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Characterization of non-O157 enterohemorrhagic Escherichia coli isolated from different sources in Egypt. 从埃及不同来源分离的非 O157 肠出血性大肠杆菌的特征。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-21 DOI: 10.1186/s12866-024-03636-3
Omnia T Bahgat, Dina E Rizk, Hany I Kenawy, Rasha Barwa

Background: Enterohemorrhagic Escherichia coli (EHEC) O157 is implicated in serious food and water-borne diseases as hemorrhagic colitis (HC), and the potentially fatal hemolytic uremic syndrome (HUS). However, new players of non-O157 EHEC have been implicated in serious infections worldwide. This work aims at analyzing serotype and genotypic-based virulence profile of EHEC local isolates.

Methods: A total of 335 samples were collected from different sources in Egypt. E. coli was isolated and subjected to serotyping. Non-O157 EHEC isolates were tested for virulence genes using PCR, phenotypic examination, phylogenetic typing, and molecular investigation by ERIC typing and MLST to disclose genetic relatedness of isolates. A heat map was used to identify potential associations between the origin of the isolates, their phenotypic and genotypic characteristics.

Results: A total of 105 out of 335 isolates were identified as E. coli. Surprisingly, 49.5% of these isolates were EHEC, where O111, O91, O26 and O55 were the most prevalent serotypes including 38.46% from stool, 21.15% urine, 23.1% cheese, 9.62% meat products, 3.85% from both yogurt and sewage water. Screening 15 different virulence genes revealed that sheA, stx2 and eae were the most prevalent with abundance rates of 85%, 75% and 36%, respectively. Fifteen profiles of virulence gene association were identified, where the most abundant one was stx2/sheA (19%) followed by stx2/stx2g/sheA/eae (11.5%). Both stx2/sheA/eae and stx2/stx2g/sheA were equally distributed in 9.6% of total isolates. Phylogenetic typing revealed that pathogenic phylogroups B2 and D were detected among clinical isolates only. Forty-six different patterns were detected by ERIC genotyping. MLST resolved three sequence types of ST70, ST120 and ST394. The heat map showed that 21 isolates were of 70% similarity, 9 groups were of 100% clonality.

Conclusions: The prevalence of non-O157 EHEC pathotype was marginally higher among the food isolates compared to the clinical ones. The endemic ST120 was detected in cheese, necessitating crucial measures to prevent the spread of this clone. Clinical EHEC isolates exhibited a higher score, and combination of virulence genes compared to food and sewage water isolates, thereby posing a significant public health concern.

背景:肠出血性大肠杆菌(EHEC)O157 与出血性结肠炎(HC)和可能致命的溶血性尿毒症(HUS)等严重食源性和水源性疾病有关。然而,非 O157 型 EHEC 的新成员也卷入了全球范围内的严重感染。本研究旨在分析当地分离出的 EHEC 的血清型和基于基因型的毒力特征:方法:从埃及不同来源共收集 335 份样本。方法:从埃及不同来源共采集 335 份样本,分离出大肠杆菌并进行血清型鉴定。利用 PCR、表型检查、系统发育分型以及 ERIC 分型和 MLST 进行分子调查,检测非 O157 型 EHEC 分离物的毒力基因,以揭示分离物的遗传亲缘关系。热图用于确定分离物的来源、表型和基因型特征之间的潜在关联:结果:在 335 个分离物中,共有 105 个被鉴定为大肠杆菌。令人惊讶的是,49.5%的分离物为大肠埃希氏菌,其中O111、O91、O26和O55是最常见的血清型,包括38.46%来自粪便、21.15%来自尿液、23.1%来自奶酪、9.62%来自肉制品、3.85%来自酸奶和污水。15 种不同毒力基因的筛选结果表明,sheA、stx2 和 eae 是最常见的毒力基因,丰度分别为 85%、75% 和 36%。发现了 15 个毒力基因关联图谱,其中含量最高的是 stx2/sheA(19%),其次是 stx2/stx2g/sheA/eae(11.5%)。在9.6%的分离物中,stx2/sheA/eae和stx2/stx2g/sheA分布相同。系统发育分型显示,仅在临床分离物中检测到致病性系统群 B2 和 D。ERIC 基因分型检测出 46 种不同的模式。MLST 分辨出 ST70、ST120 和 ST394 三种序列类型。热图显示,21 个分离物的相似度为 70%,9 个群体的克隆度为 100%:结论:与临床分离物相比,非O157 EHEC病原型在食品分离物中的流行率稍高。在奶酪中检测到了流行的 ST120,因此有必要采取重要措施防止该克隆的传播。与食品和污水分离物相比,临床大肠埃希氏菌分离物的得分更高,毒力基因的组合也更多,因此对公共卫生构成了重大威胁。
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引用次数: 0
Chemical quantification of N-acyl alanine methyl ester (NAME) production and impact on temporal gene expression patterns in Roseovarius tolerans EL-164. N-acyl alanine methyl ester (NAME) 生成的化学定量及其对 Roseovarius tolerans EL-164 中时间基因表达模式的影响。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-21 DOI: 10.1186/s12866-024-03624-7
Janina Leinberger, Diana Koteska, Judith Boldt, Jörn Petersen, Sahana Shivaramu, Jürgen Tomasch, Stefan Schulz, Thorsten Brinkhoff

Background: Previous studies have identified structurally diverse N-acyl amino acid methyl esters (NAMEs) in culture extracts of Roseovarius tolerans EL-164 (Roseobacteraceae). NAMEs are structural analogues of the common signaling compounds N-acyl homoserine lactones (AHLs), but do not participate in AHL-mediated signaling. NAMEs show minor antialgal and antimicrobial activity, but whether this activity serves as the primary ecological role remains unclear.

Results: To enable dose-dependent bioactivity-testing, we have established a chromatographic method for quantification of NAMEs in bacterial culture extracts. The concentrations determined for the two major NAMEs produced by EL-164, C16:1-NAME and C17:1-NAME, ranged between 0.685 and 5.731 mg L- 1 (2.0-16.9 µM) and 5.3-86.4 µg L- 1 (15.0-244.3 nM), respectively. Co-quantification of the C14:1-AHL showed concentrations ranging between 17.5 and 58.7 mg L- 1 (56.6-189.7 µM). We observed distinct production patterns for NAMEs and AHLs, with a continuous NAME production during the entire incubation period. We conducted a spike-in experiment, using the determined metabolite concentrations. By comparing the transcriptomes of pre- and post-metabolite-spikes, we identified three clusters of differentially expressed genes with distinct temporal expression patterns. Expression levels of stress response genes differed between NAME- and AHL-spiked EL-164 cultures in the stationary phase.

Conclusions: Our findings support previous studies suggesting an ecological role for C16:1-NAME as antibiotic, by proving that NAME concentrations in batch cultures were higher than the minimal inhibitory concentrations against Maribacter sp. 62 - 1 (Flavobacteriia) and Skeletonema costatum CCMP 1332 (Coscinodiscophyceae) reported in the literature. Our study further exemplified the broad application range of dose-dependent testing and highlighted the different biological activities of NAMEs and AHLs.

背景:先前的研究发现,玫瑰细菌(Roseobacteraceae)EL-164培养液中含有结构多样的N-酰基氨基酸甲酯(NAMEs)。NAMEs是常见信号化合物N-酰基高丝氨酸内酯(AHLs)的结构类似物,但不参与AHL介导的信号传导。NAMEs 具有轻微的抗藻类和抗菌活性,但这种活性是否作为主要的生态作用尚不清楚:为了进行剂量依赖性生物活性测试,我们建立了一种色谱法,用于定量检测细菌培养提取物中的 NAMEs。EL-164产生的两种主要NAMEs(C16:1-NAME和C17:1-NAME)的浓度分别为0.685-5.731 mg L- 1(2.0-16.9 µM)和5.3-86.4 µg L- 1(15.0-244.3 nM)。C14:1-AHL的共定量分析显示,其浓度范围为17.5-58.7 mg L- 1(56.6-189.7 µM)。我们观察到 NAMEs 和 AHLs 的不同生产模式,NAMEs 在整个培养期间持续生产。我们利用测定的代谢物浓度进行了尖峰实验。通过比较代谢物尖峰前和尖峰后的转录组,我们发现了三个具有不同时间表达模式的差异表达基因簇。在静止期,NAME和AHL加标EL-164培养物的应激反应基因表达水平不同:我们的研究结果证明,批次培养物中的 NAME 浓度高于文献中报道的对黄杆菌属(Maribacter sp.我们的研究进一步体现了剂量依赖性测试的广泛应用范围,并突出了NAMEs和AHLs的不同生物活性。
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引用次数: 0
A novel computational approach for the mining of signature pathways using species co-occurrence networks in gut microbiomes. 利用肠道微生物群中的物种共现网络挖掘特征通路的新型计算方法。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-21 DOI: 10.1186/s12866-024-03633-6
Suyeon Kim, Ishwor Thapa, Hesham Ali

Background: Advances in metagenome sequencing data continue to enable new methods for analyzing biological systems. When handling microbial profile data, metagenome sequencing has proven to be far more comprehensive than traditional methods such as 16s rRNA data, which rely on partial sequences. Microbial community profiling can be used to obtain key biological insights that pave the way for more accurate understanding of complex systems that are critical for advancing biomedical research and healthcare. However, such attempts have mostly used partial or incomplete data to accurately capture those associations.

Methods: This study introduces a novel computational approach for the identification of co-occurring microbial communities using the abundance and functional roles of species-level microbiome data. The proposed approach is then used to identify signature pathways associated with inflammatory bowel disease (IBD). Furthermore, we developed a computational pipeline to identify microbial species co-occurrences from metagenome data at various granularity levels.

Results: When comparing the IBD group to a control group, we show that certain co-occurring communities of species are enriched for potential pathways. We also show that the identified co-occurring microbial species operate as a community to facilitate pathway enrichment.

Conclusions: The obtained findings suggest that the proposed network model, along with the computational pipeline, provide a valuable analytical tool to analyze complex biological systems and extract pathway signatures that can be used to diagnose certain health conditions.

背景:元基因组测序数据的不断进步为分析生物系统提供了新的方法。在处理微生物概况数据时,元基因组测序已被证明比 16s rRNA 数据等依赖部分序列的传统方法要全面得多。微生物群落图谱分析可用于获得关键的生物学见解,为更准确地了解复杂系统铺平道路,这对推动生物医学研究和医疗保健至关重要。然而,这些尝试大多使用部分或不完整的数据来准确捕捉这些关联:本研究介绍了一种新的计算方法,利用物种级微生物组数据的丰度和功能作用来识别共生微生物群落。方法:本研究介绍了一种新的计算方法,利用物种级微生物组数据的丰度和功能作用识别共生微生物群落,然后利用该方法识别与炎症性肠病(IBD)相关的特征通路。此外,我们还开发了一个计算管道,从不同粒度水平的元基因组数据中识别微生物物种共现:结果:当将 IBD 组与对照组进行比较时,我们发现某些物种共存群落富集了潜在的通路。我们还表明,已确定的共生微生物物种作为一个群落运作,促进了通路的富集:结论:研究结果表明,所提出的网络模型和计算管道为分析复杂的生物系统和提取可用于诊断某些健康状况的通路特征提供了有价值的分析工具。
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引用次数: 0
Retraction Note: A modified multilocus sequence typing protocol to genotype Kingella kingae from oropharyngeal swabs without bacterial isolation. 撤稿说明:无需分离细菌即可对口咽拭子中的金氏菌进行基因分型的改良多焦点序列分型方案。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s12866-024-03652-3
Nawal El Houmami, Janek Bzdrenga, Jean-Christophe Pons, Philippe Minodier, Guillaume André Durand, Anis Oubraham, Dimitri Ceroni, Pablo Yagupsky, Didier Raoult, Philippe Bidet, Pierre-Edouard Fournier
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引用次数: 0
Functional and molecular characterization of millet associated probiotic bacteria. 小米相关益生菌的功能和分子特征。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s12866-024-03606-9
Bashipangu Gouthami, Alaguthevar Ramalakshmi, Murugesan Balakrishnan, Subburamu Karthikeyan, Iniyakumar Muniraj, Jeyakumar Saranya Packialakshmi

The lactic acid bacteria are one of the sustainable ways of food production. As the native lactic acid bacteria (LAB) easily manipulate the substrate, helps in production of health essential probiotics with enhancing the bioavailability of the substrate. Here also, in present study, the native LAB isolates isolated from the millets and characterize them for the functional analysis for the human health association. In the present study, fermented millet-associated lactic acid bacteria were screened and characterized for their probiotic potential, safety evaluation and antimicrobial activity. A total of 33 isolates were purified as lactic acid bacteria based on colony shape and biochemical assays. However, only 13 isolates were found to be catalase-negative. Among the 13 isolates, 5 isolates exhibited optimum growth at 6.5% and 9.5% of salt concentrations, pH of 4.5 to 8.5 and 17 °C to 40 °C of the temperature. The probiotic properties of the five isolates exhibited that the survival rates in acid and bile salt concentration ranged from 56.2 to 73.7% and 55.3 to 70.3%, respectively. Similarly, the surface hydrophobicity of the isolates was 41-75%. Antibiotic assay revealed that all five isolates were resistant to Amoxicillin, Cloxacillin, and Penicillin-V. Interestingly, all the isolates except ME26 displayed susceptibility towards Penicillin (2 units) and Tetracycline (10 µg). Further, the four isolates (ME25, ME26, ME9, and ME2) had more antifungal activity against Aspergillus flavus. However, only three, except ME1 and ME2, showed maximum antibacterial activity and produced more antimicrobial compounds compared to reference strain L. plantarum Pb3. The potential probiotic isolates were identified as Weisella cibaria ME9, Weisella cibaria ME26, and Weisella confusa ME25.

乳酸菌是一种可持续的食品生产方式。本地乳酸菌(LAB)很容易处理基质,有助于生产对健康至关重要的益生菌,提高基质的生物利用率。在本研究中,也从黍米中分离出了本地乳酸菌,并对其进行了功能分析,以确定其与人类健康的关系。在本研究中,对发酵的黍米相关乳酸菌进行了筛选,并对其益生菌潜力、安全性评价和抗菌活性进行了表征。根据菌落形状和生化测定,共有 33 个分离物被纯化为乳酸菌。然而,只有 13 个分离物被发现是过氧化氢酶阴性的。在这 13 个分离物中,5 个分离物在盐浓度为 6.5% 和 9.5%、pH 值为 4.5 至 8.5 和温度为 17 °C 至 40 °C 的条件下表现出最佳生长状态。这 5 个分离物的益生特性表明,它们在酸和胆盐浓度下的存活率分别为 56.2% 至 73.7% 和 55.3% 至 70.3%。同样,分离物的表面疏水性为 41-75%。抗生素检测显示,所有五种分离物都对阿莫西林、氯唑西林和青霉素-V 具有抗药性。有趣的是,除 ME26 外,所有分离物都对青霉素(2 单位)和四环素(10 µg)有敏感性。此外,四种分离物(ME25、ME26、ME9 和 ME2)对黄曲霉具有更强的抗真菌活性。然而,与参考菌株 L. plantarum Pb3 相比,除 ME1 和 ME2 外,只有三个菌株显示出最大的抗菌活性,并产生了更多的抗菌化合物。经鉴定,这些潜在的益生菌分离物分别为西伯利亚魏氏菌 ME9、西伯利亚魏氏菌 ME26 和 confusa 魏氏菌 ME25。
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引用次数: 0
The role of New World vultures as carriers of environmental antimicrobial resistance. 新世界秃鹫作为环境抗菌药耐药性载体的作用。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-20 DOI: 10.1186/s12866-024-03621-w
Anaïs K Tallon, Renotta K Smith, Scott Rush, Adrian Naveda-Rodriguez, John P Brooks

Background: Although antibiotics have significantly improved human and animal health, their intensive use leads to the accumulation of antimicrobial resistance (AMR) in the environment. Moreover, certain waste management practices create the ideal conditions for AMR development while providing predictable resources for wildlife. Here, we investigated the role of landfills in the potentiation of New World vultures to disseminate environmental AMR. We collected 107 samples (soil, water, and feces) between 2023 and 2024, in different bird use sites (roosts, landfills and boneyards).

Results: We isolated enterococci (EN), Escherichia coli (EC), and Salmonella spp. (SM), performed antibiotic susceptibility tests, and quantified the presence of antibiotic resistance genes (ARGs) within all samples. We identified EN, EC, and SM, in 50, 37, and 26 samples, from the three vulture use areas, respectively. AMR was mainly to aminoglycoside, cephalosporin, and tetracycline, and the prevalence of multidrug resistance (MDR) was 5.3% (EC), 78.2% (EN), and 17.6% (SM). Variations in bacterial abundance and AMR/MDR profiles were found based on the season, use site, and sample types, which was corroborated by ARG analyses.

Conclusions: Our study suggests that landfills constitute a source of zoonotic pathogens and AMR for wildlife, due to readily available refuse input. Using non-invasive molecular methods, we highlight an often-ignored ecosystem within the One Health paradigm.

背景:尽管抗生素极大地改善了人类和动物的健康,但其大量使用会导致环境中抗菌素抗药性(AMR)的积累。此外,某些废物管理方法在为野生动物提供可预测资源的同时,也为 AMR 的发展创造了理想条件。在此,我们调查了垃圾填埋场在新世界秃鹫传播环境 AMR 中的潜在作用。2023 年至 2024 年期间,我们在不同的鸟类栖息地(栖息地、垃圾填埋场和骨灰堂)收集了 107 份样本(土壤、水和粪便):我们分离了肠球菌(EN)、大肠杆菌(EC)和沙门氏菌属(SM),进行了抗生素敏感性测试,并对所有样本中存在的抗生素耐药基因(ARGs)进行了量化。我们分别在三个秃鹫使用区的 50 份、37 份和 26 份样本中发现了 EN、EC 和 SM。AMR主要针对氨基糖苷类、头孢菌素和四环素,多重耐药性(MDR)的发生率分别为5.3%(EC)、78.2%(EN)和17.6%(SM)。根据季节、使用地点和样本类型的不同,细菌丰度和AMR/MDR概况也有所不同,ARG分析也证实了这一点:我们的研究表明,垃圾填埋场是野生动物人畜共患病病原体和 AMR 的来源之一,因为这里有现成的垃圾输入。利用非侵入性分子方法,我们强调了 "同一健康 "模式中一个经常被忽视的生态系统。
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引用次数: 0
Pseudomegasporoporia margallensis sp. nov. and Microporellus subadustus (Polyporaceae, Basidiomycota) newly described and recorded from Pakistan. 巴基斯坦新描述和记录的 Pseudomegasporoporia margallensis sp.
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-19 DOI: 10.1186/s12866-024-03567-z
Fauzia Razzaq, Rimsha Murtaza, Muhammad Usman, Abdul Nasir Khalid, Shazia Ashraf, Qirui Li

In this study, we describe Pseudomegasporoporia margallensis as a novel species and report Microporellus subadustus as a new record from Pakistan based on morphology and phylogeny. Both genera are firstly reported in the country. Pseudomegasporoporia margallensis sp. nov. is characterized by light brownish gray poroid hymenphore, grayish brown subiculum surface, frequent generative hyphae in the subiculum with abundant clamp connections, and arboriform-branched skeletal hyphae in subiculum and tubes, and it forms a sister lineage of P. nerricola in the phylogeny. This novel taxon, showing 37 nucleotide differences in ITS nrDNA and 10 in LSU nrLSU sequences from P. nerricola, has GenBank accession numbers PP702233, PP702234, PP702235, PP702236, and PP70223, and is registered under MycoBank no. MB853645. This higlights the genetic distinction and importance of molecular identification in fungal taxonomy. The new species and the new record are described with detail illustrations.

在这项研究中,我们描述了 Pseudomegasporoporia margallensis 这一新种,并根据形态学和系统发育报告了 Microporellus subadustus 这一来自巴基斯坦的新记录。这两个属都是首次在巴基斯坦报道。Pseudomegasporoporia margallensis sp.nov.的特征是:膜柄呈浅棕色灰色孔状,子房表面呈灰棕色,子房内有丰富的生成菌丝,子房和管内有乔木状分枝的骨架菌丝,在系统发育中与P. nerricola是姊妹系。该新类群的 ITS nrDNA 与 P. nerricola 有 37 个核苷酸差异,LSU nrLSU 序列与 P. nerricola 有 10 个核苷酸差异,其 GenBank 登录号分别为 PP702233、PP702234、PP702235、PP702236 和 PP70223,MycoBank 注册号为 MB853645。MB853645。这凸显了分子鉴定在真菌分类学中的遗传区别和重要性。新种和新记录的描述附有详细插图。
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引用次数: 0
Brain injury in premature infants may be related to abnormal colonization of early gut microbiome. 早产儿脑损伤可能与早期肠道微生物组定植异常有关。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-19 DOI: 10.1186/s12866-024-03643-4
Li Hou, Jing Zhao, Linlin Yin, Lu Dai, Hong Deng, Lin Jiang

Background: Premature infants are more prone to brain injuries owing to incomplete nervous system development and poor adaptation outside the mother's body. Without timely intervention, premature infants with brain injuries often develop intellectual disabilities, causing significant burdens on families and the society. Multiple studies have shown that gut dysbiosis can affect the nervous system, and vice versa. This study aimed to explore the changes in gut microbiota of typical premature infants and those with brain injuries on the third and seventh days after birth using 16 S rRNA technology.

Methods: Fecal samples from typical premature infants (non-brain injury group, n = 17) and those with brain injuries (brain injury group, n = 21) were collected on days 1, 3, and 7 after birth for 16 S rRNA sequencing. Alpha diversity analysis was used to evaluate the diversity of gut microbiome. LEfSe and DESeq2 were used to analyze of the microorganisms' characteristics and differentiate the microorganisms between the two groups.

Results: At the phylum level, Firmicutes, Proteobacteria, and Actinobacteria were the dominant flora in both groups. At the genus level, the proportion of Enterococcus in fecal samples of the brain injury group was higher than that of the non-brain injury group on day three after birth; however, the opposite was observed on day seven. Rothia and Lactobacillales were characteristic bacteria of the non-brain injury group on days three and seven after birth, whereas Enterococcus and Bifidobacteria were characteristic bacteria of the brain injury group on days three and seven after birth, respectively. Three days after birth, the Shannon and Simpson indices of the non-brain injury group were significantly higher than those of the brain injury group.

Conclusion: Premature infants with brain injuries have a unique gut microbiota that is different from that of typical premature infants, indicating correlation between brain injuries and gut microbiota.

背景:早产儿由于神经系统发育不完全,对母体外的适应能力较差,容易发生脑损伤。如果不及时干预,脑损伤早产儿往往会发展成智障,给家庭和社会带来沉重负担。多项研究表明,肠道菌群失调会影响神经系统,反之亦然。本研究旨在利用 16 S rRNA 技术探讨典型早产儿和脑损伤患儿出生后第三天和第七天肠道微生物群的变化:方法:收集典型早产儿(非脑损伤组,n = 17)和脑损伤早产儿(脑损伤组,n = 21)出生后第 1、3 和 7 天的粪便样本,进行 16 S rRNA 测序。阿尔法多样性分析用于评估肠道微生物组的多样性。使用 LEfSe 和 DESeq2 分析微生物的特征,并区分两组之间的微生物:结果:在门的水平上,固缩菌、蛋白菌和放线菌是两组的优势菌群。在属一级,脑损伤组在出生后第三天粪便样本中的肠球菌比例高于非脑损伤组,但在第七天则相反。罗氏菌和乳酸杆菌是非脑损伤组在出生后第三天和第七天的特征细菌,而肠球菌和双歧杆菌则分别是脑损伤组在出生后第三天和第七天的特征细菌。出生后三天,非脑损伤组的香农指数和辛普森指数明显高于脑损伤组:结论:脑损伤早产儿的肠道微生物群与一般早产儿不同,表明脑损伤与肠道微生物群之间存在相关性。
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引用次数: 0
Isolation, characterization and genome analysis of the orphan phage Kintu infecting Xanthomonas vasicola pv. musacearum. 感染黄单胞菌 vasicola pv. musacearum 的孤儿噬菌体 Kintu 的分离、特征和基因组分析。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s12866-024-03629-2
Ritah Nakayinga, Ibrahim Ntulume, Jeroen Wagemans, Marta Vallino, Reagan Kanaabi, Ali Kajubi, Justus Kwetegyeka

Background: Xanthomonas vasicola pv. musacearum is responsible for the widespread Banana Xanthomonas Wilt in banana cultivation regions across the globe. Biocontrol measures for disease management remain limited amidst increasing antimicrobial resistance and unsustainable conventional agricultural practices. The purpose of this study is to explore a viable alternative or adjunct strategy through the use of bacteriophages for disease management.

Results: Kintu was isolated from sewage and displayed clear and circular plaques measuring 3 mm. Based on transmission electron microscopy, Kintu displays siphovirus characteristics, including an icosahedral head and a non-contractile tail. Kintu infects 78% (22 out of 28) Ugandan Xvm strains, has an optimal multiplicity of infection of 1, a 10 min adsorption and latent period, a 35 min burst period, and a burst size of 15 particles per bacterium. Phage titers remain stable for two and half months (75 days) in SM buffer at -20 oC and - 40 oC but decrease significantly (p ≤ 0.0001) at 4 oC. Kintu is active at pH 3 and 11, maintains viability at temperatures between 25 oC and 120 oC and tolerates UV irradiation for up to 2 min and 20 s. Kintu inhibits Xvm growth at MOI ratios of 0.1, 1 and 10. The genome is a double stranded DNA molecule that consists of 48,985 base pairs and a G + C content of 51.71%. Antibiotic resistance genes or genes associated with a lysogenic life cycle are absent. There is limited sequence similarity of Kintu with other phages, making it a novel phage belonging to an unclassified genus of the class Caudoviricetes.

Conclusion: Kintu is a novel bacteriophage that infects and lyses Xanthomonas vasicola pv. musacearum, the causative agent for Banana Xanthomonas Wilt. Its stability across diverse temperatures and pH conditions highlights its potential as a biocontrol agent for managing the disease.

背景:黄单胞菌 vasicola pv. musacearum 是全球香蕉种植区广泛存在的香蕉黄单胞菌枯萎病的罪魁祸首。在抗菌素抗药性不断增加和传统农业做法不可持续的情况下,用于病害管理的生物防治措施仍然有限。本研究的目的是通过使用噬菌体来控制病害,探索一种可行的替代或辅助策略:结果:Kintu 从污水中分离出来,显示出 3 毫米大小的透明圆形斑块。根据透射电子显微镜观察,Kintu 显示出虹吸式病毒的特征,包括二十面体头部和无收缩的尾部。Kintu 能感染 78% 的乌干达 Xvm 菌株(28 株中的 22 株),最佳感染倍数为 1,吸附和潜伏期为 10 分钟,爆发期为 35 分钟,爆发大小为每个细菌 15 个颗粒。噬菌体滴度在 -20 oC 和 -40 oC 的 SM 缓冲液中可保持稳定两个半月(75 天),但在 4 oC 时会显著降低(p ≤ 0.0001)。Kintu 在 pH 值为 3 和 11 时具有活性,在 25 oC 至 120 oC 的温度范围内保持活力,并能耐受长达 2 分钟 20 秒的紫外线照射。基因组为双链 DNA 分子,由 48 985 个碱基对组成,G+C 含量为 51.71%。不存在抗生素耐药性基因或与溶菌性生命周期相关的基因。Kintu 与其他噬菌体的序列相似性有限,因此是一种属于 Caudoviricetes 类未分类属的新型噬菌体:结论:Kintu 是一种新型噬菌体,能感染并裂解香蕉黄单胞菌枯萎病的病原体--黄单胞菌 vasicola pv. musacearum。它在不同温度和酸碱度条件下的稳定性突显了其作为生物控制剂控制该疾病的潜力。
{"title":"Isolation, characterization and genome analysis of the orphan phage Kintu infecting Xanthomonas vasicola pv. musacearum.","authors":"Ritah Nakayinga, Ibrahim Ntulume, Jeroen Wagemans, Marta Vallino, Reagan Kanaabi, Ali Kajubi, Justus Kwetegyeka","doi":"10.1186/s12866-024-03629-2","DOIUrl":"10.1186/s12866-024-03629-2","url":null,"abstract":"<p><strong>Background: </strong>Xanthomonas vasicola pv. musacearum is responsible for the widespread Banana Xanthomonas Wilt in banana cultivation regions across the globe. Biocontrol measures for disease management remain limited amidst increasing antimicrobial resistance and unsustainable conventional agricultural practices. The purpose of this study is to explore a viable alternative or adjunct strategy through the use of bacteriophages for disease management.</p><p><strong>Results: </strong>Kintu was isolated from sewage and displayed clear and circular plaques measuring 3 mm. Based on transmission electron microscopy, Kintu displays siphovirus characteristics, including an icosahedral head and a non-contractile tail. Kintu infects 78% (22 out of 28) Ugandan Xvm strains, has an optimal multiplicity of infection of 1, a 10 min adsorption and latent period, a 35 min burst period, and a burst size of 15 particles per bacterium. Phage titers remain stable for two and half months (75 days) in SM buffer at -20 <sup>o</sup>C and - 40 <sup>o</sup>C but decrease significantly (p ≤ 0.0001) at 4 <sup>o</sup>C. Kintu is active at pH 3 and 11, maintains viability at temperatures between 25 <sup>o</sup>C and 120 <sup>o</sup>C and tolerates UV irradiation for up to 2 min and 20 s. Kintu inhibits Xvm growth at MOI ratios of 0.1, 1 and 10. The genome is a double stranded DNA molecule that consists of 48,985 base pairs and a G + C content of 51.71%. Antibiotic resistance genes or genes associated with a lysogenic life cycle are absent. There is limited sequence similarity of Kintu with other phages, making it a novel phage belonging to an unclassified genus of the class Caudoviricetes.</p><p><strong>Conclusion: </strong>Kintu is a novel bacteriophage that infects and lyses Xanthomonas vasicola pv. musacearum, the causative agent for Banana Xanthomonas Wilt. Its stability across diverse temperatures and pH conditions highlights its potential as a biocontrol agent for managing the disease.</p>","PeriodicalId":9233,"journal":{"name":"BMC Microbiology","volume":"24 1","pages":"482"},"PeriodicalIF":4.0,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11572249/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142667296","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A dispensable SepIVA orthologue in Streptomyces venezuelae is associated with polar growth and not cell division. 委内瑞拉链霉菌中可有可无的 SepIVA 同源物与极性生长而非细胞分裂有关。
IF 4 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2024-11-18 DOI: 10.1186/s12866-024-03625-6
Beer Chakra Sen, Parminder Singh Mavi, Oihane Irazoki, Susmita Datta, Sebastian Kaiser, Felipe Cava, Klas Flärdh

Background: SepIVA has been reported to be an essential septation factor in Mycolicibacterium smegmatis and Mycobacterium tuberculosis. It is a coiled-coil protein with similarity to DivIVA, a protein necessary for polar growth in members of the phylum Actinomycetota. Orthologues of SepIVA are broadly distributed among actinomycetes, including in Streptomyces spp.

Results: To clarify the role of SepIVA and its potential involvement in cell division in streptomycetes, we generated sepIVA deletion mutants in Streptomyces venezuelae and found that sepIVA is dispensable for growth, cell division and sporulation. Further, mNeonGreen-SepIVA fusion protein did not localize at division septa, and we found no evidence of involvement of SepIVA in cell division. Instead, mNeonGreen-SepIVA was accumulated at the tips of growing vegetative hyphae in ways reminiscent of the apical localization of polarisome components like DivIVA. Bacterial two-hybrid system analyses revealed an interaction between SepIVA and DivIVA. The results indicate that SepIVA is associated with polar growth. However, no phenotypic effects of sepIVA deletion could be detected, and no evidence was observed of redundancy with the other DivIVA-like coiled-coil proteins Scy and FilP that are also associated with apical growth in streptomycetes.

Conclusions: We conclude that S. venezuelae SepIVA, in contrast to the situation in mycobacteria, is dispensable for growth and viability. The results suggest that it is associated with polar growth rather than septum formation.

背景:据报道,SepIVA 是烟曲霉分枝杆菌(Mycolicibacterium smegmatis)和结核分枝杆菌(Mycobacterium tuberculosis)的重要隔膜因子。它是一种与 DivIVA 相似的线圈蛋白,DivIVA 是放线菌门成员极性生长所必需的蛋白。SepIVA 的同源物广泛分布于放线菌中,包括链霉菌中:为了明确 SepIVA 在链霉菌中的作用及其在细胞分裂中的潜在参与,我们在委内瑞拉链霉菌中产生了 sepIVA 缺失突变体,并发现 sepIVA 对生长、细胞分裂和孢子形成是不可或缺的。此外,mNeonGreen-SepIVA 融合蛋白没有定位在分裂隔膜上,我们也没有发现 SepIVA 参与细胞分裂的证据。相反,mNeonGreen-SepIVA 在生长的无性菌丝顶端积累,其方式与 DivIVA 等极性体成分的顶端定位相似。细菌双杂交系统分析揭示了 SepIVA 与 DivIVA 之间的相互作用。结果表明,SepIVA 与极性生长有关。然而,SepIVA 基因缺失并没有产生表型效应,也没有证据表明它与其他类似 DivIVA 的线圈蛋白 Scy 和 FilP 之间存在冗余,而这两种蛋白在链霉菌中也与顶端生长有关:结论:我们得出结论认为,与分枝杆菌的情况不同,委内瑞拉链霉菌的 SepIVA 对其生长和存活是不可或缺的。结果表明,它与极性生长而非隔膜形成有关。
{"title":"A dispensable SepIVA orthologue in Streptomyces venezuelae is associated with polar growth and not cell division.","authors":"Beer Chakra Sen, Parminder Singh Mavi, Oihane Irazoki, Susmita Datta, Sebastian Kaiser, Felipe Cava, Klas Flärdh","doi":"10.1186/s12866-024-03625-6","DOIUrl":"10.1186/s12866-024-03625-6","url":null,"abstract":"<p><strong>Background: </strong>SepIVA has been reported to be an essential septation factor in Mycolicibacterium smegmatis and Mycobacterium tuberculosis. It is a coiled-coil protein with similarity to DivIVA, a protein necessary for polar growth in members of the phylum Actinomycetota. Orthologues of SepIVA are broadly distributed among actinomycetes, including in Streptomyces spp.</p><p><strong>Results: </strong>To clarify the role of SepIVA and its potential involvement in cell division in streptomycetes, we generated sepIVA deletion mutants in Streptomyces venezuelae and found that sepIVA is dispensable for growth, cell division and sporulation. Further, mNeonGreen-SepIVA fusion protein did not localize at division septa, and we found no evidence of involvement of SepIVA in cell division. Instead, mNeonGreen-SepIVA was accumulated at the tips of growing vegetative hyphae in ways reminiscent of the apical localization of polarisome components like DivIVA. Bacterial two-hybrid system analyses revealed an interaction between SepIVA and DivIVA. The results indicate that SepIVA is associated with polar growth. However, no phenotypic effects of sepIVA deletion could be detected, and no evidence was observed of redundancy with the other DivIVA-like coiled-coil proteins Scy and FilP that are also associated with apical growth in streptomycetes.</p><p><strong>Conclusions: </strong>We conclude that S. venezuelae SepIVA, in contrast to the situation in mycobacteria, is dispensable for growth and viability. The results suggest that it is associated with polar growth rather than septum formation.</p>","PeriodicalId":9233,"journal":{"name":"BMC Microbiology","volume":"24 1","pages":"481"},"PeriodicalIF":4.0,"publicationDate":"2024-11-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11571769/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142667290","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
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