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Antibacterial Activity of 1-Dodecylpyridinium Tetrafluoroborate and Its Inclusion Complex With Sulfobutyl Ether-β-Cyclodextrin Against MDR Acinetobacter baumannii Strains 1-Dodecylpyridinium Tetrafluoroborate 及其与磺丁基醚-β-环糊精包合物对 MDR 鲍曼不动杆菌菌株的抗菌活性
Pub Date : 2023-11-21 DOI: 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.
背景。细菌病原体鲍曼不动杆菌(Acinetobacter baumannii)是最危险的多重耐药(MDR)微生物之一,可导致多种细菌感染。目前,迫切需要具有特定分子作用机制的新型广谱抗菌剂。长链 1-烷基吡啶盐是一种高效的阳离子杀菌剂,可抑制参与细菌脂肪酸生物合成的酶。将这些化合物与环状寡糖 β-环糊精结合成包合复合物,可降低其相对较高的急性毒性。研究目的本研究旨在开发基于疏水性 1-烷基吡啶鎓盐及其与磺丁基醚 b-环糊精(SBECD)包合复合物的新型抗鲍曼不动杆菌药物。方法。合成了疏水性阳离子杀菌剂 1-十二烷基吡啶鎓四氟硼酸盐(PyrC12-BF4)及其与 SBECD 的包合物。通过 1H 核磁共振光谱和紫外光谱对 SBECD/PyrC12-BF4 复合物的结构进行了表征。采用标准盘扩散法评估了合成化合物对鲍曼不动杆菌 MDR 临床分离株的体外抗菌活性。对大型蚤模型水生生物进行了急性毒性研究。利用鲍曼不动杆菌 3-氧代乙酰基-[酰基载体蛋白]还原酶(FabG)的晶体结构进行了分子对接。结果。1H NMR 研究结果表明,SBECD 和 PyrC12-BF4 之间形成了包涵复合物。阳离子杀菌剂对四种经测试的耐抗生素鲍曼尼氏菌菌株具有很高的活性,而 SBECD/PyrC12-BF4 复合物仅对两种细菌菌株具有活性。1-decylpyridinium 配体与鲍曼不动杆菌(FabG)活性位点的分子对接显示,在位于亚基 C、D 之间的异构位点形成了复合物。PyrC12-BF4 及其包合物的急性毒性(LC50)分别为 0.007 和 0.033 毫升/克。结论疏水性阳离子杀菌剂 PyrC12-BF4 对 MDR 鲍曼尼氏菌具有很高的抗菌活性。对活性位点 FabG 的抑制可能是 PyrC12-BF4 具有抗鲍曼尼氏菌活性的机制之一。与 PyrC12-BF4 相比,SBECD/PyrC12-BF4 包合复合物的急性毒性降低了近 5 倍,同时还保留了对某些受试鲍曼尼氏菌菌株的活性。
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引用次数: 0
Viability of Nodule Bacteria Bradyrhizobium japonicummon Soybean Seeds by Treatment With Fungicides During Extended Storage 用杀菌剂处理长期储存的大豆种子后,大豆根瘤菌的活力
Pub Date : 2023-11-16 DOI: 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.
背景。随着大豆种植面积的增加,接种剂越来越受到人们的青睐。它们不仅与大豆种子处理的原始制剂兼容,而且还可以在播种前几天甚至几个月将其施用到土壤中。目标在接种种子的长期储存过程中,研究了用杀菌剂 Fever 和 Maxim XL 处理过的大豆种子表面 Bradyrhizobium japonicum(菌株 PC07 和菌株 B78)新菌株的活力。此外,还评估了它们在共生条件下的固氮能力。方法培养、系列稀释法、接种物细菌滴度测定、菌落形成单位(CFU)定量、气相色谱法。结果在实验室条件下,在 Almaz 和 Vasylkivska 品种的大豆种子上,纯培养物中对这些杀菌剂活性物质具有较强抗性的日本节杆菌 PC07 和 B78 的活力下降。这种降低与储存时间、使用的杀菌剂和接种菌株有关。在未使用杀真菌剂的种子上接种一天后,CFU 保留了最初数量的 68.6-75.4%。用杀菌剂 Fever 处理过的种子含有 36.5%-38.3% 的 CFU,用 Maxim XL 处理过的种子含有 26.2%-27.0% 的 CFU。使用基于抗杀菌剂菌株 PC07 和 B78 的接种剂也能提供较高的细胞活力--贮藏 5 天后,细胞活力分别为 2.41-2.8×106(未经处理的种子)、1.40-1.70×106(使用 Fever)和 0.8-1.17×106(使用 Maxim XL)CFU/种子。与对照变种(未使用杀菌剂处理的种子)相比,使用 Fever 处理的 Almaz 和 Vasylkivska 大豆品种的种子在播种前 5 天和 7 天的储藏期间,其根瘤固氮活性分别降低了 18.4-22.4% 和 32.1-39.5%,而使用 Maxim XL 处理的种子则分别降低了 24.5-33.7% 和 47.7-75.2%。结论利用抗杀菌剂的日本酵母菌 PC07 和 B78 菌株对经过杀菌剂 Fever 和 Maxim XL 处理的大豆种子进行杀菌,可在 5 天的储存期间提供较高的接种滴度。经杀菌剂处理过的种子在储藏 7-14 天后,微生物细胞的活力会明显降低。贮藏 14 天后,种子表面的日本疫霉菌株 PC07 和 B78 的 CFU 滴度不足,会严重影响生物制剂的功效。因此,寻找能增强种子上根瘤菌对杀真菌剂种子处理不利影响的抵抗力的物质,确保延长其存活期至关重要。
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引用次数: 0
Effect of Zinc Supplementation to Diluent Boar Semen on Sperm Characteristics and Activity of Antioxidant Enzymes 稀释公猪精液中添加锌对精子特征和抗氧化酶活性的影响
Pub Date : 2023-11-12 DOI: 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.
背景。锌对男性生殖功能至关重要,因为它是精子成熟、获能、顶体反应和受精的必要条件。已经证实,使用金属与氨基酸、核苷酸、肽和碳水化合物的螯合物不仅更有效,而且更经济。研究目的本研究旨在评估和比较不同浓度的谷氨酸锌螯合物(ZnGlu)在体外培养过程中对公猪精子活力、存活率参数和前氧化-抗氧化平衡的影响。研究方法将加入 "Ecosperm "稀释剂后的新鲜射精公猪精液分为对照组和三个实验组。实验组样品中添加的谷氨酸锌浓度分别为 1.0、2.0 和 5.0 µg/ml。精液样本在 18 °C 下储存 4 天。每隔 24 小时,通过二烯共轭物(DC)水平、丙二醛(MDA)浓度以及过氧化氢酶和超氧化物歧化酶的活性来评估精子的活力和存活率以及抗氧化状态。结果研究发现,公猪精子的运动能力随着储存时间的延长而下降,而添加 2.0 和 5.0 µg ZnGlu 则能显著提高精子在 48-96 小时孵育期间的总运动能力和活力精子百分比。我们的研究还表明,ZnGlu 在减轻公猪精子体外氧化应激方面具有保护作用。与对照组相比,添加 ZnGlu 能明显降低所有实验组精液样本中 MDA 和 DC 的含量。此外,在公猪精液中添加锌谷氨酸后,超氧化物歧化酶和过氧化氢酶的活性也有所提高,尤其是在浓度为 2.0 和 5.0 毫克/毫升时(P < 0.05)。结论这些数据表明,补充谷氨酸锌可增强精子的抗氧化防御系统,提高公猪精液的体外质量。
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引用次数: 0
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Innovative Biosystems and Bioengineering
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