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Bioprospecting for polyesterase activity relevant for PET degradation in marine Enterobacterales isolates. 海洋肠杆菌分离物中与PET降解相关的聚酯酶活性的生物勘探。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2023-01-01 DOI: 10.3934/microbiol.2023027
Denisse Galarza-Verkovitch, Onur Turak, Jutta Wiese, Tanja Rahn, Ute Hentschel, Erik Borchert

Plastics have quickly become an integral part of modern life. Due to excessive production and improper waste disposal, they are recognized as contaminants present in practically all habitat types. Although there are several polymers, polyethylene terephthalate (PET) is of particular concern due to its abundance in the environment. There is a need for a solution that is both cost-effective and ecologically friendly to address this pollutant. The use of microbial depolymerizing enzymes could offer a biological avenue for plastic degradation, though the full potential of these enzymes is yet to be uncovered. The purpose of this study was to use (1) plate-based screening methods to investigate the plastic degradation potential of marine bacteria from the order Enterobacterales collected from various organismal and environmental sources, and (2) perform genome-based analysis to identify polyesterases potentially related to PET degradation. 126 bacterial isolates were obtained from the strain collection of RD3, Research Unit Marine Symbioses-GEOMAR-and sequentially tested for esterase and polyesterase activity, in combination here referred to as PETase-like activity. The results show that members of the microbial families Alteromonadaceae, Shewanellaceae, and Vibrionaceae, derived from marine sponges and bryozoans, are the most promising candidates within the order Enterobacterales. Furthermore, 389 putative hydrolases from the α/β superfamily were identified in 23 analyzed genomes, of which 22 were sequenced for this study. Several candidates showed similarities with known PETases, indicating underlying enzymatic potential within the order Enterobacterales for PET degradation.

塑料已迅速成为现代生活中不可或缺的一部分。由于过度生产和不当的废物处理,它们被认为是几乎所有生境类型中存在的污染物。虽然有几种聚合物,但聚对苯二甲酸乙二醇酯(PET)由于其在环境中的丰度而受到特别关注。需要一种既具有成本效益又对生态友好的解决方案来处理这种污染物。微生物解聚酶的使用可以为塑料降解提供生物途径,尽管这些酶的全部潜力尚未被发现。本研究的目的是使用(1)基于平板的筛选方法来研究从各种有机和环境来源中收集的肠杆菌目海洋细菌的塑料降解潜力;(2)进行基因组分析以鉴定可能与PET降解相关的聚酯酶。从海洋共生研究单元(Research Unit Marine symbioses - geomar)的RD3菌株收集中获得126株细菌,并依次检测酯酶和聚酯酶活性,这里将两者结合称为pase样活性。结果表明,从海绵和苔藓虫中分离出来的Alteromonadaceae、Shewanellaceae和Vibrionaceae是肠杆菌目中最有希望的候选微生物。此外,在分析的23个基因组中鉴定出389个来自α/β超家族的推定水解酶,其中22个测序用于本研究。一些候选物显示出与已知的PET酶的相似性,表明肠杆菌目中潜在的PET降解酶潜力。
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引用次数: 1
Prospect of nanomaterials as antimicrobial and antiviral regimen. 纳米材料在抗菌和抗病毒治疗中的应用前景。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2023-01-01 DOI: 10.3934/microbiol.2023024
Ashok Chakraborty, Anil Diwan, Jayant Tatake

In recent years studies of nanomaterials have been explored in the field of microbiology due to the increasing evidence of antibiotic resistance. Nanomaterials could be inorganic or organic, and they may be synthesized from natural products from plant or animal origin. The therapeutic applications of nano-materials are wide, from diagnosis of disease to targeted delivery of drugs. Broad-spectrum antiviral and antimicrobial activities of nanoparticles are also well evident. The ratio of nanoparticles surface area to their volume is high and that allows them to be an advantageous vehicle of drugs in many respects. Effective uses of various materials for the synthesis of nanoparticles impart much specificity in them to meet the requirements of specific therapeutic strategies. The potential therapeutic use of nanoparticles and their mechanisms of action against infections from bacteria, fungi and viruses were the focus of this review. Further, their potential advantages, drawbacks, limitations and side effects are also included here. Researchers are characterizing the exposure pathways of nano-medicines that may cause serious toxicity to the subjects or the environment. Indeed, societal ethical issues in using nano-medicines pose a serious question to scientists beyond anything.

近年来,由于抗生素耐药性的证据越来越多,纳米材料的研究在微生物学领域得到了探索。纳米材料可以是无机的也可以是有机的,它们可以从植物或动物的天然产物中合成。纳米材料的治疗应用非常广泛,从疾病诊断到药物靶向递送。纳米颗粒的广谱抗病毒和抗菌活性也很明显。纳米颗粒的表面积与体积之比很高,这使得它们在许多方面成为一种有利的药物载体。有效地利用各种材料合成纳米粒子赋予它们许多特异性,以满足特定治疗策略的要求。纳米颗粒的潜在治疗用途及其对细菌、真菌和病毒感染的作用机制是本综述的重点。此外,它们的潜在优点,缺点,局限性和副作用也包括在这里。研究人员正在描述可能对受试者或环境造成严重毒性的纳米药物的暴露途径。事实上,使用纳米药物的社会伦理问题给科学家们提出了一个严重的问题。
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引用次数: 0
Chemical structure, properties and potential applications of surfactin, as well as advanced strategies for improving its microbial production. 表面素的化学结构、性质和潜在应用,以及改善其微生物生产的先进策略。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2023-01-01 DOI: 10.3934/microbiol.2023012
Cheng Zhen, Xian-Feng Ge, Yi-Ting Lu, Wen-Zheng Liu

Surfactin, a cyclic lipopeptide produced by microbes belonging to the genus Bacillus, is one of the most effective biosurfactants available in many industrial fields. However, its low production and high cost have intensively constrained its commercial applications. In this review, we first summarize the molecular structure, biological properties, beneficial roles and potential applications of surfactin in the fields of medical care and food safety, highlighting the great medical and commercial values of making its industrial production into reality. Further, genetic regulation for surfactin biosynthesis and advanced strategies for enhancing its microbial production, including optimizing fermentation conditions, rational genetic engineering and synthetic biology combined with metabolic engineering approaches, are elucidated. Finally, prospects for improving surfactin biosynthesis are discussed, and the establishment of suitable chassis hosts for exogenous production of surfactin might serve as an important strategy in future research.

表面素是一种由芽孢杆菌属微生物产生的环状脂肽,是许多工业领域中最有效的生物表面活性剂之一。然而,它的低产量和高成本严重制约了其商业应用。本文首先综述了表面素的分子结构、生物学特性、有益作用及其在医疗和食品安全领域的潜在应用,强调了其工业化生产的巨大医学价值和商业价值。此外,本文还阐述了表面素生物合成的基因调控机制,以及优化发酵条件、合理的基因工程、合成生物学与代谢工程相结合等提高表面素微生物产量的先进策略。最后,对改善表面素生物合成的前景进行了展望,并指出建立适合外源生产表面素的底盘宿主可能是未来研究的重要策略。
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引用次数: 3
Impact of grape polyphenols on Akkermansia muciniphila and the gut barrier. 葡萄多酚对嗜粘阿克曼氏菌和肠道屏障的影响。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2022-12-22 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022035
Esther Mezhibovsky, Yue Wu, Fiona G Bawagan, Kevin M Tveter, Samantha Szeto, Diana Roopchand

A healthy gastrointestinal tract functions as a highly selective barrier, allowing the absorption of nutrients and metabolites while preventing gut bacteria and other xenobiotic compounds from entering host circulation and tissues. The intestinal epithelium and intestinal mucus provide a physical first line of defense against resident microbes, pathogens and xenotoxic compounds. Prior studies have indicated that the gut microbe Akkermansia muciniphila, a mucin-metabolizer, can stimulate intestinal mucin thickness to improve gut barrier integrity. Grape polyphenol (GP) extracts rich in B-type proanthocyanidin (PAC) compounds have been found to increase the relative abundance of A. muciniphila, suggesting that PACs alter the gut microbiota to support a healthy gut barrier. To further investigate the effect of GPs on the gut barrier and A. muciniphila, male C57BL/6 mice were fed a high-fat diet (HFD) or low-fat diet (LFD) with or without 1% GPs (HFD-GP, LFD-GP) for 12 weeks. Compared to the mice fed unsupplemented diets, GP-supplemented mice showed increased relative abundance of fecal and cecal A. muciniphila, a reduction in total bacteria, a diminished colon mucus layer and increased fecal mucus content. GP supplementation also reduced the presence of goblet cells regardless of dietary fat. Compared to the HFD group, ileal gene expression of lipopolysaccharide (LPS)-binding protein (Lbp), an acute-phase protein that promotes pro-inflammatory cytokine expression, was reduced in the HFD-GP group, suggesting reduced LPS in circulation. Despite depletion of the colonic mucus layer, markers of inflammation (Ifng, Il1b, Tnfa, and Nos2) were similar among the four groups, with the exception that ileal Il6 mRNA levels were lower in the LFD-GP group compared to the LFD group. Our findings suggest that the GP-induced increase in A. muciniphila promotes redistribution of the intestinal mucus layer to the intestinal lumen, and that the GP-induced decrease in total bacteria results in a less inflammatory intestinal milieu.

健康的胃肠道作为一个高度选择性的屏障,允许营养物质和代谢物的吸收,同时防止肠道细菌和其他外来化合物进入宿主循环和组织。肠上皮和肠粘液提供了抵抗常驻微生物、病原体和异种毒性化合物的物理第一道防线。先前的研究表明,肠道微生物Akkermansia muciniphila是一种黏液代谢物质,可以刺激肠道黏液厚度,提高肠道屏障的完整性。富含b型原花青素(PAC)化合物的葡萄多酚(GP)提取物已被发现可以增加嗜粘杆菌的相对丰度,这表明PAC可以改变肠道微生物群以支持健康的肠道屏障。为了进一步研究gp对肠道屏障和嗜粘单歧杆菌的影响,将雄性C57BL/6小鼠分别饲喂高脂饲粮(HFD)和低脂饲粮(LFD),分别添加或不添加1% gp (HFD- gp, LFD- gp) 12周。与未添加gp的小鼠相比,添加gp的小鼠粪便和盲肠嗜粘杆菌的相对丰度增加,细菌总数减少,结肠黏液层减少,粪便黏液含量增加。无论饮食中是否含有脂肪,补充GP也能减少杯状细胞的存在。与HFD组相比,HFD- gp组回肠中脂多糖(LPS)结合蛋白(Lbp)的基因表达减少,Lbp是一种促进促炎细胞因子表达的急性期蛋白,提示循环中的LPS减少。尽管结肠黏液层耗竭,炎症标志物(Ifng、Il1b、Tnfa和Nos2)在四组之间相似,除了LFD- gp组的回肠Il6 mRNA水平低于LFD组。我们的研究结果表明,gp诱导的嗜粘杆菌的增加促进了肠黏液层向肠腔的重新分布,gp诱导的总细菌的减少导致肠道炎症环境的减少。
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引用次数: 2
SARS-CoV-2 vaccines: What we know, what we can do to improve them and what we could learn from other well-known viruses. SARS-CoV-2 疫苗:我们知道什么,我们能做些什么来改进它们,以及我们能从其他知名病毒中学到什么。
IF 2.7 Q3 MICROBIOLOGY Pub Date : 2022-11-16 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022029
Sirio Fiorino, Andrea Carusi, Wandong Hong, Paolo Cernuschi, Claudio Giuseppe Gallo, Emanuele Ferrara, Thais Maloberti, Michela Visani, Federico Lari, Dario de Biase, Maddalena Zippi

In recent weeks, the rate of SARS-CoV-2 infections has been progressively increasing all over the globe, even in countries where vaccination programs have been strongly implemented. In these regions in 2021, a reduction in the number of hospitalizations and deaths compared to 2020 was observed. This decrease is certainly associated with the introduction of vaccination measures. The process of the development of effective vaccines represents an important challenge. Overall, the breakthrough infections occurring in vaccinated subjects are in most cases less severe than those observed in unvaccinated individuals. This review examines the factors affecting the immunogenicity of vaccines against SARS-CoV-2 and the possible role of nutrients in modulating the response of distinct immune cells to the vaccination.

最近几周,SARS-CoV-2 感染率在全球范围内逐步上升,即使在大力实施疫苗接种计划的国家也是如此。与 2020 年相比,这些地区 2021 年的住院人数和死亡人数都有所减少。这种减少肯定与疫苗接种措施的实施有关。开发有效疫苗的过程是一项重要挑战。总体而言,在大多数情况下,接种过疫苗的受试者发生的突破性感染的严重程度低于未接种疫苗者。本综述探讨了影响 SARS-CoV-2 疫苗免疫原性的因素,以及营养物质在调节不同免疫细胞对疫苗接种的反应中可能发挥的作用。
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引用次数: 0
Antimicrobials cetylpyridinium-chloride and miramistin demonstrate non-inferiority and no "protein-error" compared to established wound care antiseptics in vitro. 抗菌剂 cetylpyridinium-chloride 和 miramistin 在体外实验中与现有的伤口护理抗菌剂相比没有劣势,也没有 "蛋白质错误"。
IF 2.7 Q3 MICROBIOLOGY Pub Date : 2022-10-24 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022026
Julian-Dario Rembe, Vivian-Denise Thompson, Ewa Klara Stuermer

Concern about microbial tolerance and resistance to established antimicrobials drives research into alternatives for local antiseptic wound treatment. Precise efficacy profiles are thereby important in the evaluation of potential alternative antimicrobials, and protein interference ("protein error") is a key factor. Here, the antimicrobial efficacy of cetylpyridinium chloride (CPC) and miramistin (MST) was compared to the established antimicrobials octenidine (OCT), povidon-iodine (PVP-I), polyhexamethylene-biguanide (PHMB) and chlorhexidine (CHX). Efficacy was evaluated after 0.5, 1, 3, 5 and 10 min against Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Enterococcus faecium and Candida albicans using an in vitro quantitative suspension method (based on DIN EN 13727). To investigate protein interference, 0.3% or 3% bovine albumin was used as the challenge. OCT and PVP-I demonstrated a significant efficacy within 0.5 min, regardless of the microbial organism and protein challenge (p < 0.01). CPC and MST showed no inferiority in efficacy, with only MST needing up to 3 min to achieve the same microbial reduction. PHMB and CHX also achieved significant reduction rates over the tested time-course, yet demonstrated a necessity for prolonged exposure (up to 10 min) for comparable reduction. A protein interference was predominantly observed for PHMB against S. aureus, but without statistically significant differences in antimicrobial efficacy between the 0.3% and 3% protein challenges. All other tested agents showed no relevant interference with the presence of protein. CPC and MST proved to be non-inferior to established wound antiseptics agents in vitro. In fact, CPC showed a more efficient reduction than PHMB and CHX despite there being an introduced protein challenge. Both agents demonstrated no significant "protein error" under challenging conditions (3% albumin), posing them as valid potential candidates for alternative antimicrobials in wound management.

由于担心微生物对既有抗菌剂产生耐受性和抗药性,人们开始研究局部伤口消毒处理的替代品。因此,在评估潜在的替代抗菌剂时,精确的药效曲线非常重要,而蛋白质干扰("蛋白质误差")是一个关键因素。在此,我们将十六烷基氯化吡啶(CPC)和米拉米星(MST)的抗菌功效与已有的抗菌剂辛烯胺(OCT)、聚维碘(PVP-I)、聚六亚甲基双胍(PHMB)和洗必泰(CHX)进行了比较。采用体外定量悬浮法(基于 DIN EN 13727),在 0.5、1、3、5 和 10 分钟后对金黄色葡萄球菌、铜绿假单胞菌、大肠杆菌、粪肠球菌和白色念珠菌的功效进行了评估。为了研究蛋白质的干扰,使用了 0.3% 或 3% 的牛白蛋白作为挑战。在 0.5 分钟内,OCT 和 PVP-I 显示出明显的疗效,与微生物和蛋白质挑战无关(p < 0.01)。CPC 和 MST 的疗效并不逊色,只有 MST 需要长达 3 分钟才能达到同样的微生物减少效果。PHMB 和 CHX 在测试的时间过程中也达到了显著的减少率,但需要长时间接触(长达 10 分钟)才能达到相同的减少率。PHMB 对金黄色葡萄球菌的抗菌效果主要受到蛋白质的干扰,但在 0.3% 和 3% 蛋白质挑战中,抗菌效果没有明显的统计学差异。所有其他测试药剂均未显示出与蛋白质存在相关的干扰。事实证明,CPC 和 MST 在体外测试中并不逊色于已有的伤口抗菌剂。事实上,尽管引入了蛋白质挑战,但 CPC 比 PHMB 和 CHX 更有效地减少了蛋白质。在具有挑战性的条件下(3% 的白蛋白),这两种制剂都没有表现出明显的 "蛋白质误差",因此它们有可能成为伤口管理中的替代抗菌剂。
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引用次数: 0
Co-fermentation involving Lysinibacillus sp. and Aspergillus flavus for simultaneous palm oil waste treatment and renewable biomass fuel production. 赖氨酸芽孢杆菌和黄曲霉共同发酵棕榈油废弃物处理和可再生生物质燃料生产。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2022-09-16 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022025
Nurul Alia Syufina Abu Bakar, Nur Aliyyah Khuzaini, Siti Baidurah

Biomass fuel is one of the renewable energy sources that can be produced by valorization of palm oil mill effluent (POME) and empty fruit bunch (EFB). POME and EFB are available abundantly in Malaysia and only small portion is utilized to produce other value-added products. The objective of this study is to: (1) utilize the wastes from agro-industrial sector especially palm oil wastes and bio-valorize into value-added product of biomass fuel with high CEV, and simultaneously (2) reduce the waste accumulated in the palm oil factory. In this study, co-fermentation of bacteria (Lysinibacillus sp.) and fungus (Aspergillus flavus) at 37 °C, 180 rpm for 5 days, followed by overnight oven-dry at 85 °C was conducted utilizing a mixture of POME and EFB with the ratio of 7:3 at laboratory scale. Three fermentation medium conditions were performed, namely: (1) Group 1: autoclaved POME and EFB without addition of any microorganisms, (2) Group 2: autoclaved POME and EFB with the addition of Lysinibacillus sp. LC 556247 and Aspergillus flavus, and (3) Group 3: POME and EFB as it is (non-sterile). Among all condition, Group 2 with co-fermentation evinced the highest calorific energy value (CEV) of 26.71 MJ/kg, highest biochemical oxygen demand (BOD) removal efficiency of 61.11%, chemical oxygen demand (COD) removal efficiency at 48.47%, and total suspended solid (TSS) reduction of 37.12%. Overall, this study successfully utilized abundant POME and EFB waste and turn into value added product of renewable biomass fuel with high CEV percentage and simultaneously able to reduce abundant liquid waste.

生物质燃料是利用棕榈油厂废水(POME)和空果束(EFB)进行再生利用的可再生能源之一。POME和EFB在马来西亚供应充足,只有一小部分用于生产其他增值产品。本研究的目的是:(1)利用农业工业部门的废物,特别是棕榈油废物并生物增值为高CEV的生物质燃料增值产品,同时(2)减少棕榈油工厂积累的废物。在本研究中,在实验室规模下,使用POME和EFB的混合物,以7:3的比例在37℃,180转/分的条件下,对细菌(Lysinibacillus sp.)和真菌(Aspergillus flavus)进行了5天的共发酵,然后在85℃的烤箱干燥过夜。采用三种发酵培养基条件,即:(1)组1:高压灭菌POME和EFB,不添加任何微生物;(2)组2:高压灭菌POME和EFB,添加Lysinibacillus sp. LC 556247和Aspergillus flavus;(3)组3:POME和EFB原汁原味(非无菌)。其中,共发酵组2的热能值(CEV)最高,为26.71 MJ/kg,生化需氧量(BOD)去除率最高,为61.11%,化学需氧量(COD)去除率最高,为48.47%,总悬浮固体(TSS)减少率为37.12%。总体而言,本研究成功地利用了大量的POME和EFB废物,将其转化为高CEV百分比的可再生生物质燃料的增值产品,同时减少了大量的液体废物。
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引用次数: 5
Probiotics for immunomodulation in prevention against respiratory viral infections with special emphasis on COVID-19. 免疫调节益生菌预防呼吸道病毒感染,特别是COVID-19。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2022-09-15 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022024
T Amrouche, M L Chikindas

COVID-19 pandemic is currently causing high mortality and economic crisis, and several drugs-based therapeutic strategies and vaccines are unfortunately used with little efficiency. Therefore, here is an urgent need to provide additives therapies that prevent or improve symptoms in populations infected by SARS-CoV-2 variants. This review aimed to examine relevant scientific information related to SARS-CoV-2 and host antiviral immunity, as well the possible role of probiotics in gut-lung cross talk pathways to promote lung immune response to COVID-19 infection. We searched online databases such as PubMed, Embase, Chinese databases, and selected articles and studies with relevant data reported on COVID-19 and other respiratory viral infections. Recent research highlighted potential immunomodulatory activities of probiotics assessed in animal models and clinical trials. However, the role of probiotics and gut microbiome in COVID-19 management, and approaches with significant understanding in molecular mechanism of probiotic action remain poorly investigated. Clinical investigations as well as animal model studies published have demonstrated that probiotics such as Lactobacillus rhamnosus and Bifidumbacterium lactis HN019, may influence positively not only microbiota balance but also antiviral immunity by improving both innate and adaptive responses and controlling inflammatory reaction in respiratory viral infection. Given the immunological interactions in gut-lung axis and the crucial role of probiotics in modulating immune responses by promoting dendritic cells (DCs) to regulate T cell responses, we hypothesized that application of probiotics may be successful in prevention or treatment of both intestinal disorders and airway diseases in patients with COVID-19.

COVID-19大流行目前正在造成高死亡率和经济危机,遗憾的是,几种基于药物的治疗策略和疫苗的使用效率很低。因此,迫切需要提供预防或改善SARS-CoV-2变体感染人群症状的添加剂疗法。本综述旨在探讨SARS-CoV-2与宿主抗病毒免疫的相关科学信息,以及益生菌在肠-肺串扰通路中促进肺部对COVID-19感染免疫反应的可能作用。我们检索了PubMed、Embase、中文数据库等在线数据库,并选择了与COVID-19及其他呼吸道病毒感染相关数据的文章和研究。最近的研究强调了益生菌在动物模型和临床试验中潜在的免疫调节活性。然而,益生菌和肠道微生物组在COVID-19治疗中的作用,以及益生菌作用的分子机制的研究仍然很少。临床研究和已发表的动物模型研究表明,鼠李糖乳杆菌和乳酸双歧杆菌HN019等益生菌不仅可以积极影响微生物群平衡,还可以通过改善呼吸道病毒感染的先天和适应性反应以及控制炎症反应来改善抗病毒免疫。鉴于肠-肺轴的免疫相互作用以及益生菌通过促进树突状细胞(dc)调节T细胞反应来调节免疫反应的关键作用,我们假设益生菌的应用可能成功预防或治疗COVID-19患者的肠道疾病和气道疾病。
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引用次数: 4
Potential use of Bacillus paramycoides for the production of the biopolymer polyhydroxybutyrate from leftover carob fruit agro-waste. 副芽孢杆菌在角豆果农业废弃物中生产聚羟基丁酸生物聚合物的潜在用途。
IF 4.8 Q3 MICROBIOLOGY Pub Date : 2022-08-08 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022023
Leila Djerrab, Zohra Chekroud, Amer Rouabhia, Mohamed Abdesselem Dems, Imane Attailia, Luis Isidoro Romero Garcia, Mustapha Adnane Smadi

This study was designed to investigate, at a laboratory scale, the possibility of valorizing the leftover carob fruits to produce the eco-friendly biopolymer polyhydroxybutyrate (PHB) by using the bacterial strain Bacillus paramycoides, which has been isolated from the botanical garden of Skikda University in Algeria. The PHB production was tested under various conditions: a pH of 3-8, temperature range of 30-44 °C, carob extracted molasses concentration of 2-8% v/v, an incubation time of 24-96 h and an agitation speed of 150-300 rpm. The effects of different nitrogen sources and carob extracted molasses treatment types were also investigated. The PHB concentration was determined quantitatively as crotonic acid by measuring the absorbance at 300 nm. Cell growth was quantified by measuring the density of the culture at 600 nm. The presence of PHB was confirmed by applying high-performance liquid chromatography (HPLC) using an Aminex HPX-87H and implementing gas chromatography analysis. The best yield of PHB synthesis was obtained by using 6% v/v of 5 M H2SO4 treated with carob molasses as a carbon source, with peptone as a nitrogen source; incubation was conducted at 37 °C for 96 h at an agitation speed of 300 rpm (114.95 mg/L). The HPLC analysis confirmed the synthesis of PHB by B. paramycoides to have a chromatogram retention time of 22.5 min. Carob waste was successfully valorized to PHB.

本研究的目的是在实验室规模上,利用从阿尔及利亚Skikda大学植物园分离到的副芽孢杆菌菌株,研究对角豆果剩余果实进行增殖以生产环保型生物聚合物聚羟基丁酸酯(PHB)的可能性。pH值为3-8,温度范围为30-44℃,角豆提取物糖蜜浓度为2-8% v/v,孵育时间为24-96 h,搅拌速度为150-300 rpm,对PHB的生产进行了测试。研究了不同氮源和角豆糖蜜处理方式对其生长的影响。在300 nm处测定吸光度,定量测定PHB浓度为巴豆酸。在600 nm处测定培养密度,定量细胞生长情况。采用Aminex HPX-87H高效液相色谱(HPLC)和气相色谱分析证实了PHB的存在。以6% v/v的5 M H2SO4为碳源,以角豆糖蜜为碳源,以蛋白胨为氮源,合成PHB的收率最高;37℃孵育96 h,搅拌速度300 rpm (114.95 mg/L)。通过高效液相色谱分析,证实了B. parycoides合成了PHB,色谱保留时间为22.5 min,成功地将角豆废料转化为PHB。
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引用次数: 0
Evaluation of the use of CRISPR loci for discrimination of Salmonella enterica subsp. enterica serovar Enteritidis strains recovered in Canada and comparison with other subtyping methods. 评估使用 CRISPR 基因位点对在加拿大发现的肠炎沙门氏菌亚种肠炎血清肠炎菌株进行鉴别的情况,并与其他亚型鉴定方法进行比较。
IF 2.7 Q3 MICROBIOLOGY Pub Date : 2022-07-15 eCollection Date: 2022-01-01 DOI: 10.3934/microbiol.2022022
Susan Nadin-Davis, Louise Pope, John Devenish, Ray Allain, Dele Ogunremi

Salmonella enterica subsp. enterica serovar Enteritidis remains one of the most important foodborne pathogens worldwide. To minimise its public health impact when outbreaks of the disease occur, timely investigation to identify and recall the contaminated food source is necessary. Central to this approach is the need for rapid and accurate identification of the bacterial subtype epidemiologically linked to the outbreak. While traditional methods of S. Enteritidis subtyping, such as pulsed field gel electrophoresis (PFGE) and phage typing (PT), have played an important role, the clonal nature of this organism has spurred efforts to improve subtyping resolution and timeliness through molecular based approaches. This study uses a cohort of 92 samples, recovered from a variety of sources, to compare these two traditional methods for S. Enteritidis subtyping with recently developed molecular techniques. These latter methods include the characterisation of two clustered regularly interspaced short palindromic repeats (CRISPR) loci, either in isolation or together with sequence analysis of virulence genes such as fimH. For comparison, another molecular technique developed in this laboratory involved the scoring of 60 informative single nucleotide polymorphisms (SNPs) distributed throughout the genome. Based on both the number of subtypes identified and Simpson's index of diversity, the CRISPR method was the least discriminatory and not significantly improved with the inclusion of fimH gene sequencing. While PT analysis identified the most subtypes, the SNP-PCR process generated the greatest index of diversity value. Combining methods consistently improved the number of subtypes identified, with the SNP/CRISPR typing scheme generating a level of diversity comparable with that of PT/PFGE. While these molecular methods, when combined, may have significant utility in real-world situations, this study suggests that CRISPR analysis alone lacks the discriminatory capability required to support investigations of foodborne disease outbreaks.

肠炎沙门氏菌肠炎亚种仍是全球最重要的食源性致病菌之一。为了在疾病暴发时将其对公众健康的影响降至最低,有必要及时开展调查,以确定和召回受污染的食物来源。这种方法的核心是需要快速准确地识别与疫情流行有关的细菌亚型。虽然传统的肠炎双球菌亚型鉴定方法(如脉冲场凝胶电泳 (PFGE) 和噬菌体分型 (PT))发挥了重要作用,但这种生物的克隆性促使人们努力通过分子方法提高亚型鉴定的分辨率和及时性。本研究使用从不同来源采集的 92 份样本,将这两种传统的肠炎双球菌亚型鉴定方法与最新开发的分子技术进行比较。后一种方法包括对两个簇状规则间隔短回文重复序列(CRISPR)基因座进行单独或与 fimH 等毒力基因的序列分析一起使用。作为比较,该实验室开发的另一项分子技术涉及对分布在整个基因组中的 60 个信息量丰富的单核苷酸多态性(SNPs)进行评分。从鉴定出的亚型数量和辛普森多样性指数来看,CRISPR 方法的鉴别力最低,加入 fimH 基因测序后也没有明显改善。虽然 PT 分析鉴定出的亚型最多,但 SNP-PCR 过程产生的多样性指数值最大。将各种方法结合起来能不断提高亚型鉴定的数量,SNP/CRISPR 分型方案产生的多样性水平与 PT/PFGE 相当。虽然这些分子方法结合起来在实际情况中可能有很大的用途,但本研究表明,仅靠 CRISPR 分析缺乏支持食源性疾病爆发调查所需的鉴别能力。
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AIMS Microbiology
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