首页 > 最新文献

Microbiology Research最新文献

英文 中文
Antibiotic-Resistant Bacteria in Environmental Water Sources from Southern Chile: A Potential Threat to Human Health 智利南部环境水源中的耐抗生素细菌:对人类健康的潜在威胁
Q3 MICROBIOLOGY Pub Date : 2023-11-01 DOI: 10.3390/microbiolres14040121
Matías Jofré Bartholin, Boris Barrera Vega, Liliana Berrocal Silva
Antimicrobial resistance (AMR) is a critical global issue affecting public and animal health. The overuse of antibiotics in human health, animal production, agriculture, and aquaculture has led to the selection of antibiotic-resistant strains, particularly in Gram-negative bacteria. Mutations and horizontal gene transfer play a significant role in the development of antimicrobial resistance, leading to the reduced efficacy of current antibiotics. Today, AMR in bacteria and antibiotic-resistance genes (ARGs) are increasingly recognized in multiple environmental sources, including recreational and irrigation waters. This study aims to identify Gram-negative bacteria from surface aquatic reservoirs in southern Chile and assess their susceptibility to clinically relevant antibiotics. Water samples were collected from four lakes, five rivers, one waterfall, and one watershed in southern Chile to isolate environmental Gram-negative bacilli (GNB). API-20E and MALDI–TOF were employed for bacterial identification. Kirby–Bauer disc diffusion tests and multiplex PCR were performed to determine their susceptibility profile. A total of 26 GNB strains were isolated from environmental water samples, predominantly belonging to the Pseudomonas (n = 9) and Acinetobacter (n = 7) genera. Among these strains, 96.2% were resistant to ampicillin and cefazoline, while 26.9% and 34.6% showed resistance to ceftazidime and cefepime, respectively. Additionally, 38.5% exhibited resistance to colistin. Two Enterobacter cloacae strains obtained from Cachapoal River (sixth region) and Villarrica Lake (ninth region), respectively, presented a multidrug-resistant (MDR) phenotype and carried at least two extended-spectrum β-lactamase (ESBL) genes. Thus, antibiotic-resistant GNB and ARGs were found in natural water reservoirs, raising concerns about the dissemination of resistance determinants among potentially pathogenic bacteria in environmental microbial communities.
抗微生物药物耐药性(AMR)是一个影响公众和动物健康的重大全球性问题。人类健康、动物生产、农业和水产养殖中抗生素的过度使用导致了抗生素耐药菌株的选择,特别是革兰氏阴性菌。突变和水平基因转移在抗菌素耐药性的发展中起着重要作用,导致当前抗生素的疗效降低。今天,细菌和抗生素耐药基因(ARGs)的抗菌素耐药性越来越多地在多种环境来源中得到认识,包括娱乐和灌溉用水。本研究旨在鉴定智利南部地表水生水库中的革兰氏阴性菌,并评估其对临床相关抗生素的敏感性。在智利南部4个湖泊、5条河流、1个瀑布和1个流域采集水样,分离环境革兰氏阴性杆菌(GNB)。采用API-20E和MALDI-TOF进行细菌鉴定。采用Kirby-Bauer圆盘扩散试验和多重PCR测定其敏感性谱。从环境水样中共分离到GNB菌株26株,主要属假单胞菌属(n = 9)和不动杆菌属(n = 7)。其中,对氨苄西林和头孢唑啉的耐药率为96.2%,对头孢他啶和头孢吡肟的耐药率分别为26.9%和34.6%。此外,38.5%的人表现出对粘菌素的耐药性。分别从cachhapoal River(第6区)和Villarrica Lake(第9区)获得的2株阴沟肠杆菌呈现多重耐药表型,并携带至少2个扩展谱β-内酰胺酶(ESBL)基因。因此,在天然水库中发现了耐药GNB和ARGs,引起了人们对环境微生物群落中潜在致病菌中耐药决定因素传播的关注。
{"title":"Antibiotic-Resistant Bacteria in Environmental Water Sources from Southern Chile: A Potential Threat to Human Health","authors":"Matías Jofré Bartholin, Boris Barrera Vega, Liliana Berrocal Silva","doi":"10.3390/microbiolres14040121","DOIUrl":"https://doi.org/10.3390/microbiolres14040121","url":null,"abstract":"Antimicrobial resistance (AMR) is a critical global issue affecting public and animal health. The overuse of antibiotics in human health, animal production, agriculture, and aquaculture has led to the selection of antibiotic-resistant strains, particularly in Gram-negative bacteria. Mutations and horizontal gene transfer play a significant role in the development of antimicrobial resistance, leading to the reduced efficacy of current antibiotics. Today, AMR in bacteria and antibiotic-resistance genes (ARGs) are increasingly recognized in multiple environmental sources, including recreational and irrigation waters. This study aims to identify Gram-negative bacteria from surface aquatic reservoirs in southern Chile and assess their susceptibility to clinically relevant antibiotics. Water samples were collected from four lakes, five rivers, one waterfall, and one watershed in southern Chile to isolate environmental Gram-negative bacilli (GNB). API-20E and MALDI–TOF were employed for bacterial identification. Kirby–Bauer disc diffusion tests and multiplex PCR were performed to determine their susceptibility profile. A total of 26 GNB strains were isolated from environmental water samples, predominantly belonging to the Pseudomonas (n = 9) and Acinetobacter (n = 7) genera. Among these strains, 96.2% were resistant to ampicillin and cefazoline, while 26.9% and 34.6% showed resistance to ceftazidime and cefepime, respectively. Additionally, 38.5% exhibited resistance to colistin. Two Enterobacter cloacae strains obtained from Cachapoal River (sixth region) and Villarrica Lake (ninth region), respectively, presented a multidrug-resistant (MDR) phenotype and carried at least two extended-spectrum β-lactamase (ESBL) genes. Thus, antibiotic-resistant GNB and ARGs were found in natural water reservoirs, raising concerns about the dissemination of resistance determinants among potentially pathogenic bacteria in environmental microbial communities.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"35 11","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135270789","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: 0
Phaeohyphomycosis: A Review 褐丝酵母菌病:综述
Q3 MICROBIOLOGY Pub Date : 2023-10-31 DOI: 10.3390/microbiolres14040120
Carlos D. Sánchez-Cárdenas, Mariel Isa-Pimentel, Roberto Arenas
Phaeohyphomycosis, a worldwide fungal infection, refers to uncommon superficial, subcutaneous, or systemic infections caused by melanized fungi. Currently, about 70 genera and 150 species have been described. It can be confirmed by mycological, histopathological, or molecular methods. Treatment can be performed by surgery or systemic antifungals.
褐丝菌病是一种世界性的真菌感染,是指由黑化真菌引起的罕见的浅表、皮下或全身感染。目前已发现的有70属150种。它可以通过真菌学、组织病理学或分子方法证实。治疗可通过手术或全身抗真菌药物进行。
{"title":"Phaeohyphomycosis: A Review","authors":"Carlos D. Sánchez-Cárdenas, Mariel Isa-Pimentel, Roberto Arenas","doi":"10.3390/microbiolres14040120","DOIUrl":"https://doi.org/10.3390/microbiolres14040120","url":null,"abstract":"Phaeohyphomycosis, a worldwide fungal infection, refers to uncommon superficial, subcutaneous, or systemic infections caused by melanized fungi. Currently, about 70 genera and 150 species have been described. It can be confirmed by mycological, histopathological, or molecular methods. Treatment can be performed by surgery or systemic antifungals.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"54 4","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135871307","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: 0
Exploring the Variability in Antibacterial Testing of Resin Dental Composites among Investigators: A Narrative Review 在调查人员中探索树脂牙科复合材料抗菌测试的变异性:叙述综述
Q3 MICROBIOLOGY Pub Date : 2023-10-31 DOI: 10.3390/microbiolres14040119
Ashlesha Pal, Noorulain Altaf, Anum Mahmood, Shazia Akbar, Nazrah Maher, Naresh Kumar, Muhammad Sohail Zafar, Laura Murcia, Ricardo E. Oñate-Sánchez
Caries is a common dental problem brought on by factors like excessive sugar consumption, poor oral hygiene, and the presence of microorganisms in the mouth. This dental pathology is treated with a variety of filling materials, including tooth-colored direct resin dental composite (RDC), glass ionomer cement (GIC), and dental amalgam (also known as silver filling). RDC is the most preferred filling material in dental clinics due to its excellent esthetics and minimal tooth preparation, making it the need of the modern era. However, antimicrobial agents were added to this material in order to enhance its ability to prevent secondary caries. The antibacterial activity of RDC has been tested using a variety of methods, but testing protocols have been found to vary. Thusly, the point of this article is to examine the disparity in the strategy involved by specialists for testing the antibacterial properties of RDCs.
蛀牙是一种常见的牙齿问题,由摄入过多的糖、口腔卫生不良和口腔中存在微生物等因素引起。这种牙齿病变可以用各种填充物来治疗,包括牙色直接树脂牙科复合材料(RDC)、玻璃离子水合物(GIC)和牙科汞合金(也称为银填充物)。RDC因其优异的美观性和最少的牙齿准备而成为牙科诊所最首选的填充材料,使其成为现代时代的需要。然而,抗菌药物被添加到这种材料中,以提高其预防继发性龋齿的能力。RDC的抗菌活性已使用多种方法进行了测试,但测试方案各不相同。因此,本文的重点是检查专家测试rdc抗菌特性所涉及的策略的差异。
{"title":"Exploring the Variability in Antibacterial Testing of Resin Dental Composites among Investigators: A Narrative Review","authors":"Ashlesha Pal, Noorulain Altaf, Anum Mahmood, Shazia Akbar, Nazrah Maher, Naresh Kumar, Muhammad Sohail Zafar, Laura Murcia, Ricardo E. Oñate-Sánchez","doi":"10.3390/microbiolres14040119","DOIUrl":"https://doi.org/10.3390/microbiolres14040119","url":null,"abstract":"Caries is a common dental problem brought on by factors like excessive sugar consumption, poor oral hygiene, and the presence of microorganisms in the mouth. This dental pathology is treated with a variety of filling materials, including tooth-colored direct resin dental composite (RDC), glass ionomer cement (GIC), and dental amalgam (also known as silver filling). RDC is the most preferred filling material in dental clinics due to its excellent esthetics and minimal tooth preparation, making it the need of the modern era. However, antimicrobial agents were added to this material in order to enhance its ability to prevent secondary caries. The antibacterial activity of RDC has been tested using a variety of methods, but testing protocols have been found to vary. Thusly, the point of this article is to examine the disparity in the strategy involved by specialists for testing the antibacterial properties of RDCs.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"61 4","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135809955","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: 0
In Vitro Cytotoxicity and Hemolysis Effect of Poly-Gamma-Glutamic Acid Nano-Polymer Biosynthesized Using Some Isolates of Bacillus spp. 利用芽孢杆菌分离物合成的聚γ -谷氨酸纳米聚合物的体外细胞毒性和溶血作用。
Q3 MICROBIOLOGY Pub Date : 2023-10-29 DOI: 10.3390/microbiolres14040118
Eman M. Elsayed, Ahmed A. Farghali, Mohamed I. Zanaty, Medhat Abdel-Fattah, Dalal Hussien M. Alkhalifah, Wael N. Hozzein, Ahmed M. Mahmoud
Natural polymers have good biological, biocompatible properties and a low degradation rate, so they can be used in medical applications. This study demonstrates an optimized biosynthesis for poly-gamma-glutamic acid nano-polymer (ɣ-PGA NP) using three bacterial isolates encoded as B4, B5, and B6. The 16S rRNA gene sequence of the isolates showed 98.5, 99, and 99.8% similarity with Bacillus sp., with accession numbers MZ976778, MZ976779, and MZ956153, respectively. The optimal conditions of the biosynthesis were 35 °C, followed by incubation for 30 h, 7 pH, 0.7 O.D600 of bacterial broth, carbon sources of glutamic acid and glucose, and ammonium sulfate as the nitrogen source. Biosynthesized ɣ-PGA NPs were characterized using X-ray diffraction (XRD), Zeta sizer, Fourier transform infrared (FT-IR), transmission electron microscopy (TEM), and nuclear magnetic resonance (1HNMR). The TEM study showed the formation of oval-shaped ɣ-PGA NPs with sizes 31.75–32.69 nm, 18.63–21.66 nm, and 37.57–39.22 nm for B4, B5, and B6, respectively. XRD, FT-IR, and 1HNMR confirmed the purity of the polymer. The hemolysis rates of the biosynthesized ɣ-PGA NPs were below the acceptable value. Moreover, the cytotoxicity test conducted using an MTT assay on peripheral blood mononuclear cells (PBMCs) revealed no significant effect on the cell viability within a dose range of up to 512 μg/mL.
天然聚合物具有良好的生物相容性和低降解率,可用于医疗领域。本研究利用编码为B4、B5和B6的三种细菌分离物优化了聚γ -谷氨酸纳米聚合物(γ -PGA NP)的生物合成。分离菌株的16S rRNA基因序列与芽孢杆菌相似度分别为98.5%、99%和99.8%,登录号分别为MZ976778、MZ976779和MZ956153。生物合成的最佳条件为35℃,7 pH, 0.7 O.D600菌液,谷氨酸和葡萄糖为碳源,硫酸铵为氮源,孵育30 h。利用x射线衍射(XRD)、Zeta粒度仪、傅里叶变换红外(FT-IR)、透射电子显微镜(TEM)和核磁共振(1HNMR)对生物合成的α -PGA NPs进行了表征。透射电镜研究表明,B4、B5和B6分别形成了尺寸为31.75 ~ 32.69 nm、18.63 ~ 21.66 nm和37.57 ~ 39.22 nm的椭圆形的α -PGA NPs。XRD, FT-IR和1HNMR证实了该聚合物的纯度。生物合成的α -PGA NPs溶血率低于可接受值。此外,采用MTT法对外周血单个核细胞(PBMCs)进行的细胞毒性试验显示,在高达512 μg/mL的剂量范围内,对细胞活力无显著影响。
{"title":"In Vitro Cytotoxicity and Hemolysis Effect of Poly-Gamma-Glutamic Acid Nano-Polymer Biosynthesized Using Some Isolates of Bacillus spp.","authors":"Eman M. Elsayed, Ahmed A. Farghali, Mohamed I. Zanaty, Medhat Abdel-Fattah, Dalal Hussien M. Alkhalifah, Wael N. Hozzein, Ahmed M. Mahmoud","doi":"10.3390/microbiolres14040118","DOIUrl":"https://doi.org/10.3390/microbiolres14040118","url":null,"abstract":"Natural polymers have good biological, biocompatible properties and a low degradation rate, so they can be used in medical applications. This study demonstrates an optimized biosynthesis for poly-gamma-glutamic acid nano-polymer (ɣ-PGA NP) using three bacterial isolates encoded as B4, B5, and B6. The 16S rRNA gene sequence of the isolates showed 98.5, 99, and 99.8% similarity with Bacillus sp., with accession numbers MZ976778, MZ976779, and MZ956153, respectively. The optimal conditions of the biosynthesis were 35 °C, followed by incubation for 30 h, 7 pH, 0.7 O.D600 of bacterial broth, carbon sources of glutamic acid and glucose, and ammonium sulfate as the nitrogen source. Biosynthesized ɣ-PGA NPs were characterized using X-ray diffraction (XRD), Zeta sizer, Fourier transform infrared (FT-IR), transmission electron microscopy (TEM), and nuclear magnetic resonance (1HNMR). The TEM study showed the formation of oval-shaped ɣ-PGA NPs with sizes 31.75–32.69 nm, 18.63–21.66 nm, and 37.57–39.22 nm for B4, B5, and B6, respectively. XRD, FT-IR, and 1HNMR confirmed the purity of the polymer. The hemolysis rates of the biosynthesized ɣ-PGA NPs were below the acceptable value. Moreover, the cytotoxicity test conducted using an MTT assay on peripheral blood mononuclear cells (PBMCs) revealed no significant effect on the cell viability within a dose range of up to 512 μg/mL.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"40 6","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136135712","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: 0
Short-Term PE Generation Processes in the Soils of a Farmer Plots Network in the Madagascar Highlands: Actors and Drivers 马达加斯加高原农民小区网络土壤中短期PE生成过程:行动者和驱动因素
Q3 MICROBIOLOGY Pub Date : 2023-10-27 DOI: 10.3390/microbiolres14040117
Kanto Razanamalala, Tantely Razafimbelo, Pierre-Alain Maron, Lionel Ranjard, Nicolas Chemidlin Prévost-Bouré, Samuel Dequiedt, Thierry Becquer, Jean Trap, Eric Blanchart, Laetitia Bernard
Carbon sequestration in agricultural soils, through the accumulation of high-quality organic matter, represents great potential to mitigate climate change and simultaneously improve soil fertility. Such a challenge is particularly important and relevant in developing tropical countries like Madagascar, where soil carbon storage is vulnerable to climatic variations and where fertilization is generally applied through amendments in organic matter of various origins. The priming effect (PE) is considered here as the stimulation of the mineralization of soil organic matter (SOM) by a supply of fresh organic matter (FOM). PE results from different microbial processes driven by specific biotic and abiotic parameters. Depending on the processes involved, it has been suggested that PE could either counteract SOM accumulation or promote it. The objective of the present study was to explore the relationships between certain agricultural practices (type of crop, quality of fertilization, association with trees), the potential intensity of PE, as well as several abiotic (texture, quantity and quality SOM, nutrient enrichment) a1nd biotic (biomass and phylogenetic composition of microbial communities) factors which have been proposed in the literature as specific determinants of the different PE generation mechanisms. The soils for this study come from a network of farms in a commune in the Highlands of Madagascar. The PE, generated by a supply of 13C-enriched wheat straw, could not directly correlate with agricultural treatments. However, several indirect correlations could be found via several specific abiotic and microbial determinants that are discussed in terms of soil fertility restoration.
农业土壤中的碳固存通过高质量有机质的积累,在减缓气候变化的同时提高土壤肥力方面具有巨大潜力。在马达加斯加这样的热带发展中国家,这样的挑战尤其重要和相关。在这些国家,土壤碳储量很容易受到气候变化的影响,而且施肥通常是通过对各种来源的有机质进行修正来实施的。启动效应(PE)被认为是由新鲜有机质(FOM)的供应刺激土壤有机质(SOM)的矿化。PE是由特定的生物和非生物参数驱动的不同微生物过程产生的。根据所涉及的过程,有人认为PE可以抵消或促进SOM的积累。本研究的目的是探讨某些农业实践(作物类型、施肥质量、与树木的关联)、潜在PE强度之间的关系,以及文献中提出的几种非生物(SOM的质地、数量和质量、养分富集)和生物(生物量和微生物群落的系统发育组成)因素之间的关系,这些因素是不同PE产生机制的具体决定因素。这项研究的土壤来自马达加斯加高地一个公社的农场网络。由富含13c的麦秆产生的PE与农业处理没有直接关系。然而,可以通过几种特定的非生物和微生物决定因素发现一些间接相关性,这些决定因素在土壤肥力恢复方面进行了讨论。
{"title":"Short-Term PE Generation Processes in the Soils of a Farmer Plots Network in the Madagascar Highlands: Actors and Drivers","authors":"Kanto Razanamalala, Tantely Razafimbelo, Pierre-Alain Maron, Lionel Ranjard, Nicolas Chemidlin Prévost-Bouré, Samuel Dequiedt, Thierry Becquer, Jean Trap, Eric Blanchart, Laetitia Bernard","doi":"10.3390/microbiolres14040117","DOIUrl":"https://doi.org/10.3390/microbiolres14040117","url":null,"abstract":"Carbon sequestration in agricultural soils, through the accumulation of high-quality organic matter, represents great potential to mitigate climate change and simultaneously improve soil fertility. Such a challenge is particularly important and relevant in developing tropical countries like Madagascar, where soil carbon storage is vulnerable to climatic variations and where fertilization is generally applied through amendments in organic matter of various origins. The priming effect (PE) is considered here as the stimulation of the mineralization of soil organic matter (SOM) by a supply of fresh organic matter (FOM). PE results from different microbial processes driven by specific biotic and abiotic parameters. Depending on the processes involved, it has been suggested that PE could either counteract SOM accumulation or promote it. The objective of the present study was to explore the relationships between certain agricultural practices (type of crop, quality of fertilization, association with trees), the potential intensity of PE, as well as several abiotic (texture, quantity and quality SOM, nutrient enrichment) a1nd biotic (biomass and phylogenetic composition of microbial communities) factors which have been proposed in the literature as specific determinants of the different PE generation mechanisms. The soils for this study come from a network of farms in a commune in the Highlands of Madagascar. The PE, generated by a supply of 13C-enriched wheat straw, could not directly correlate with agricultural treatments. However, several indirect correlations could be found via several specific abiotic and microbial determinants that are discussed in terms of soil fertility restoration.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"38 2","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136262979","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: 0
Evaluation of Predicted siRNA as an Antiviral against MERS-CoV Targeting the Membrane Gene in the Vero Cell Line 预测siRNA在Vero细胞系中作为MERS-CoV膜基因抗病毒药物的评价
Q3 MICROBIOLOGY Pub Date : 2023-10-17 DOI: 10.3390/microbiolres14040116
Amany Y. El-Sayed, Mahmoud Shehata, Sara H. Mahmoud, Mahmoud ElHefnawi, Dina M. Seoudi, Mohamed A. Ali
The Middle East respiratory syndrome coronavirus (MERS-CoV) was first isolated from a patient with acute pneumonia and renal failure in Saudi Arabia in 2012. By July 2023, MERS-CoV had resulted in 2605 human cases worldwide, causing a fatality rate of 36%, with 90.2% of cases being located in the Arabian Peninsula. The dromedary camel (Camelus dromedarius) is presumed to be an intermediate host for viral transmission to humans. So far, no prophylactic vaccines or effective antiviral treatments have been approved for MERS-CoV. RNA silencing is a novel approach for treating several diseases. A web-based bioinformatics tool (i-Score Designer) with integrative computational methods was used to predict and evaluate the designed siRNAs. This approach enabled the targeting of a highly conserved region of the MERS-CoV membrane (M) gene to inhibit virus replication. siRNA-M1, -M2, and -M3 were selected as the best of 559 designed siRNA candidates for an in vitro validation based on 2nd generation algorithm scoring, thermodynamic properties, off-target filtration, position-specific nucleotide preferences, and a free immune-stimulatory motifs. siRNAs were evaluated in Vero cells for their cytotoxicity and antiviral efficacy in vitro. Our results showed that the predicted siRNAs had no apparent cytotoxicity observed in Vero cells. The obtained results from the plaque reduction assay and RT-qPCR indicated that siRNA-M3 was the best candidate to inhibit MERS-CoV replication with a defined concentration of 400 picoMolar (pM). The computational methods used, and the in vitro evaluation, may provide an insight for a new antiviral strategy against MERS-CoV, a further in vivo study will nevertheless be required.
中东呼吸综合征冠状病毒(MERS-CoV)于2012年首次从沙特阿拉伯的一名急性肺炎和肾衰竭患者身上分离出来。截至2023年7月,中东呼吸综合征冠状病毒已在全球造成2605例人间病例,致死率为36%,其中90.2%的病例发生在阿拉伯半岛。单峰骆驼(Camelus dromedarius)被认为是病毒传播给人类的中间宿主。到目前为止,尚未批准针对中东呼吸综合征冠状病毒的预防性疫苗或有效的抗病毒治疗。RNA沉默是一种治疗多种疾病的新方法。使用基于网络的生物信息学工具(i-Score Designer)和综合计算方法来预测和评估设计的sirna。这种方法能够靶向MERS-CoV膜(M)基因的高度保守区域来抑制病毒复制。基于第二代算法评分、热力学性质、脱靶过滤、位置特异性核苷酸偏好和自由免疫刺激基序,siRNA- m1、-M2和-M3被选为559个设计的siRNA候选物中最好的,用于体外验证。在Vero细胞中评估sirna的细胞毒性和体外抗病毒效果。我们的研究结果表明,预测的sirna在Vero细胞中没有明显的细胞毒性。斑块减少实验和RT-qPCR结果表明,siRNA-M3是抑制MERS-CoV复制的最佳候选物,限定浓度为400皮摩尔(pM)。所使用的计算方法和体外评估可能为针对MERS-CoV的新抗病毒策略提供见解,但仍需要进一步的体内研究。
{"title":"Evaluation of Predicted siRNA as an Antiviral against MERS-CoV Targeting the Membrane Gene in the Vero Cell Line","authors":"Amany Y. El-Sayed, Mahmoud Shehata, Sara H. Mahmoud, Mahmoud ElHefnawi, Dina M. Seoudi, Mohamed A. Ali","doi":"10.3390/microbiolres14040116","DOIUrl":"https://doi.org/10.3390/microbiolres14040116","url":null,"abstract":"The Middle East respiratory syndrome coronavirus (MERS-CoV) was first isolated from a patient with acute pneumonia and renal failure in Saudi Arabia in 2012. By July 2023, MERS-CoV had resulted in 2605 human cases worldwide, causing a fatality rate of 36%, with 90.2% of cases being located in the Arabian Peninsula. The dromedary camel (Camelus dromedarius) is presumed to be an intermediate host for viral transmission to humans. So far, no prophylactic vaccines or effective antiviral treatments have been approved for MERS-CoV. RNA silencing is a novel approach for treating several diseases. A web-based bioinformatics tool (i-Score Designer) with integrative computational methods was used to predict and evaluate the designed siRNAs. This approach enabled the targeting of a highly conserved region of the MERS-CoV membrane (M) gene to inhibit virus replication. siRNA-M1, -M2, and -M3 were selected as the best of 559 designed siRNA candidates for an in vitro validation based on 2nd generation algorithm scoring, thermodynamic properties, off-target filtration, position-specific nucleotide preferences, and a free immune-stimulatory motifs. siRNAs were evaluated in Vero cells for their cytotoxicity and antiviral efficacy in vitro. Our results showed that the predicted siRNAs had no apparent cytotoxicity observed in Vero cells. The obtained results from the plaque reduction assay and RT-qPCR indicated that siRNA-M3 was the best candidate to inhibit MERS-CoV replication with a defined concentration of 400 picoMolar (pM). The computational methods used, and the in vitro evaluation, may provide an insight for a new antiviral strategy against MERS-CoV, a further in vivo study will nevertheless be required.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"19 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136033425","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: 0
Serum Level of Ceruloplasmin, Angiotensin-Converting Enzyme and Transferrin as Markers of Severity in SARS-CoV-2 Infection in Patients with Type 2 Diabetes 2型糖尿病患者血清铜蓝蛋白、血管紧张素转换酶和转铁蛋白水平与SARS-CoV-2感染严重程度的关系
Q3 MICROBIOLOGY Pub Date : 2023-10-16 DOI: 10.3390/microbiolres14040115
Patricia-Andrada Reștea, Ștefan Țigan, Laura Grațiela Vicaș, Luminița Fritea, Eleonora Marian, Tunde Jurca, Annamaria Pallag, Iulius Liviu Mureșan, Corina Moisa, Otilia Micle, Mariana Eugenia Mureșan
The aim of this study was to analyze other possible new markers of severity, at hospital admission, that can be assessed in patients with type 2 diabetes and a SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) infection. Our study included 90 subjects: 45 patients with type 2 diabetes mellitus infected with the SARS-CoV-2 virus, and 45 healthy persons as controls. The serum level of ceruloplasmin at hospital admission was statistically significant and directly correlated with the severity of COVID-19 (coronavirus disease) (p = 0.037) and with the serum level of IL-6 (interleukin 6) (p = 0.0001). Ceruloplasmin was statistically significant and inversely correlated with the serum iron level (p = 0.0001). However, we observed that ACE (angiotensin-converting enzyme) decreased in severe forms of SARS-CoV-2 infections in patients with type 2 diabetes (p = 0.001). Moreover, the decrease in ACE levels was correlated with an increase in IL-6 levels in these patients (p = 0.001). IL-6 increases were statistically significant and inversely correlated with serum iron, transferrin, and ACE levels. There was a noticed decreasing tendency of the transferrin depending on the severity of the COVID-19 infection (p = 0.0001). In addition to the known severity factors in the context of infection with the new coronavirus, increased concentrations of ceruloplasmin and decreased concentrations of ACE and transferrin may represent new markers of COVID-19 severity in patients with type 2 diabetes. These parameters, if analyzed upon admission to the hospital, could better inform health professionals about the evolution towards more severe forms of SARS-CoV-2 infections.
本研究的目的是分析其他可能的新的严重程度指标,在入院时,可以评估2型糖尿病患者和SARS-CoV-2(严重急性呼吸综合征冠状病毒2)感染。本研究纳入90例受试者:感染SARS-CoV-2病毒的2型糖尿病患者45例,健康对照45例。入院时血清铜蓝蛋白水平与COVID-19(冠状病毒病)严重程度直接相关(p = 0.037),与血清IL-6(白细胞介素6)水平直接相关(p = 0.0001)。铜蓝蛋白与血清铁水平呈显著负相关(p = 0.0001)。然而,我们观察到,在严重的SARS-CoV-2感染的2型糖尿病患者中,ACE(血管紧张素转换酶)下降(p = 0.001)。此外,ACE水平的降低与这些患者IL-6水平的升高相关(p = 0.001)。IL-6升高具有统计学意义,且与血清铁、转铁蛋白和ACE水平呈负相关。转铁蛋白随感染程度的不同有明显的下降趋势(p = 0.0001)。除了感染新型冠状病毒的已知严重程度因素外,铜蓝蛋白浓度升高、ACE和转铁蛋白浓度降低可能是2型糖尿病患者COVID-19严重程度的新标志物。如果在入院时分析这些参数,可以更好地向卫生专业人员通报向更严重形式的SARS-CoV-2感染的演变。
{"title":"Serum Level of Ceruloplasmin, Angiotensin-Converting Enzyme and Transferrin as Markers of Severity in SARS-CoV-2 Infection in Patients with Type 2 Diabetes","authors":"Patricia-Andrada Reștea, Ștefan Țigan, Laura Grațiela Vicaș, Luminița Fritea, Eleonora Marian, Tunde Jurca, Annamaria Pallag, Iulius Liviu Mureșan, Corina Moisa, Otilia Micle, Mariana Eugenia Mureșan","doi":"10.3390/microbiolres14040115","DOIUrl":"https://doi.org/10.3390/microbiolres14040115","url":null,"abstract":"The aim of this study was to analyze other possible new markers of severity, at hospital admission, that can be assessed in patients with type 2 diabetes and a SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) infection. Our study included 90 subjects: 45 patients with type 2 diabetes mellitus infected with the SARS-CoV-2 virus, and 45 healthy persons as controls. The serum level of ceruloplasmin at hospital admission was statistically significant and directly correlated with the severity of COVID-19 (coronavirus disease) (p = 0.037) and with the serum level of IL-6 (interleukin 6) (p = 0.0001). Ceruloplasmin was statistically significant and inversely correlated with the serum iron level (p = 0.0001). However, we observed that ACE (angiotensin-converting enzyme) decreased in severe forms of SARS-CoV-2 infections in patients with type 2 diabetes (p = 0.001). Moreover, the decrease in ACE levels was correlated with an increase in IL-6 levels in these patients (p = 0.001). IL-6 increases were statistically significant and inversely correlated with serum iron, transferrin, and ACE levels. There was a noticed decreasing tendency of the transferrin depending on the severity of the COVID-19 infection (p = 0.0001). In addition to the known severity factors in the context of infection with the new coronavirus, increased concentrations of ceruloplasmin and decreased concentrations of ACE and transferrin may represent new markers of COVID-19 severity in patients with type 2 diabetes. These parameters, if analyzed upon admission to the hospital, could better inform health professionals about the evolution towards more severe forms of SARS-CoV-2 infections.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"81 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136115171","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: 0
Control of Biofilm and Virulence in Pseudomonas aeruginosa by Green-Synthesized Titanium–Cerium Nanocomposites 绿色合成钛-铈纳米复合材料对铜绿假单胞菌生物膜及毒力的控制
Q3 MICROBIOLOGY Pub Date : 2023-10-13 DOI: 10.3390/microbiolres14040114
Mohammad Altaf, Nagma Parveen, Faizan Abul Qais, K. M. Abdullah, Iqbal Ahmad
Antimicrobial resistance (AMR) has become a critical global health challenge. Infections, particularly those caused by multidrug-resistant (MDR) pathogens, rank among the top causes of human mortality worldwide. Pseudomonas aeruginosa occupies a prominent position among pathogens responsible for opportunistic infections in humans. P. aeruginosa stands as a primary cause of chronic respiratory infections, significantly contributing to the burden of these chronic diseases. In the medical domain, nanotechnologies offer significant potential, spanning various applications, including advanced imaging, diagnostic devices, drug delivery systems, implants, tissue-engineered structures, and pharmaceutical treatments. Given the challenges associated with AMR and the limited discovery of new drugs to combat MDR microbes, there is a critical need for alternative strategies to address the problem of AMR. In this study, we synthesized titanium–cerium nanocomposites (Ti–Ce–NCs) using an eco-friendly green synthesis approach. X-ray diffraction (XRD) analysis confirmed the crystalline nature of the Ti–Ce–NCs and determined the particle size to be 17.07 nm. Electron microscopy revealed the size range of the particles to be 13 to 54 nm, where the majority of the particles were in the 20 to 25 nm range. Upon examining the composition, the Ti–Ce–NCs were determined to be composed of cerium, oxygen, and titanium, whose relative abundance were 36.86, 36.6, and 24.77% by weight, respectively. These nanocomposites were then evaluated for their effectiveness against various virulent traits and biofilms in P. aeruginosa. Out of six tested virulence factors, more than 50% inhibition of five virulence factors of P. aeruginosa was found. Roughly 60% inhibition of biofilm was also found in the presence of 400 µg/mL Ti–Ce–NCs. The nanocomposites also altered the biofilm architecture of the test bacterium. The success of this research opens doors for the potential use of such nanomaterials in the discovery of new antibacterial agents to combat drug-resistant bacteria.
抗微生物药物耐药性(AMR)已成为一项重大的全球卫生挑战。感染,特别是由耐多药病原体引起的感染,是全世界人类死亡的主要原因之一。铜绿假单胞菌在导致人类机会性感染的病原体中占有突出地位。铜绿假单胞菌是慢性呼吸道感染的主要原因,是造成这些慢性疾病负担的主要原因。在医疗领域,纳米技术提供了巨大的潜力,跨越各种应用,包括先进的成像、诊断设备、药物输送系统、植入物、组织工程结构和药物治疗。鉴于与耐多药耐药性相关的挑战以及对抗耐多药微生物的新药发现有限,迫切需要寻找替代策略来解决耐多药耐药性问题。在本研究中,我们采用环保绿色合成方法合成了钛-铈纳米复合材料(Ti-Ce-NCs)。x射线衍射(XRD)分析证实了Ti-Ce-NCs的结晶性质,确定了其粒径为17.07 nm。电子显微镜显示,颗粒的尺寸范围为13 ~ 54 nm,其中大部分颗粒在20 ~ 25 nm范围内。经成分分析,Ti-Ce-NCs由铈、氧和钛组成,其相对丰度分别为36.86、36.6%和24.77%(重量计)。然后评估了这些纳米复合材料对铜绿假单胞菌各种毒力性状和生物膜的有效性。在检测的6种毒力因子中,发现铜绿假单胞菌对5种毒力因子的抑制作用超过50%。400µg/mL的ti - ce - nc对生物膜的抑制作用约为60%。纳米复合材料也改变了测试细菌的生物膜结构。这项研究的成功为这种纳米材料在发现新的抗菌剂以对抗耐药细菌方面的潜在应用打开了大门。
{"title":"Control of Biofilm and Virulence in Pseudomonas aeruginosa by Green-Synthesized Titanium–Cerium Nanocomposites","authors":"Mohammad Altaf, Nagma Parveen, Faizan Abul Qais, K. M. Abdullah, Iqbal Ahmad","doi":"10.3390/microbiolres14040114","DOIUrl":"https://doi.org/10.3390/microbiolres14040114","url":null,"abstract":"Antimicrobial resistance (AMR) has become a critical global health challenge. Infections, particularly those caused by multidrug-resistant (MDR) pathogens, rank among the top causes of human mortality worldwide. Pseudomonas aeruginosa occupies a prominent position among pathogens responsible for opportunistic infections in humans. P. aeruginosa stands as a primary cause of chronic respiratory infections, significantly contributing to the burden of these chronic diseases. In the medical domain, nanotechnologies offer significant potential, spanning various applications, including advanced imaging, diagnostic devices, drug delivery systems, implants, tissue-engineered structures, and pharmaceutical treatments. Given the challenges associated with AMR and the limited discovery of new drugs to combat MDR microbes, there is a critical need for alternative strategies to address the problem of AMR. In this study, we synthesized titanium–cerium nanocomposites (Ti–Ce–NCs) using an eco-friendly green synthesis approach. X-ray diffraction (XRD) analysis confirmed the crystalline nature of the Ti–Ce–NCs and determined the particle size to be 17.07 nm. Electron microscopy revealed the size range of the particles to be 13 to 54 nm, where the majority of the particles were in the 20 to 25 nm range. Upon examining the composition, the Ti–Ce–NCs were determined to be composed of cerium, oxygen, and titanium, whose relative abundance were 36.86, 36.6, and 24.77% by weight, respectively. These nanocomposites were then evaluated for their effectiveness against various virulent traits and biofilms in P. aeruginosa. Out of six tested virulence factors, more than 50% inhibition of five virulence factors of P. aeruginosa was found. Roughly 60% inhibition of biofilm was also found in the presence of 400 µg/mL Ti–Ce–NCs. The nanocomposites also altered the biofilm architecture of the test bacterium. The success of this research opens doors for the potential use of such nanomaterials in the discovery of new antibacterial agents to combat drug-resistant bacteria.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"7 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135855853","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: 0
Grass Meal Acts as a Probiotic in Chicken 草粉在鸡体内起益生菌的作用
Q3 MICROBIOLOGY Pub Date : 2023-10-13 DOI: 10.3390/microbiolres14040113
Elena S. Bogdanova, Maria A. Danilova, Maria S. Smirnova, Elena V. Trubnikova, Hoa T. Nguyen, Svetlana N. Petrova, Sergey V. Apanasenok, Alexei B. Shevelev
Probiotics can act as an alternative to antibiotics in animal feeding, but their use is minimal due to their expensive production. Dry grass is rich with bacteria beneficial for animal feeding and can be used as a probiotic. However, data about the quantitative dependence of the grass microbiome on environmental factors and seasons remain insufficient for preparing “grass-meal-based probiotics”. Four grass samples were collected in two geographically remote regions of Russia; their microbiome was characterized by metagenomic sequencing of 16S rDNA libraries and microbiological seeding, and biological testing of the grass meal was carried out on 6 groups of birds containing 20 Ross 308 cross broilers each for a period of 42 days. The positive control group (PC) obtained 16–25 mg/mL toltrazuril (coccidiostatic agent) and 0.5 mL/L liquid antibiotic enrostin (100 mg/mL ciprofloxacin and 106 MU/mL colistin sulfate in the commercial preparation) within the drinking water, while the negative control group (NC) obtained no medicines. Four experimental groups were fed the diet supplemented with 1% grass meal over the period of 7–42 days of life; no commercial medicines were used here. A spontaneous infection with Eimeria was registered in the NC control groups, which caused the loss of 7 chickens. No losses were registered in the PC group or the two experimental groups. In two other experimental groups, losses of coccidiosis amounted to 10% and 15%, respectively. All specimens of the grass meal demonstrated a significant effect on the average body weight gain compared to NC. Taken together, these observations support the hypothesis that the grass meal may substitute toltrazuril for protecting the chickens from parasitic invasion and increase average daily weight gain (ADG) as effectively as the antibiotic enrostin.
在动物饲养中,益生菌可以作为抗生素的替代品,但由于生产成本高昂,它们的使用很少。干草富含有益动物饲养的细菌,可作为益生菌使用。然而,关于草菌群对环境因子和季节的定量依赖的数据仍然不足,无法用于制备“草源益生菌”。在俄罗斯两个地理位置偏远的地区收集了四个草样本;采用16S rDNA文库宏基因组测序和微生物播种技术对其微生物组进行了鉴定,并对6组鸡(每组20只罗斯308杂交肉鸡)进行了草粕生物学试验,试验时间为42 d。阳性对照组(PC)在饮用水中添加16 ~ 25 mg/mL的托曲祖利(防球虫剂)和0.5 mL/L的液体抗生素恩柔素(市售制剂中环丙沙星100 mg/mL,硫酸粘菌素106 MU/mL),阴性对照组(NC)不添加任何药物。4个试验组在饲粮中添加1%草粕,试验期为7 ~ 42 d;这里没有使用商业药品。NC对照组发生自发性艾美耳球虫感染,死亡7只鸡。PC组和两个实验组均未见损失。在另外两个实验组中,球虫病的损失分别达到10%和15%。与NC相比,所有草粉对平均体重增加都有显著影响。综上所述,这些观察结果支持了这样一种假设,即草粉可能取代托曲祖利来保护鸡免受寄生虫入侵,并像抗生素enrostin一样有效地增加平均日增重(ADG)。
{"title":"Grass Meal Acts as a Probiotic in Chicken","authors":"Elena S. Bogdanova, Maria A. Danilova, Maria S. Smirnova, Elena V. Trubnikova, Hoa T. Nguyen, Svetlana N. Petrova, Sergey V. Apanasenok, Alexei B. Shevelev","doi":"10.3390/microbiolres14040113","DOIUrl":"https://doi.org/10.3390/microbiolres14040113","url":null,"abstract":"Probiotics can act as an alternative to antibiotics in animal feeding, but their use is minimal due to their expensive production. Dry grass is rich with bacteria beneficial for animal feeding and can be used as a probiotic. However, data about the quantitative dependence of the grass microbiome on environmental factors and seasons remain insufficient for preparing “grass-meal-based probiotics”. Four grass samples were collected in two geographically remote regions of Russia; their microbiome was characterized by metagenomic sequencing of 16S rDNA libraries and microbiological seeding, and biological testing of the grass meal was carried out on 6 groups of birds containing 20 Ross 308 cross broilers each for a period of 42 days. The positive control group (PC) obtained 16–25 mg/mL toltrazuril (coccidiostatic agent) and 0.5 mL/L liquid antibiotic enrostin (100 mg/mL ciprofloxacin and 106 MU/mL colistin sulfate in the commercial preparation) within the drinking water, while the negative control group (NC) obtained no medicines. Four experimental groups were fed the diet supplemented with 1% grass meal over the period of 7–42 days of life; no commercial medicines were used here. A spontaneous infection with Eimeria was registered in the NC control groups, which caused the loss of 7 chickens. No losses were registered in the PC group or the two experimental groups. In two other experimental groups, losses of coccidiosis amounted to 10% and 15%, respectively. All specimens of the grass meal demonstrated a significant effect on the average body weight gain compared to NC. Taken together, these observations support the hypothesis that the grass meal may substitute toltrazuril for protecting the chickens from parasitic invasion and increase average daily weight gain (ADG) as effectively as the antibiotic enrostin.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"58 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135918090","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: 0
Strategies to Develop Aptamer Probes to Detect MRSA and Study of Antibacterial Activity MRSA适体探针检测策略及抑菌活性研究
Q3 MICROBIOLOGY Pub Date : 2023-10-13 DOI: 10.3390/microbiolres14040112
Jagath C. Kasturiarachchi
This study investigated the development of aptamer-based molecular probes to detect Methicillin-Resistant Staphylococcus aureus (MRSA) and evaluated the antibacterial activity. Early detection of MRSA infection will improve patients’ recovery and reduce the cost for treating patients. S. aureus can become resistant to methicillin and other β-lactam antibiotics through the expression of PBP2A protein, which is resistant to the action of methicillin. We have developed two aptamer molecular probes against PBP2A protein and whole bacterial cell (MRSA) under optimized in vitro conditions using SELEX approach. Target aptamer sequences were identified, and chemically synthesized aptamer probes were evaluated using fluorescently-labelled aptamer probes using flow cytometry and confocal imaging. Antibacterial activities of those aptamers were also evaluated using a bacterial killing assay. The results showed that high specific aptamers were developed against purified PBP2A protein. However, these aptamers showed less specificity to detect MRSA under in vitro condition. These aptamers showed no cytotoxic effect on 3T3 cells and no antibacterial activity against MRSA. The results suggested that the specific aptamer development and the in vitro selection methodology require further refinement to improve the diagnostic and therapeutic utility of these aptamers.
本研究研究了基于适配体的分子探针检测耐甲氧西林金黄色葡萄球菌(MRSA),并对其抑菌活性进行了评价。MRSA感染的早期发现将提高患者的康复,降低治疗患者的成本。金黄色葡萄球菌可通过表达PBP2A蛋白对甲氧西林及其他β-内酰胺类抗生素产生耐药性,该蛋白对甲氧西林具有耐药性。在优化的体外条件下,采用SELEX方法开发了两种针对PBP2A蛋白和MRSA的适体分子探针。鉴定靶适体序列,利用流式细胞术和共聚焦成像技术,利用荧光标记适体探针对化学合成的适体探针进行评估。这些适体的抗菌活性也用细菌杀灭试验进行了评价。结果表明,针对纯化的PBP2A蛋白,获得了高特异性适配体。然而,这些适体在体外条件下检测MRSA的特异性较低。这些适体对3T3细胞无细胞毒作用,对MRSA无抗菌活性。结果表明,特异性适配体的开发和体外选择方法需要进一步完善,以提高这些适配体的诊断和治疗效用。
{"title":"Strategies to Develop Aptamer Probes to Detect MRSA and Study of Antibacterial Activity","authors":"Jagath C. Kasturiarachchi","doi":"10.3390/microbiolres14040112","DOIUrl":"https://doi.org/10.3390/microbiolres14040112","url":null,"abstract":"This study investigated the development of aptamer-based molecular probes to detect Methicillin-Resistant Staphylococcus aureus (MRSA) and evaluated the antibacterial activity. Early detection of MRSA infection will improve patients’ recovery and reduce the cost for treating patients. S. aureus can become resistant to methicillin and other β-lactam antibiotics through the expression of PBP2A protein, which is resistant to the action of methicillin. We have developed two aptamer molecular probes against PBP2A protein and whole bacterial cell (MRSA) under optimized in vitro conditions using SELEX approach. Target aptamer sequences were identified, and chemically synthesized aptamer probes were evaluated using fluorescently-labelled aptamer probes using flow cytometry and confocal imaging. Antibacterial activities of those aptamers were also evaluated using a bacterial killing assay. The results showed that high specific aptamers were developed against purified PBP2A protein. However, these aptamers showed less specificity to detect MRSA under in vitro condition. These aptamers showed no cytotoxic effect on 3T3 cells and no antibacterial activity against MRSA. The results suggested that the specific aptamer development and the in vitro selection methodology require further refinement to improve the diagnostic and therapeutic utility of these aptamers.","PeriodicalId":43788,"journal":{"name":"Microbiology Research","volume":"125 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135918837","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: 0
期刊
Microbiology Research
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1