Pub Date : 2023-01-17DOI: 10.15407/microbiolj84.04.077
O. V. Hadzevych, A. Paliy, B. T. Stehniĭ, A.B. Stehnii, О.N. Chechet, D. V. Hadzevych, A. Palii, O. Pavlichenko, R. Severyn, R. Petrov, L. P. Livoshchenko
The aquatic environment is an integral part of biocenosis that directly affects its condition and safety in terms of epidemiology and epizootology. The study of the aquatic environment for the presence of pathogens and the quantitative characteristics of sanitary-indicative microorganisms is extremely important. The obtained data allow us to assess and predict the risks of infections, and to develop a plan of measures to prevent the spread of certain pathogens. The aim of the work. To analyze the microbial state of the aquatic environment in different hydro ecosystems of fish farms in the Kharkiv region and to assess the presence of microbiological risks to public health. Methods. The research objects were 150 samples of water taken from different hydro ecosystems in the Kharkiv region. Water was taken from closed water supply systems (n=30) and from ponds (n=120), where commercial fish is bred for sale. The presence and number of sanitary-indicative microorganisms and pathogenic bacteria were determined by the bacteriological (cultural) method. Resistances to antibacterial drugs in selected sanitary-indicative microorganisms were determined using the Agar disk-diffusion method. Estimation of the reliability of the difference between the compared indicators was determined using Student’s t-test. Results. The dominant sanitary-indicative microorganisms in the aquatic environment of fish farming were bacteria of the genus Citrobacter spp., Aeromonas spp., and Pseudomonas spp. The total bacterial contamination of water bodies ranged from 1.9±0.50×104 to 2.1±1.20×105 CFU in 1 cm3 of water. No pathogenic to humans bacteria have been detected. Isolated sanitary-indicative microorganisms had significant resistance to antibacterial drugs. Resistance to penicillins, sulfonamides, and nitrofurans was the highest (p=0.0001). The percentage of penicillin resistance strains ranged from 81.5% to 87.0%, sulfonamide — from 74.1% to 94.4%, and nitrofuran — from 55.5% to 66.7%. Fluoroquinolone and cephalosporin resistance varied depending on the type of antibacterial substance, but it did not exceed 29.6%. Conclusions. According to the research results for the aquatic environment of fish farms in the Kharkiv region, no pathogenic microorganisms were detected. However, it has been established that sanitary-indicating microorganisms (Citrobacter spp., Aeromonas spp., Pseudomonas spp.), which were dominant and had polyresistance to antibacterial drugs, may be risk factors for human health. Thus, the hydro ecosystems of fish farms have favorable conditions for the accumulation of bacterial strains resistant to antibiotics. Therefore, the use of antibacterial drugs should be scientifi cally justifi ed and strictly controlled.
{"title":"Antibiotic Resistance of Microbiotas of Fishery Enterprises Hydro Ecosystems","authors":"O. V. Hadzevych, A. Paliy, B. T. Stehniĭ, A.B. Stehnii, О.N. Chechet, D. V. Hadzevych, A. Palii, O. Pavlichenko, R. Severyn, R. Petrov, L. P. Livoshchenko","doi":"10.15407/microbiolj84.04.077","DOIUrl":"https://doi.org/10.15407/microbiolj84.04.077","url":null,"abstract":"The aquatic environment is an integral part of biocenosis that directly affects its condition and safety in terms of epidemiology and epizootology. The study of the aquatic environment for the presence of pathogens and the quantitative characteristics of sanitary-indicative microorganisms is extremely important. The obtained data allow us to assess and predict the risks of infections, and to develop a plan of measures to prevent the spread of certain pathogens. The aim of the work. To analyze the microbial state of the aquatic environment in different hydro ecosystems of fish farms in the Kharkiv region and to assess the presence of microbiological risks to public health. Methods. The research objects were 150 samples of water taken from different hydro ecosystems in the Kharkiv region. Water was taken from closed water supply systems (n=30) and from ponds (n=120), where commercial fish is bred for sale. The presence and number of sanitary-indicative microorganisms and pathogenic bacteria were determined by the bacteriological (cultural) method. Resistances to antibacterial drugs in selected sanitary-indicative microorganisms were determined using the Agar disk-diffusion method. Estimation of the reliability of the difference between the compared indicators was determined using Student’s t-test. Results. The dominant sanitary-indicative microorganisms in the aquatic environment of fish farming were bacteria of the genus Citrobacter spp., Aeromonas spp., and Pseudomonas spp. The total bacterial contamination of water bodies ranged from 1.9±0.50×104 to 2.1±1.20×105 CFU in 1 cm3 of water. No pathogenic to humans bacteria have been detected. Isolated sanitary-indicative microorganisms had significant resistance to antibacterial drugs. Resistance to penicillins, sulfonamides, and nitrofurans was the highest (p=0.0001). The percentage of penicillin resistance strains ranged from 81.5% to 87.0%, sulfonamide — from 74.1% to 94.4%, and nitrofuran — from 55.5% to 66.7%. Fluoroquinolone and cephalosporin resistance varied depending on the type of antibacterial substance, but it did not exceed 29.6%. Conclusions. According to the research results for the aquatic environment of fish farms in the Kharkiv region, no pathogenic microorganisms were detected. However, it has been established that sanitary-indicating microorganisms (Citrobacter spp., Aeromonas spp., Pseudomonas spp.), which were dominant and had polyresistance to antibacterial drugs, may be risk factors for human health. Thus, the hydro ecosystems of fish farms have favorable conditions for the accumulation of bacterial strains resistant to antibiotics. Therefore, the use of antibacterial drugs should be scientifi cally justifi ed and strictly controlled.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"33 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84760233","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 : 2023-01-17DOI: 10.15407/microbiolj84.04.009
S. López-Pazos, F.M. Chavarrio Cañas, A. C. Rojas Arias
Bacillus thuringiensis (Bt) produces Cry toxins against pest insects. Cry proteins are conformed by domains related to pore formation and recognition of protein receptors. Plant-induced systemic resistance (ISR) is triggered due to pest attack, it could be activated by Bacillus sp. Tecia solanivora (Ts) is a potato pest, susceptible to Cry1Ac and Cry1B proteins. This paper indicates the endorsement of Bt kurstaki HD-1 (BtkHD1) in relation to Ts control (Cry1Ac and Cry1B proteins), potato growth promotion, and plant ISR due to pests related to the BtkHD1-potato system. To ensure that ongoing quality control of BtkHD1 was maintained, crystal synthesis (microscopy), cry1 genes presence, and Cry protein production were checked. Bioassays Ts larvae and potato plantlets and an in silico analysis of the hybrid Cry1Ac-Cry1Ba protein and potato ISR related to the BtkHD1 infl uence were performed. Bioassay on Ts larvae shows an LC50 of 536 ng/cm2 of diet. A potato growth promotion assay revealed the effect of BtkHD1 on the length and dry weight of stems. The prospective analysis took into account relevant factors affecting the biological function of the hybrid protein focused on domain II. In silico identification of 15 BtkHD1 proteins and 68 potato proteins related to plant ISR due to pests was completed. This project serves to validation of toxicity on Ts larvae and potato growth effect based on BtkHD1, including a forward analysis of the hybrid Cry1Ac1-Cry1Ba1, and proteins associated with this strain and potato for eliciting plant ISR due to pests.
{"title":"Insecticidal and Potato Growth Stimulation Activity of Bacillus thuringiensis kurstaki HD-1","authors":"S. López-Pazos, F.M. Chavarrio Cañas, A. C. Rojas Arias","doi":"10.15407/microbiolj84.04.009","DOIUrl":"https://doi.org/10.15407/microbiolj84.04.009","url":null,"abstract":"Bacillus thuringiensis (Bt) produces Cry toxins against pest insects. Cry proteins are conformed by domains related to pore formation and recognition of protein receptors. Plant-induced systemic resistance (ISR) is triggered due to pest attack, it could be activated by Bacillus sp. Tecia solanivora (Ts) is a potato pest, susceptible to Cry1Ac and Cry1B proteins. This paper indicates the endorsement of Bt kurstaki HD-1 (BtkHD1) in relation to Ts control (Cry1Ac and Cry1B proteins), potato growth promotion, and plant ISR due to pests related to the BtkHD1-potato system. To ensure that ongoing quality control of BtkHD1 was maintained, crystal synthesis (microscopy), cry1 genes presence, and Cry protein production were checked. Bioassays Ts larvae and potato plantlets and an in silico analysis of the hybrid Cry1Ac-Cry1Ba protein and potato ISR related to the BtkHD1 infl uence were performed. Bioassay on Ts larvae shows an LC50 of 536 ng/cm2 of diet. A potato growth promotion assay revealed the effect of BtkHD1 on the length and dry weight of stems. The prospective analysis took into account relevant factors affecting the biological function of the hybrid protein focused on domain II. In silico identification of 15 BtkHD1 proteins and 68 potato proteins related to plant ISR due to pests was completed. This project serves to validation of toxicity on Ts larvae and potato growth effect based on BtkHD1, including a forward analysis of the hybrid Cry1Ac1-Cry1Ba1, and proteins associated with this strain and potato for eliciting plant ISR due to pests.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"30 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76012986","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 : 2023-01-17DOI: 10.15407/microbiolj84.04.098
O. Shevchenko, A. Kharina, H. Snihur, V. Holovan, T. Shevchenko, I. Budzanivska, Hao Liping
This work covers important aspects of the occurrence and viability of various viruses in the two most common reusable waste resources: wastewater and biomass waste. Detection of human, bacterial and plant viruses in these wastes are summarized. Historically, human viruses have been monitored in wastewater for decades. Evidence suggests that wastewater mostly contains fecal-orally transmitted viruses, which are abundant and diverse. Recently, an increasing occurrence of SARS-CoV2 in sewage water with the spreading epidemics has been confirmed but lacking biological proof of infectivity yet. Besides human pathogens, wastewater is shown to be rich in bacteriophages and plant viruses as well, which supposedly enter the water from human guts. Viruses serving as water quality indicators are also discussed here. Lastly, we focus on biomass waste treatment, showing the presence of some common and stable plant viruses which may supposedly survive the technological process.
{"title":"Virus Occurrence and Survival in Reusable Resources: A Minireview","authors":"O. Shevchenko, A. Kharina, H. Snihur, V. Holovan, T. Shevchenko, I. Budzanivska, Hao Liping","doi":"10.15407/microbiolj84.04.098","DOIUrl":"https://doi.org/10.15407/microbiolj84.04.098","url":null,"abstract":"This work covers important aspects of the occurrence and viability of various viruses in the two most common reusable waste resources: wastewater and biomass waste. Detection of human, bacterial and plant viruses in these wastes are summarized. Historically, human viruses have been monitored in wastewater for decades. Evidence suggests that wastewater mostly contains fecal-orally transmitted viruses, which are abundant and diverse. Recently, an increasing occurrence of SARS-CoV2 in sewage water with the spreading epidemics has been confirmed but lacking biological proof of infectivity yet. Besides human pathogens, wastewater is shown to be rich in bacteriophages and plant viruses as well, which supposedly enter the water from human guts. Viruses serving as water quality indicators are also discussed here. Lastly, we focus on biomass waste treatment, showing the presence of some common and stable plant viruses which may supposedly survive the technological process.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"151 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"77759276","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 : 2023-01-17DOI: 10.15407/microbiolj84.04.003
O. Gudzenko, K. V. Avdiyuk, N. Borzova, V. Ivanytsia, L. Varbanets
For a long time, the main interest in the marine environment, considered extreme, was the isolation and identification of natural products with biological properties, and for that, numerous organisms and chemical structures have been studied. Thus, marine bacteria isolated from various substrates, such as sediments, seawater, and mangrove detritus, are producers of enzymes with different activities, i.e., amylase, cellulase, alginate lyase, chitinase, glucosidase, inulinase, keratinase, ligninase, xylanase, and others. Nowadays, researchers are also focusing on the enzymes produced in the marine environment that can present special properties. Therefore, the aim of this study was to investigate the ability of marine strains of microorganisms to exhibit cellulase, β-mannanase, keratinase, and caseinolytic activities. Methods. Enzymatic activities were studied in the culture liquid supernatant. To determine β-mannanase and cellulase activities, guar gum galactomannan and Na-carboxymethylcellulose respectively were used as substrates. Casein and crushed defatted feathers served as substrates for the determination of proteolytic activity. Results. Growing 10 cultures of microorganisms on a nutrient medium containing chicken feathers as the sole source of carbon and nitrogen (nutrient medium 1) did not give positive results. When using medium 2, active growth was observed in four of the studied strains (51, 52, 54, 247) in the supernatant of culture liquid (CLS), the activity of which both to keratin (6.0—16.0 U/mL) and casein (0.025—0.33 U/mL) was found. In the CLS of only six of the 10 studied cultures (7, 20, 51, 52, 50, 247), cellulase and β-mannanase activities were observed. The highest cellulase activity was found in culture 20 (1.8 U/mL). The activity of culture 7 was somewhat lower (1.0 U/mL). An insignificant activity was noted in cultures 54 (0.06 U/mL), 56, and 50 (0.05 U/mL). Trace levels of activity were observed in culture 247. Conclusions. Strains 7, 20, 247, and 51, for the first time isolated from the Black Sea, are promising for further studies as producers of cellulase, β-mannanase, keratinase, and caseinolytic enzymes.
{"title":"Keratinase, Caseinolitic, Cellulase and β-Mananase Activities of Bacteria Isolated from the Black Sea","authors":"O. Gudzenko, K. V. Avdiyuk, N. Borzova, V. Ivanytsia, L. Varbanets","doi":"10.15407/microbiolj84.04.003","DOIUrl":"https://doi.org/10.15407/microbiolj84.04.003","url":null,"abstract":"For a long time, the main interest in the marine environment, considered extreme, was the isolation and identification of natural products with biological properties, and for that, numerous organisms and chemical structures have been studied. Thus, marine bacteria isolated from various substrates, such as sediments, seawater, and mangrove detritus, are producers of enzymes with different activities, i.e., amylase, cellulase, alginate lyase, chitinase, glucosidase, inulinase, keratinase, ligninase, xylanase, and others. Nowadays, researchers are also focusing on the enzymes produced in the marine environment that can present special properties. Therefore, the aim of this study was to investigate the ability of marine strains of microorganisms to exhibit cellulase, β-mannanase, keratinase, and caseinolytic activities. Methods. Enzymatic activities were studied in the culture liquid supernatant. To determine β-mannanase and cellulase activities, guar gum galactomannan and Na-carboxymethylcellulose respectively were used as substrates. Casein and crushed defatted feathers served as substrates for the determination of proteolytic activity. Results. Growing 10 cultures of microorganisms on a nutrient medium containing chicken feathers as the sole source of carbon and nitrogen (nutrient medium 1) did not give positive results. When using medium 2, active growth was observed in four of the studied strains (51, 52, 54, 247) in the supernatant of culture liquid (CLS), the activity of which both to keratin (6.0—16.0 U/mL) and casein (0.025—0.33 U/mL) was found. In the CLS of only six of the 10 studied cultures (7, 20, 51, 52, 50, 247), cellulase and β-mannanase activities were observed. The highest cellulase activity was found in culture 20 (1.8 U/mL). The activity of culture 7 was somewhat lower (1.0 U/mL). An insignificant activity was noted in cultures 54 (0.06 U/mL), 56, and 50 (0.05 U/mL). Trace levels of activity were observed in culture 247. Conclusions. Strains 7, 20, 247, and 51, for the first time isolated from the Black Sea, are promising for further studies as producers of cellulase, β-mannanase, keratinase, and caseinolytic enzymes.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"38 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90381322","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.051
G. Fadieienko, I. Kushnir, V. Chernova, T. Solomentseva, Y. Nikiforova, O. Kurinna, V.Yu. Galchynska, T. Bondar'
Nonalcoholic fatty liver disease (NAFLD) pathogenesis displays a close relation with intestinal dysbiosis. Thus, the aim of this study was to investigate the intestinal microbiota (IM) composition and to determine the correlation of changes in its main phylotypes with the amount and activity of adipose tissue in NAFLD patients. Methods. The prospective study enrolled 114 NAFLD patients with metabolic disorders and 30 healthy subjects as the control group. Along with routine examination, the authors assessed intestinal microbiota composition by identifying total bacterial DNA and DNA of Bacteroidetes, Firmicutes, and Actinobacteria by means of a quantitative real-time PCR. Results. NAFLD patients showed a signifi cant decrease in the relative amount of Bacteroidetes with a simultaneous increase in the Firmicutes and an increase in Firmicutes/Bacteroidetes ratio compared with healthy subjects (p<0.05). NAFLD patients with concomitant overweight and obesity displayed a more significant imbalance of IM with an increase in the Firmicutes/Bacteroidetes ratio due to the inhibition of Bacteroidetes, compared with patients of normal body mass index. The revealed changes in the main phylotypes of IM in the examined patients were proven linked not only to an increase in body weight but also to the amount and activity of visceral adipose tissue. Furthermore, deviations in the gut microbiota composition had an impact on the formation and severity of steatosis. Conclusions. The study revealed an imbalance of IM in NAFLD patients. Further research in gut microbiota will help to elucidate their role in NAFLD pathogenesis and to lay a foundation for the development of individualized treatment.
{"title":"Dependence of Intestinal Microbiota Composition on Distribution and Activity of Adipose Tissue in Nonalcoholic Fatty Liver Disease","authors":"G. Fadieienko, I. Kushnir, V. Chernova, T. Solomentseva, Y. Nikiforova, O. Kurinna, V.Yu. Galchynska, T. Bondar'","doi":"10.15407/microbiolj84.03.051","DOIUrl":"https://doi.org/10.15407/microbiolj84.03.051","url":null,"abstract":"Nonalcoholic fatty liver disease (NAFLD) pathogenesis displays a close relation with intestinal dysbiosis. Thus, the aim of this study was to investigate the intestinal microbiota (IM) composition and to determine the correlation of changes in its main phylotypes with the amount and activity of adipose tissue in NAFLD patients. Methods. The prospective study enrolled 114 NAFLD patients with metabolic disorders and 30 healthy subjects as the control group. Along with routine examination, the authors assessed intestinal microbiota composition by identifying total bacterial DNA and DNA of Bacteroidetes, Firmicutes, and Actinobacteria by means of a quantitative real-time PCR. Results. NAFLD patients showed a signifi cant decrease in the relative amount of Bacteroidetes with a simultaneous increase in the Firmicutes and an increase in Firmicutes/Bacteroidetes ratio compared with healthy subjects (p<0.05). NAFLD patients with concomitant overweight and obesity displayed a more significant imbalance of IM with an increase in the Firmicutes/Bacteroidetes ratio due to the inhibition of Bacteroidetes, compared with patients of normal body mass index. The revealed changes in the main phylotypes of IM in the examined patients were proven linked not only to an increase in body weight but also to the amount and activity of visceral adipose tissue. Furthermore, deviations in the gut microbiota composition had an impact on the formation and severity of steatosis. Conclusions. The study revealed an imbalance of IM in NAFLD patients. Further research in gut microbiota will help to elucidate their role in NAFLD pathogenesis and to lay a foundation for the development of individualized treatment.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"26 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"84338478","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.017
O. Kisten, K. Hetman, E. V. Koval, I. Hretskyi, L.F. Zyryanova, L. Tyshchenko, N. Fedosova, N. Cheremshenko, A. Chumak
The level of oxygen mass transfer (KV) is an important parameter influencing the growth rate of aerobic microorganisms and the synthesis of metabolites. It is mainly determined by the agitation and the aeration rates in the fermenter. Aim. To study changes in pH, optical density (OD), and hemagglutinating (lectin) activity (HAA) of culture fluid (CF) of Bacillus subtilis strain IMV B-7724, a producer of extracellular cytotoxic lectin (ECL), during its cultivation in a laboratory fermenter at different agitation and aeration rates as well as to determine and compare the HAA, carbohydrate specifi city, and cytotoxic properties of the corresponding samples of the preparation isolated from CF. Methods. Batch antifoam-free fermentations were performed by culturing the strain in the modified Gause medium with galactose in two identical lab-scale fermenters with a working volume of 2.5 L at 37ºC for 48—72 h according to three fermentation variants. Variant 1: n — 400 rpm for the whole cultivation, the air supply to the CF — through a sparger at 0.1 vvm until the 39th h with further gradual decrease, KV — 4.2±0.3 g O2·L−1·h−1. Variant 2: n — 400 rpm for the first 24 h, then a gradual decrease to 200 rpm, air supply — through a sparger at 0.1 rpm for the first 12 h, followed by its switching into the fermenter free space, corresponding KV — from 4.2±0.3 to 0.3±0.1 g O2·L−1·h−1. Variant 3: n — 400 rpm and air supply to the fermenter free space during the whole cultivation, KV — 4.0±0.3 g O2·L−1·h−1. A number of biological properties of strain CF and isolated lectin samples were evaluated by biochemical, spectrophotometric, immunological, and culture methods. Statistical analysis was performed using Student’s t-test. Results. The maximum increase in the OD of CF relative to the initial values (28 and 21-fold) at the end of the period of the rapid growth of the strain (at 9th h), the μmax values of 0.33 and 0.41 h−1, and pH not lower than 6.7 and 6.3 units were observed for fermentation variants 1 and 2, respectively. In the case of variant 2, the HAA of CF reached 32 hemagglutinating units (HAU), and the samples isolated from it had a lectin activity of 512±64 HAU, whereas for variant 1 such values were lower:16 and 32±8 HAA, respectively; carbohydrate specificity of preparations to bovine submandibular gland mucin was the same, i.e. 0.2±0.1 mg/mL. In contrast to the above, a slower increase in the OD of the CF, a decrease in μmax, and significant acid formation (15-fold at the 9th h, 0.25 h−1, and pH decrease to 5.8 units, respectively) were observed for variant 3; in this case, the level of HAA of CF was minimal (2—4 HAU) and was absent in the corresponding isolated samples. The probable reason for such differences was the limited mass transfer in the CF due to the isolating effect of the foam layer on its surface formed as a result of intensive agitation. Conclusions. The rapid growth of the strain and an increase in the HAA of CF were observed during cu
氧传质水平(KV)是影响好氧微生物生长速度和代谢产物合成的重要参数。这主要取决于发酵箱内的搅拌和曝气率。的目标。研究产生细胞外毒性凝集素(ECL)的枯草芽孢杆菌(Bacillus subtilis)菌株IMV B-7724培养液(CF)在不同搅拌和通风率下的pH、光密度(OD)和血凝(凝集素)活性(HAA)的变化，并测定和比较从枯草芽孢杆菌中分离的相应样品的HAA、碳水化合物特异性和细胞毒性。根据三种发酵方式，将菌株在含半乳糖的改良Gause培养基中，在两个相同的实验室规模发酵罐中，工作体积为2.5 L, 37ºC下培养48-72 h，进行批量无泡沫发酵。变异1:在整个培养过程中，以0.1 vvm的速度通过分散器给CF供气，直至第39 h, KV - 4.2±0.3 g O2·L−1·h−1。变体2:前24小时n - 400转/分，然后逐渐降低到200转/分，供气-在前12小时以0.1转/分的速度通过一个分散器，然后将其切换到发酵罐自由空间，相应的KV -从4.2±0.3到0.3±0.1 g O2·L−1·h−1。变体3:在整个培养过程中，n - 400rpm并向发酵罐自由空间供气，KV - 4.0±0.3 g O2·L−1·h−1。通过生化、分光光度、免疫学和培养等方法，对菌株CF和分离的凝集素样品的生物学特性进行了评价。统计学分析采用Student’s t检验。结果。菌株快速生长结束时(第9 h)， CF的OD值较初始值(28倍和21倍)最大，μmax值分别为0.33和0.41 h−1,pH值分别不低于6.7和6.3个单位。在变异2中，CF的HAA达到32个血凝素单位(HAU)，其凝集素活性为512±64个HAU，而变异1的这一数值较低，分别为16和32±8个HAA;各制剂对牛颌下腺黏液的糖特异性相同，均为0.2±0.1 mg/mL。与此相反，变异3的CF OD增加较慢，μmax降低，酸生成显著(第9 h 15倍，0.25 h−1,pH降至5.8单位);在这种情况下，CF的HAA水平极低(2-4 HAU)，并且在相应的分离样品中不存在。产生这种差异的可能原因是CF中的传质有限，这是由于剧烈搅拌形成的泡沫层在其表面上的隔离作用。结论。在实验室规模的发酵罐培养过程中，通过分散器向CF供气，保持最大的氧传质水平，直到达到最大OD，然后在进一步培养过程中逐渐降低到规定的水平，观察到菌株的快速生长和CF的HAA的增加。
{"title":"Features of the Synthesis of Extracellular Cytotoxic Lectin Bacillus subtilis IMV B-7724, Depending on the Cultivation Conditions in the Laboratory Fermenter","authors":"O. Kisten, K. Hetman, E. V. Koval, I. Hretskyi, L.F. Zyryanova, L. Tyshchenko, N. Fedosova, N. Cheremshenko, A. Chumak","doi":"10.15407/microbiolj84.03.017","DOIUrl":"https://doi.org/10.15407/microbiolj84.03.017","url":null,"abstract":"The level of oxygen mass transfer (KV) is an important parameter influencing the growth rate of aerobic microorganisms and the synthesis of metabolites. It is mainly determined by the agitation and the aeration rates in the fermenter. Aim. To study changes in pH, optical density (OD), and hemagglutinating (lectin) activity (HAA) of culture fluid (CF) of Bacillus subtilis strain IMV B-7724, a producer of extracellular cytotoxic lectin (ECL), during its cultivation in a laboratory fermenter at different agitation and aeration rates as well as to determine and compare the HAA, carbohydrate specifi city, and cytotoxic properties of the corresponding samples of the preparation isolated from CF. Methods. Batch antifoam-free fermentations were performed by culturing the strain in the modified Gause medium with galactose in two identical lab-scale fermenters with a working volume of 2.5 L at 37ºC for 48—72 h according to three fermentation variants. Variant 1: n — 400 rpm for the whole cultivation, the air supply to the CF — through a sparger at 0.1 vvm until the 39th h with further gradual decrease, KV — 4.2±0.3 g O2·L−1·h−1. Variant 2: n — 400 rpm for the first 24 h, then a gradual decrease to 200 rpm, air supply — through a sparger at 0.1 rpm for the first 12 h, followed by its switching into the fermenter free space, corresponding KV — from 4.2±0.3 to 0.3±0.1 g O2·L−1·h−1. Variant 3: n — 400 rpm and air supply to the fermenter free space during the whole cultivation, KV — 4.0±0.3 g O2·L−1·h−1. A number of biological properties of strain CF and isolated lectin samples were evaluated by biochemical, spectrophotometric, immunological, and culture methods. Statistical analysis was performed using Student’s t-test. Results. The maximum increase in the OD of CF relative to the initial values (28 and 21-fold) at the end of the period of the rapid growth of the strain (at 9th h), the μmax values of 0.33 and 0.41 h−1, and pH not lower than 6.7 and 6.3 units were observed for fermentation variants 1 and 2, respectively. In the case of variant 2, the HAA of CF reached 32 hemagglutinating units (HAU), and the samples isolated from it had a lectin activity of 512±64 HAU, whereas for variant 1 such values were lower:16 and 32±8 HAA, respectively; carbohydrate specificity of preparations to bovine submandibular gland mucin was the same, i.e. 0.2±0.1 mg/mL. In contrast to the above, a slower increase in the OD of the CF, a decrease in μmax, and significant acid formation (15-fold at the 9th h, 0.25 h−1, and pH decrease to 5.8 units, respectively) were observed for variant 3; in this case, the level of HAA of CF was minimal (2—4 HAU) and was absent in the corresponding isolated samples. The probable reason for such differences was the limited mass transfer in the CF due to the isolating effect of the foam layer on its surface formed as a result of intensive agitation. Conclusions. The rapid growth of the strain and an increase in the HAA of CF were observed during cu","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"28 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87064912","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.003
O. Gudzenko, V. Ivanytsia, L. Varbanets
Despite the fact that in recent years there has been a certain enhancing interest in the study of marine microorganisms, nevertheless, marine bacteria as producers of biologically active substances, in particular enzymes, are still poorly studied. The marine biota is significantly different from the terrestrial one; therefore, there is a high probability of detecting in the marine environment different from terrestrial bacteria producers of enzymes with unique specificity and activity, for the needs of modern biotechnology. Proteolytic enzymes play an important role in these studies. Since the majority of microbial producers are characterized by a number of serious deficiencies, in particular, most of the elastase producers described in the literature are pathogenic for humans, the search for new, effective producers continues to be an urgent problem, given that highly active producers of proteolytic enzymes, in particular elastase, are generally absent in Ukraine. In this regard, the purpose of this work was to screen microorganisms isolated from the Black Sea for the presence of effective producers of proteolytic enzymes. Methods. We used methods of determining proteolytic (caseinilytic, elastolytic, fibrinolytic, fibrinogenolytic) activity. Results. The study of the enzymatic activity of the isolates showed that on the 10th day of cultivation in the supernatant of the culture liquid, caseinolytic activity was not detected only in one isolate 56, whereas very insignificant activity was observed in isolates 7, 20, and 50. The maximum activity was detected in isolate 247 (0.2 units/mL), and lower one - in isolates 46 (0.16 U/mL), 52 (0.15 U/mL), 51 (0.135 U/mL), 54 (0.08 U/mL), and 44 (0.05 U/mL). Of the 10 studied isolates, elastase activity was found only in four of them. The highest activity was found in isolates 51 and 54 (20.83 and 19.96 U/mL, respectively). Lower levels of activity (15.62 U/mL and 12.15 U/mL, respectively) were shown by isolates 52 and 247. The studied isolates also differed in their ability to hydrolyze fibrin and fibrinogen. T e highest fi brinolytic activity (2.33 U/mL) was found in isolates 46 and 54, significantly lower in isolate 20 (0.5 U/mL) and isolate 44 (0.33 U/mL). The rest isolates did not show fibrinolytic activity. As for fibrinogenolytic activity, it was noted in 6 studied cultures. The highest levels of activity were observed in isolate 51 (1.16 U/mL). Lower activity was found in isolates 54 (0.66 U/mL), 7 (0.5 U/mL), and 247 (0.33 U/mL). In isolate 50, it was minimal (0.083 U/mL). Conclusions. No correlation was found between elastase, fibrinolytic and fibrinogenic activity in the studied isolates. Thus, isolates 51, 54 and, to a lesser extent, 52 and 247 synthesize elastase activity. The highest fibrinolytic activity was in isolates 46 and 54, and fibrinogenolytic activity was in isolate 51. It was shown that the Black Sea is rich in marine bacterial species, which can be effective producers of a number o
{"title":"Bacteria of the Black Sea Are Producers of Proteolytic Enzymes","authors":"O. Gudzenko, V. Ivanytsia, L. Varbanets","doi":"10.15407/microbiolj84.03.003","DOIUrl":"https://doi.org/10.15407/microbiolj84.03.003","url":null,"abstract":"Despite the fact that in recent years there has been a certain enhancing interest in the study of marine microorganisms, nevertheless, marine bacteria as producers of biologically active substances, in particular enzymes, are still poorly studied. The marine biota is significantly different from the terrestrial one; therefore, there is a high probability of detecting in the marine environment different from terrestrial bacteria producers of enzymes with unique specificity and activity, for the needs of modern biotechnology. Proteolytic enzymes play an important role in these studies. Since the majority of microbial producers are characterized by a number of serious deficiencies, in particular, most of the elastase producers described in the literature are pathogenic for humans, the search for new, effective producers continues to be an urgent problem, given that highly active producers of proteolytic enzymes, in particular elastase, are generally absent in Ukraine. In this regard, the purpose of this work was to screen microorganisms isolated from the Black Sea for the presence of effective producers of proteolytic enzymes. Methods. We used methods of determining proteolytic (caseinilytic, elastolytic, fibrinolytic, fibrinogenolytic) activity. Results. The study of the enzymatic activity of the isolates showed that on the 10th day of cultivation in the supernatant of the culture liquid, caseinolytic activity was not detected only in one isolate 56, whereas very insignificant activity was observed in isolates 7, 20, and 50. The maximum activity was detected in isolate 247 (0.2 units/mL), and lower one - in isolates 46 (0.16 U/mL), 52 (0.15 U/mL), 51 (0.135 U/mL), 54 (0.08 U/mL), and 44 (0.05 U/mL). Of the 10 studied isolates, elastase activity was found only in four of them. The highest activity was found in isolates 51 and 54 (20.83 and 19.96 U/mL, respectively). Lower levels of activity (15.62 U/mL and 12.15 U/mL, respectively) were shown by isolates 52 and 247. The studied isolates also differed in their ability to hydrolyze fibrin and fibrinogen. T e highest fi brinolytic activity (2.33 U/mL) was found in isolates 46 and 54, significantly lower in isolate 20 (0.5 U/mL) and isolate 44 (0.33 U/mL). The rest isolates did not show fibrinolytic activity. As for fibrinogenolytic activity, it was noted in 6 studied cultures. The highest levels of activity were observed in isolate 51 (1.16 U/mL). Lower activity was found in isolates 54 (0.66 U/mL), 7 (0.5 U/mL), and 247 (0.33 U/mL). In isolate 50, it was minimal (0.083 U/mL). Conclusions. No correlation was found between elastase, fibrinolytic and fibrinogenic activity in the studied isolates. Thus, isolates 51, 54 and, to a lesser extent, 52 and 247 synthesize elastase activity. The highest fibrinolytic activity was in isolates 46 and 54, and fibrinogenolytic activity was in isolate 51. It was shown that the Black Sea is rich in marine bacterial species, which can be effective producers of a number o","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"111 12 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"76134241","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.039
N. Hrynchuk, L. Zelena, T. Bukhtiarova, N. Vrynchanu, L. Ishchenko, E. Vazhnichaya
Staphylococcus aureus is a widespread opportunistic pathogen, causing community-acquired and nosocomial infections with both acute and chronic recurrent courses. The process of chronicity of the disease is provided by biofilms. Features of the structure and functioning of biofilms, in particular the presence of matrix, quorum sensing systems, persistent cells, and efflux pumps, provide microbial communities with resistance to antimicrobial drugs under their action in therapeutic concentrations. The insufficient eff ectiveness of modern antimicrobial chemotherapy against biofi lm microorganisms indicates the urgency of the problem to search for compounds with antibiofilm activity that can affect various stages of the biofilm formation and the formed biofilm. The aim of the study is to establish the antibiofilm activity of 4-(adamantyl-1)-1-(1-aminobutyl) benzol against methicillin-resistant S. aureus (MRSA) and to determine the mechanism of its action. Methods. The ability of adamantane-containing compound 4-(adamantyl-1)-1-(1-aminobutyl) benzol (AM-166) to prevent biofilm formation and destroy the formed biofilm of S. aureus was investigated on polystyrene plates by the sorption of gentian violet on its structures followed with desorption of the dye into the organic solvent. The viability of S. aureus cells at the first stage of biofilm formation and in the composition of mature biofilms was evaluated using specific dyes for living (acridine orange) and non-viable (propidium iodide) cells. Detection of genes responsible for antibiotic resistance and biofi lm formation was performed by the polymerase chain reaction (PCR) with detection of PCR products in agarose gel. Evaluation of the effect of AM-166 on the expression of genes regulating the biofilm formation (ica, agrA, sarA, and sigB) was investigated by the real-time PCR and semi-quantitative PCR. Results. It was found that the compound AM-166 shows activity against S. aureus biofilm formation. The most pronounced effect was registered at a concentration of 5.0 minimum inhibitory concentration (MIC) (92.3%.) Under the action of AM-166 on the formed 2-day biofilms, their destruction was marked: the biomass decreases by 30.9% at 5.0 MIC. According to the results of fluorescence microscopy, the adamantane derivative at 5.0 MIC helps to reduce the number of viable cells at different stages of formation of the S. aureus biofilm. The results of molecular genetic studies indicate that the ica gene expression is significantly inhibited by the action of subinhibitory concentrations of the compound AM-116. No significant changes in the expression of sarA, agrA, and sigB genes were registered. Conclusions. Experiments on the effect of adamantane derivative on S. aureus biofilms showed that the most pronounced activity of AM-116 was observed at the stage of biofilm formation, as evidenced by the inhibition of transcriptional activity of the ica gene responsible for early stages of the biofilm formation, i
{"title":"Antibiofilm Activity of 4-(Adamantyl-1)-1-(1-Aminobutyl) Benzol against Methicillin-Resistant Staphylococcus aureus","authors":"N. Hrynchuk, L. Zelena, T. Bukhtiarova, N. Vrynchanu, L. Ishchenko, E. Vazhnichaya","doi":"10.15407/microbiolj84.03.039","DOIUrl":"https://doi.org/10.15407/microbiolj84.03.039","url":null,"abstract":"Staphylococcus aureus is a widespread opportunistic pathogen, causing community-acquired and nosocomial infections with both acute and chronic recurrent courses. The process of chronicity of the disease is provided by biofilms. Features of the structure and functioning of biofilms, in particular the presence of matrix, quorum sensing systems, persistent cells, and efflux pumps, provide microbial communities with resistance to antimicrobial drugs under their action in therapeutic concentrations. The insufficient eff ectiveness of modern antimicrobial chemotherapy against biofi lm microorganisms indicates the urgency of the problem to search for compounds with antibiofilm activity that can affect various stages of the biofilm formation and the formed biofilm. The aim of the study is to establish the antibiofilm activity of 4-(adamantyl-1)-1-(1-aminobutyl) benzol against methicillin-resistant S. aureus (MRSA) and to determine the mechanism of its action. Methods. The ability of adamantane-containing compound 4-(adamantyl-1)-1-(1-aminobutyl) benzol (AM-166) to prevent biofilm formation and destroy the formed biofilm of S. aureus was investigated on polystyrene plates by the sorption of gentian violet on its structures followed with desorption of the dye into the organic solvent. The viability of S. aureus cells at the first stage of biofilm formation and in the composition of mature biofilms was evaluated using specific dyes for living (acridine orange) and non-viable (propidium iodide) cells. Detection of genes responsible for antibiotic resistance and biofi lm formation was performed by the polymerase chain reaction (PCR) with detection of PCR products in agarose gel. Evaluation of the effect of AM-166 on the expression of genes regulating the biofilm formation (ica, agrA, sarA, and sigB) was investigated by the real-time PCR and semi-quantitative PCR. Results. It was found that the compound AM-166 shows activity against S. aureus biofilm formation. The most pronounced effect was registered at a concentration of 5.0 minimum inhibitory concentration (MIC) (92.3%.) Under the action of AM-166 on the formed 2-day biofilms, their destruction was marked: the biomass decreases by 30.9% at 5.0 MIC. According to the results of fluorescence microscopy, the adamantane derivative at 5.0 MIC helps to reduce the number of viable cells at different stages of formation of the S. aureus biofilm. The results of molecular genetic studies indicate that the ica gene expression is significantly inhibited by the action of subinhibitory concentrations of the compound AM-116. No significant changes in the expression of sarA, agrA, and sigB genes were registered. Conclusions. Experiments on the effect of adamantane derivative on S. aureus biofilms showed that the most pronounced activity of AM-116 was observed at the stage of biofilm formation, as evidenced by the inhibition of transcriptional activity of the ica gene responsible for early stages of the biofilm formation, i","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"1 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"89553855","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.029
T. V. Bulyhina, L.D. Varbanest
Azospirillum brasilense is a gram-negative, nitrogen-fixing bacterium that colonizes the rhizosphere of various types of grasses and cereals. Lipopolysaccharides (LPS) are a class of complex glycolipids present in the cell membrane of gramnegative bacteria and mediate plant-bacteria interactions. Although the effects of LPS of pathogenic plant bacteria on the induction of plant defense mechanisms have been characterized, the role of LPS of beneficial rhizobacteria on plant growth is less clear. Therefore, a very important point is the study of the chemical, biological, and functional activities of A. brasilense LPS, which was the aim of this work. Methods. A. brasilense LPSs were isolated from dry bacterial mass by the phenol-water method. The carbohydrates were analyzed by the Dubois method, nucleic acids — by Spirin, protein content — by Lowry and 2-keto-3-deoxyoctonic acid (KDO) — by Osborn. Pyrogenicity of LPS was tested observing the rules of bioethics in rabbits. Serological studies were performed by the Ouchterlony method. The identification of monosaccharides and fatty acids in LPS preparations was carried out on an Agilent 6890N/5973 inert chromatomass spectrometry system. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAAG electrophoresis) was performed according to Laemmli. Results. LPS of 3 strains of A. brasilense were isolated from dry bacterial mass and purifi ed from nucleic acids by ultracentrifugation. The purified LPSs were characterized by different relative yields from 2.44% to 4.75%, which is slightly higher than other strains of the A. brasilense (1—3%). The studied preparations were characterized by a rather high content of carbohydrates from 50.1% to 72.1%. All LPS contained up to 0.17% KDO, which is a specific component of the LPS of gram-negative bacteria. Analysis of the monosaccharide composition indicates that the LPSs of the studied A. brasilense strains turned out to be heterogeneous. At the same time, such monosaccharides as mannose, galactose, glucose, and heptose were recorded in the LPS of all tested strains. The study of the fatty acid composition of LPS showed the presence of fatty acids containing from 14 to 18 carbon atoms. Нydroxylated, saturated, and monounsaturated acids and their cis isomers were found. In the investigated LPS, the dominant fatty acids were 16:0, 18:1, 14:0(3-OH), and 16:0(3-OH), which coincides with the literature data. The research of the pyrogenic effect of LPS of A. brasilense studied strains showed that LPS solutions are apyrogenic. The double immunodiffusion reaction in Ouchterlon agar showed that all tested LPS in homologous systems exhibited ancultitigenic activity. Serological cross-reactions can be used as an approach in classifying different bacteria. Thus, we found that antisera to A. brasilense 18-2 and 61 react with all LPSs of the studied strains, which may indicate the presence of common antigenic determinants in them and that these str
{"title":"Characterization of Azospirillum brasilense Lipopolysaccharides","authors":"T. V. Bulyhina, L.D. Varbanest","doi":"10.15407/microbiolj84.03.029","DOIUrl":"https://doi.org/10.15407/microbiolj84.03.029","url":null,"abstract":"Azospirillum brasilense is a gram-negative, nitrogen-fixing bacterium that colonizes the rhizosphere of various types of grasses and cereals. Lipopolysaccharides (LPS) are a class of complex glycolipids present in the cell membrane of gramnegative bacteria and mediate plant-bacteria interactions. Although the effects of LPS of pathogenic plant bacteria on the induction of plant defense mechanisms have been characterized, the role of LPS of beneficial rhizobacteria on plant growth is less clear. Therefore, a very important point is the study of the chemical, biological, and functional activities of A. brasilense LPS, which was the aim of this work. Methods. A. brasilense LPSs were isolated from dry bacterial mass by the phenol-water method. The carbohydrates were analyzed by the Dubois method, nucleic acids — by Spirin, protein content — by Lowry and 2-keto-3-deoxyoctonic acid (KDO) — by Osborn. Pyrogenicity of LPS was tested observing the rules of bioethics in rabbits. Serological studies were performed by the Ouchterlony method. The identification of monosaccharides and fatty acids in LPS preparations was carried out on an Agilent 6890N/5973 inert chromatomass spectrometry system. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS-PAAG electrophoresis) was performed according to Laemmli. Results. LPS of 3 strains of A. brasilense were isolated from dry bacterial mass and purifi ed from nucleic acids by ultracentrifugation. The purified LPSs were characterized by different relative yields from 2.44% to 4.75%, which is slightly higher than other strains of the A. brasilense (1—3%). The studied preparations were characterized by a rather high content of carbohydrates from 50.1% to 72.1%. All LPS contained up to 0.17% KDO, which is a specific component of the LPS of gram-negative bacteria. Analysis of the monosaccharide composition indicates that the LPSs of the studied A. brasilense strains turned out to be heterogeneous. At the same time, such monosaccharides as mannose, galactose, glucose, and heptose were recorded in the LPS of all tested strains. The study of the fatty acid composition of LPS showed the presence of fatty acids containing from 14 to 18 carbon atoms. Нydroxylated, saturated, and monounsaturated acids and their cis isomers were found. In the investigated LPS, the dominant fatty acids were 16:0, 18:1, 14:0(3-OH), and 16:0(3-OH), which coincides with the literature data. The research of the pyrogenic effect of LPS of A. brasilense studied strains showed that LPS solutions are apyrogenic. The double immunodiffusion reaction in Ouchterlon agar showed that all tested LPS in homologous systems exhibited ancultitigenic activity. Serological cross-reactions can be used as an approach in classifying different bacteria. Thus, we found that antisera to A. brasilense 18-2 and 61 react with all LPSs of the studied strains, which may indicate the presence of common antigenic determinants in them and that these str","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"92 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"90424036","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 : 2022-12-17DOI: 10.15407/microbiolj84.03.092
H. Tsekhmister, А.S. Kyslynska
Fungal diseases cause signifi cant damage to agriculture. Plectosphaerella melonis (syn. Acremonium cucurbitacearum and Nodulisporium melonis) is a pathogen of cultivated plant diseases in Spain, Italy, Japan, USA, Egypt, and Ukraine. This review discusses the main results of research related to this phytopathogen. By morphological and cultural features, P. melonis is a morphologically intermediate species between A. strictum and A. charticola, however, 5.8S-ITS regionbased phylogenetic analysis showed that P. melonis is a monophyletic taxon more closely related to Plectosphaerella than to other species of the genus Acremonium. The most susceptible plants are at the stage of germination; however, the development of the disease is manifested in the fruiting period. For a comprehensive assessment of virulence, real leaf area (RLA) of the first two leaves, lesion of hypocotyl (RH), root collar (RSR), primary (R1R) and secondary roots (R2R) are measured. P. melonis affects the root system, in particular the root collar and hypocotyl, and colonizes the epidermis and cortex of the root centrographically towards the stem. The range of host plants includes Cucurbitaceae, however, peppers, tomatoes, basil, and parsley are infected as well. Plants vary in susceptibility depending on the species and even variety. The pathogenic response of plants differs depending on the growing conditions (protected and open soil), the interaction between the pathogen and competing microorganisms, and other ecological and trophic relationships. The main means of control are the use of long-term crop rotations and the selection of resistant varieties. In Ukraine, a strain of the antagonist fungus Trichoderma viride was selected, which is an effective means for controlling P. melonis 502. The aim of our work was to establish the role of P. melonis in the development of diseases of cultivated plants.
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