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Detection of Biofilm Formation and Some Virulence Factors in Pseudomonas aeruginosa, and the Effect of Some Antibiotics 铜绿假单胞菌生物膜形成及部分毒力因子的检测及抗生素的作用
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2022-11-28 DOI: 10.15407/microbiolj84.02.033
J. H. Makhrmash
Objective. Pseudomonas aeruginosa is a present everywhere and opportunistic bacterium pathogen. The existence of numerous virulence factors i.e. exo-toxin, exo-enzyme genes, and biofi lm may be contributed in the pathogenesis and pathogenicity of the bacterium. The goals of the present work were to detect biofilm formation, some biofilm genes, and the effect of antibiotics against P. aeruginosa. Methods. All isolates were identified using API 20E and 16S rRNA techniques. The microtiter plate method (MTPM) was used to detect biofi lm formation. Th e polymerase chain reaction (PCR) was used to fi nd some virulence genes e.g. pelA, pslA. Results. A total of 64 P. aeruginosa isolates were identified as P. aeruginosa. The majority of infection belonged to burn infections — 27 (42.2%), followed by ear — 17 (26.5%), and urine — 20 (31.3%). The results of biofilm detection using MTPM showed that all P. aeruginosa isolates were able to produce biofilm but at different levels. PCR technique was used to detect biofilm genes. Studies showed that 61 (95.30%) and 63 (99.32%) isolates carried pelA and pslA genes, respectively. Moreover, a susceptibility test was used to select 10 antibiotics. P. aeruginosa isolates were resistant to cefotaxime — 61 (95.3%), carbenicillin — 59 (92.2%), ampicillin — 38 (59.4%), piperacilin/tazobactam — 29 (45.3%), streptomycin — 28 (43.8%), moxifloxacin — 27 (42.4%), ticarcilin — 26 (40.6%), ciprofloxacin — 24 (37.5%), gentamicin — 20 (31.3%), and neomycin — 13 (20.3%). Conclusions. Biofilm is produced by P. aeruginosa at different levels. The molecular technique showed that the pelA and pslA genes are associated with the form of biofilm in P. aeruginosa isolates. The susceptibility tests showed that the most active antibiotics against P. aeruginosa were neomycin, gentamycin, and ciprofloxacin, respectively.
目标。铜绿假单胞菌是一种普遍存在的机会致病菌。许多毒力因子如外毒素、外酶基因和生物膜的存在可能在细菌的发病和致病性中起作用。本研究的目的是检测铜绿假单胞菌的生物膜形成、部分生物膜基因以及抗生素对铜绿假单胞菌的作用。方法。所有分离株均采用API 20E和16S rRNA技术进行鉴定。采用微滴板法(MTPM)检测生物膜的形成。采用聚合酶链反应(PCR)技术对pelA、pslA等毒力基因进行检测。结果。64株铜绿假单胞菌被鉴定为铜绿假单胞菌。烧伤感染27例(42.2%),耳部感染17例(26.5%),尿部感染20例(31.3%)。MTPM生物膜检测结果表明,所有铜绿假单胞菌分离株均能产生生物膜,但水平不同。采用PCR技术检测生物膜基因。研究表明,61株(95.30%)和63株(99.32%)分别携带pelA和pslA基因。并进行药敏试验,筛选出10种抗生素。铜绿假单胞菌对头孢噻肟61(95.3%)、卡比西林59(92.2%)、氨苄西林38(59.4%)、哌拉西林/他唑巴坦29(45.3%)、链霉素28(43.8%)、莫西沙星27(42.4%)、替卡西林26(40.6%)、环丙沙星24(37.5%)、庆大霉素20(31.3%)、新霉素13(20.3%)耐药。结论。铜绿假单胞菌在不同水平上产生生物膜。分子技术表明,pslA和pelA基因与铜绿假单胞菌生物膜的形成有关。药敏试验显示,对铜绿假单胞菌最有效的抗生素分别是新霉素、庆大霉素和环丙沙星。
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引用次数: 0
Selection of Optimal Conditions for Cultivation of Bacillus megaterium UCM B-5710 – Producer of Keratinase 角化酶产生菌巨芽孢杆菌UCM B-5710的最佳培养条件选择
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.032
K. V. Avdiyuk, Anisha Roy
Every year the volume of production of poultry products all over the world is growing steadily. This contributes to a constant increase in the amount of by-products of poultry processing in the form of down and feather waste, which are dangerous for the environment due to the hard-to-degrade keratin protein and a large number of microbial pathogens. Therefore, the use of environmentally friendly methods for the destruction of keratin substrates due to keratinases of microorganisms is an urgent area of research. The aim of this work was to select the optimal cultivation conditions for the Bacillus megaterium strain UCM B-5710 to increase the activity of the keratinase synthesized by it. Methods. The culture was grown at 28°C, 201 rpm for 7 days on a basic nutrient medium containing defatted chicken feathers as the only source of carbon and nitrogen. The selection of optimal cultivation conditions was carried out according to the following parameters: temperature (21°C, 28°C, 42°C), stirring speed (201 rpm, 212 rpm), amount of inoculum (5%, 10%, 15% , 20%, 25%), the initial pH value of the nutrient medium (4.0–11.0), concentration of keratin-containing substrate (0.1%, 0.2%, 0.5%, 1.0%, 1.5%, 2.0%), additional carbon source (glucose, galactose, lactose, maltose, sucrose, mannitol, potato and corn starch, soluble starch, soybean meal) and nitrogen (NH4Cl, NH4NO3, (NH4)2SO4, NaNO3, urea, peptone, tryptone, yeast extract and soybean meal) at a concentration of 1%. Keratinase activity was assessed by the UV absorption at 280 nm of the hydrolysis products of keratin-containing raw materials. Protein was determined by the Lowry method. Results. The dynamics of the enzyme synthesis showed that the culture of B. megaterium UCM B-5710 exhibited the highest keratinase activity on the 3rd day, and complete splitting of feathers was observed on the 4–5th days. The selection of the concentration of the keratin-containing substrate showed that 0.5% is the optimal concentration. The study of the influence of the initial pH value of the nutrient medium indicates that the culture grew well at pH 6.0–7.0 and pH 9.0–11.0, but at pH 8.0 its growth was very weak. The culture exhibited the maximum keratinase activity at pH 10.0. In addition, at this pH value, complete splitting of feathers was visually observed. The influence of such a key factor as temperature on the growth and synthesis of the enzyme by B. megaterium UCM B-5710 culture demonstrated complete splitting of feathers already on the 2nd day of cultivation at 42°C, at 21°C the culture split feathers very poorly. The introduction of the inoculum into the composition of the nutrient medium in an amount of 15% of the volume of the medium and the mixing intensity of 212 rpm turned out to be optimal. Besides, it was shown that the introduction of an additional source of carbon or nitrogen had an ambiguous effect on the level of keratinase activity of B. megaterium UCM B-5710. Complete inhibition of enzyme synthes
全世界家禽产品的产量每年都在稳步增长。这导致以羽绒和羽毛废料形式出现的家禽加工副产品的数量不断增加,由于难以降解的角蛋白和大量微生物病原体,这些副产品对环境是危险的。因此,利用环境友好的方法来破坏微生物角蛋白酶引起的角蛋白底物是一个迫切的研究领域。为提高巨芽孢杆菌UCM B-5710合成的角化酶活性,选择最佳培养条件。方法。在含脱脂鸡毛作为唯一碳氮来源的基本营养培养基上,在28°C, 201 rpm下培养7天。根据以下参数选择最佳培养条件:温度(21°C, 28°C, 42°C),搅拌速度(201 rpm, 212 rpm),接种量(5%,10%,15%,20%,25%),营养培养基的初始pH值(4.0-11.0),含角蛋白底物浓度(0.1%,0.2%,0.5%,1.0%,1.5%,2.0%),附加碳源(葡萄糖,半乳糖,乳糖,麦芽糖,蔗糖,甘露醇,马铃薯和玉米淀粉,可溶性淀粉,豆粕)和氮(NH4Cl, NH4NO3, (NH4)2SO4, NaNO3,尿素,蛋白胨,色氨酸,酵母浸膏和豆粕),浓度为1%。利用含角蛋白原料的水解产物在280 nm处的紫外吸收来评价角蛋白酶的活性。蛋白质用Lowry法测定。结果。酶合成动力学结果表明,培养后第3天角化酶活性最高,第4 ~ 5天羽毛完全分裂。对含角蛋白底物浓度的选择表明,含角蛋白底物的最佳浓度为0.5%。对营养培养基初始pH值影响的研究表明,培养物在pH 6.0 ~ 7.0和pH 9.0 ~ 11.0时生长良好,但在pH 8.0时生长非常弱。培养物在pH 10.0时角化酶活性最高。此外,在这个pH值下,可以直观地观察到羽毛的完全分裂。温度等关键因素对megaterium UCM B-5710培养基生长和酶合成的影响表明,在42°C培养的第2天,羽毛已经完全分裂,在21°C培养的羽毛分裂非常差。结果表明,将接种菌以培养基体积的15%加入营养培养基,搅拌强度为212转/分时,接种菌的混合效果最佳。此外,研究表明,引入额外的碳源或氮源对巨型白僵菌UCM B-5710的角化酶活性水平有不明确的影响。在营养培养基中加入硫酸铵可以完全抑制酶的合成,在葡萄糖、乳糖和麦芽糖的情况下可以部分抑制酶的合成。马铃薯、玉米和可溶性淀粉刺激角化酶的合成。大多数无机氮源(氯化铵和硝酸盐)对大褐藻UCM B-5710角化酶的合成没有影响,而有机氮源(尿素、蛋白胨、色氨酸、酵母提取物)使角化酶活性水平提高了20-50%。其中以豆粕的效果最好,在营养培养基中添加豆粕可使角化酶活性提高2.5倍。结论。研究结果确定了大芽孢杆菌UCM B-5710菌株的最佳培养条件:培养物生长发育的最佳温度为42℃,初始pH值为10.0,搅拌速度为212转/分,接种量为15%,外加浓度为0.5%的豆粕形式的碳氮源。这使得角化酶的活性提高了4倍。
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引用次数: 1
Growth and Phytase Activities of Bacillus subtilis IMV B-7023 During Cultivation with Sodium Phytate 植酸钠培养枯草芽孢杆菌IMV B-7023的生长和植酸酶活性
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.013
N. Chuiko, A. Chobotarov, I. Kurdish
Bacteria of the genus Bacillus are known for their ability to mineralize organic phosphorus compounds. Phytates constitute up to 60–80% of the total plant phosphorus and almost 50% of soil organic phosphorus. Phytates phosphorus is unavailable for plants. Bacillus can synthesize phosphatases both wide spectrum of action, and highly specific phytases that catalyze the hydrolysis of phytates. Therefore, the aim of this work was to study the growth and phytase activity of Bacillus subtilis IMV B-7023, which is the component of the ”Azogran” complex bacterial preparation for plant growing. Methods. The growth activity of bacteria was studied by cultivation methods, the phytase activity – by measuring the amount of phosphate released from sodium phytate during the enzymatic reaction. Results. It was shown that B. subtilis IMV B-7023 assimilated phytate as the source of phosphorus nutrition during cultivation in media with 0.5, 1.0 and 2.0 g/L of sodium phytate. The highest growth activity of these bacteria was observed after two days of cultivation in medium with 1.0 g/L of phytate. The number of bacteria was (3.91±0.32)×109 CFU/mL under these conditions. At the same time, B. subtilis IMV B-7023 demonstrated a low level of phytate assimilation as a source of carbon nutrition. Thus, after two days of cultivation the number of bacteria increased from (4.12±0.09)×106 CFU/mL to (1.07±0.07–3.11±0.51)×107 CFU/mL in the presence of 0.5–2.0 g/L phytate in the medium and the absence of another carbon source. It was determined that strain B. subtilis IMV B-7023 had phytase activity, the highest activity (221.85±0.12 U/g) was on the first day of their cultivation in medium with inorganic phosphates. It should be noted that B. subtilis IMV B-7023 phytase activity was lower during cultivating in medium with sodium phytate as a source of phosphorus nutrition, than in medium with inorganic phosphates. The obtained fact may be due to phytate hydrolysis by widespecific phosphatases. Higher rates of phytase activity obtained on the first and third days compared to the second and fourth days of bacterial cultivation may indicate the expression of phosphatases genes only in the period required for maximum bacterial development, in the absence of these proteins in the media. At the same time, the phytase activity of B. subtilis IMV B-7023 after 2 days cultivation in a media with 0.5 and 1.0 g/L of sodium phytate (194.80±0.15 U/g and 160.90±0.13 U/g, respectively) as the source of carbon and phosphorus was higher compared to the activity of bacteria on medium with inorganic phosphates (137.79±0.10 U/g). This may be caused by the synthesis of a larger number of highly specific phosphatases (phytases) in bacterial cells at the presence of only phytate in the medium as a substrate. Conclusions. B. subtilis IMV B-7023 strain is characterized by growth on nutrient medium with sodium phytate and phytase activity. Because they are soil microorganisms used as the component of the
芽孢杆菌属的细菌以其矿化有机磷化合物的能力而闻名。植酸盐占植物总磷的60-80%,占土壤有机磷的近50%。植物无法获得植酸磷。芽孢杆菌既能合成作用谱广的磷酸酶,又能合成催化植酸水解的高度特异性的植酸酶。因此,本研究的目的是研究枯草芽孢杆菌IMV B-7023的生长和植酸酶活性,IMV B-7023是植物生长用“偶氮”复合菌制剂的组成部分。方法。采用培养法研究了细菌的生长活性,通过测定酶促反应中植酸钠释放的磷酸盐量来测定植酸酶的活性。结果。结果表明,枯草芽孢杆菌IMV B-7023在植酸钠浓度分别为0.5、1.0和2.0 g/L的培养基中均能吸收植酸作为磷的营养来源。在植酸浓度为1.0 g/L的培养基中培养2天后,这些细菌的生长活性达到最高。在此条件下细菌数量为(3.91±0.32)×109 CFU/mL。与此同时,枯草芽孢杆菌IMV B-7023作为碳营养来源的植酸同化水平较低。因此,在培养基中添加0.5 ~ 2.0 g/L植酸且不添加其他碳源的情况下,培养2天后细菌数量从(4.12±0.09)×106 CFU/mL增加到(1.07±0.07-3.11±0.51)×107 CFU/mL。结果表明,枯草芽孢杆菌IMV B-7023具有一定的植酸酶活性,在无机磷酸盐培养基中培养第1天的植酸酶活性最高(221.85±0.12 U/g)。在植酸钠为磷营养源的培养基中培养枯草芽孢杆菌IMV B-7023植酸酶活性低于无机磷酸盐培养基。所获得的事实可能是由于广泛的磷酸酶水解植酸盐。与细菌培养的第二天和第四天相比,第一天和第三天获得的植酸酶活性较高,这可能表明磷酸酶基因只在细菌最大发育所需的时期表达,而培养基中没有这些蛋白质。同时,以0.5和1.0 g/L植酸钠(分别为194.80±0.15 U/g和160.90±0.13 U/g)为碳源和磷源的培养基培养2 d后,枯草芽孢杆菌IMV B-7023的植酸酶活性高于以137.79±0.10 U/g为无机磷酸盐的培养基。这可能是由于在培养基中仅存在植酸盐作为底物时,细菌细胞中合成了大量高度特异性的磷酸酶(植酸酶)。结论。枯草芽孢杆菌IMV B-7023菌株在具有植酸钠和植酸酶活性的营养培养基上生长。由于它们是用于植物生长的“固氮”复合细菌制剂的土壤微生物,因此水解和同化植酸的能力对该菌株在根际的功能很重要。研究结果拓展了枯草芽孢杆菌IMV B-7023对植物磷营养和发育影响的认识。
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引用次数: 1
Biotechnology of Newly Created Bacterial Composition for Siloing Based on Lactic Acid Bacteria 基于乳酸菌的新型筒仓菌群生物技术研究
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.020
S. Danylenko, O. V. Naumenko, A. S. Onishchenko, S. Teterina, M. Khonkiv, S.O. Skrotskyi
Peculiarities of high-quality silage production are the use of biological products based on lactic acid bacteria. The composition of such starters varies greatly according to the use of bacterial cultures, so among the starters available on the market, the range of their effectiveness is also different. It is very common to use a one-sided approach to the choice of bacterial components, which in combination with imperfect production technology have low preservative activity. The study of combined preparations, which combine homo- and heterofermentative types of lactic acid fermentation, allows to stabilize the preservative properties throughout the ensiling time, and increase the aerobic stability of the silage after access of oxygen. Aim. Development of biotechnology of bacterial preparation for corn ensiling, optimization of cultivation conditions of newly created bacterial composition, and selection of cryoprotectants for its lyophilization. Methods. The combined preparation was created on the basis of heterofermentative strain Lactobacillus buchneri 3806 combining it in two- and three-strain compositions with other representatives of lactic acid bacteria, which are characterized by obligate homofermentative and facultative heterofermentative types of metabolism. Optimization of the environment and technological parameters was carried out using a central-compositional plan, further statistical analysis of the obtained data and determination of optimal values of input parameters according to the created mathematical model of optical density response. The effectiveness of the selected protective media was tested for the survival of bacteria after lyophilization. Results. The most effective bacterial composition was found during experiments: L. buchneri 3806, Enterococcus faecium C-8-12, L. plantarum 3216. The effectiveness of the obtained composition was tested by laboratory silage of corn. Tests of the drug based on the selected bacterial composition showed an improvement in the chemical composition of the silage compared to the untreated control and treated only with monoculture L. buchneri 3806, namely: there was a decrease in dry matter loss by 2.21% and 2.04%, 22 due to the increase of lactic acid content, and increase of aerobic stability of silage – 341 h against 57 h of the control sample, and 313 h in case of using monoculture. For the obtained bacterial composition, the culture medium of the following composition was optimized: base (hydrolyzed milk with the addition of the following components: monosubstituted potassium phosphate – 2 g/L; 5-aqueous manganese sulfate – 0.05 g/L; 7-aqueous magnesium sulfate – 0.2 g/L; twin-80 – 1.0 g/L); glucose – 19.7 g/L; yeast extract – 7.8 g/L; corn extract – 23.6 g/L; peptone – 9.1 g/L; sodium citrate – 6.6 g/L; sodium acetate – 3,4 g/L. Cultivation of the bacterial composition on an optimized medium made it possible to obtain the maximum biomass yield, at which the optical density was 2.01 units,
优质青贮饲料生产的特点是使用以乳酸菌为基础的生物制品。这类发酵剂的成分根据细菌培养物的用途有很大的不同,所以在市场上可买到的发酵剂中,它们的效力范围也不同。使用片面的方法来选择细菌成分是非常常见的,这与不完善的生产技术相结合,具有低的防腐活性。联合制剂的研究,将同种和异种发酵类型的乳酸发酵结合起来,可以在青贮的整个时间内稳定其防腐性能,并增加青贮在获得氧气后的有氧稳定性。的目标。玉米青贮细菌制备的生物技术进展、新细菌组合培养条件的优化及其冻干保护剂的选择。方法。该组合制剂是在杂交菌布氏乳杆菌3806的基础上,将其与其他具有专性同发酵和兼性异发酵代谢类型的乳酸菌代表菌组合成2株和3株组合而成。采用中心组成方案对环境参数和工艺参数进行优化,根据建立的光密度响应数学模型对获得的数据进行统计分析,并确定输入参数的最优值。所选择的保护介质在冻干后对细菌的存活率进行了有效性测试。结果。实验发现,最有效的细菌组成为布氏乳杆菌3806、屎肠球菌C-8-12、植物乳杆菌3216。通过室内玉米青贮试验,验证了所获组合物的有效性。根据所选细菌组成进行的药物试验表明,与未处理对照和仅单培养布氏乳杆菌3806相比,青贮的化学成分有所改善,即:干物质损失量分别减少2.21%和2.04%,22由于乳酸含量的增加,青贮的有氧稳定性增加- 341 h,比对照样品的57 h,而使用单培养时为313 h。对于所得的细菌组合物,优化了以下组合物的培养基:碱水解乳,添加以下组分:单取代磷酸钾- 2 g/L;5-水硫酸锰- 0.05 g/L;7-水溶液硫酸镁- 0.2 g/L;twin-80 - 1.0 g/L);葡萄糖- 19.7 g/L;酵母膏- 7.8 g/L;玉米提取物23.6 g/L;蛋白胨- 9.1 g/L;柠檬酸钠6.6 g/L;醋酸钠- 3,4 g/L。在优化的培养基上培养细菌组合物可以获得最大的生物量,其光密度为2.01个单位,几乎是在MRS培养基中培养相同组合物的两倍。确定了培养细菌组成的最佳工艺参数,即在36.4±0.4℃的温度下,培养基的pH值保持在6.5±0.1单位的水平上,生长效果最佳。此外,选择了柠檬酸钠、蔗糖和琼脂组成的最佳保护培养基,保证了乳酸菌冻干后的存活率为98.4%。结论。新形成的细菌组成可用于生产玉米青贮饲料的制剂,并在其他原料上进行试验,特别是在一些多年生豆科植物(苜蓿、三叶草)上进行试验,其生产条件可用于扩大该技术的规模。
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引用次数: 0
Inhibitors of Corrosion Induced by Sulfate-Reducing Bacteria 硫酸盐还原菌腐蚀抑制剂的研究
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.095
L. Purish, D. Abdulina, G. Iutynska
Currently, a lot of researcher’s attention is devoted to the problem of microbiologically influenced corrosion (MIC), since it causes huge damages to the economy, initiating the destruction of oil and gas pipelines and other underground constructions. To protect industrial materials from MIC effects an organic chemical inhibitors are massively used. However, the problem of their use is associated with toxicity, dangerous for the environment that caused the need for development the alternative methods of MIC repression. At the review, the data about different types of inhibitors-biocides usage has provided. The chemical inhibitors features are given and the mechanisms of their protective action are considered. The screening results and use of alternative and eco-friendly methods for managing the effect of corrosion caused by sulfate-reducing bacteria (SRB) are highlighted. Methods of joint application of chemical inhibitors and enhancers, such as chelators, biosurfactants, which contribute to reducing the concentration of chemical inhibitors, are discussed. The possibility of disruption of the quorum sensing interaction in the bacterial community to prevent the biofilm formation is considered. The information about the use of natural plant extracts, food waste, as well as by-products of agro-industrial production to combat MIC is provided. The development of biological corrosion control methods (to combat MIC) is of great importance for creating the best alternative and eco-friendly approaches to managing the effect of corrosion caused by SRB. The analysis of the literature data indicates the need to find the best alternatives and environmentally friendly solutions.
微生物影响腐蚀(MIC)对经济造成了巨大的损害,引发了石油和天然气管道和其他地下建筑的破坏,目前引起了许多研究者的关注。为了保护工业材料不受MIC效应的影响,有机化学抑制剂被大量使用。然而,它们的使用问题与毒性有关,对环境有危险,因此需要开发抑制MIC的替代方法。在综述中,提供了不同类型的抑制剂-杀菌剂的使用数据。给出了化学抑制剂的特点,并对其保护作用机理进行了探讨。重点介绍了硫酸盐还原菌(SRB)腐蚀影响的筛选结果和替代环保方法的使用。讨论了化学抑制剂和增强剂(如螯合剂、生物表面活性剂)联合应用的方法,这些方法有助于降低化学抑制剂的浓度。考虑了破坏细菌群落中群体感应相互作用以防止生物膜形成的可能性。提供了有关利用天然植物提取物、食物废物以及农用工业生产副产品来防治传染性疾病的信息。生物腐蚀控制方法的发展(以对抗MIC)对于创造最佳替代和生态友好的方法来管理SRB引起的腐蚀影响非常重要。对文献数据的分析表明,需要找到最佳的替代方案和环境友好的解决方案。
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引用次数: 4
Assessment of Phenol Detoxification by Rhodococcus aetherivorans UСM Ac-602 Using the Phytotesting Method 用植物试验法评价嗜热红球菌UСM Ac-602对苯酚的解毒作用
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.003
L. Khomenko, T. Nogina, V. Pidgorskyi
Monoaromatic compounds are related to widespread pollutants of soil and groundwater. Among them phenol is one of the most toxic and carcinogenic compounds. Therefore biodestruction of phenol is of much importance for environment protection. The use of metabolic potential of microorganisms for depolluting environment is a safe and economical alternative to widely used physicochemical methods. Aim. To assess efficacy of phenol detoxification with strain Rhodococcus aetherivorans UСM Ac-602 using the phytotesting method. Methods. Bacteria were cultivated in liquid mineral medium with initial concentration of phenol 500, 750 and 1000 mg/L as a single source of carbon and energy. Cultivation time was 24 h, 48 h and 72 h respectively. Phytotoxicity was determined in express-test with use of seeds of spring wheat variety “Pecheryanka” (Triticum aestivum L.). Plant seeds were incubated by temperature 20±2°C during 7 days in Petri dishes with filter paper treated with respective phenol aqueous solutions or post-fermentative cultural fluids (PFCFs). PFCFs were obtained after cultivation of strain in growth medium with same concentration of phenol. Morphometric parameters of wheat were assessed against control plants cultivated on distilled water. Comparative analysis of samples toxicity and toxicity class determination was performed according to Kabirov method by calculation of index of test factor toxicity (ITF). Results. Phenol aqueous solutions and PFCFs were much different in effect on wheat. Phenol solutions 500 and 700 mg/L have shown significant inhibitory effect on all initial growth parameters of test plants. The weakest growth inhibition was induced by phenol concentration of 500 mg/L which caused decrease in number of germinated seeds by 59.6%, shoot length – by 59.7%, root length – by 84.5%, sprout dry weight – by 35.0%. In the presence of phenol concentration of 750 mg/L these indicators increased by 7−30%; roots of test plants were the most sensitive to effect of phenol. Phenol concentration of 1000 mg/L caused total seed mortality. Unlike phenol aqueous solutions PFCFs have shown insignificant effect on all morphometric indicators of plants compared to control. Similar effects on plants were observed in the presence of PFCFs obtained from cultivation of strain R. aetherivorans UСM Ac-602 in the growth medium with initial concentrations of phenol of 500 and 750 mg/L. Under the influence of these PFCFs, the number of germinated seeds decreased on average by 15.8%, root length decreased by 19.8%, at the same time shoot length and their dry weight increased by 17.8% and 7.2% respectively. More negative effect on wheat was shown by PFCF obtained after strain cultivation on medium with phenol concentration 1000 mg/L. It caused reduction in number of germinated seeds by 18.0 %, shoot length – by 25.3%, root length – by 29.0%, sprout dry weight – by 7.2%. For phenol aqueous solutions ITFs had much lower values 0–0.40 than for PFCFs (0.71–1.0). Concl
单芳香族化合物是土壤和地下水中广泛存在的污染物。其中苯酚是毒性和致癌性最强的化合物之一。因此,苯酚的生物降解对环境保护具有重要意义。利用微生物的代谢潜能对环境进行净化是一种安全、经济的方法,可以替代目前广泛使用的物理化学方法。的目标。采用植物试验法评价嗜热红球菌UСM Ac-602对苯酚的解毒作用。方法。细菌在初始浓度为500、750和1000 mg/L的液体矿物培养基中培养,作为碳和能量的单一来源。培养时间分别为24 h、48 h和72 h。以春小麦品种“Pecheryanka”(Triticum aestivum L.)种子为材料,通过表达试验测定了其植物毒性。植物种子在温度20±2℃的培养皿中培养7天,滤纸分别用苯酚水溶液或发酵后培养液(pfcf)处理。菌株在相同苯酚浓度的培养基中培养后可得到pfcf。以蒸馏水栽培的对照植株为对照,对小麦的形态计量学参数进行了评价。根据卡比洛夫法计算试验因子毒性指数(ITF),对样品毒性进行对比分析,并确定毒性等级。结果。苯酚水溶液和全氟氯烃对小麦的影响差异较大。500和700 mg/L苯酚溶液对试验植物的所有初始生长参数均有显著的抑制作用。苯酚浓度为500 mg/L时的生长抑制作用最弱,使发芽种子数减少59.6%,芽长减少59.7%,根长减少84.5%,芽干重减少35.0%。苯酚浓度为750 mg/L时,这些指标提高了7 ~ 30%;受试植物的根对苯酚的影响最为敏感。1000mg /L苯酚浓度导致种子总死亡率。与苯酚水溶液不同,与对照相比,全氟氯烃对植物所有形态计量指标的影响均不显著。菌株R. aetherivorans UСM Ac-602在苯酚初始浓度为500 mg/L和750 mg/L的培养基中培养得到的pfcf对植物也有类似的影响。在这些pfcf的影响下,种子发芽数平均减少15.8%,根长平均减少19.8%,茎长和干重平均增加17.8%和7.2%。在苯酚浓度为1000 mg/L的培养基上进行菌种培养后得到的PFCF对小麦的负面影响更大。处理后种子发芽数减少18.0%,茎长减少25.3%,根长减少29.0%,芽干重减少7.2%。苯酚水溶液的ITFs值0 ~ 0.40远低于全氟氯烃(0.71 ~ 1.0)。结论。根据本研究获得的数据,可以得出结论,菌株R. aetherivorans UCM Ac-602对高浓度含酚培养基具有活性解毒作用。对ITF培养基值(ITFm)计算结果的分析表明,在所研究菌株的影响下,苯酚溶液(500、750和1000 mg/L)的毒性有所降低。根据Kabirov毒性量表,对初始值为II级(高)和I级(非常高)的苯酚溶液进行毒性评估,将其毒性降至IV级(低)和V级(正常)。本研究结果证明了该菌株UCM AC-602对苯酚破坏的最终产物具有生态安全性,并展望了其在苯酚污染环境解毒生物技术中的应用前景。
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引用次数: 0
The Effect of Surfactants of Microbial Origin on Phytopathogenic Microorganisms 微生物源表面活性剂对植物病原微生物的影响
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.075
T. Pirog, D. Piatetska, H. Yarova, G. Iutynska
Biodegradable non-toxic surfactants of microbial origin are multifunctional preparations, which due to antimicrobial activity are promising for use in crop production to control phytopathogenic microorganisms. Studies on the prospects of using microbial surfactants to control the number of phytopathogenic microorganisms are conducted in three directions: laboratory studies of antimicrobial activity of surfactants in vitro, determination of the effect of surfactants on phytopathogens in vegetative experiments in the process of plants growing in a laboratory or greenhouse, post-harvest treatment of fruits and vegetables with solutions of microbial surfactants to extend their shelf life. The review presents literature data on antimicrobial activity of surfactants against phytopathogenic bacteria and fungi in vitro. Antimicrobial activity of surfactants is evaluated by three main parameters: minimum inhibitory concentration, zones of growth retardation of test cultures on agar media and inhibition of growth of test cultures on agar or liquid media. The vast majority of available publications relate to the antifungal activity of surfactant lipopeptides and rhamnolipids, while data on the effect of these microbial surfactants on phytopathogenic bacteria (representatives of the genera Ralstonia, Xanthomonas, Pseudomonas, Agrobacterium, Pectobacterium) are few. The researchers determined the antimicrobial activity of either total lipopeptides extracted with organic solvents from the culture broth supernatant, or individual lipopeptides (iturin, surfactin, fengycin, etc.) isolated from a complex of surfactants, or culture broth supernatant. Lipopeptides synthesized by members of the genus Bacillus exhibit antimicrobial activity on phytopathogenic fungi of the genera Alternaria, Verticillium, Aspergillus, Aureobasidium, Botrytis, Rhizoctonia, Fusarium, Penicillium, Phytophora, Sclerotinia, Curvularia, Colletotrichum, etc. in sufficiently high concentrations. Thus, the minimum inhibitory concentrations of lipopeptides against phytopathogenic fungi are orders of magnitude higher (in average 0.04–8.0 mg/mL, or 40–8000 μg/mL) than against phytopathogenic bacteria (3–75 μg/mL). However, the antifungal activity of lipopeptidecontaining supernatants is not inferior by the efficiency to the activity of lipopeptides isolated from them, and therefore, to control the number of phytopathogenic fungi in crop production, the use of lipopeptidecontaining supernatants is more appropriate. Rhamnolipids synthesized by bacteria of the genus Pseudomonas are more effective antimicrobial agents comparing to lipopeptides: the minimum inhibitory concentrations of rhamnolipids against phytopathogenic fungi are 4–276 μg/mL, which is an order of magnitude lower than lipopeptides. In contrast to the data on the antifungal activity of rhamnolipids against phytopathogens, there are only a few reports in the literature on the effect of these surfactants on phytopathogenic bacteria, whil
可生物降解的无毒微生物源表面活性剂是一种多功能制剂,由于其抗菌活性,在作物生产中具有控制植物病原微生物的前景。利用微生物表面活性剂控制植物病原微生物数量的前景研究从三个方面展开:体外表面活性剂抗菌活性的实验室研究、植物在实验室或温室生长过程中营养实验中表面活性剂对植物病原作用的测定、果蔬采后用微生物表面活性剂溶液处理以延长其货架期。综述了表面活性剂对植物病原菌和真菌体外抑菌活性的研究进展。表面活性剂的抗菌活性是通过三个主要参数来评估的:最低抑制浓度、琼脂培养基上培养物的生长迟滞区和琼脂或液体培养基上培养物的生长抑制。绝大多数现有的出版物都与表面活性剂脂肽和鼠李糖脂的抗真菌活性有关,而关于这些微生物表面活性剂对植物致病菌(Ralstonia属、黄单胞菌属、假单胞菌属、农杆菌属、胸腺杆菌属的代表)的影响的数据很少。研究人员测定了用有机溶剂从培养液上清液中提取的总脂肽,或从表面活性剂复合物或培养液上清液中分离的单个脂肽(iturin, surfactin, fengycin等)的抗菌活性。由芽孢杆菌属成员合成的脂肽在足够高浓度的情况下对Alternaria、Verticillium、Aspergillus、Aureobasidium、Botrytis、Rhizoctonia、Fusarium、Penicillium、Phytophora、Sclerotinia、Curvularia、Colletotrichum等植物病原真菌具有抗菌活性。因此,脂肽对植物致病真菌的最低抑制浓度(平均0.04-8.0 mg/mL,或40-8000 μg/mL)比对植物致病细菌的最低抑制浓度(3-75 μg/mL)高几个数量级。然而,含脂肽上清液的抗真菌活性并不低于从其中分离的脂肽的活性,因此,为了控制作物生产中植物病原真菌的数量,使用含脂肽上清液更为合适。假单胞菌属细菌合成的鼠李糖脂是比脂肽更有效的抗菌药物:鼠李糖脂对植物病原真菌的最低抑制浓度为4-276 μg/mL,比脂肽低一个数量级。与鼠李糖脂对植物病原菌的抗真菌活性数据相比,文献中关于鼠李糖脂对植物病原菌作用的报道较少,但其最低抑菌浓度却相当高(可达5000 μg/mL)。与脂肽相比,鼠李糖脂作为抗菌剂的优势在于其合成水平高、成本低且可从大量工业废弃物中获得。目前文献中关于微生物源的表面活性槐脂对植物病原真菌作用的研究较少,主要是关于槐脂的抗真菌作用。我们注意到,与表面活性剂脂肽和鼠李糖脂相比,大多数槐油脂的有效浓度更高,对植物病原体具有最高的抗菌活性,达到10,000 μg/mL。
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引用次数: 0
First Report of Potato Viruses Infecting Lamium purpureum in Ukraine 乌克兰马铃薯病毒感染紫斑Lamium pupureum的首次报告
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.055
A. Kyrychenko, M. Bohdan, H. Snihur, I. Shcherbatenko, I. Antipov
Weeds as reservoirs for destructive plant pathogens have a significant impact on the viral epidemiology, ecology and, as a result, on local economy, and are therefore being investigated in many parts of the world. Thus, the aim of this study was to investigate virus occurrence in red dead-nettle plants (Lamium purpureum L.) widespread in urban and field conditions throughout the in the Kyiv region of Ukraine. Methods. Field crop observations, visual diagnosis, biological testing of the virus, immunoassay (ELISA), polymerase chain reaction with reverse transcription (RT-PCR), sanger sequencing of partial genome sequences of PVX, PVY, PVS, PVM. Results. The results obtained in the study indicate that Lamium plants could be alternative weed hosts of number important viral diseases including potatoes and other vegetables. Serological and molecular test results evidence plants were infected by Potato virus X, Potato virus Y, Potato virus M, Potato virus S and therefore Lamium L. species can serve as a potential source of inoculum for wide range of vegetables and ornamentals. This study is the first report of Lamium plants being naturally infected with Potato virus M and Potato virus S in central Europe. Conclusions. These plants are alternative host of mixed infection with viruses belonging to different families: Alphaflexiviridae, Betaflexiviridae and Potyviridae.
杂草作为破坏性植物病原体的储存库,对病毒流行病学、生态学以及当地经济产生重大影响,因此正在世界许多地方进行调查。因此,本研究的目的是调查病毒在乌克兰基辅地区广泛分布于城市和田野条件下的红死荨麻植物(Lamium purpureum L.)中的发生情况。方法。田间作物观察、目测诊断、病毒生物学检测、免疫测定(ELISA)、逆转录聚合酶链反应(RT-PCR)、PVX、PVY、PVS、PVM部分基因组序列的sanger测序。结果。研究结果表明,叶状叶属植物可作为包括马铃薯和其他蔬菜在内的许多重要病毒病害的替代杂草寄主。血清学和分子检测结果表明,植物感染了马铃薯X病毒、马铃薯Y病毒、马铃薯M病毒和马铃薯S病毒,因此Lamium L.可以作为广泛的蔬菜和观赏植物的潜在接种源。本研究是中欧地区首次报道马铃薯M病毒和马铃薯S病毒自然感染的植物。结论。这些植物是不同科病毒混合感染的替代宿主:甲屈病毒科、倍屈病毒科和波蒂病毒科。
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引用次数: 5
Detection and Investigation of Some Properties of the Regulators of Antibiotic Biosynthesis Produced by Streptomyces Strains S. sp. AN26 and S. sp. B35 链霉菌S. sp. AN26和S. sp. B35产抗生素生物合成调控因子的检测与特性研究
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.049
B. Matselyukh
The aim of this work was the isolation, purification and some properties investigation of two regulators of antibiotic biosynthesis of streptomycetes. Methods includes extraction of regulators from agar cultures and their concentration by vacuum rotary evaporator, thin layer chromatography and spectrophotometry. Results. Two strains of streptomycetes AN26 and B35 isolated from soils of different regions of Ukraine produce the regulators restoring the landomycin E biosynthesis and sporulation in mutant strain Streptomyces globispoprus 1912-B2. Both regulators were purified by thin layer chromatography and have the same Rf 0.69. Absorption curves of regulators were established by means of spectrophotometry. Maxima of absorption of regulators were 232.5 nm. The next study of the isolated regulators by means of NMR will give the possibility to elucidate their molecular structures. Conclusions. It is shown that two strains of streptomycetes isolated from the soils of Askania Nova and Brovary produce transcriptional regulators such as signaling molecules, which, like A-factor, restore the biosynthesis of antibiotics landomycin E and streptomycin in test strains S. globisporus 1912-B2 and S. griseis 1439, respectively. In terms of absorption maxima, they are similar and differ from similar indicators of known regulators of streptomycetes. It is possible that these compounds belong to new, not yet described signaling molecules, and the answer to this question will give future studies of their molecular structure by NMR spectroscopy.
本文对链霉菌合成抗生素的两种调节因子进行了分离纯化和性质研究。方法包括从琼脂培养物中提取调节剂,并通过真空旋转蒸发器、薄层色谱和分光光度法进行浓度测定。结果。从乌克兰不同地区土壤中分离的两株链霉菌AN26和B35产生了恢复突变株全球链霉菌1912-B2中陆霉素E生物合成和产孢的调节因子。两种调节剂均经薄层色谱纯化,具有相同的Rf 0.69。用分光光度法建立了调节剂的吸收曲线。调节剂的最大吸光度为232.5 nm。下一步对分离调控分子的核磁共振研究将为阐明其分子结构提供可能。结论。结果表明,从Askania Nova和Brovary土壤中分离的两株链霉菌产生信号分子等转录调节因子,与a因子一样,恢复了测试菌株S. globisporus 1912-B2和S. griseis 1439中抗生素陆霉素E和链霉素的生物合成。就吸收最大值而言,它们与已知链霉菌调节剂的类似指标相似,但又不同。这些化合物可能属于新的,尚未被描述的信号分子,这个问题的答案将为未来通过核磁共振光谱研究它们的分子结构提供帮助。
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引用次数: 0
Species Composition and Susceptibility to Antibiotics of Microorganisms Isolated from Tooth Sockets of Extracted Teeth in Cases of Alveolar Osteitis 牙槽骨炎拔牙牙槽中分离微生物的种类组成及对抗生素的敏感性
Q4 Biochemistry, Genetics and Molecular Biology Pub Date : 2021-12-17 DOI: 10.15407/microbiolj83.06.041
V. Malanchuk, A. Javadiasl, A. Rybachuk, M. Oblap, V. Potochilova
Alveolar osteitis (AO) is one of the most common infectious complications after dental extractions. The data on the species composition of AO pathogens and their susceptibility to antimicrobial drugs can be the basis for their empirical use in case of inflammatory process aggravation. Objective. To determine the species composition and susceptibility to the antimicrobial agents of microorganisms, which were detected in patients with AO, who sought medical help in the oral surgery department of the dental medical center of Bogomolets National Medical University. Methods. Throughout 2018–2021, microbiological examination of tooth sockets from 30 patients with AO and 20 patients without AO was performed. The studied biological material was plated on appropriate nutrient media for isolation of aerobic, facultative and obligate-anaerobic microorganisms. Anaerobic conditions were achieved in GENbox 7.0 L and GENbox 2.5 L aerostats using GENbox anaerobic packages (“Biomerieux”, France). The genus and species identity of the bacteria were determined according to Bergey. Antibiotic susceptibility of the isolated strains was determined by disk diffusion method. Results. It was found that most commonly microorganisms from tooth sockets in case of AO are: Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp., Escherichia coli, Bacteroides spp., Clostridium spp., and Candida species, as well as their mixed cultures of 3–5 species of microorganisms. These aerobic and facultative anaerobic bacteria were susceptible to amoxicillin, ceftriaxone and ciprofloxacin in 92.6–100% of cases. The growth of anaerobic bacteria in 100% of cases was inhibited by colistin and meropenem. Conclusions. AO developing is caused by pathological colonization of socket of the extracted tooth by representatives of endogenous microbiota, namely Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus spp., Escherichia coli, which are present mainly in the mixed cultures with Candida albicans. For empirical antibiotic therapy of complicated forms of AO, amoxicillin or ceftriaxone or ciprofloxacin in complex with colistin or meropenem should be used, since these drugs suppress the growth of 92.6–100% of strains of aerobic, facultative and obligate anaerobic microorganisms, which are potential pathogens of the purulent forms of AO.
牙槽骨炎是拔牙后最常见的感染性并发症之一。有关AO病原菌种类组成及其对抗菌药物敏感性的数据可作为炎症过程加重时经验性用药的依据。目标。目的确定在Bogomolets国立医科大学牙科医疗中心口腔外科就诊的AO患者中检测到的微生物的种类组成和对抗菌药物的敏感性。方法。2018-2021年,对30例AO患者和20例无AO患者的牙槽进行微生物检查。将所研究的生物材料镀在适当的营养培养基上,用于分离好氧、兼性和专性厌氧微生物。采用GENbox厌氧包(“Biomerieux”,法国),在GENbox 7.0 L和GENbox 2.5 L浮空器中实现厌氧条件。根据Bergey确定了细菌的属和种身份。采用纸片扩散法测定分离菌株的药敏。结果。发现AO患者牙槽中最常见的微生物有:金黄色葡萄球菌、表皮葡萄球菌、链球菌、大肠杆菌、拟杆菌、梭菌、念珠菌等,以及它们混合培养的3-5种微生物。其中需氧和兼性厌氧菌对阿莫西林、头孢曲松和环丙沙星敏感的占92.6 ~ 100%。粘菌素和美罗培南均能抑制100%病例的厌氧菌生长。结论。AO的发生是由内源性菌群的代表,即金黄色葡萄球菌、表皮葡萄球菌、链球菌、大肠杆菌在拔牙槽内的病理性定植引起的,这些微生物主要存在于与白色念珠菌混合培养的菌群中。由于阿莫西林、头孢曲松或环丙沙星与粘菌素或美罗培南联合使用可抑制92.6-100%的好氧、兼性和专门性厌氧微生物的生长,这些微生物是化脓性AO的潜在病原体,因此对于复杂AO的经经验抗生素治疗应采用阿莫西林或头孢曲松或环丙沙星。
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Mikrobiolohichnyi zhurnal
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