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Resistance to Tetracycline and Oleandomycin of a Number of Streptomycetes — Producers of Polyketide Antibiotics 一些链霉菌对四环素和夹竹桃霉素的耐药性——聚酮类抗生素的生产者
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-12-17 DOI: 10.15407/microbiolj84.03.060
L. Polishchuk, O. Bambura
Recently, antibiotic resistance of pathogenic and opportunistic microorganisms is one of the primary problems of medicine. Scientists pay considerable attention to the study of genes for resistance of strains of streptomycetes as sources of such genes for microorganisms. The aim of this study was to determine the sensitivity of 9 strains of streptomycetes producing polyketide antibiotics to tetracycline and oleandomycin and to identify possible correlations in resistant and sensitive strains between the level of their resistance and the presence of resistance genes in chromosomes. Methods. 9 strains of producers of polyketide antibiotics were studied: Streptomyces cyanogenus S136, S. fradiae Tu2717, S. glaucescens Tu49, S. olivaceus Tu2353, S. antibioticus 35, S. globisporus 1912, S. aureofaciens 019, S. coelicolor A3(2), S. lividans TK24. Appropriate microbiological (method of serial dilution in agar) and biotechnological (method of computerized analysis of sequences) methods were used. Results. According to the sensitivity to oleandomycin and tetracycline, the studied strains of streptomycetes can be divided into 3 groups. The first group includes strains resistant to both antibiotics — S. coelicolor A3(2) and S. lividans TK24, the second group includes strains resistant to only one of the antibiotics: more resistant to oleandomycin — S. globisporus 1912, S. glaucescens Tu49, S antibiotic 35-1; more resistant to tetracycline — S. olivaceus Tu2353, S. fradiae Tu2717, S. aureofaciens 019. Strain S. cyanogenus S136 is sensitive to both antibiotics. Conclusions. A correlation was found between the level of tetracycline resistance and the presence (the number and similarity of structures) in the genomes of strains S. lividans TK24, S. globisporus 1912, and S. cyanogenus S136 sequences, which are similar to the sequences of tetracycline resistance genes of strain S. coelicolor A3(2).
近年来,病原微生物和机会微生物的抗生素耐药性是医学的主要问题之一。科学家们相当重视链霉菌菌株耐药基因的研究,将其作为微生物耐药基因的来源。本研究的目的是测定9株生产聚酮类抗生素的链霉菌对四环素和夹竹桃霉素的敏感性,并确定耐药菌株和敏感菌株的耐药水平与染色体中耐药基因的存在之间可能存在的相关性。方法对产生多酮类抗生素的9株菌株进行了研究:蓝色链霉菌S136、传统链球菌Tu2717、青绿链球菌Tu49、橄榄链球菌Tu2353、抗生素链球菌35、全球链球菌1912、金黄色葡萄球菌019、深蓝链球菌A3(2)、lividans TK24。采用适当的微生物学(琼脂连续稀释法)和生物技术(计算机化序列分析法)方法。结果。根据对夹竹桃霉素和四环素的敏感性,将所研究的链菌分为3组。第一组包括对两种抗生素都耐药的菌株——S. colelicolor A3(2)和S. lividans TK24;第二组包括只对一种抗生素耐药的菌株:对夹竹桃霉素更耐药的菌株——S. globisporus 1912、S. glaucescens Tu49、S. antibiotic 35-1;对四环素更耐药的有:橄榄链球菌Tu2353、传统链球菌Tu2717、金黄色葡萄球菌019。菌株S. cyanogenus S136对这两种抗生素都敏感。结论。菌株S. lividans TK24、S. globisporus 1912和S. cyanogenus S136的基因组序列与菌株S. coelicolor A3的四环素抗性基因序列相似,发现菌株对四环素的抗性水平与其基因组中存在的序列(数量和结构相似性)存在相关性(2)。
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引用次数: 0
Modernization of the Pseudomonas syringae Pathovars Serogrouping Scheme 丁香假单胞菌病原菌血清分型方案的现代化
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-12-17 DOI: 10.15407/microbiolj84.03.009
L. Pasichnyk, L. Butsenko
In 1979, L.T. Pastushenko and I.D. Symonovych developed a scheme of serogrouping phytopathogenic bacteria of the Pseudomonas genus, which is still used now. However, today’s using this serogrouping scheme is complicated by the lack of all data accumulated over the years of its application. Moreover, the scheme does not correspond to the modern taxonomy of phytopathogenic bacteria of the Pseudomonas genus. Aim. On the basis of own experimental results and data of scientific literature, to carry out modernization of the serogrouping scheme of phytopathogenic bacteria of the Pseudomonas genus. Methods. The strains of Pseudomonas syringae pathovars such as atrofaciens, coronafaciens, tabaci, which were isolated from plants of wheat, rye, oats, tobacco, and various species of affected weeds in different regions of Ukraine have been studied in the work. Antigenic properties of bacterial strains were studied by agglutination and precipitation reactions (the Ouchterlony double immunodiffusion techniques) using antisera to P. syringae strains of nine serological groups (I, II, III, IV, V, VI, VII, VIII, IX). To carry out the precipitation reaction, O- and OH-antigens were obtained by a modified Grasse’s method. The presence of the same number of precipitation lines of the studied antigens as the number of lines with homologous antiserum of the corresponding serogroup testified to their belonging to this serogroup according to the known serogrouping scheme of phytopathogenic bacteria developed in 1979 by L.T. Pastushenko and I.D. Symonovych. Results. It has been proved that strains of P. syringae pathovars isolated from different cereals (rye, wheat, oats) and segetal vegetation differ in antigenic composition. The antigenic composition of P. syringae strains depends on the host plant from which the pathogen was isolated. Strains of the causative agent of basal glume rot P. syringae pv. atrofaciens isolated from wheat belong to four serological groups (II, IV, V, VI), from rye — to five serological groups (I, II, IV, V, VI), as well as strains of this pathogen isolated from segetal vegetation of wheat agrophytocenosis. Strains of the halo bligth P. syringae pv. coronafaciens from affected oat plants belong to two serological groups (I, V). The serogrouping scheme has been supplemented by new data on the antigenic properties of P. syringae pv. tomato, the causative agent of the bacterial speck disease of tomatoes (Solanum lycopersicum), which is classified as serogroup IV. It has been found that P. syringae pv. tabaci strains, which cause wildfire of tobacco, are part of three serogroups — VII, VIII, IX, and not of two ones, as was presented in the known scheme (1979). Conclusions. Therefore, on the basis of our own research and literature data, the serogrouping scheme of phytopathogenic bacteria of the Pseudomonas genus developed by L.T. Pastushenko and I.D. Symonovych has been modernized. In the renovated scheme, current species’ names of phytopathogens a
1979年,L.T. Pastushenko和i.d ymonovych提出了假单胞菌属植物致病菌血清分型方案,该方案沿用至今。然而,由于缺乏多年来积累的所有数据,今天使用这种血清分组方案变得复杂。此外,该方案不符合假单胞菌属植物致病菌的现代分类。的目标。在自身实验结果和科学文献资料的基础上,对假单胞菌属植物致病菌的血清分型方案进行现代化的研究。方法。从乌克兰不同地区的小麦、黑麦、燕麦、烟草和各种受影响的杂草中分离出的丁香假单胞菌病原菌株,如atrofaciens、coronafaciens、tabaci菌株。用抗血清对丁香假单胞菌9个血清学组(I、II、III、IV、V、VI、VII、VIII、IX)进行凝集和沉淀反应(Ouchterlony双免疫扩散技术),研究菌株的抗原性。沉淀反应采用改进的Grasse法获得O-和oh -抗原。根据L.T. Pastushenko和i.d ymonovych于1979年建立的已知植物致病菌血清分型方案,所研究抗原的沉淀线数目与相应血清群的同源抗血清的沉淀线数目相同,证明它们属于该血清群。结果。已经证明,从不同谷物(黑麦、小麦、燕麦)和部分植物中分离的丁香假单胞菌病原菌的抗原组成不同。丁香假单胞菌菌株的抗原组成取决于其分离的寄主植物。丁香腐病病原菌的研究。从小麦中分离到的atrofaciens可分为4个血清学类群(II、IV、V、VI),从黑麦中分离到5个血清学类群(I、II、IV、V、VI),以及从小麦农藓病的分生植被中分离到的菌株。丁香晕疫病菌株的研究。来自受感染燕麦植株的冠状病毒分属于两个血清学类群(I, V)。该血清分型方案得到了syringae pv抗原特性新数据的补充。番茄细菌性斑点病(Solanum lycopersicum)的病原,被归类为血清IV群。引起烟草野火的烟粉虱菌株属于三种血清群——VII、VIII、IX的一部分,而不是像已知方案(1979年)中提出的那样属于两种血清群。结论。因此,我们在自己的研究和文献资料的基础上,对L.T. Pastushenko和i.d ymonovych开发的假单胞菌属植物致病菌的血清分型方案进行了现代化的研究。在修改后的方案中,现有的植物病原体的物种名称,该方案引入了新的信息,关于P. syringae pv血清群。从黑麦种子和植物、杂草中分离得到atrofaciens;冠状病毒-来自燕麦植物,紫丁香P.;烟草(tabaci) -来自烟草;番茄-取自受影响的番茄植株。
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引用次数: 0
A Survey of Grapevine Leafroll-Associated Virus 1 and 3 in the South of Ukraine and Development of Primers for GLRaV-3 Identification 乌克兰南部葡萄叶卷相关病毒1和3的调查及glrav3鉴定引物的研制
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-12-17 DOI: 10.15407/microbiolj84.03.082
A. Kyrychenko, K. Hrynchuk, I. Antipov, A. Konup
Viticulture is one of the most intensive and complex branches of Ukraine agriculture. Grapevine virus diseases are responsible for considerable economic losses to grape productivity and wine industries. One of the most notable and widespread viruses associated with vine leafroll disease is grapevine leafroll-associated viruses (GLRaV), belonging to the genus Ampelovirus, family Closteroviridae. Thus, the aim of this study was to conduct a survey targeting two viruses involved in the grapevine leafroll, namely grapevine leafroll-associated virus 1 (GLRaV-1) and virus 3(GLRaV-3) distributed in commercial wine grapes growing in the vineyards of the Ovidiopol and Bolhrad districts of the Odesa region. For efficient and accurate virus detection, we aimed to design universal primers based on conserved nucleotide sequences. Methods. Virus surveys of vineyards, visual diagnosis, immunoassay (ELISA), polymerase chain reaction with reverse transcription (RT-PCR), and Sanger sequencing of partial genome sequences of GLRaV. Results. The results obtained indicate that grapevine leafroll disease symptoms in field-grown grapevines in the south of Ukraine are caused by GLRaV-3. GLRaV-1 was not detected in any of the samples tested. To confirm the presence of GLRaV-3 in the samples, specific primers were designed targeting the coat protein region (GLRaV3-7f 5’-AGTAGGGGATGCAGCACAAG-3’; GLRaV3-7r 5’-ATCCAAAGCTATTCCCTTGC-3’) of the virus. A new set of primers (GLRaV3-7f / GLRaV3-7r) has been validated for sensitive detection of GLRaV-3 by RT-PCR and may be useful for routine virus detection in the laboratory as well as for large-scale testing. The partial coat protein gene of the isolate, GLRaV-3 ukr, distributed in the south of Ukraine, was sequenced, and the obtained sequence was deposited in GenBank under Acc. No. ON015835. The phylogenetic study demonstrated that GLRaV-3 ukr was closely related to isolates from Russia (MZ065372 and MZ065370).
葡萄栽培是乌克兰农业中最密集和最复杂的分支之一。葡萄病毒病对葡萄生产和葡萄酒工业造成了相当大的经济损失。与葡萄叶卷病相关的最显著和最广泛的病毒之一是葡萄叶卷相关病毒(GLRaV),属于Closteroviridae Ampelovirus属。因此,本研究的目的是对敖德萨地区Ovidiopol和Bolhrad地区的商业酿酒葡萄中分布的葡萄叶卷相关病毒1 (glrav1)和病毒3(glrav3)两种与葡萄叶卷相关的病毒进行调查。为了高效准确地检测病毒,我们的目标是设计基于保守核苷酸序列的通用引物。方法。葡萄园病毒调查、视觉诊断、免疫测定(ELISA)、反转录聚合酶链反应(RT-PCR)和GLRaV部分基因组序列的Sanger测序。结果。结果表明,乌克兰南部田间种植的葡萄叶卷病症状是由glrav3引起的。在所有检测样本中均未检测到glrav1。为了确认样品中是否存在glrav3,我们设计了针对其表皮蛋白区域的特异性引物(glrav3 - 7f5 ' -AGTAGGGGATGCAGCACAAG-3 ';GLRaV3-7r 5 ' -ATCCAAAGCTATTCCCTTGC-3 ')一组新的引物(GLRaV3-7f / GLRaV3-7r)已被验证可用于RT-PCR对glrav3的敏感检测,可用于实验室常规病毒检测以及大规模检测。对分布于乌克兰南部的分离物glrav3 ukr的部分外壳蛋白基因进行测序,并将测序结果存放在GenBank中。不。ON015835。系统发育研究表明,glrav3 ukr与俄罗斯分离株MZ065372和MZ065370亲缘关系较近。
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引用次数: 0
Antimicrobial and Antiviral Activity of Silver-Containing Nanocomposites Formed by 3D Printing Technology 3D打印技术制备的含银纳米复合材料的抗菌和抗病毒活性
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-12-17 DOI: 10.15407/microbiolj84.03.069
N. Rybalchenko, К.S. Naumenko, L. Artiukh, V. Demchenko, О.P. Masiuchok, V. Ovsyankina, T. Rybalchenko, V.V. Yurzhenko
Silver nanoparticles have become the focus of numerous researchers to create efficient antimicrobial and antiviral agents due to their pronounced pharmacological effects, low toxicity to the human body and the environment, and high stability in extreme conditions. To create antimicrobial drugs with silver nanoparticles, matrices from polymers of both synthetic and natural origin are used. Biopolymer polylactide (PLA) is one of the most promising materials for 3D printing (additive production) due to its physicochemical and technological properties. The aim of the work was to study the antimicrobial and antiviral activity of silver-containing nanocomposites formed on the basis of PLA with the addition of chitosan or polyethyleneimine (PEI) by 3D printing technology. Methods. Peculiarities of the structural organization of silver-containing materials were investigated by the method of wide-angle radiography on an XRD-7000 diffractometer. The morphology of the samples was studied by transmission electron microscopy on a JEM-1230 instrument. Filament formation proceeded through melting PLA-AgPalm-PEI or PLA-AgPalm-chitosan films in an extruder at a temperature of T=160±1°С. Samples were formed from the obtained filaments using a 3D printer. Th e antimicrobial activity of silvercontaining nanocomposites was determined by the disk diffusion method against opportunistic pathogens S. aureus and E. coli. Cytotoxicity and antiviral activity were investigated using the MTT method and staining by gentian violet. Results. Analysis of wide-angle X-ray diffraction patterns of products formed by 3D printing technology at 160°C showed that PLA-Ag-PEI and PLA-Ag-chitosan materials contain metallic silver. Analysis of microphotographs of PLA-4%Ag-PEI and PLA-4%Ag-chitosan products formed by 3D printing technology showed that silver nanoparticles formed by adding chitosan as a reducing agent and stabilizer to the polymer matrix of PLA are much smaller than when using PEI. Silver-containing nanocomposite samples, such as filaments and products formed from PLA-4%Ag-PEI and PLA-4%Agchitosan films subjected to heat treatment and without heat treatment, show antimicrobial activity against test cultures of S. aureus and E. coli. It was found that nanocomposites based on PEI and chitosan do not show cytotoxic effects in MDCK and HEP-2 cultures. Nanocomposites of both types show a weak antiviral effect against adenovirus serotype 2; the reduction of infectious titer was 0.5 lgTCID50/mL. None of the studied nanocomposites showed antiviral action on the influenza virus model. Conclusions. The investigated silver-containing nanocomposites with a silver concentration of 4% by weight, formed by 3D printing technology from PLA-4%Ag-PEI and PLA-4%Ag-chitosan films, show antimicrobial activity against S. aureus and E. coli test cultures and antiviral activity on influenza A virus and human adenovirus and do not show a cytotoxic effect on cells. The obtained data allow us to state t
银纳米颗粒具有药理作用显著、对人体和环境毒性低、在极端条件下具有较高的稳定性等特点,已成为众多研究人员开发高效抗菌和抗病毒药物的热点。要用银纳米颗粒制造抗菌药物,需要使用合成聚合物和天然聚合物的基质。生物聚合物聚乳酸(PLA)由于其物理化学和工艺性能而成为3D打印(增材生产)最有前途的材料之一。本工作的目的是通过3D打印技术研究在聚乳酸的基础上添加壳聚糖或聚乙烯亚胺(PEI)形成的含银纳米复合材料的抗菌和抗病毒活性。方法。用XRD-7000衍射仪广角射线照相法研究了含银材料的结构组织特性。在JEM-1230型透射电镜上观察样品的形貌。在T=160±1°С的温度下,将PLA-AgPalm-PEI或pla - agpalm -壳聚糖薄膜熔化成丝。用3D打印机将获得的细丝制成样品。采用纸片扩散法测定含银纳米复合材料对条件致病菌金黄色葡萄球菌和大肠杆菌的抑菌活性。采用MTT法和龙胆紫染色法研究其细胞毒性和抗病毒活性。结果。在160℃下对3D打印技术形成的产品进行广角x射线衍射图分析,发现PLA-Ag-PEI和pla - ag -壳聚糖材料中含有金属银。对3D打印技术制备的PLA-4% ag -PEI和PLA-4% ag -壳聚糖产品的显微照片分析表明,在PLA的聚合物基体中加入壳聚糖作为还原剂和稳定剂形成的银纳米粒子比使用PEI时要小得多。含银纳米复合材料样品,如由PLA-4%Ag-PEI和pla -4% ag壳聚糖薄膜经热处理和不经热处理形成的细丝和产物,对金黄色葡萄球菌和大肠杆菌的试验培养物显示出抗菌活性。我们发现PEI和壳聚糖纳米复合材料在MDCK和HEP-2培养中没有细胞毒性作用。两种类型的纳米复合材料对血清2型腺病毒的抗病毒作用较弱;感染滴度降低0.5 lgTCID50/mL。所研究的纳米复合材料均未对流感病毒模型显示抗病毒作用。结论。采用3D打印技术,将PLA-4%Ag-PEI和pla -4% ag -壳聚糖薄膜制成银浓度为重量比4%的含银纳米复合材料,对金黄色葡萄球菌和大肠杆菌试验培养物具有抗菌活性,对甲型流感病毒和人类腺病毒具有抗病毒活性,并且对细胞没有细胞毒性作用。所得数据表明,所研究的含银纳米复合材料是一种很有前途的抗菌剂,可用于医药和食品工业的各个领域。
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引用次数: 0
Properties of Microorganisms Isolated from Soils under Conventional and Organic Farming 常规耕作与有机耕作土壤中分离微生物的特性
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.012
I. Gumeniuk, A. Levishko, O. Demyanyuk, O. Sherstoboeva
Оbjective. The article presents the results of research aimed at determining the influence of different methods of tillage on the functional diversity of the soil microbiota. Soil samples containing plant residues from agricultural plots under conventional and organic farming in the Kyiv oblast were used for the study. Methods. Analysis of soil microbiota using differential diagnostic nutrient media by serial dilutions of soil suspension was performed. To quantify the phosphate-mobilizing properties of the isolated microorganisms, the concentration of phosphorus in the solution was measured (grown in NBRIP liquid medium) and detected by the Arenius spectrophotometric method on a Ulab 102UV Spectrophotometer. Results. Th e soil of the plots under organic agrotechnology of cultivation was marked by a greater number of microorganisms of all ecological and trophic groups, except oligonitrophilic and phosphate-solubilizing bacteria. The vast majority of phosphate-transforming bacteria were isolated from the soil of agricultural plots under convection farming. The largest number of cellulose-degrading isolates was isolated from the soil under organic farming plots. Five isolates have the widest range of agronomically useful properties, in particular, the ability to mobilize organic and inorganic phosphates and cellulosolytic activity: 6b, 13b, 18b, 19b, and 8m. After incubation of the isolates on an NBRIP medium at 28°C and 200 rpm for 72 hr, special analyzes for dissolved phosphorus content and pH level in the culture fluid were performed. Isolate 8m selected from chornozem (black soil) under convection agriculture and classified by us as Trichoderma sp. exhibited the highest phosphate-mobilizing activity. The vast majority of bacteria capable of phosphate transformation were isolated from the soil of agricultural areas affected by convection agriculture; and isolates capable of dissolving cellulose — from the soil of organic farming. Conclusions. The initial identification of certain isolates allowed us to classify them as Bacillus and Trichoderma. These isolates are important for further research with the prospect of creating a complex biological preparation with fungicidal properties and the ability to mobilize organic and inorganic phosphorus compounds.
Оbjective。本文介绍了旨在确定不同耕作方式对土壤微生物群功能多样性影响的研究结果。研究使用了基辅州传统农业和有机农业中含有植物残留物的土壤样本。方法。通过对土壤悬浮液进行连续稀释,采用鉴别诊断营养培养基对土壤微生物群进行了分析。为了量化分离微生物的磷酸盐动员特性,测定了溶液中磷的浓度(在NBRIP液体培养基中培养),并在Ulab 102UV分光光度计上用Arenius分光光度法检测。结果。有机农业技术耕作的样地土壤中除寡氮菌和增磷菌外,所有生态和营养类群的微生物数量都较多。绝大多数转化磷酸盐的细菌是从对流耕作的农田土壤中分离出来的。从有机耕作地块土壤中分离出的纤维素降解菌株数量最多。5个分离株具有最广泛的农艺有用特性,特别是调动有机和无机磷酸盐的能力和纤维素水解活性:6b, 13b, 18b, 19b和8m。分离株在NBRIP培养基上28°C和200 rpm孵育72小时后,对培养液中的溶解磷含量和pH值进行特殊分析。在对流农业条件下,从黑土土壤中分离出8m的木霉(Trichoderma sp.),其磷素动员活性最高。绝大多数能转化磷酸盐的细菌是从受对流农业影响的农业区土壤中分离出来的;从有机农业的土壤中分离出能够溶解纤维素的细菌。结论。对某些分离株的初步鉴定使我们能够将它们分类为芽孢杆菌和木霉。这些分离物对于进一步研究具有重要意义,具有创造具有杀真菌特性和调动有机和无机磷化合物能力的复杂生物制剂的前景。
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引用次数: 0
Fatty Acid Composition of Comamonas testosteroni under Hexachlorobenzene Loading Conditions 六氯苯负载条件下睾酮单胞菌的脂肪酸组成
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.024
M. Dimova, G. Iutynska
Changes in the lipid composition in bacterial membranes are considered to be the most important adaptation mechanisms to adverse chemical factors. The aim of the study was to compare the hexachlorobenzene effects on the fatty acid composition of total lipids Comamonas testosteroni. Methods. The study was performed with C. testosteroni UCM B-400 and B-401, B-213 strains. Bacteria were grown in the Luria-Bertrani (LB) liquid medium containing 10 and 20 mg/L of hexachlorobenzene (HCB). After cultivation, the biomass was separated by centrifugation and the fatty acid composition of total lipids was determined through analyzing its methyl esters. To assess the cell membrane properties, such parameters as the lipid unsaturation index, the average carbon chain length of fatty acids, and the membrane viscosity index were determined. Results. In the fatty acids spectrum of C. testosteroni B-400 after cultivation in a medium containing 20 mg/L of HCB, the contents of unsaturated hexadecenoic (C16:1) and octadecenoic (C18:1) acids were lower by 10.6 and 5.5%, respectively, and that of saturated hexadecanoic (C16:0) acid was higher by 8.4%, compared to the control. The fatty acid composition of C. testosteroni B-401 was more stable compared to strain B-400. Collection strain C. testosteroni B-213 compared to strains isolated from soil with high HCB load, in the presence of 10 and 20 mg/L of HCB had the highest relative content of saturated hexadecanoic acid (C16:0) up to 38.33—40.7%. Unsaturated octadecenoic acid decreased at the doses 10 and 20 mg/L to 1.5—2% compared to the control. In all strains under the HCB impact, there was an increase in the relative content of C17-cyclopropanoic acid compared to control variants. Conclusions. C. testosteroni UCM B-400, B-401, and B-213 bacteria under cultivation conditions in HCB-containing medium, decreasing the degree of lipid unsaturation and increasing the relative content of C17-cyclopropanoic acid can be considered as the main mechanisms regulating the cytoplasmic membrane fluidity; the displaying of these protective reactions had a strain trait and did not depend on the adaptation in natural isolating places.
细菌膜中脂质组成的变化被认为是对不良化学因素最重要的适应机制。本研究的目的是比较六氯苯对睾酮单胞菌总脂质脂肪酸组成的影响。方法。研究对象为睾酮C. UCM B-400和B-401、B-213菌株。细菌在含有10和20 mg/L六氯苯(HCB)的Luria-Bertrani (LB)液体培养基中生长。培养后,离心分离生物量,通过分析其甲酯来确定总脂质的脂肪酸组成。通过测定脂质不饱和指数、脂肪酸平均碳链长度、膜粘度指数等参数来评价细胞膜的性能。结果。在含有20 mg/L HCB的培养基中培养后,睾酮c -400的脂肪酸谱中,不饱和十六烯酸(C16:1)和十八烯酸(C18:1)的含量分别比对照降低了10.6和5.5%,饱和十六烯酸(C16:0)的含量比对照提高了8.4%。与菌株B-400相比,C. testosterone B-401的脂肪酸组成更稳定。收集菌株C. testosterone -213与高负荷土壤分离菌株相比,在HCB浓度为10和20 mg/L时饱和六酸(C16:0)的相对含量最高,达38.33 ~ 40.7%。与对照组相比,不饱和十八烯酸在10和20 mg/L剂量下下降至1.5-2%。在所有受HCB影响的菌株中,与对照变体相比,c17 -环丙酸的相对含量都有所增加。结论。C.睾酮UCM B-400、B-401和B-213细菌在含hcb培养基培养条件下,降低脂质不饱和程度和增加c17 -环丙酸相对含量可被认为是调节细胞质膜流动性的主要机制;这些保护反应的表现具有品系特征,不依赖于自然隔离地的适应。
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引用次数: 1
Ways of Auxin Biosynthesis in Microorganisms 生长素在微生物中的生物合成途径
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.057
T. Pirog, D. Piatetska, N. Klymenko, G. Iutynska
Among plant hormones, auxins, in particular indole-3-acetic acid (IAA), are the most studied and researched. Almost all groups of soil microorganisms, both plant-associated and non-plant-associated bacteria, fungi, and phytopathogenic microorganisms are capable of producing auxins. The development of preparations for crop production is directly related to the production of bacterial strains with high auxin-synthesizing potential, which is possible only with a full understanding of the ways of regulation and synthesis of auxins in bacteria. The synthesis of auxins in microorganisms can take place in two ways: by the gradual conversion of tryptophan to IAA (tryptophan-dependent pathway) or by the use of other intermediates (tryptophan-independent pathway). The latter is poorly clarified, and in the literature available today, there is only a small amount of information on the functioning of this pathway in microorganisms. The review presents literature data on the ways of auxin biosynthesis in different groups of microorganisms, as well as approaches to the intensification of indole-3-acetic acid synthesis. The formation of IAA from tryptophan can be carried out in the following ways: through indole-3-pyruvate, through indole-3-acetamide, and through indole-3-acetonitrile. The vast majority of available publications are related to the assimilation of tryptophan through the formation of indole-3-pyruvate as this pathway is the most common among microorganisms. Thus, it functions in rhizospheric, symbiotic, endophytic, and free-living bacteria. The concentration of synthesized IAA among natural strains is in the range from 260 to 1130 μg/mL. Microorganisms in which the indole-3-acetamide pathway functions are characterized by lower auxin-synthesizing ability compared to those that assimilate tryptophan through indole-3-pyruvate. These include bacteria of the genera Streptomyces, Pseudomonas, and Bradyrhizobium and fungi of the genus Fusarium. The level of synthesis of IAA in such microorganisms is from 1.17×10−4 to 255.6 μg/mL. To date, only two strains that assimilate tryptophan via the indole-3-acetonitrile pathway and form up to 31.5 μg/mL IAA have been described in the available literature. To intensify the synthesis of indole-3-acetic acid, researchers use two main approaches: the first consists in introducing into the culture medium of exogenous precursors of biosynthesis (usually tryptophan, less often indole-3-pyruvate, indole-3-acetamide, and indole-3-acetonitrile); the second — in increasing the expression of the corresponding genes and creating recomindolebinant strains-supersynthetics of IAA. The largest number of publications is devoted to increasing the synthesis of IAA in the presence of biosynthesis precursors. Depending on the type of bacteria, the composition of the nutrient medium, and the amount of exogenously introduced precursor, the synthesis of the final product was increased by 1.2—27 times compared to that before the intensi
在植物激素中,生长素,尤其是吲哚-3-乙酸(IAA)是研究最多的。几乎所有种类的土壤微生物,包括植物相关的和非植物相关的细菌、真菌和植物病原微生物都能够产生生长素。作物制剂的发展直接关系到生长素合成潜力高的菌株的产生,只有充分了解生长素在细菌中的调控和合成途径才有可能。微生物中生长素的合成可以通过两种方式进行:通过色氨酸逐渐转化为IAA(色氨酸依赖途径)或通过使用其他中间体(色氨酸独立途径)。后者尚不清楚,并且在目前可用的文献中,只有少量关于该途径在微生物中的功能的信息。本文综述了生长素在不同微生物群中合成途径的文献资料,以及加强吲哚-3-乙酸合成的途径。色氨酸可通过吲哚-3-丙酮酸、吲哚-3-乙酰胺和吲哚-3-乙腈三种途径生成IAA。绝大多数现有的出版物都与色氨酸通过形成吲哚-3-丙酮酸的同化有关,因为这一途径在微生物中最常见。因此,它在根际、共生、内生和自由生活的细菌中起作用。天然菌株合成IAA浓度范围为260 ~ 1130 μg/mL。与那些通过吲哚-3-丙酮酸吸收色氨酸的微生物相比,具有吲哚-3-乙酰胺途径功能的微生物的生长素合成能力较低。这些细菌包括链霉菌属的细菌、假单胞菌、慢生根瘤菌和镰刀菌属的真菌。这些微生物中IAA的合成水平为1.17×10−4 ~ 255.6 μg/mL。迄今为止,文献中只报道了两种通过吲哚-3-乙腈途径吸收色氨酸并形成高达31.5 μg/mL IAA的菌株。为了加强吲哚-3-乙酸的合成,研究人员主要采用两种方法:第一种是在培养基中引入外源性生物合成前体(通常是色氨酸,较少使用的是吲哚-3-丙酮酸酯、吲哚-3-乙酰胺和吲哚-3-乙腈);二是增加相应基因的表达,创造出IAA的超合成菌株。最多的出版物致力于在生物合成前体存在的情况下增加IAA的合成。根据细菌种类、营养培养基组成和外源前体引入量的不同,最终产物的合成比强化前提高了1.2 ~ 27倍。因此,在11 g/L色氨酸存在的情况下,肠杆菌sp. DMKU-RP206合成了5.56 g/L的IAA,而在没有前体的培养基中,IAA的产量仅为0.45 g/L。当向培养基中添加色氨酸(10 g/L)时,重组菌株谷氨酸棒状杆菌ATCC 13032和大肠杆菌MG165分别形成7.1和7.3 g/L的IAA。在胁迫条件下(温度、pH、生物和非生物胁迫因素),微生物的生长素合成水平可能会增加,但在这种情况下,IAA浓度不超过100 mg/L,因此这种强化方法不能与上述其他方法竞争。
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引用次数: 0
The Effect of Ag-Doping on the Cytotoxicity of ZnO Nanostructures Grown on Ag/Si Substrates by APMOCVD Ag掺杂对APMOCVD在Ag/Si衬底上生长的ZnO纳米结构细胞毒性的影响
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.047
K. Naumenko, A. Ievtushenko, V. Karpyna, O. Bykov, L. Myroniuk
The search and development of new nanostructures and nanomaterials are very important for the progress of nanotechnology and modern microbiology. Due to the unique properties of silver and zinc oxide, these nanoparticles are the optimal basis for creating nanostructures with potential antiviral activity. An important issue in these studies is the establishment of cytotoxicity of these nanoparticles and their composites. Aim. To define the influence of substrate temperature and Ag concentration in ZnO lattice on the microstructure and cytotoxicity of zinc oxide nanostructures. Methods. Pure and Ag-doped ZnO nanostructures were grown on Ag/Si substrates by atmospheric pressure metalorganic chemical vapor deposition method using a mixture of zinc acetylacetonate and silver acetylacetonate powders as a precursor. Argentum thin films were deposited on Si substrates by a thermal evaporation method. MTT-assay was used for the analysis of MDBK and MDCK cell viability in the definition of zinc oxide nanostructure cytotoxicity. Results. Ag-doped zinc oxide nanostructures were grown and characterized by X-ray diff raction, scanning electron microscopy, and energy dispersive X-ray spectroscopy. It was found that Si substrate and pure zinc oxide do not inhibit the cell viability of both epithelial cultures whereas Ag-doped ZnO nanostructures inhibit the cell viability because of all-time exposure in a sample without dilution. The cytotoxic effect was not observed at higher dilutions for Ag-doped zinc oxide nanostructures. Conclusions. The investigation of the effect of Ag-doping on the morphology and cytotoxicity of zinc oxide nanostructures is very important for implementing zinc oxide nanostructures into the current optoelectronics and photocatalysis.
寻找和开发新的纳米结构和纳米材料对纳米技术和现代微生物学的发展具有重要意义。由于银和氧化锌的独特性质,这些纳米颗粒是创建具有潜在抗病毒活性的纳米结构的最佳基础。这些研究中的一个重要问题是建立这些纳米颗粒及其复合材料的细胞毒性。的目标。研究底物温度和ZnO晶格中Ag浓度对氧化锌纳米结构的微观结构和细胞毒性的影响。方法。以乙酰丙酮锌和乙酰丙酮银混合粉末为前驱体,采用常压金属有机化学气相沉积法在Ag/Si衬底上生长出纯ZnO和掺银ZnO纳米结构。采用热蒸发法在硅衬底上沉积了银薄膜。在氧化锌纳米结构细胞毒性定义中,采用mtt法分析MDBK和MDCK细胞活力。结果。采用x射线衍射、扫描电镜和能量色散x射线能谱等方法对掺杂银氧化锌纳米结构进行了生长和表征。研究发现,硅衬底和纯氧化锌不会抑制两种上皮细胞培养物的细胞活力,而ag掺杂ZnO纳米结构由于长期暴露在未稀释的样品中而抑制细胞活力。在较高的稀释度下,对银掺杂的氧化锌纳米结构没有观察到细胞毒性作用。结论。研究ag掺杂对氧化锌纳米结构形貌和细胞毒性的影响,对于将氧化锌纳米结构应用于光电子和光催化领域具有重要意义。
{"title":"The Effect of Ag-Doping on the Cytotoxicity of ZnO Nanostructures Grown on Ag/Si Substrates by APMOCVD","authors":"K. Naumenko, A. Ievtushenko, V. Karpyna, O. Bykov, L. Myroniuk","doi":"10.15407/microbiolj84.02.047","DOIUrl":"https://doi.org/10.15407/microbiolj84.02.047","url":null,"abstract":"The search and development of new nanostructures and nanomaterials are very important for the progress of nanotechnology and modern microbiology. Due to the unique properties of silver and zinc oxide, these nanoparticles are the optimal basis for creating nanostructures with potential antiviral activity. An important issue in these studies is the establishment of cytotoxicity of these nanoparticles and their composites. Aim. To define the influence of substrate temperature and Ag concentration in ZnO lattice on the microstructure and cytotoxicity of zinc oxide nanostructures. Methods. Pure and Ag-doped ZnO nanostructures were grown on Ag/Si substrates by atmospheric pressure metalorganic chemical vapor deposition method using a mixture of zinc acetylacetonate and silver acetylacetonate powders as a precursor. Argentum thin films were deposited on Si substrates by a thermal evaporation method. MTT-assay was used for the analysis of MDBK and MDCK cell viability in the definition of zinc oxide nanostructure cytotoxicity. Results. Ag-doped zinc oxide nanostructures were grown and characterized by X-ray diff raction, scanning electron microscopy, and energy dispersive X-ray spectroscopy. It was found that Si substrate and pure zinc oxide do not inhibit the cell viability of both epithelial cultures whereas Ag-doped ZnO nanostructures inhibit the cell viability because of all-time exposure in a sample without dilution. The cytotoxic effect was not observed at higher dilutions for Ag-doped zinc oxide nanostructures. Conclusions. The investigation of the effect of Ag-doping on the morphology and cytotoxicity of zinc oxide nanostructures is very important for implementing zinc oxide nanostructures into the current optoelectronics and photocatalysis.","PeriodicalId":18628,"journal":{"name":"Mikrobiolohichnyi zhurnal","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2022-11-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"73525085","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}
引用次数: 1
Isolation and Diagnosis of Methicillin-Resistant Staphylococcus aureus in Meat and Butcher Tool Surfaces in Baghdad 巴格达地区肉制品和刀具表面耐甲氧西林金黄色葡萄球菌的分离与诊断
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.040
H. K. Yakob, L. R. Hamad, M. B. Farhan
Due to the medical and epidemiological importance of the spread of Methicillin-Resistant Staphylococcus aureus, this study was conducted for the purpose of isolating and diagnosing these bacteria from local sheep meat and butcher's tools in Baghdad. Methods. 200 samples were collected. Mannitol salt agar and Staph.110 medium were used to isolate the bacteria. The isolates were identified using standard cultivation methods, biochemical tests, the GP24 diagnostic system, and an integrated Vitek 2 device. The isolates were tested for sensitivity to methicillin by the disk diff usion method. DNA was extracted and the mecA gene was detected in the isolates that showed methicillin resistance by polymerase chain reaction. Results. S. aureus was diagnosed in 83 (41.5%) of the samples. Of them, 35 (42.2%) were methicillin-resistant. Out of these, 24 (68.6%) were found to have the mecA gene. Conclusions. Methicillin-Resistant S. aureus strains were detected with a high prevalence due to the underdeveloped reality of slaughter places.
由于耐甲氧西林金黄色葡萄球菌的传播在医学和流行病学上具有重要意义,因此进行了这项研究,以便从巴格达当地的羊肉和屠夫工具中分离和诊断这些细菌。方法:采集标本200份。采用甘露醇盐琼脂和葡萄球菌110培养基进行分离。采用标准培养方法、生化试验、GP24诊断系统和集成的Vitek 2装置对分离株进行鉴定。采用纸片扩散法检测分离菌株对甲氧西林的敏感性。提取甲氧西林耐药菌株DNA,采用聚合酶链反应检测mecA基因。结果。检出金黄色葡萄球菌83例(41.5%)。其中耐甲氧西林35例(42.2%)。其中,24人(68.6%)携带mecA基因。结论。由于屠宰场实际条件不发达,检测到耐甲氧西林金黄色葡萄球菌的流行率较高。
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引用次数: 0
Double Coordination Compounds of Fe(II)/Co(II)/Ni(II)/Cu(II) 1,10-Phenanthroline/2,2ʹ-Bipyridine Cations with Tartratogermanate(IV) Anions as Novel Nonresistant Antimicrobial Agents Fe(II)/Co(II)/Ni(II)/Cu(II) 1,10-菲罗啉/2,2′-联吡啶阳离子与酒石酸根酸(IV)阴离子的双配位化合物研究
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.003
E. Afanasenko, I. Seifullina, O. E. Martsinko, L. Konup, M. Kyryk
Objective. To study the antimicrobial activity of double coordination compounds with 1,10-phenanthroline/2,2ʹ-bipyridine complexes of Fe(II)/Co(II)/Ni(II)/Cu(II) as cations and diff erent tartratogermanate(IV) anions, reveal the main factors of their effi ciency and establish relations between their composition, structure features, and biological properties. Methods. The developed synthesis method allowed us to obtain three diff erent tartratogermanate anions, which exist together in the solution and can be selectively recognized by the certain type of 1,10-phenanthroline/2,2ʹ-bipyridine cation. Th e antimicrobial activity of the compound was investigated by a rapid twofold dilution method in a standard liquid nutrient medium (Hottinger digestion) to determine the minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC). Results. Th e complex nature of studied compounds, synergism of their biologically active structural units, and the presence of diff erent types of intermolecular bonds result in the high antimicrobial activity against a wide range of microorganisms such as gram-positive Planococcus citreus, Microcoсcus luteus, Bacillus cereus, Staphylococcus aureus, Streptococcus lactis, and, in a less degree, gram-negative Escherichia coli and Agrobacterium tumefaciens. Compounds (1)—(8) show a high antimicrobial activity because all of them belong to the type of double coordination compounds and contain similar structural units. Nevertheless, complexes (1) (23.44 μg/mL), (3) (46.9 μg/mL), (4) (23.44 μg/mL), and (8) (46.9 μg/mL) turned out to be the most eff ective, while (6) (>500 μg/mL) and (7) (>500 μg/mL) are less productive. Complexes that have anions [Ge2(OH)(H2Tart)(μ-Tart)2]3- (1), (8) and [Ge2(OH)(HTart)(μ-Tart)2]4- (4) with free hydroxyl and carboxyl groups of the terminal tartaric acid are able to interact with metals in the enzymes of microorganisms and appear to be better antimicrobial drugs because they show lower inhibitory and bactericidal concentrations. Conclusions. Structural features such as the cation-anionic type of compounds, variability of intermolecular interactions, joint of diff erent biologically active units and free chelating groups in tartaric ligands lead to the combination of different action mechanisms and exclude the possibility of strain resistance.
目标。研究以Fe(II)/Co(II)/Ni(II)/Cu(II)为阳离子和不同酒石酸根酸(IV)为阴离子的1,10-菲罗啉/2,2′-联吡啶配合物双配位化合物的抑菌活性,揭示其抑菌效果的主要影响因素,建立其组成、结构特征和生物学特性之间的关系。方法。该合成方法使我们得到了三种不同的酒石酸盐阴离子,它们一起存在于溶液中,可以被某种类型的1,10-菲罗啉/2,2′-联吡啶选择性识别。采用标准营养液(Hottinger消化法)快速倍稀释法测定化合物的最小抑菌浓度(MIC)和最小杀菌浓度(MBC)。结果。所研究化合物的复杂性质,其生物活性结构单元的协同作用,以及不同类型分子间键的存在,导致其对多种微生物具有很高的抗菌活性,如革兰氏阳性的柑橘平球菌、黄体微芽孢杆菌、蜡样芽孢杆菌、金黄色葡萄球菌、乳酸链球菌,以及在较小程度上对革兰氏阴性的大肠杆菌和农杆菌。化合物(1)-(8)均为双配位化合物,结构单元相似,具有较高的抗菌活性。然而,配合物(1)(23.44 μg/mL)、(3)(46.9 μg/mL)、(4)(23.44 μg/mL)和(8)(46.9 μg/mL)的效率最高,而(6)(bbb500 μg/mL)和(7)(>500 μg/mL)的效率较低。具有阴离子[Ge2(OH)(H2Tart)(μ-Tart)2]3-(1)、(8)和[Ge2(OH)(H2Tart)(μ-Tart)2] 4-(4)与末端酒石酸的游离羟基和羧基的配合物能够与微生物酶中的金属相互作用,表现出较低的抑制和杀菌浓度,是较好的抗菌药物。结论。化合物的正阴离子类型、分子间相互作用的可变性、酒石酸配体中不同生物活性单元的结合以及游离螯合基团等结构特征导致了不同作用机制的结合,排除了菌株抗性的可能性。
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引用次数: 1
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Mikrobiolohichnyi zhurnal
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