Pub Date : 2023-11-21DOI: 10.20535/ibb.2023.7.4.288529
S. Rogalsky, D. Hodyna, I. Semenyuta, Mykhaylo Frasinyuk, O. Tarasyuk, Sergii Riabov, Larisa Kobrina, Igor Tetko, L. Metelytsia
Background. The bacterial pathogen Acinetobacter baumannii is one of the most dangerous multi-drug-resistant (MDR) microorganisms, which causes numerous bacterial infections. Nowadays, there is an urgent need for new broad-spectrum antibacterial agents with specific molecular mechanisms of action. Long-chain 1-alkylpyridinium salts are efficient cationic biocides which can inhibit enzymes involved in the biosynthesis of bacterial fatty acids. Incorporating these compounds into inclusion complexes with cyclic oligosaccharide β-cyclodextrin can reduce their relatively high acute toxicity. Objective. The aim of this research was to develop new anti-A. baumannii agents based on hydrophobic 1-alkylpyridinium salt and its inclusion complex with sulfobutyl ether b-cyclodextrin (SBECD). Methods. Hydrophobic cationic biocide 1-dodecylpyridinium tetrafluoroborate (PyrC12-BF4) and its inclusion complex with SBECD have been synthesized. The structure of the SBECD/PyrC12-BF4 complex was characterized by 1H Nuclear Magnetic Resonance spectroscopy, as well as UV spectroscopy. In vitro antibacterial activity of the synthesized compounds was estimated against MDR clinical isolates of A. baumannii using standard disc diffusion method. Acute toxicity studies were performed on Daphnia magna model hydrobiont. Molecular docking was performed using the crystal structure of the A. baumannii 3-oxoacyl-[acyl-carrier-protein] reductase (FabG). Results. The results of 1H NMR study revealed the formation of an inclusion complex between SBECD and PyrC12-BF4. The cationic biocide demonstrated high activity against four tested antibiotic-resistant strains of A. baumannii, whereas the SBECD/PyrC12-BF4 complex was active against only two bacterial strains. Molecular docking of 1-dodecylpyridinium ligand into the active site of the A. baumannii (FabG) showed complex formation at an allosteric site located between subunits C, D. The acute toxicity (LC50) of PyrC12-BF4 and its inclusion complex was found to be 0.007 and 0.033 ml/g, respectively. Conclusions. Hydrophobic cationic biocide PyrC12-BF4 has high antibacterial activity against MDR A. baumannii. The inhibition of the active site FabG may be one of the possible mechanisms of anti-A. baumannii activity of the PyrC12-BF4. The SBECD/PyrC12-BF4 inclusion complex showed an almost 5-fold reduction in acute toxicity compared to PyrC12-BF4, while retaining activity against certain tested A. baumannii bacterial strains.
{"title":"Antibacterial Activity of 1-Dodecylpyridinium Tetrafluoroborate and Its Inclusion Complex With Sulfobutyl Ether-β-Cyclodextrin Against MDR Acinetobacter baumannii Strains","authors":"S. Rogalsky, D. Hodyna, I. Semenyuta, Mykhaylo Frasinyuk, O. Tarasyuk, Sergii Riabov, Larisa Kobrina, Igor Tetko, L. Metelytsia","doi":"10.20535/ibb.2023.7.4.288529","DOIUrl":"https://doi.org/10.20535/ibb.2023.7.4.288529","url":null,"abstract":"Background. The bacterial pathogen Acinetobacter baumannii is one of the most dangerous multi-drug-resistant (MDR) microorganisms, which causes numerous bacterial infections. Nowadays, there is an urgent need for new broad-spectrum antibacterial agents with specific molecular mechanisms of action. Long-chain 1-alkylpyridinium salts are efficient cationic biocides which can inhibit enzymes involved in the biosynthesis of bacterial fatty acids. Incorporating these compounds into inclusion complexes with cyclic oligosaccharide β-cyclodextrin can reduce their relatively high acute toxicity. Objective. The aim of this research was to develop new anti-A. baumannii agents based on hydrophobic 1-alkylpyridinium salt and its inclusion complex with sulfobutyl ether b-cyclodextrin (SBECD). Methods. Hydrophobic cationic biocide 1-dodecylpyridinium tetrafluoroborate (PyrC12-BF4) and its inclusion complex with SBECD have been synthesized. The structure of the SBECD/PyrC12-BF4 complex was characterized by 1H Nuclear Magnetic Resonance spectroscopy, as well as UV spectroscopy. In vitro antibacterial activity of the synthesized compounds was estimated against MDR clinical isolates of A. baumannii using standard disc diffusion method. Acute toxicity studies were performed on Daphnia magna model hydrobiont. Molecular docking was performed using the crystal structure of the A. baumannii 3-oxoacyl-[acyl-carrier-protein] reductase (FabG). Results. The results of 1H NMR study revealed the formation of an inclusion complex between SBECD and PyrC12-BF4. The cationic biocide demonstrated high activity against four tested antibiotic-resistant strains of A. baumannii, whereas the SBECD/PyrC12-BF4 complex was active against only two bacterial strains. Molecular docking of 1-dodecylpyridinium ligand into the active site of the A. baumannii (FabG) showed complex formation at an allosteric site located between subunits C, D. The acute toxicity (LC50) of PyrC12-BF4 and its inclusion complex was found to be 0.007 and 0.033 ml/g, respectively. Conclusions. Hydrophobic cationic biocide PyrC12-BF4 has high antibacterial activity against MDR A. baumannii. The inhibition of the active site FabG may be one of the possible mechanisms of anti-A. baumannii activity of the PyrC12-BF4. The SBECD/PyrC12-BF4 inclusion complex showed an almost 5-fold reduction in acute toxicity compared to PyrC12-BF4, while retaining activity against certain tested A. baumannii bacterial strains.","PeriodicalId":502500,"journal":{"name":"Innovative Biosystems and Bioengineering","volume":"67 4","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-11-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139251846","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 : 2023-11-16DOI: 10.20535/ibb.2023.7.4.278905
N. Vorobey, K. Kukol, S. Kots, P. Pukhtaievych, Volodymyr Patyka
Background. With the increase in soybean cultivation areas, inoculants are becoming increasingly sought after. They are not only compatible with the original preparations for seed treatment of soybean but also offer the possibility of applying them for several days or even months before sowing into the soil. Objective. The viability of new strains of nodule bacteria Bradyrhizobium japonicum (strain PC07 and strain B78) was investigated on the surface of soybean seeds treated with fungicides Fever and Maxim XL during the extended storage of inoculated seeds. Additionally, their ability for nitrogen fixation under symbiotic conditions was evaluated. Methods. Сultivation, serial dilution method, determination of bacterial titer of inoculants, quantification of colony forming units (CFU), gas chromatography. Results. In laboratory conditions, on the soybean seeds of Almaz and Vasylkivska varieties, the viability of the nodule bacteria B. japonicum PC07 and B78, which are more resistant to the active substances of these fungicides in pure culture, decreased. This reduction was dependent on the storage period, the fungicide used, and the strain of inoculant. One day after inoculation on the seeds without the use of fungicide, 68.6–75.4% CFU of the initial number was retained. Seeds treated with the fungicide Fever contained 36.5–38.3% CFU, and those treated with Maxim XL contained 26.2–27.0% CFU. The use of inoculants based on fungicide-resistant strains of B. japonicum PC07 and B78 also provided high cell viability – 2.41–2.8×106 (on the seeds without treatment), 1.40–1.70×106 (with Fever) and 0.8–1.17×106 (with Maxim XL) CFU/seed after 5 days of storage. The nitrogen-fixing activity of root nodules in soybean plants of Almaz and Vasylkivska varieties, whose seeds were treated with Fever, decreased on both varieties by 18.4–22.4% and 32.1–39.5%, respectively and for treatment with Maxim XL, the reduction was 24.5–33.7% and 47.7–75.2%, respectively, during the storage of seeds for 5 and 7 days before sowing, compared to control variants (seeds without fungicide treatment). Conclusions. The utilization of fungicide-resistant strains of B. japonicum PC07 and B78 for bacterization of soybean seeds treated with fungicides Fever and Maxim XL provides a high inoculation titer during 5 days of storage. Viability of microbial cells on seeds treated with fungicides significantly diminishes during storage for more than 7–14 days. Insufficient titer of CFU of B. japonicum strains PC07 and B78 on the seed surface after 14 days of storage can considerably impede the efficacy of biopreparations. Therefore, it is crucial to seek substances that can enhance the resistance of rhizobia on seeds to the adverse effects of fungicide seed treatment, ensuring a longer period of their viability.
{"title":"Viability of Nodule Bacteria Bradyrhizobium japonicummon Soybean Seeds by Treatment With Fungicides During Extended Storage","authors":"N. Vorobey, K. Kukol, S. Kots, P. Pukhtaievych, Volodymyr Patyka","doi":"10.20535/ibb.2023.7.4.278905","DOIUrl":"https://doi.org/10.20535/ibb.2023.7.4.278905","url":null,"abstract":"Background. With the increase in soybean cultivation areas, inoculants are becoming increasingly sought after. They are not only compatible with the original preparations for seed treatment of soybean but also offer the possibility of applying them for several days or even months before sowing into the soil. Objective. The viability of new strains of nodule bacteria Bradyrhizobium japonicum (strain PC07 and strain B78) was investigated on the surface of soybean seeds treated with fungicides Fever and Maxim XL during the extended storage of inoculated seeds. Additionally, their ability for nitrogen fixation under symbiotic conditions was evaluated. Methods. Сultivation, serial dilution method, determination of bacterial titer of inoculants, quantification of colony forming units (CFU), gas chromatography. Results. In laboratory conditions, on the soybean seeds of Almaz and Vasylkivska varieties, the viability of the nodule bacteria B. japonicum PC07 and B78, which are more resistant to the active substances of these fungicides in pure culture, decreased. This reduction was dependent on the storage period, the fungicide used, and the strain of inoculant. One day after inoculation on the seeds without the use of fungicide, 68.6–75.4% CFU of the initial number was retained. Seeds treated with the fungicide Fever contained 36.5–38.3% CFU, and those treated with Maxim XL contained 26.2–27.0% CFU. The use of inoculants based on fungicide-resistant strains of B. japonicum PC07 and B78 also provided high cell viability – 2.41–2.8×106 (on the seeds without treatment), 1.40–1.70×106 (with Fever) and 0.8–1.17×106 (with Maxim XL) CFU/seed after 5 days of storage. The nitrogen-fixing activity of root nodules in soybean plants of Almaz and Vasylkivska varieties, whose seeds were treated with Fever, decreased on both varieties by 18.4–22.4% and 32.1–39.5%, respectively and for treatment with Maxim XL, the reduction was 24.5–33.7% and 47.7–75.2%, respectively, during the storage of seeds for 5 and 7 days before sowing, compared to control variants (seeds without fungicide treatment). Conclusions. The utilization of fungicide-resistant strains of B. japonicum PC07 and B78 for bacterization of soybean seeds treated with fungicides Fever and Maxim XL provides a high inoculation titer during 5 days of storage. Viability of microbial cells on seeds treated with fungicides significantly diminishes during storage for more than 7–14 days. Insufficient titer of CFU of B. japonicum strains PC07 and B78 on the seed surface after 14 days of storage can considerably impede the efficacy of biopreparations. Therefore, it is crucial to seek substances that can enhance the resistance of rhizobia on seeds to the adverse effects of fungicide seed treatment, ensuring a longer period of their viability.","PeriodicalId":502500,"journal":{"name":"Innovative Biosystems and Bioengineering","volume":"53 3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-11-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139268393","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 : 2023-11-12DOI: 10.20535/ibb.2023.7.4.284774
O. Slyvchuk, Oksana Shtapenko, Iryna Yaremchuk, Serhiy Kornyat, Y. Dzen
Background. Zinc is essential for male reproductive function as it is necessary for spermatozoa maturation, capacitation, acrosome reaction, and fertilization. It has been established that the use of chelates compounds of metals with amino acids, nucleotides, peptides, and carbohydrates is not only more effective but also economically justified. Objective. The aim of this study was to evaluate and compare the effect of different concentrations of Zn glutamic amino acid chelate (ZnGlu) on boar spermatozoa motility, viability parameters, and prooxidant-antioxidant homeostasis during in vitro incubation. Methods. Freshly ejaculated boar semen, after the addition of "Ecosperm" diluent, was divided into groups: a control group and three experimental groups. Zinc glutamate was supplemented to the experimental samples at concentrations of 1.0, 2.0, and 5.0 µg/ml, respectively. Semen samples were stored at 18 °C for 4 days. Every 24 hours, sperm motility and viability and the antioxidant status were assessed by the level of diene conjugates (DC), concentration of malondialdehydere (MDA), and the activities of catalase and superoxide dismutase. Results. It was observed that boar sperm motility decreased with the extension of storage time, while the addition of 2.0 and 5.0 µg ZnGlu significantly improved sperm total motility and the percentage of vitality spermatozoa during 48–96 hours of incubation. Our studies also demonstrate that ZnGlu possess a protective effect in alleviating oxidative stress in boar sperm in vitro. The addition of ZnGlu significantly reduced the content of MDA and DC in ejaculate samples in all experimental groups during incubation, compared to the control group. Moreover, the activity of superoxide dismutase and catalase increased after adding ZnGlu to the boar semen, especially at concentrations of 2.0 and 5.0 mg/ml (P < 0.05). Conclusions. These data demonstrate that the supplementation of zinc glutamate enhances the antioxidant defence system of sperm and improves quality of boar semen in vitro.
{"title":"Effect of Zinc Supplementation to Diluent Boar Semen on Sperm Characteristics and Activity of Antioxidant Enzymes","authors":"O. Slyvchuk, Oksana Shtapenko, Iryna Yaremchuk, Serhiy Kornyat, Y. Dzen","doi":"10.20535/ibb.2023.7.4.284774","DOIUrl":"https://doi.org/10.20535/ibb.2023.7.4.284774","url":null,"abstract":"Background. Zinc is essential for male reproductive function as it is necessary for spermatozoa maturation, capacitation, acrosome reaction, and fertilization. It has been established that the use of chelates compounds of metals with amino acids, nucleotides, peptides, and carbohydrates is not only more effective but also economically justified. Objective. The aim of this study was to evaluate and compare the effect of different concentrations of Zn glutamic amino acid chelate (ZnGlu) on boar spermatozoa motility, viability parameters, and prooxidant-antioxidant homeostasis during in vitro incubation. Methods. Freshly ejaculated boar semen, after the addition of \"Ecosperm\" diluent, was divided into groups: a control group and three experimental groups. Zinc glutamate was supplemented to the experimental samples at concentrations of 1.0, 2.0, and 5.0 µg/ml, respectively. Semen samples were stored at 18 °C for 4 days. Every 24 hours, sperm motility and viability and the antioxidant status were assessed by the level of diene conjugates (DC), concentration of malondialdehydere (MDA), and the activities of catalase and superoxide dismutase. Results. It was observed that boar sperm motility decreased with the extension of storage time, while the addition of 2.0 and 5.0 µg ZnGlu significantly improved sperm total motility and the percentage of vitality spermatozoa during 48–96 hours of incubation. Our studies also demonstrate that ZnGlu possess a protective effect in alleviating oxidative stress in boar sperm in vitro. The addition of ZnGlu significantly reduced the content of MDA and DC in ejaculate samples in all experimental groups during incubation, compared to the control group. Moreover, the activity of superoxide dismutase and catalase increased after adding ZnGlu to the boar semen, especially at concentrations of 2.0 and 5.0 mg/ml (P < 0.05). Conclusions. These data demonstrate that the supplementation of zinc glutamate enhances the antioxidant defence system of sperm and improves quality of boar semen in vitro.","PeriodicalId":502500,"journal":{"name":"Innovative Biosystems and Bioengineering","volume":"7 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-11-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139279409","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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