Pub Date : 2023-08-16DOI: 10.15407/microbiolj85.04.046
R. Fedoruk, I. Kovalchuk, A. Pylypets, M. Tsap, Y. Lesyk, R. L. Androshulik, O. A. Demchenko, N. Tymoshok, L. Babenko
Recently, there has been a trend toward the use of new effective natural preparations to fight diseases and improve the health of honey bees. It is also known that a well-balanced structure of the intestinal microbiota of honey bees is the basis for their growth, development, strengthening of the immune response, and resistance to infections. It has been established that some strains of lactic acid bacteria that have antibacterial, anti-inflammatory, and immunomodulatory properties, are promising for the development of broad-spectrum probiotic preparations based on them. Therefore, the aim of the work was to determine the effect of probiotic strains Lactobacillus сasei IMV B-7280 and L. plantarum IMV B-7679 on catalase activity, protein content and protein profile of hemolymph, as well as microbiota spectrum of different parts of the intestines of Apis mellifera honey bees. Methods. To conduct the research, a control and two experimental groups of 60-90 bees each were formed. The bees of the control group were fed 60% sugar syrup + 1 mL of distilled H2O for 28 days. The experimental group of bees D1 received 1 mL of 60% sugar syrup + 1 mL of aqueous suspension containing cells of the L. casei IMV B-7280 strain at a concentration of 1 ∙ 106 CFU/mL every day; experimental group of bees D2, in addition to 1 mL of 60% sugar syrup, received 1 mL of aqueous suspension containing cells of L. plantarum IMV B-7976 strain at a concentration of 1 ∙ 104 CFU/mL. Catalase activity of the whole organism tissues was determined using the ability of hydrogen peroxide to form a stable colored complex with molybdenum salts on a spectrophotometer at a wavelength of 410 nm against water. The amount of protein in the whole organism tissues was determined by the Lowry method. The content of total protein in the body of bees was carried out according to the Kjeldahl method. Determination of the content of individual fractions of soluble proteins of the hemolymph was carried out by the method of vertical electrophoresis in a 7.5% polyacrylamide gel. The relative content of protein fractions was determined using the TotalLab TL120 program and expressed as a percentage of the total pool. To determine the qualitative and quantitative spectrum of the gut microbiota of bees, the hindgut and midgut were sampled (separately) from bees of control and experimental groups. The obtained samples were plated on eight selective solid media for cultivation of different groups of microorganisms. Results. A tendency to increase the catalase activity of bee tissues after 28 days of L. casei IMV B-7280 strain use and a consistently higher activity of this enzyme throughout the experimental period under the action of L. plantarum IMV B-7679 strain was established. In the control group of 28th days, the content of bees and catalase activity remained at a constant level. It was shown that on the 14th day and total protein in the body of bees that received L. casei IMV B-7280 strain increased s
{"title":"Effect of Probiotic Microorganisms on Catalase Activity, Fractional Composition of Soluble Proteins, and Intestinal Microbiota of Honey Bee","authors":"R. Fedoruk, I. Kovalchuk, A. Pylypets, M. Tsap, Y. Lesyk, R. L. Androshulik, O. A. Demchenko, N. Tymoshok, L. Babenko","doi":"10.15407/microbiolj85.04.046","DOIUrl":"https://doi.org/10.15407/microbiolj85.04.046","url":null,"abstract":"Recently, there has been a trend toward the use of new effective natural preparations to fight diseases and improve the health of honey bees. It is also known that a well-balanced structure of the intestinal microbiota of honey bees is the basis for their growth, development, strengthening of the immune response, and resistance to infections. It has been established that some strains of lactic acid bacteria that have antibacterial, anti-inflammatory, and immunomodulatory properties, are promising for the development of broad-spectrum probiotic preparations based on them. Therefore, the aim of the work was to determine the effect of probiotic strains Lactobacillus сasei IMV B-7280 and L. plantarum IMV B-7679 on catalase activity, protein content and protein profile of hemolymph, as well as microbiota spectrum of different parts of the intestines of Apis mellifera honey bees. Methods. To conduct the research, a control and two experimental groups of 60-90 bees each were formed. The bees of the control group were fed 60% sugar syrup + 1 mL of distilled H2O for 28 days. The experimental group of bees D1 received 1 mL of 60% sugar syrup + 1 mL of aqueous suspension containing cells of the L. casei IMV B-7280 strain at a concentration of 1 ∙ 106 CFU/mL every day; experimental group of bees D2, in addition to 1 mL of 60% sugar syrup, received 1 mL of aqueous suspension containing cells of L. plantarum IMV B-7976 strain at a concentration of 1 ∙ 104 CFU/mL. Catalase activity of the whole organism tissues was determined using the ability of hydrogen peroxide to form a stable colored complex with molybdenum salts on a spectrophotometer at a wavelength of 410 nm against water. The amount of protein in the whole organism tissues was determined by the Lowry method. The content of total protein in the body of bees was carried out according to the Kjeldahl method. Determination of the content of individual fractions of soluble proteins of the hemolymph was carried out by the method of vertical electrophoresis in a 7.5% polyacrylamide gel. The relative content of protein fractions was determined using the TotalLab TL120 program and expressed as a percentage of the total pool. To determine the qualitative and quantitative spectrum of the gut microbiota of bees, the hindgut and midgut were sampled (separately) from bees of control and experimental groups. The obtained samples were plated on eight selective solid media for cultivation of different groups of microorganisms. Results. A tendency to increase the catalase activity of bee tissues after 28 days of L. casei IMV B-7280 strain use and a consistently higher activity of this enzyme throughout the experimental period under the action of L. plantarum IMV B-7679 strain was established. In the control group of 28th days, the content of bees and catalase activity remained at a constant level. It was shown that on the 14th day and total protein in the body of bees that received L. casei IMV B-7280 strain increased s","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-08-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73149490","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.061
O. Povnitsa, S. Zahorodnia, L. Artiukh, M. Zahornyi, A. Ievtushenko
Today, the search for safe ways to inactivate pathogens is becoming especially relevant in connection with the coronavirus pandemic. Standard methods using chlorides and ultraviolet irradiation have disadvantages related to toxicity and low efficiency. Photodynamic inactivation involving nanoparticles is already used to disinfect water and air from microorganisms and enveloped viruses such as human herpes simplex virus, vesicular stomatitis virus, human immunodeficiency virus, and hepatitis B and C viruses. The aim of this work was to evaluate the possibility of the inactivation of human adenovirus type 5 in an organic medium using titanium dioxide irradiated with ultraviolet light. Methods. The nanosized titanium dioxide material was obtained by the thermal decomposition of a suspension of hydrated titanium dioxide TiO(OH)2 (metatitanic acid). The analysis of the morphology of the TiO2 nanopowder was carried out using electron scanning microscopy (SEM), which showed that TiO2 nanopowder contains soft aggregates of nanoparticles mostly 20‒30 nm in size. Cytotoxicity, virulicidal and antiviral action of titanium dioxide were determined by standard methods using (3-(4,5-dimathylthiazol-2-yl)-2,5-dipheniltetrazolium bromide (MTT). The titanium dioxide suspension was irradiated at a distance of 20 cm from 1 to 30 min with a bactericidal UV lamp (OBB15P, BactoSfera, Poland (254 nm)). The concentration of nanoparticles for irradiation was 1.0 mg/mL. Adenovirus suspension with titer 6.0 log10 TCID50 /mL was added to the nanoparticles immediately after irradiation. The titer of virus synthesized in the presence of titanium dioxide was determined by the end of the virus dilution, which causes 50% of the cytopathic effect of the virus on cells. All studies were performed in three replicates; the number of parallel determinations was three. Results. A dose-dependent effect of titanium dioxide nanoparticles on the viability of Hep-2 cells was revealed. At the NPs concentration of 1 mg/mL, quite a low cell viability was observed (32—39%), with a decrease in concentration to 0.1 and 0.01 mg/mL, the NPs were less toxic (cell viability was in the range of 62—90%). The TiO2 NPs dissolved in glycerin-water had no virulicidal effect, as the virus titer was similar to the control values. Instead, NPs dissolved in propanediol-ethanol reduced the infectious titer of the virus by 6.0 log10, which indicates their high virulicidal effect. The absence of an antiviral effect was shown when NPs were added to infected cells. A decrease in the virus titer by 4.5‒5.0 log10 was recorded uponitsinteracting with irradiated NPs for 1‒30 min. The effect persisted for 3 h after exposure to NPs. Conclusions. The cytotoxic, virulicidal, and antiviral effects of optically active TiO2 nanoparticles were determined in optimal conditions. Regardless of the solvent, NPs had low toxicity at a concentration of 0.1 mg/mL. The TiO2 NPs dissolved in glycerin-water had no virulicidal effect; but
{"title":"Photodynamic Treatment of Titanium Dioxide Nanoparticles is a Convenient Method of Adenoviral Inactivation","authors":"O. Povnitsa, S. Zahorodnia, L. Artiukh, M. Zahornyi, A. Ievtushenko","doi":"10.15407/microbiolj85.03.061","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.061","url":null,"abstract":"Today, the search for safe ways to inactivate pathogens is becoming especially relevant in connection with the coronavirus pandemic. Standard methods using chlorides and ultraviolet irradiation have disadvantages related to toxicity and low efficiency. Photodynamic inactivation involving nanoparticles is already used to disinfect water and air from microorganisms and enveloped viruses such as human herpes simplex virus, vesicular stomatitis virus, human immunodeficiency virus, and hepatitis B and C viruses. The aim of this work was to evaluate the possibility of the inactivation of human adenovirus type 5 in an organic medium using titanium dioxide irradiated with ultraviolet light. Methods. The nanosized titanium dioxide material was obtained by the thermal decomposition of a suspension of hydrated titanium dioxide TiO(OH)2 (metatitanic acid). The analysis of the morphology of the TiO2 nanopowder was carried out using electron scanning microscopy (SEM), which showed that TiO2 nanopowder contains soft aggregates of nanoparticles mostly 20‒30 nm in size. Cytotoxicity, virulicidal and antiviral action of titanium dioxide were determined by standard methods using (3-(4,5-dimathylthiazol-2-yl)-2,5-dipheniltetrazolium bromide (MTT). The titanium dioxide suspension was irradiated at a distance of 20 cm from 1 to 30 min with a bactericidal UV lamp (OBB15P, BactoSfera, Poland (254 nm)). The concentration of nanoparticles for irradiation was 1.0 mg/mL. Adenovirus suspension with titer 6.0 log10 TCID50 /mL was added to the nanoparticles immediately after irradiation. The titer of virus synthesized in the presence of titanium dioxide was determined by the end of the virus dilution, which causes 50% of the cytopathic effect of the virus on cells. All studies were performed in three replicates; the number of parallel determinations was three. Results. A dose-dependent effect of titanium dioxide nanoparticles on the viability of Hep-2 cells was revealed. At the NPs concentration of 1 mg/mL, quite a low cell viability was observed (32—39%), with a decrease in concentration to 0.1 and 0.01 mg/mL, the NPs were less toxic (cell viability was in the range of 62—90%). The TiO2 NPs dissolved in glycerin-water had no virulicidal effect, as the virus titer was similar to the control values. Instead, NPs dissolved in propanediol-ethanol reduced the infectious titer of the virus by 6.0 log10, which indicates their high virulicidal effect. The absence of an antiviral effect was shown when NPs were added to infected cells. A decrease in the virus titer by 4.5‒5.0 log10 was recorded uponitsinteracting with irradiated NPs for 1‒30 min. The effect persisted for 3 h after exposure to NPs. Conclusions. The cytotoxic, virulicidal, and antiviral effects of optically active TiO2 nanoparticles were determined in optimal conditions. Regardless of the solvent, NPs had low toxicity at a concentration of 0.1 mg/mL. The TiO2 NPs dissolved in glycerin-water had no virulicidal effect; but","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77734264","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.032
H. Mykhailyshyn, S. I. Klumnyuk, M. Spivak, A. Sverstiuk, L. Lazarenko
The aim of the research is to determine the effect of the probiotic preparation «Dialak» (dietary supplement), which includes the strain Lactobacillus casei IMV B-7280, on the vaginal microbiota and humoral immunity in women with bacterial vaginosis (BV). Methods. 40 female patients aged 20—45 years with disturbed vaginal microbiota and 10 healthy individuals were examined. The verification of 3 types of vaginal biocenosis states, namely normocenosis, intermediate type, and vaginal dysbiosis, was carried out on the basis of the Recommendations for the Treatment of Sexually Transmitted Infections Weekly Morbidity and Mortality Report (2021) and laboratory diagnostic methods according to the well-known criteria proposed by R. Amsel. Female patients with an intermediate type of BV (group 1) received suppositories and capsules of the probiotic (once daily) for 10 days. Women with vaginal dysbiosis (group 2) received metronidazole in a dosage of 500 mg twice a day for 7 days during the first stage, and then 1 suppository at night and oral capsules of the probiotic in the morning for 10 days during the second stage. The studied vaginal secretion was stained by the Gram method in the Kopeloff modification and also sown on nutrient media to determine facultatively anaerobic and obligately anaerobic microorganisms. Microorganism identification was carried out on the basis of morphological, cultural, biochemical, and antigenic properties according to the classification of D. H. Bergey (2009). The activity of humoral immunity was determined by evaluating the number of B-lymphocytes in the peripheral blood of patients using flow cytometry, as well as the levels of serum Ig A, M, and G before treatment and aft er 1 month using the immunoturbidimetric method and the Cobas 6000 test system from Roche Diagnostics (Switzerland). Results. When analyzing the vaginal microbiota in two groups of patients before treatment, a decrease in the number of Lactobacillus spp. and Bifidobacterium spp. and a significant increase in the number of obligate anaerobic microorganisms, including Gardnerella vaginalis, were found compared to the control group. Before treatment, the number of Lactobacillus spp. in women of group 2 was lower compared to group 1. In patients with vaginal dysbiosis before treatment, the number of obligate anaerobic microorganisms was higher than in patients with bacterial vaginosis, except for Eubacterium spp. At the same time, in women in both comparison groups, the indicators of the humoral immune response were partially disrupted, as evidenced by a decrease in the level of IgG and IgA (in women of group 2) in the serum against the normal level of B lymphocytes (CD19+ cells). However, these patients showed an increase in the IgM level in the serum, which may be due to the development of anaerobic microflora. After treatment, the number of Lactobacillus spp. and Bifidobacterium spp. in the vagina of women in both comparison groups increased compared to
{"title":"Effect of Probiotic Therapy on the Vagina Microbiota and the Humoral Link of Immunity in Bacterial Vaginosis","authors":"H. Mykhailyshyn, S. I. Klumnyuk, M. Spivak, A. Sverstiuk, L. Lazarenko","doi":"10.15407/microbiolj85.03.032","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.032","url":null,"abstract":"The aim of the research is to determine the effect of the probiotic preparation «Dialak» (dietary supplement), which includes the strain Lactobacillus casei IMV B-7280, on the vaginal microbiota and humoral immunity in women with bacterial vaginosis (BV). Methods. 40 female patients aged 20—45 years with disturbed vaginal microbiota and 10 healthy individuals were examined. The verification of 3 types of vaginal biocenosis states, namely normocenosis, intermediate type, and vaginal dysbiosis, was carried out on the basis of the Recommendations for the Treatment of Sexually Transmitted Infections Weekly Morbidity and Mortality Report (2021) and laboratory diagnostic methods according to the well-known criteria proposed by R. Amsel. Female patients with an intermediate type of BV (group 1) received suppositories and capsules of the probiotic (once daily) for 10 days. Women with vaginal dysbiosis (group 2) received metronidazole in a dosage of 500 mg twice a day for 7 days during the first stage, and then 1 suppository at night and oral capsules of the probiotic in the morning for 10 days during the second stage. The studied vaginal secretion was stained by the Gram method in the Kopeloff modification and also sown on nutrient media to determine facultatively anaerobic and obligately anaerobic microorganisms. Microorganism identification was carried out on the basis of morphological, cultural, biochemical, and antigenic properties according to the classification of D. H. Bergey (2009). The activity of humoral immunity was determined by evaluating the number of B-lymphocytes in the peripheral blood of patients using flow cytometry, as well as the levels of serum Ig A, M, and G before treatment and aft er 1 month using the immunoturbidimetric method and the Cobas 6000 test system from Roche Diagnostics (Switzerland). Results. When analyzing the vaginal microbiota in two groups of patients before treatment, a decrease in the number of Lactobacillus spp. and Bifidobacterium spp. and a significant increase in the number of obligate anaerobic microorganisms, including Gardnerella vaginalis, were found compared to the control group. Before treatment, the number of Lactobacillus spp. in women of group 2 was lower compared to group 1. In patients with vaginal dysbiosis before treatment, the number of obligate anaerobic microorganisms was higher than in patients with bacterial vaginosis, except for Eubacterium spp. At the same time, in women in both comparison groups, the indicators of the humoral immune response were partially disrupted, as evidenced by a decrease in the level of IgG and IgA (in women of group 2) in the serum against the normal level of B lymphocytes (CD19+ cells). However, these patients showed an increase in the IgM level in the serum, which may be due to the development of anaerobic microflora. After treatment, the number of Lactobacillus spp. and Bifidobacterium spp. in the vagina of women in both comparison groups increased compared to ","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"17 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76213700","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.022
O. M. Zahrychuk, H. Mykhailyshyn, I. Volch, S. I. Klumnyuk, L. Romanyuk
In pediatric surgery, acute appendicitis is considered one of the most common problems requiring surgical intervention. Among the causes of this disease, microorganisms are of primary importance. The specificity of postoperative treatment depends both on the degree of virulence of the pathogen and on the microbial load that caused the inflammatory process. The increase in the use of antimicrobial agents is of great concern because of the emergence of antibiotic-resistant bacteria. Therefore, the issue of rational postoperative antibiotic therapy remains relevant, as excessive, often unjustified use and incorrect dosage of drugs have become the cause of many medical problems. The aim of the research was to determine the species structure and analyze antibiotic resistance of microorganisms in biomaterial obtained from children after appendectomy for acute appendicitis. Methods. We studied biomaterial obtained from 74 patients aged 2—18 years who were treated at the MNCE Ternopil Regional Children Clinical Hospital TRC in the period from September 2021 to March 2022. After appendectomy, the samples were placed in a transport medium for further laboratory research, which involved staining smears according to the Gram method, sowing microorganisms on nutrient media such as blood agar, salt agar, sugar broth and serum agar, and Endo medium for enterobacteria as well as for anaerobic pathogens — thioglycolic medium and Kitta-Tarozzi medium, and identifying by morphological, tinctorial, cultural and biochemical properties. The sensitivity of selected pathogenic microorganisms to antibiotics was determined using the Kirby-Bauer method. Statistical processing of digital data was carried out using Excel software (Microsoft, USA) and the Statistica 10.0 program. Results. 74 children aged from 2 to 18 years were involved in the study. E. coli (28.4% of all examined), S. aureus (21.6%), and P. aeruginosa (14.9%) were found during the laboratory study of biomaterial. E. faecalis, Klebsiella spp., S. epidermidis, and S. viridans occurred much less often (from 9.4% to 4.5%). The study of antibiotic resistance showed that the isolated microorganisms differed significantly in their sensitivity both to different groups of antimicrobial agents and to generations of drugs within the same group. Ceftriaxone was the most effective inhibitor of all detected microorganisms. E. coli, S. aureus, P. aeruginosa, and S. epidermidis showed 100% sensitivity to it, and the others — within 75—50%. Bacteria Klebsiella spp. and S. epidermidis were sensitive to amikacin, strains of E. coli — 90.5%, P. aeruginosa and S. aureus — 81.8% and 81.2%, respectively. Amoxiclav and ampisulbin had weak inhibitory activity, except for 100% of Klebsiella spp. and 75% of E. faecalis, which were inhibited only by amoxiclav. However, almost all studied microorganisms were partially sensitive to azithromycin. The activity of this antibiotic ranged from 100—81.8% (S. epidermidis, S. aureus) to 36.4% (
{"title":"Species Characteristics of Causative Agents of Acute Appendicitis in Children and Determination of Their Susceptibility to Antibiotics","authors":"O. M. Zahrychuk, H. Mykhailyshyn, I. Volch, S. I. Klumnyuk, L. Romanyuk","doi":"10.15407/microbiolj85.03.022","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.022","url":null,"abstract":"In pediatric surgery, acute appendicitis is considered one of the most common problems requiring surgical intervention. Among the causes of this disease, microorganisms are of primary importance. The specificity of postoperative treatment depends both on the degree of virulence of the pathogen and on the microbial load that caused the inflammatory process. The increase in the use of antimicrobial agents is of great concern because of the emergence of antibiotic-resistant bacteria. Therefore, the issue of rational postoperative antibiotic therapy remains relevant, as excessive, often unjustified use and incorrect dosage of drugs have become the cause of many medical problems. The aim of the research was to determine the species structure and analyze antibiotic resistance of microorganisms in biomaterial obtained from children after appendectomy for acute appendicitis. Methods. We studied biomaterial obtained from 74 patients aged 2—18 years who were treated at the MNCE Ternopil Regional Children Clinical Hospital TRC in the period from September 2021 to March 2022. After appendectomy, the samples were placed in a transport medium for further laboratory research, which involved staining smears according to the Gram method, sowing microorganisms on nutrient media such as blood agar, salt agar, sugar broth and serum agar, and Endo medium for enterobacteria as well as for anaerobic pathogens — thioglycolic medium and Kitta-Tarozzi medium, and identifying by morphological, tinctorial, cultural and biochemical properties. The sensitivity of selected pathogenic microorganisms to antibiotics was determined using the Kirby-Bauer method. Statistical processing of digital data was carried out using Excel software (Microsoft, USA) and the Statistica 10.0 program. Results. 74 children aged from 2 to 18 years were involved in the study. E. coli (28.4% of all examined), S. aureus (21.6%), and P. aeruginosa (14.9%) were found during the laboratory study of biomaterial. E. faecalis, Klebsiella spp., S. epidermidis, and S. viridans occurred much less often (from 9.4% to 4.5%). The study of antibiotic resistance showed that the isolated microorganisms differed significantly in their sensitivity both to different groups of antimicrobial agents and to generations of drugs within the same group. Ceftriaxone was the most effective inhibitor of all detected microorganisms. E. coli, S. aureus, P. aeruginosa, and S. epidermidis showed 100% sensitivity to it, and the others — within 75—50%. Bacteria Klebsiella spp. and S. epidermidis were sensitive to amikacin, strains of E. coli — 90.5%, P. aeruginosa and S. aureus — 81.8% and 81.2%, respectively. Amoxiclav and ampisulbin had weak inhibitory activity, except for 100% of Klebsiella spp. and 75% of E. faecalis, which were inhibited only by amoxiclav. However, almost all studied microorganisms were partially sensitive to azithromycin. The activity of this antibiotic ranged from 100—81.8% (S. epidermidis, S. aureus) to 36.4% (","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"26 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"86806304","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.003
O. Gudzenko, N. Borzova, L. Varbanets, I. Seifullina, E. Martsinko, O. Buchko, А. Pesaroglo
The intensive development of biotechnology in the last decade is largely determined by the growing requirement needs of both medicine and various industries for products of microbial synthesis, including glycosidases, in particular α-L-rhamnosidases. Their wide use to solve current biological-medical and chemical-technological problems stimulates researchers to search for compounds capable of influencing their catalytic activity. Therefore, the purpose of this work was to isolate and purify α-L-rhamnosidase from a new producer of Penicillium restrictum and to investigate multi-ligand germanium-3d-metal complexes with citric acid, phenanthroline, and bipyridine as effectors of its activity. Methods. The object of the study was α-L-rhamnosidase of P. restrictum. Its purification was carried out by gel filtration and ion exchange chromatography on TSK-gels and Sepharose 6B. The activity of α-L-rhamnosidase was determined using the Davis method with naringin as a substrate. As modifiers of enzyme activity, purposefully synthesized multiligand germanium-3d-metal complexes with citric acid, phenanthroline, and bipyridine ([Ni(bipy)3][Ge(HCit)2]·3H2O (1); [Ni(phen)3][Ge(HCit)2]·2H2O (2); [{Cu(bipy)2}2Ge(m-Cit)2]·12Н2О (3); [{Cu(phen)2}2Ge(m-Cit)2]·13H2O (4); [Zn(bipy)3][Ge(HCit)2]·2H2O (5); [Zn(phen)3][Ge(HCit)2]·3H2O (6)), were used. Results. From the supernatant of culture fluid of P. restrictum, α-L-rhamnosidase was isolated and purified 23.1 times with a yield of 0.09%. The specific activity of the enzyme was 27.8 units/mL. The enzyme was homogeneous according to gel filtration on Sepharose 6B and had a molecular mass of 50 kDa. It was established that the considered coordination compounds are able to regulate the catalytic activity of α-L-rhamnosidase of P. restrictum. All of them manifest themselves either as activators or as inert substances, no inhibition was observed. In addition, the dependence of the degree of enzyme activation by the compounds on their concentration is traced and corresponds to the following series: at a concentration of 0.01% — 1 > 6 ≈ 5 > 3 >2 ≈ 4 and at a concentration of 0.1% — 1 > 4 > 2 > 5 ≈ 6. 3. The catalytic activity is also significantly affected by the time of exposure to the compounds: at a concentration of 0.01% for 1h, the activity of the enzyme at the control level was observed for all compounds, whereas at a concentration of 0.1% for 24 h, the activity increased sharply in the presence of compounds 1 (300%), 6 (153%), and 2 (134%). The action of the others was at the control level. Conclusions. The obtained data on new complex metal compounds with an activating effect on microbial α-L-rhamnosidases. It has been established that compounds whose structural organization ensures the synergism of the action of all components are the most promising enzyme effectors in a series of coordination compounds of biologically active metals and ligands.
近十年来生物技术的迅猛发展，在很大程度上是由于医药和各种工业对微生物合成产物，包括糖苷酶，特别是α- l -鼠李糖苷酶的需求不断增长。它们在解决当前生物医学和化学技术问题上的广泛应用促使研究人员寻找能够影响其催化活性的化合物。因此，本研究的目的是从一种新的限制性青霉中分离纯化α- l -鼠李糖苷酶，并研究与柠檬酸、菲罗啉和联吡啶配合的多配体锗-3d金属配合物对其活性的影响。方法。本研究的对象是限制草的α- l -鼠李糖苷酶。用tsk -凝胶和Sepharose 6B进行凝胶过滤和离子交换层析纯化。以柚皮苷为底物，采用Davis法测定α- l -鼠李糖苷酶活性。作为酶活性调节剂，有目的地与柠檬酸、菲罗啉、联吡啶合成了多配体锗-3d金属配合物([Ni(bipy)3][Ge(HCit)2]·3H2O (1);[倪(苯酚的)3][通用电气(职业)2]·2水(2);[{铜(bipy) 2} 2通用电气(m-Cit) 2]·12Н2О(3);[{铜(苯酚的)2}2通用电气(m-Cit) 2] 13·h2o (4);[锌(bipy) 3][通用电气(职业)2]·2水(5);[Zn(phen)3][Ge(HCit)2]·3H2O(6))。结果。从鲎培养液上清液中分离得到α- l -鼠李糖苷酶23.1次，产率为0.09%。酶的比活性为27.8单位/mL。经Sepharose 6B凝胶过滤，酶均相，分子量为50 kDa。结果表明，所考虑的配位化合物能够调节限制草α- l -鼠李糖苷酶的催化活性。它们都表现为活化剂或惰性物质，未观察到抑制作用。此外，化合物对酶的激活程度随其浓度的变化规律为:0.01% - 1 > 6≈5 > 3 >2≈4和0.1% - 1 > 4 >2 > 5≈6。3.化合物暴露时间对催化活性也有显著影响:在浓度为0.01%时，所有化合物的酶活性均为对照水平，而在浓度为0.1%时，化合物1(300%)、6(153%)和2(134%)存在时，酶活性急剧增加。其他人的行为处于控制水平。结论。获得了对微生物α- l -鼠李糖苷酶具有活化作用的新型络合金属化合物。在一系列生物活性金属与配体的配位化合物中，结构组织保证各组分作用协同的化合物是最有前途的酶效应器。
{"title":"Influence of New Types of Biscitratogermanates on Penicillium restrictum α-L-Rhamnosidase","authors":"O. Gudzenko, N. Borzova, L. Varbanets, I. Seifullina, E. Martsinko, O. Buchko, А. Pesaroglo","doi":"10.15407/microbiolj85.03.003","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.003","url":null,"abstract":"The intensive development of biotechnology in the last decade is largely determined by the growing requirement needs of both medicine and various industries for products of microbial synthesis, including glycosidases, in particular α-L-rhamnosidases. Their wide use to solve current biological-medical and chemical-technological problems stimulates researchers to search for compounds capable of influencing their catalytic activity. Therefore, the purpose of this work was to isolate and purify α-L-rhamnosidase from a new producer of Penicillium restrictum and to investigate multi-ligand germanium-3d-metal complexes with citric acid, phenanthroline, and bipyridine as effectors of its activity. Methods. The object of the study was α-L-rhamnosidase of P. restrictum. Its purification was carried out by gel filtration and ion exchange chromatography on TSK-gels and Sepharose 6B. The activity of α-L-rhamnosidase was determined using the Davis method with naringin as a substrate. As modifiers of enzyme activity, purposefully synthesized multiligand germanium-3d-metal complexes with citric acid, phenanthroline, and bipyridine ([Ni(bipy)3][Ge(HCit)2]·3H2O (1); [Ni(phen)3][Ge(HCit)2]·2H2O (2); [{Cu(bipy)2}2Ge(m-Cit)2]·12Н2О (3); [{Cu(phen)2}2Ge(m-Cit)2]·13H2O (4); [Zn(bipy)3][Ge(HCit)2]·2H2O (5); [Zn(phen)3][Ge(HCit)2]·3H2O (6)), were used. Results. From the supernatant of culture fluid of P. restrictum, α-L-rhamnosidase was isolated and purified 23.1 times with a yield of 0.09%. The specific activity of the enzyme was 27.8 units/mL. The enzyme was homogeneous according to gel filtration on Sepharose 6B and had a molecular mass of 50 kDa. It was established that the considered coordination compounds are able to regulate the catalytic activity of α-L-rhamnosidase of P. restrictum. All of them manifest themselves either as activators or as inert substances, no inhibition was observed. In addition, the dependence of the degree of enzyme activation by the compounds on their concentration is traced and corresponds to the following series: at a concentration of 0.01% — 1 > 6 ≈ 5 > 3 >2 ≈ 4 and at a concentration of 0.1% — 1 > 4 > 2 > 5 ≈ 6. 3. The catalytic activity is also significantly affected by the time of exposure to the compounds: at a concentration of 0.01% for 1h, the activity of the enzyme at the control level was observed for all compounds, whereas at a concentration of 0.1% for 24 h, the activity increased sharply in the presence of compounds 1 (300%), 6 (153%), and 2 (134%). The action of the others was at the control level. Conclusions. The obtained data on new complex metal compounds with an activating effect on microbial α-L-rhamnosidases. It has been established that compounds whose structural organization ensures the synergism of the action of all components are the most promising enzyme effectors in a series of coordination compounds of biologically active metals and ligands.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"2 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90244742","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.070
A. Buzun, B. Stegniy, A. Paliy, M. Spivak, M. Bogach, M. Stegniy, A. V. Kuzminov, O. Pavlichenko
African swine fever (ASF) remains an urgent problem of pig farming in Ukraine, the solution of which is possible only on the basis of deep scientific knowledge about the specific driving forces of the epizootic in its specific nozoareal. This is necessary in order to target anti-epizootic measures on the most vulnerable link of the epizootic chain in a specific nozoareal. The aim of the work was to develop a low-budget methodological base for experimental epizootology of low-virulent ASFV variants in Ukraine, in particular, to study the mechanisms of the formation of enzootic areas, quality control of anti-epizootic measures, and evaluation of the effectiveness of the antiviral drugs against them in Ukraine. Methods. Experimental and epizootological studies in the adaptation of suckling piglets to keeping in the biosecurity-level BSL-3 for laboratory animals (Patent UA No. 133248 dated 03/25/2019) were conducted at the laboratory base of the Odesa branch of NSC «IECVM». All procedures with infectious active biological materials in the current order were carried out in the BSL-3 module, built and certified with the assistance of the US Government in UAPRI (Odesa). The ASF agent strain «IECVM/Ternopil/2017» (infectious activity 4.0—7.5lg HAdU50/cm 3) circulating in the Ternopil region in 2017—2020 was used as a test virus. The presence of low-virulence variants of the ASFV pathogen in the studied samples was determined by a bioassay on suckling piglets, followed by three consecutive passages on a stable Vero line of the baby green monkey kidney cells. The isolated ASF virus was identified according to the methods and reagents recommended by the OIE Manual. Results. Intermittent passages «by the founder’s method» of dilutions 10-1 and 10-2 of the ASF virus strain «IECVM/Ternopil/2017» on piglets (n=20) and the culture of porcine alveolar macrophages («ASFVPAM») allowed us to identify highly-, moderately-, and low-virulent variants/clones in its composition. Verifi cation by bioassay on suckling piglets (n=5) of low-virulent clones of the agent, which were stabilized in Vero cell culture («ASFVVero»), showed that after intraperitoneal infection at a dose of 4.25 lgHAdU50/cm 3, they are capable of causing only a non-lethal (within 2 weeks) viral infection with a maximum daily rectal temperature of 39.4±0.22 °C and duration of fever on average 1.6±0.14 days (5 of 5 piglets). Clones with greater virulence («ASFVPAM») under similar conditions were able to cause a lethal infection with a maximum temperature of 40.7±0.37°C and duration of fever on average 3.9±0.27 days (17 of 20 piglets). Low-virulent clones were revealed by direct immunofluorescence in pulmonary and spleen smears of clinically healthy piglets on days 14 and 17 post-infection (p.i.); their antigens were visualized in Vero cells by indirect immunoperoxidase method after 48 h p.i. at dose about 0.01 lg HAdU50/cm 3. They caused «crumbly» hemadsorption of infected Vero cells and their virions
非洲猪瘟(ASF)仍然是乌克兰养猪业面临的一个紧迫问题，只有在深入了解该动物流行病在其特定疫源地的具体驱动力的基础上，才有可能解决这一问题。这是必要的，以便针对特定疫区动物流行病链中最脆弱的环节采取抗动物流行病措施。这项工作的目的是为乌克兰低毒性ASFV变异的动物流行病实验建立一个低预算的方法基础，特别是研究乌克兰动物流行病区形成的机制、抗动物流行病措施的质量控制以及抗病毒药物对它们的有效性评估。方法。哺乳仔猪适应实验动物生物安全水平BSL-3的实验和流行病学研究(专利号133248，日期为2019年3月25日)在NSC«IECVM»敖德萨分公司的实验室基地进行。当前订单中所有具有传染性活性生物材料的程序都在美国政府在UAPRI (Odesa)的协助下建造和认证的BSL-3模块中进行。采用2017 - 2020年在捷尔诺皮尔地区流行的非洲猪瘟病原毒株“IECVM/Ternopil/2017”(感染活性4.0 - 7.5 g HAdU50/ cm3)作为试验病毒。通过在哺乳仔猪上进行生物测定，然后在稳定的Vero细胞系上连续三次传代，确定研究样本中存在ASFV病原体的低毒力变体。根据世界动物卫生组织手册推荐的方法和试剂鉴定分离出的非洲猪瘟病毒。结果。通过“创始人方法”将ASF病毒株“IECVM/Ternopil/2017”稀释10-1和10-2在仔猪(n=20)和猪肺泡巨噬细胞(“ASFVPAM”)的培养中间歇性传递，使我们能够鉴定其组成中的高、中、低毒力变体/克隆。通过对5只哺乳仔猪(n=5)进行生物试验验证，该制剂的低毒力克隆在Vero细胞培养中稳定(“ASFVVero”)，结果表明，在4.25 lgHAdU50/ cm3的剂量下腹腔感染后，它们只能引起非致死(2周内)病毒感染，最高日直肠温度为39.4±0.22°C，平均发烧时间为1.6±0.14天(5头仔猪中有5头)。在类似条件下，毒力更强的克隆(“ASFVPAM”)能够引起致命感染，最高温度为40.7±0.37°C，平均发烧时间为3.9±0.27天(20头仔猪中有17头)。感染后第14天和第17天，临床健康仔猪肺和脾涂片采用直接免疫荧光法检测出低毒力克隆;在剂量为0.01 lg HAdU50/ cm3时，经间接免疫过氧化物酶法在Vero细胞中观察48 h。它们引起受感染的Vero细胞的“易碎”吸附，其病毒粒子具有典型的Asfarvirus视图和大小(210-220 nm)。所得数据可为分析非洲猪瘟病原在西凤梨地方性疫源地的生根机制、提出抗疫措施低成本质量控制概念和评价抗病毒药物抗非洲猪瘟活性提供依据。结论。已经开发出低成本的操作程序，使人们能够使用实验动物进行非洲猪瘟生物测定，并满足在实验非洲猪瘟流行病学的重要方面进行基于科学的研究的主要要求。在他们的帮助下，确认了乌克兰西部波迪利亚流行疫源地非洲猪瘟病毒种群的异质性(p<0.05, n=25)。该方法既适用于非洲猪瘟流行病学基础问题的研究，也适用于非洲猪瘟防控措施的质量控制。特别是，建议将其用于改善乌克兰农业出口计划的生物安全，乌克兰是一个受非洲猪瘟影响不利的国家。
{"title":"Experimental Epizotology of Low-Virulent Variants of African Swine Fever Virus","authors":"A. Buzun, B. Stegniy, A. Paliy, M. Spivak, M. Bogach, M. Stegniy, A. V. Kuzminov, O. Pavlichenko","doi":"10.15407/microbiolj85.03.070","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.070","url":null,"abstract":"African swine fever (ASF) remains an urgent problem of pig farming in Ukraine, the solution of which is possible only on the basis of deep scientific knowledge about the specific driving forces of the epizootic in its specific nozoareal. This is necessary in order to target anti-epizootic measures on the most vulnerable link of the epizootic chain in a specific nozoareal. The aim of the work was to develop a low-budget methodological base for experimental epizootology of low-virulent ASFV variants in Ukraine, in particular, to study the mechanisms of the formation of enzootic areas, quality control of anti-epizootic measures, and evaluation of the effectiveness of the antiviral drugs against them in Ukraine. Methods. Experimental and epizootological studies in the adaptation of suckling piglets to keeping in the biosecurity-level BSL-3 for laboratory animals (Patent UA No. 133248 dated 03/25/2019) were conducted at the laboratory base of the Odesa branch of NSC «IECVM». All procedures with infectious active biological materials in the current order were carried out in the BSL-3 module, built and certified with the assistance of the US Government in UAPRI (Odesa). The ASF agent strain «IECVM/Ternopil/2017» (infectious activity 4.0—7.5lg HAdU50/cm 3) circulating in the Ternopil region in 2017—2020 was used as a test virus. The presence of low-virulence variants of the ASFV pathogen in the studied samples was determined by a bioassay on suckling piglets, followed by three consecutive passages on a stable Vero line of the baby green monkey kidney cells. The isolated ASF virus was identified according to the methods and reagents recommended by the OIE Manual. Results. Intermittent passages «by the founder’s method» of dilutions 10-1 and 10-2 of the ASF virus strain «IECVM/Ternopil/2017» on piglets (n=20) and the culture of porcine alveolar macrophages («ASFVPAM») allowed us to identify highly-, moderately-, and low-virulent variants/clones in its composition. Verifi cation by bioassay on suckling piglets (n=5) of low-virulent clones of the agent, which were stabilized in Vero cell culture («ASFVVero»), showed that after intraperitoneal infection at a dose of 4.25 lgHAdU50/cm 3, they are capable of causing only a non-lethal (within 2 weeks) viral infection with a maximum daily rectal temperature of 39.4±0.22 °C and duration of fever on average 1.6±0.14 days (5 of 5 piglets). Clones with greater virulence («ASFVPAM») under similar conditions were able to cause a lethal infection with a maximum temperature of 40.7±0.37°C and duration of fever on average 3.9±0.27 days (17 of 20 piglets). Low-virulent clones were revealed by direct immunofluorescence in pulmonary and spleen smears of clinically healthy piglets on days 14 and 17 post-infection (p.i.); their antigens were visualized in Vero cells by indirect immunoperoxidase method after 48 h p.i. at dose about 0.01 lg HAdU50/cm 3. They caused «crumbly» hemadsorption of infected Vero cells and their virions ","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79135549","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.048
L. Lazarenko, L. Babenko, L. A. Safronova, O. Demchenko, V. V. Bila, G. M. Zaitseva, M. Spivak
The purpose of this study was to investigate the antimicrobial and immunomodulatory action of a probiotic composition of Bacillus subtilis and B. megatherium strains (UnicaUro, Sirion (Ukraine)) for experimental bacterial vaginitis. Methods. Experimental studies were conducted on female BALB/c mice; we used Staphylococcus aureus strain B-918 (ATCC 6538) to induce bacterial vaginitis. The strain was vaginally introduced into mice before treatment with probiotic bacteria. In the vagina of mice, aerobic and optionally anaerobic bacteria, including representatives of the genera Staphylococcus, Streptococcus, Lactobacillus, Bifidobacterium, Pseudomonas, coliform bacteria, and microscopic fungi were identified in different periods of observation using generally accepted microbiological methods. Serum antibody titer to S. aureus was determined by the bacterial agglutination reaction. The phagocytic activity and oxygen-dependent bactericidal activity of peritoneal exudate macrophages (PEM) were evaluated using generally accepted immunological methods. Results. The formation of bacterial vaginitis in the BALB/c mice line infected with S. aureus B-918 (ATCC 6538) was evidenced by the appearance of external clinical manifestations of the infectious and inflammatory process against the background of the increased number of aerobic and optionally anaerobic microorganisms, including representatives of the genus Staphylococcus and Streptococcus, microscopic fungi, and decreased number of lactobacilli in different observation periods. The probiotic introduction to mice with bacterial vaginitis led to a dynamic change in the vaginal microbiota: the number of aerobic and optionally anaerobic microorganisms decreased, primarily due to the normalization of the number of representatives of Staphylococcus genus accompanied by a decrease in the antibody titer to staphylococcus in the blood serum. The effective therapeutic action of the probiotic was confirmed by the gradual disappearance of the external clinical signs of the infectious-inflammatory process in the vagina against the background of the functional activity of PEM. Conclusions. The probiotic composition of B. subtilis and B. megatherium (UnicaUro, Sirion, Ukraine) is a promising antimicrobial formulation that may be used in the treatment of bacterial vaginitis; however, further studies are required to confirm its therapeutic, antimicrobial, and immunomodulatory efficacy.
{"title":"Antimicrobial and Immunomodulatory Action of Probiotic Composition of Bacilli on Bacterial Vaginitis in Mice","authors":"L. Lazarenko, L. Babenko, L. A. Safronova, O. Demchenko, V. V. Bila, G. M. Zaitseva, M. Spivak","doi":"10.15407/microbiolj85.03.048","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.048","url":null,"abstract":"The purpose of this study was to investigate the antimicrobial and immunomodulatory action of a probiotic composition of Bacillus subtilis and B. megatherium strains (UnicaUro, Sirion (Ukraine)) for experimental bacterial vaginitis. Methods. Experimental studies were conducted on female BALB/c mice; we used Staphylococcus aureus strain B-918 (ATCC 6538) to induce bacterial vaginitis. The strain was vaginally introduced into mice before treatment with probiotic bacteria. In the vagina of mice, aerobic and optionally anaerobic bacteria, including representatives of the genera Staphylococcus, Streptococcus, Lactobacillus, Bifidobacterium, Pseudomonas, coliform bacteria, and microscopic fungi were identified in different periods of observation using generally accepted microbiological methods. Serum antibody titer to S. aureus was determined by the bacterial agglutination reaction. The phagocytic activity and oxygen-dependent bactericidal activity of peritoneal exudate macrophages (PEM) were evaluated using generally accepted immunological methods. Results. The formation of bacterial vaginitis in the BALB/c mice line infected with S. aureus B-918 (ATCC 6538) was evidenced by the appearance of external clinical manifestations of the infectious and inflammatory process against the background of the increased number of aerobic and optionally anaerobic microorganisms, including representatives of the genus Staphylococcus and Streptococcus, microscopic fungi, and decreased number of lactobacilli in different observation periods. The probiotic introduction to mice with bacterial vaginitis led to a dynamic change in the vaginal microbiota: the number of aerobic and optionally anaerobic microorganisms decreased, primarily due to the normalization of the number of representatives of Staphylococcus genus accompanied by a decrease in the antibody titer to staphylococcus in the blood serum. The effective therapeutic action of the probiotic was confirmed by the gradual disappearance of the external clinical signs of the infectious-inflammatory process in the vagina against the background of the functional activity of PEM. Conclusions. The probiotic composition of B. subtilis and B. megatherium (UnicaUro, Sirion, Ukraine) is a promising antimicrobial formulation that may be used in the treatment of bacterial vaginitis; however, further studies are required to confirm its therapeutic, antimicrobial, and immunomodulatory efficacy.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"40 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"79988826","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 : 2023-06-21DOI: 10.15407/microbiolj85.03.012
L. Polishchuk, V. Lukyanchuk
The aim of the work is to identify strains of streptomycetes in the genomes of which there are nucleotide sequences similar to the gene cluster determining the synthesis of landomycin A (lan-cluster) and establish the level of similarity of their primary structures and organizations. Methods. Information on the sequences of the lan-cluster of Streptomyces cyanogenus S136 and chromosomal DNAs of S. cyanogenus S136, Streptomyces laculatispora NRRL B-24909, and Streptomyces griseoluteus JCM 4765 and their annotations are presented in the GenBank database on the NSBI server. A computerized analysis of the nucleotide sequences of streptomycetes was done using the program BLASTN from the server NSBI. Results. The localization of the lan-cluster in the terminal region of the S. cyanogenus S136 genome has been shown. The nucleotide sequences similar to the lan-cluster sequence of S. cyanogenus S136 were found in the genomes of two strains (S. laculatispora NRRL B-24909 and S. griseoluteus JCM 4765). Streptomycetes (S. cyanogenus S136, S. laculatispora NRRL B-24909, and S. griseoluteus JCM 4765) are not genetically related strains. Conclusions. There are newly found probable lan-clusters in the genomes of two streptomycetes strains (S. laculatispora NRRL B-24909 and S. griseoluteus JCM 4765). Landomycin clusters of three strains are organized according to the same scheme. The clusters of lan-genes are present in the genomes of genetically unrelated streptomycetes.
{"title":"Sequences Similar to the lan-Cluster (Streptomyces cyanogenus S136) Were Found in the Genomes of Other Streptomycetes","authors":"L. Polishchuk, V. Lukyanchuk","doi":"10.15407/microbiolj85.03.012","DOIUrl":"https://doi.org/10.15407/microbiolj85.03.012","url":null,"abstract":"The aim of the work is to identify strains of streptomycetes in the genomes of which there are nucleotide sequences similar to the gene cluster determining the synthesis of landomycin A (lan-cluster) and establish the level of similarity of their primary structures and organizations. Methods. Information on the sequences of the lan-cluster of Streptomyces cyanogenus S136 and chromosomal DNAs of S. cyanogenus S136, Streptomyces laculatispora NRRL B-24909, and Streptomyces griseoluteus JCM 4765 and their annotations are presented in the GenBank database on the NSBI server. A computerized analysis of the nucleotide sequences of streptomycetes was done using the program BLASTN from the server NSBI. Results. The localization of the lan-cluster in the terminal region of the S. cyanogenus S136 genome has been shown. The nucleotide sequences similar to the lan-cluster sequence of S. cyanogenus S136 were found in the genomes of two strains (S. laculatispora NRRL B-24909 and S. griseoluteus JCM 4765). Streptomycetes (S. cyanogenus S136, S. laculatispora NRRL B-24909, and S. griseoluteus JCM 4765) are not genetically related strains. Conclusions. There are newly found probable lan-clusters in the genomes of two streptomycetes strains (S. laculatispora NRRL B-24909 and S. griseoluteus JCM 4765). Landomycin clusters of three strains are organized according to the same scheme. The clusters of lan-genes are present in the genomes of genetically unrelated streptomycetes.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"49 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-06-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"88234071","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 : 2023-04-25DOI: 10.15407/microbiolj85.02.013
H. Huliaieva, N. Zhytkevych, T. Hnatiuk, M. Bohdan, I.P. Tokovenkov, V. Patyka
The search for effective and environmentally sound measures to fight against plant diseases caused by phytopathogenic microorganisms is of great importance. It is especially important to investigate alternative measures to protect cultivated plants that combine elements beneficial to human health such as iodine and selenium. Purpose. The study of physiological and biochemical changes in soybean leaves after artificial inoculation of plants with different strains of phytopathogenic bacteria on the background of pre-sowing treatment of seeds with a solution of iodine-selenium (I-Se) chelates. Methods. Soybean plants of the Artemis variety were grown in field conditions. Before sowing, the seeds were treated with a 1% I-Se chelated solution (I — 80 mg/L and Se citrate — 0.05 mg/L). The experimental plants were inoculated with phytopathogenic bacteria from the microbiological collection of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences, namely Pseudomonas savastanoi pv. glycinea IMВ B-9190, P. agglomerans IMВ B-9185, and P. syringae pv. syringae IMВ B-8531. The contents of chlorophyll-a, b and carotenoids in the leaves were determined by extraction in DMSO followed by spectrophotometry. Catalase activity was determined by the method of titrimetric permanganatometry with a 0.01M solution of KMnO4, the activity to non-specific peroxidases — according to Boyarkin’s method. Evaluation of the photochemical activity of photosynthesis according to the parameters F0, Fv/Fm, and RFd was carried out by the method of induction of chlorophyll fluorescence using a portable device «Floratest». Statistical processing of experimental data was carried out using the built-in functions of the Microsoft Excel program. Results. The increase of peroxidase activity of leaves was revealed after both pre-sowing treatment with 1% I-Se solution of intact plants and inoculation of them with different strains of bacterial pathogens in the following order: I-Se > I-Se+P. syringae pv. syringae 8531 > I-Se+P. agglomerans 9185 > I-Se+P. savastanoi pv. glycinea 9190. The catalase activity of leaves tissues increased only when infected with a specific pathogen P. savastanoi pv. glycinea 9190 (by 20.6%). After artificial inoculation with strains of both specific and facultative bacterial pathogens and the pre-sowing treatment with I-Se, there was observed an increase in the quantum efficiency of PSII (Fv /Fm) and fluorescence in decline index (Rfd). An increase in the content of chlorophyll-a (by 18%), b and carotenoids (by 7%) in the leaves after the pre-sowing treatment with I-Se has been shown. The content of chlorophyll-a in soybean leaves due to pre-sowing treatment with I-Se had the most significant increase after inoculation of P. agglomerans 9185 (20%). Due to the inoculation with a specific pathogen P. savastanoi pv. glycinea 9190 (after the pre-sowing treatment with I-Se), the content of chlorophyll-a tended to decrease, and the conte
{"title":"Physiological and Biochemical Changes in Soybean Plants Caused by Iodine-Selenium Chelates and Phytopathogenic Bacteria","authors":"H. Huliaieva, N. Zhytkevych, T. Hnatiuk, M. Bohdan, I.P. Tokovenkov, V. Patyka","doi":"10.15407/microbiolj85.02.013","DOIUrl":"https://doi.org/10.15407/microbiolj85.02.013","url":null,"abstract":"The search for effective and environmentally sound measures to fight against plant diseases caused by phytopathogenic microorganisms is of great importance. It is especially important to investigate alternative measures to protect cultivated plants that combine elements beneficial to human health such as iodine and selenium. Purpose. The study of physiological and biochemical changes in soybean leaves after artificial inoculation of plants with different strains of phytopathogenic bacteria on the background of pre-sowing treatment of seeds with a solution of iodine-selenium (I-Se) chelates. Methods. Soybean plants of the Artemis variety were grown in field conditions. Before sowing, the seeds were treated with a 1% I-Se chelated solution (I — 80 mg/L and Se citrate — 0.05 mg/L). The experimental plants were inoculated with phytopathogenic bacteria from the microbiological collection of the Zabolotny Institute of Microbiology and Virology of the National Academy of Sciences, namely Pseudomonas savastanoi pv. glycinea IMВ B-9190, P. agglomerans IMВ B-9185, and P. syringae pv. syringae IMВ B-8531. The contents of chlorophyll-a, b and carotenoids in the leaves were determined by extraction in DMSO followed by spectrophotometry. Catalase activity was determined by the method of titrimetric permanganatometry with a 0.01M solution of KMnO4, the activity to non-specific peroxidases — according to Boyarkin’s method. Evaluation of the photochemical activity of photosynthesis according to the parameters F0, Fv/Fm, and RFd was carried out by the method of induction of chlorophyll fluorescence using a portable device «Floratest». Statistical processing of experimental data was carried out using the built-in functions of the Microsoft Excel program. Results. The increase of peroxidase activity of leaves was revealed after both pre-sowing treatment with 1% I-Se solution of intact plants and inoculation of them with different strains of bacterial pathogens in the following order: I-Se > I-Se+P. syringae pv. syringae 8531 > I-Se+P. agglomerans 9185 > I-Se+P. savastanoi pv. glycinea 9190. The catalase activity of leaves tissues increased only when infected with a specific pathogen P. savastanoi pv. glycinea 9190 (by 20.6%). After artificial inoculation with strains of both specific and facultative bacterial pathogens and the pre-sowing treatment with I-Se, there was observed an increase in the quantum efficiency of PSII (Fv /Fm) and fluorescence in decline index (Rfd). An increase in the content of chlorophyll-a (by 18%), b and carotenoids (by 7%) in the leaves after the pre-sowing treatment with I-Se has been shown. The content of chlorophyll-a in soybean leaves due to pre-sowing treatment with I-Se had the most significant increase after inoculation of P. agglomerans 9185 (20%). Due to the inoculation with a specific pathogen P. savastanoi pv. glycinea 9190 (after the pre-sowing treatment with I-Se), the content of chlorophyll-a tended to decrease, and the conte","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"8 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76903917","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 : 2023-04-25DOI: 10.15407/microbiolj85.02.026
M. Bouacha, S. Besnaci, I. Boudiar
Objective. Honey is an extremely promising agent in the treatment of infected wounds of burned patients. This study aims to evaluate the antibacterial activity of 14 Algerian honey samples in comparison to Manuka honey towards pathogenic bacteria isolated from burn wound infections. Methods. The antibacterial effect of 14 Algerian honey samples and the Manuka honey was assessed against six multidrug-resistant bacteria: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus saprophyticus, and Enterococcus faecalis. Well agar diffusion, microdilution broth assay, and time-kill assay were used to evaluate the effects of honey samples on the growth of pathogenic bacteria. Results. The results obtained show that all tested honey samples have good antibacterial effects and there is no significant difference between Algerian honey samples and Manuka honey, except honey samples H12 and H13. The Gram-positive bacteria were more susceptible to honey samples than Gram-negative bacteria. The inhibitory diameters were between 14 to 38 mm for Gram-positive bacteria and from 8 to 28 mm for Gram-negative bacteria. The minimal inhibitory concentration of Algerian honey was between 5 and 80% (v/v) and minimal bactericidal concentration was between 10 and 80 % (v/v). However, the minimal inhibitory concentration of Manuka honey was between 5 and 40% (v/v) and minimal bactericidal concentration was between 10 and 80% (v/v). The MBC/MIC ratio was from 1 to 2, which proves that both Algeria honeys and Manuka honey have a bactericidal effect rather than a bacteriostatic effect. A time-kill assay showed that the inhibition effect of honey samples started after the first 3 hours of incubation. Honey samples 3 and 7 inhibited the growth of S. aureus and S. saprophyticus in 15 hours; however, they inhibited the growth of the other pathogenic bacteria in 18 hours. Conclusions. This study proposes honey as an extremely promising treatment against multidrug-resistant bacteria from burn infections.
{"title":"Comparative Study of the Antibacterial Activity of Algerian Honeys and Manuka Honey Toward Pathogenic Bacteria from Burn Wound Infections","authors":"M. Bouacha, S. Besnaci, I. Boudiar","doi":"10.15407/microbiolj85.02.026","DOIUrl":"https://doi.org/10.15407/microbiolj85.02.026","url":null,"abstract":"Objective. Honey is an extremely promising agent in the treatment of infected wounds of burned patients. This study aims to evaluate the antibacterial activity of 14 Algerian honey samples in comparison to Manuka honey towards pathogenic bacteria isolated from burn wound infections. Methods. The antibacterial effect of 14 Algerian honey samples and the Manuka honey was assessed against six multidrug-resistant bacteria: Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Staphylococcus aureus, Staphylococcus saprophyticus, and Enterococcus faecalis. Well agar diffusion, microdilution broth assay, and time-kill assay were used to evaluate the effects of honey samples on the growth of pathogenic bacteria. Results. The results obtained show that all tested honey samples have good antibacterial effects and there is no significant difference between Algerian honey samples and Manuka honey, except honey samples H12 and H13. The Gram-positive bacteria were more susceptible to honey samples than Gram-negative bacteria. The inhibitory diameters were between 14 to 38 mm for Gram-positive bacteria and from 8 to 28 mm for Gram-negative bacteria. The minimal inhibitory concentration of Algerian honey was between 5 and 80% (v/v) and minimal bactericidal concentration was between 10 and 80 % (v/v). However, the minimal inhibitory concentration of Manuka honey was between 5 and 40% (v/v) and minimal bactericidal concentration was between 10 and 80% (v/v). The MBC/MIC ratio was from 1 to 2, which proves that both Algeria honeys and Manuka honey have a bactericidal effect rather than a bacteriostatic effect. A time-kill assay showed that the inhibition effect of honey samples started after the first 3 hours of incubation. Honey samples 3 and 7 inhibited the growth of S. aureus and S. saprophyticus in 15 hours; however, they inhibited the growth of the other pathogenic bacteria in 18 hours. Conclusions. This study proposes honey as an extremely promising treatment against multidrug-resistant bacteria from burn infections.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"131 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-04-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87616192","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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