neutral red and trypan blue dyeing were used to study cell viability. Antiviral activity was estimated by the real-time polymerase chain reaction (RT-PCR). Transmissive electron microscopy was used to visualize nanoparticle-virus binding. Results. It was found that nanoparticles of silver and gold stabilized by tryptophan and citrate were low-toxic for the used cell cultures; the vitality of the cells was in the range of 65—100%. Silver nanoparticles in a citrate buffer were more effective against EBV because the used concentrations inhibited replication of the virus up to 70%. Gold nanoparticles reduced the amount of EBV DNA by a maximum of 16% at the lowest concentration of 0.00001 μg/mL, indicating a dose-dependent effect. The virucidal effect of gold nanoparticles against EBV was shown using transmissive electron microscopy. It was found that the interaction of the virus with 5 nm gold nanoparticles for 2 hr leads to damage of EBV virion, which indicates their virus-static effect. Conclusions. Thus, the cytotoxic and antiviral activity of silver and gold nanoparticles in different stabilizers was analyzed. Citrate buffer-stabilized silver and gold NPs were more effective against EBV.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-02-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"72857317","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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