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Novel and rare β-lactamase genes of Bacteroides fragilis group species: Detection of the genes and characterization of their genetic backgrounds 脆弱拟杆菌属物种的新型和罕见β-内酰胺酶基因:基因检测及其遗传背景特征。
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-02-13 DOI: 10.1016/j.anaerobe.2024.102832
Bakhtiyar Mahmood , Károly Péter Sárvári , Laszló Orosz , Elisabeth Nagy , József Sóki

Objectives

This study screened the prevalence of rare β-lactamase genes in Bacteroides fragilis group strains from clinical specimens and normal microbiota and examined the genetic properties of the strains carrying these genes.

Methods

blaHGD1, blaOXA347, cblA, crxA, and pbbA were detected by real-time polymerase chain reaction in collections of Bacteroides strains from clinical (n = 406) and fecal (n = 184) samples. To examine the genetic backgrounds of the samples, end-point PCR, FT-IR, and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry were used.

Results

All B. uniformis isolates were positive for cblA in both collections. Although crxA was B. xylanisolvens-specific and associated with carbapenem resistance, it was only found in six fecal and three clinical B. xylanisolvens strains. Moreover, the crxA-positive strains were not clonal among B. xylanisolvens (contrary to cfiA in B. fragilis), implicating a rate of mobility or emergence by independent evolutionary events. The Phocaeicola (B.) vulgatus/P. dorei-specific gene blaHGD1 was detected among all P. vulgatus/P. dorei isolates from fecal (n = 36) and clinical (n = 26) samples. No blaOXA347-carrying isolate was found from European collections, but all US samples (n = 6) were positive. For three clinical isolates belonging to B. thetaiotaomicron (n = 2) and B. ovatus (n = 1), pbbA was detected on mobile genetic elements, and pbbA-positive strains displayed non-susceptibility to piperacillin or piperacillin/tazobactam phenotypically.

Conclusions

Based on these observations, β-lactamases produced by rare β-lactamase genes in B. fragilis group strains should not be overlooked because they could encode important resistance phenotypes.

研究目的方法:通过实时聚合酶链反应检测临床样本(406株)和粪便样本(184株)中脆弱拟杆菌菌株中的blaaHGD1、blaOXA347、cblA、crxA和pbbA。为检测样本的遗传背景,采用了终点 PCR、傅立叶变换红外光谱和基质辅助激光解吸/电离飞行时间质谱法:结果:在这两个样本集中,所有分离出的均匀芽孢杆菌的 cblA 都呈阳性。尽管crxA是木聚糖杆菌特异性的,并且与碳青霉烯耐药性有关,但它只在6株粪便和3株临床木聚糖杆菌中发现。此外,crxA 阳性菌株在 B. xylanisolvens 中没有克隆(与 B. fragilis 中的 cfiA 相反),这意味着其流动性或出现于独立的进化事件中。从粪便样本(36 个)和临床样本(26 个)中分离出的所有 P. vulgatus/P. dorei 都检测到了 Phocaeicola (B.) vulgatus/P. dorei 特异基因 blaHGD1。欧洲采集的样本中没有发现携带 blaOXA347 的分离株,但所有美国样本(n = 6)均呈阳性。在属于 B. thetaiotaomicron(n = 2)和 B. ovatus(n = 1)的三个临床分离株中,pbbA 在移动遗传因子上被检测到,pbbA 阳性菌株在表型上对哌拉西林或哌拉西林/他唑巴坦无敏感性:基于这些观察结果,不应忽视脆弱拟杆菌属菌株中罕见的β-内酰胺酶基因所产生的β-内酰胺酶,因为它们可能编码重要的耐药表型。
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引用次数: 0
Microscopy methods for Clostridioides difficile 艰难梭菌的显微镜检查方法。
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-02-08 DOI: 10.1016/j.anaerobe.2024.102822
Chenlin Hu, Kevin W. Garey

Microscopic technologies including light and fluorescent, transmission electron microscopy (TEM), scanning electron microscopy (SEM), and cryo-electron microscopy have been widely utilized to visualize Clostridioides difficile at the molecular, cellular, community, and structural biology level. This comprehensive review summarizes the microscopy tools (fluorescent and reporter system) in their use to study different aspects of C. difficile life cycle and virulence (sporulation, germination) or applications (detection of C. difficile or use of antimicrobials). With these developing techniques, microscopy tools will be able to find broader applications and address more challenging questions to study C. difficile and C. difficile infection.

包括光和荧光、透射电子显微镜(TEM)、扫描电子显微镜(SEM)和低温电子显微镜在内的显微镜技术已被广泛用于从分子、细胞、群落和结构生物学层面观察艰难梭菌。本综述总结了用于研究艰难梭菌生命周期和毒力(孢子、发芽)或应用(艰难梭菌检测或抗菌药物的使用)不同方面的显微镜工具(荧光和报告系统)。有了这些不断发展的技术,显微镜工具将能够找到更广泛的应用,并解决研究艰难梭菌和艰难梭菌感染的更具挑战性的问题。
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引用次数: 0
The burden of Clostridioides difficile infections in South-East Asia and the Western Pacific: A narrative review 东南亚和西太平洋地区艰难梭菌感染的负担:叙述性综述。
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-02-07 DOI: 10.1016/j.anaerobe.2024.102821
Frederick J. Angulo , Canna Ghia , Mark A. Fletcher , Egemen Ozbilgili , Graciela del Carmen Morales

Background

Clostridioides difficile (formerly Clostridium difficile) is well-documented in Europe and North America to be a common cause of healthcare-associated gastrointestinal tract infections. In contrast, C difficile infection (CDI) is infrequently reported in literature from Asia, which may reflect a lack of clinician awareness. We conducted a narrative review to better understand CDI burden in Asia.

Methods

We searched the PubMed database for English language articles related to C difficile, Asia, epidemiology, and molecular characteristics (eg, ribotype, antimicrobial resistance).

Results

Fifty-eight articles that met eligibility criteria were included. C difficile prevalence ranged from 7.1% to 45.1 % of hospitalized patients with diarrhea, and toxigenic strains among all C difficile in these patients ranged from 68.2% to 91.9 % in China and from 39.0% to 60.0 % outside of China. Widespread C difficile ribotypes were RT017, RT014/020, RT012, and RT002. Recurrence in patients with CDI ranged from 3.0% to 17.2 %. Patients with CDI typically had prior antimicrobial use recently. High rates of resistance to ciprofloxacin, clindamycin, and erythromycin were frequently reported.

Conclusion

The regional CDI burden in Asia is still incompletely documented, seemingly due to low awareness and limited laboratory testing. Despite this apparent under recognition, the current CDI burden highlights the need for broader surveillance and for application of preventative measures against CDI in Asia.

背景:在欧洲和北美,艰难梭状芽孢杆菌(原艰难梭状芽孢杆菌)已被证实是医源性胃肠道感染的常见病因。相比之下,艰难梭菌感染(CDI)在亚洲的文献中鲜有报道,这可能反映了临床医生缺乏这方面的意识。我们进行了一项叙述性综述,以更好地了解亚洲的艰难梭菌感染负担:我们在 PubMed 数据库中搜索了与艰难梭菌、亚洲、流行病学和分子特征(如核型、抗菌药耐药性)相关的英文文章:结果:共收录了58篇符合资格标准的文章。在住院的腹泻患者中,艰难梭菌的感染率从 7.1% 到 45.1%不等,在这些患者的所有艰难梭菌中,毒性菌株在中国从 68.2% 到 91.9%不等,在中国以外从 39.0% 到 60.0%不等。广泛存在的艰难梭菌核型为 RT017、RT014/020、RT012 和 RT002。CDI 患者的复发率从 3.0% 到 17.2% 不等。CDI 患者近期一般都曾使用过抗菌药物。对环丙沙星、克林霉素和红霉素的耐药率很高:结论:亚洲地区的 CDI 负担仍未得到完整记录,这似乎是由于认知度低和实验室检测有限造成的。尽管认识明显不足,但目前的 CDI 负担凸显了在亚洲进行更广泛监测和采取 CDI 预防措施的必要性。
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引用次数: 0
Bacteroides and related species: The keystone taxa of the human gut microbiota Bacteroides 和相关物种:人类肠道微生物群的关键类群
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-02-01 DOI: 10.1016/j.anaerobe.2024.102819
Jae Hyun Shin , Glenn Tillotson , Tiffany N. MacKenzie , Cirle A. Warren , Hannah M. Wexler , Ellie J.C. Goldstein

Microbial communities play a significant role in maintaining ecosystems in a healthy homeostasis. Presently, in the human gastrointestinal tract, there are certain taxonomic groups of importance, though there is no single species that plays a keystone role. Bacteroides spp. are known to be major players in the maintenance of eubiosis in the human gastrointestinal tract. Here we review the critical role that Bacteroides play in the human gut, their potential pathogenic role outside of the gut, and their various methods of adapting to the environment, with a focus on data for B. fragilis and B. thetaiotaomicron. Bacteroides are anaerobic non-sporing gram-negative organisms that are also resistant to bile acids, generally thriving in the gut and having a beneficial relationship with the host. While they are generally commensal organisms, some Bacteroides spp. can be opportunistic pathogens in scenarios of GI disease, trauma, cancer, or GI surgery, and cause infection, most commonly intra-abdominal infection. B. fragilis can develop antimicrobial resistance through multiple mechanisms in large part due to its plasticity and fluid genome. Bacteroidota (formerly, Bacteroidetes) have a very broad metabolic potential in the GI microbiota and can rapidly adapt their carbohydrate metabolism to the available nutrients. Gastrointestinal Bacteroidota species produce short-chain fatty acids such as succinate, acetate, butyrate, and occasionally propionate, as the major end-products, which have wide-ranging and many beneficial influences on the host. Bacteroidota, via bile acid metabolism, also play a role in in colonization-resistance of other organisms, including Clostridioides difficile, and maintenance of gut integrity.

微生物群落在维持生态系统健康平衡方面发挥着重要作用。目前,在人类胃肠道中,虽然没有单一物种发挥关键作用,但某些分类群具有重要意义。众所周知,嗜酸乳杆菌属(Bacteroides spp.)是维持人类胃肠道优生的主要角色。在此,我们回顾了乳杆菌在人类肠道中的关键作用、它们在肠道外的潜在致病作用以及它们适应环境的各种方法,并重点介绍了脆弱拟杆菌(B. fragilis)和泰奥米克隆杆菌(B. thetaiotaomicron)的数据。Bacteroides 是厌氧的无芽孢革兰氏阴性菌,对胆汁酸也有抗性,通常在肠道中生长,与宿主之间存在有益的关系。虽然它们通常是共生生物,但在消化道疾病、创伤、癌症或消化道手术的情况下,一些杆菌属可能会成为机会性病原体,并引起感染,最常见的是腹腔内感染。脆弱拟杆菌可通过多种机制产生抗菌药耐药性,这在很大程度上是由于其可塑性和基因组的流动性。类杆菌(原为类杆菌)在消化道微生物群中具有非常广泛的新陈代谢潜能,可根据可用的营养物质迅速调整其碳水化合物新陈代谢。胃肠道类杆菌产生的主要终产物是短链脂肪酸,如琥珀酸、乙酸、丁酸,偶尔也会产生丙酸,这些脂肪酸对宿主有广泛而有益的影响。类杆菌通过胆汁酸代谢,还对其他生物(包括艰难梭状芽孢杆菌)的定植-抗性以及肠道完整性的维护起到一定作用。
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引用次数: 0
Polyphasic molecular approach to the characterization of methanogens in the saliva of Tunisian adults 突尼斯成年人唾液中甲烷菌特征的多相分子方法
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-02-01 DOI: 10.1016/j.anaerobe.2024.102820
Fériel Bouzid , Imen Gtif , Salma Charfeddine , Leila Abid , Najla Kharrat

Background

Methanogenic archaea are a minor component of human oral microbiota. Due to their relatively low abundance, the detection of these neglected microorganisms is challenging.

Objective

This study concerns the presence of methanogens in salivary samples collected from Tunisian adults to evaluate their prevalence and burden using a polyphasic molecular approach.

Methods

A total of 43 saliva samples were included. Metagenomic and standard 16S rRNA sequencing were performed as an initial screening to detect the presence of methanogens in the oral microbiota of Tunisian adults. Further investigations were performed using specific quantitative real-time PCR targeting Methanobrevibacter oralis and Methanobrevibacter smithii.

Results

Methanobrevibacter was detected in 5/43 (11.62 %) saliva samples after metagenomic 16S rRNA data analysis. The presence of M. oralis was confirmed in 6/43 samples by standard 16S rRNA sequencing. Using real-time PCR, methanogens were detected in 35/43 (81.39 %) samples, including 62.79 % positive for M. oralis and 76.74 % positive for M. smithii. These findings reflect the high prevalence of both methanogens, revealed by the high sensitivity of the real-time PCR approach. Interestingly, we also noted a significant statistical association between the detection of M. smithii and poor adherence to a Mediterranean diet, indicating the impact of diet on M. smithii prevalence.

Conclusion

Our study showed the presence of methanogens in the oral microbiota of Tunisian adults with an unprecedented relatively high prevalence. Choice of methodology is also central to picturing the real prevalence and diversity of such minor taxa in the oral microbiota.

背景产甲烷古菌是人类口腔微生物群的一个次要组成部分。本研究关注突尼斯成年人唾液样本中甲烷菌的存在,采用多相分子方法评估甲烷菌的流行率和负担。方法共纳入 43 份唾液样本,进行了元基因组和标准 16S rRNA 测序,作为初步筛选,以检测突尼斯成年人口腔微生物群中是否存在甲烷菌。结果 经过元基因组 16S rRNA 数据分析,在 5/43 份(11.62 %)唾液样本中检测到甲烷杆菌。通过标准 16S rRNA 测序,在 6/43 个样本中证实了口腔甲烷杆菌的存在。通过实时 PCR,在 35/43 个样本(81.39 %)中检测到了甲烷菌,其中 62.79 % 对口腔甲烷菌呈阳性,76.74 % 对 smithii 呈阳性。这些发现反映了这两种甲烷菌的高流行率,而实时 PCR 方法的高灵敏度也揭示了这一点。有趣的是,我们还注意到 smithii 甲型杆菌的检测结果与地中海饮食习惯不良之间存在显著的统计学关联,这表明饮食习惯对 smithii 甲型杆菌的流行有影响。方法的选择也是了解口腔微生物群中此类次要类群的真正流行率和多样性的关键。
{"title":"Polyphasic molecular approach to the characterization of methanogens in the saliva of Tunisian adults","authors":"Fériel Bouzid ,&nbsp;Imen Gtif ,&nbsp;Salma Charfeddine ,&nbsp;Leila Abid ,&nbsp;Najla Kharrat","doi":"10.1016/j.anaerobe.2024.102820","DOIUrl":"10.1016/j.anaerobe.2024.102820","url":null,"abstract":"<div><h3>Background</h3><p>Methanogenic archaea are a minor component of human oral microbiota. Due to their relatively low abundance, the detection of these neglected microorganisms is challenging.</p></div><div><h3>Objective</h3><p>This study concerns the presence of methanogens in salivary samples collected from Tunisian adults to evaluate their prevalence and burden using a polyphasic molecular approach.</p></div><div><h3>Methods</h3><p>A total of 43 saliva samples were included. Metagenomic and standard 16S rRNA sequencing were performed as an initial screening to detect the presence of methanogens in the oral microbiota of Tunisian adults. Further investigations were performed using specific quantitative real-time PCR targeting <em>Methanobrevibacter oralis</em> and <em>Methanobrevibacter smithii.</em></p></div><div><h3>Results</h3><p><em>Methanobrevibacter</em> was detected in 5/43 (11.62 %) saliva samples after metagenomic 16S rRNA data analysis. The presence of <em>M. oralis</em> was confirmed in 6/43 samples by standard 16S rRNA sequencing. Using real-time PCR, methanogens were detected in 35/43 (81.39 %) samples, including 62.79 % positive for <em>M. oralis</em> and 76.74 % positive for <em>M. smithii</em>. These findings reflect the high prevalence of both methanogens, revealed by the high sensitivity of the real-time PCR approach. Interestingly, we also noted a significant statistical association between the detection of <em>M. smithii</em> and poor adherence to a Mediterranean diet, indicating the impact of diet on <em>M. smithii</em> prevalence.</p></div><div><h3>Conclusion</h3><p>Our study showed the presence of methanogens in the oral microbiota of Tunisian adults with an unprecedented relatively high prevalence. Choice of methodology is also central to picturing the real prevalence and diversity of such minor taxa in the oral microbiota.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"85 ","pages":"Article 102820"},"PeriodicalIF":2.3,"publicationDate":"2024-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139658685","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Clinical and microbiological characteristics of Ruminococcus gnavus bacteremia and intra-abdominal infection 小反刍球菌菌血症和腹腔内感染的临床和微生物学特征
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-01-10 DOI: 10.1016/j.anaerobe.2024.102818
Naoki Watanabe , Tomohisa Watari , Yoshihito Otsuka , Naoto Hosokawa , Kazufumi Yamagata , Miyuki Fujioka

Objectives

Ruminococcus gnavus is a rare human pathogen, and clinical data on R. gnavus infection are insufficient. This retrospective study aimed to investigate the clinical characteristics of R. gnavus infections.

Methods

This study included 13 cases of bacteremia and three cases of non-bacteremia infections caused by R. gnavus. We evaluated the patient data, infection source, clinical outcomes, and antimicrobial susceptibility of R. gnavus isolates for these cases.

Results

The median age of patients was 75 years (range 47–95), and eight patients were female. Twelve cases were presumed to have an intra-abdominal infection source, and the remaining four cases had an unknown infection source. The most common underlying conditions were immunosuppression (seven cases), solid tumors (seven cases), and history of gastrointestinal surgery (five cases). Thirteen patients exhibited gastrointestinal problems (dysfunction, bleeding, intra-abdominal infection, or inflammation). Multiple pathogens were observed in six cases, and fatal outcomes were recorded in three cases. Antimicrobial susceptibility data were available for eight isolates, all of which exhibited low minimum inhibitory concentrations to penicillin (≤0.03 μg/mL), ampicillin–sulbactam (≤0.5 μg/mL), piperacillin–tazobactam (≤4 μg/mL), and metronidazole (≤0.5–1 μg/mL).

Conclusion

Ruminococcus gnavus is frequently associated with an intra-abdominal infection source, and treatment strategies should consider the possibility of multiple pathogens.

目的 gnavus反刍球菌是一种罕见的人类病原体,有关R. gnavus感染的临床数据不足。本回顾性研究旨在调查地衣刍球菌感染的临床特征。方法 本研究包括 13 例地衣刍球菌引起的菌血症和 3 例非菌血症感染。结果患者的中位年龄为 75 岁(47-95 岁),8 名患者为女性。12例推测感染源为腹腔内感染,其余4例感染源不明。最常见的基础疾病是免疫抑制(7 例)、实体瘤(7 例)和胃肠道手术史(5 例)。13 例患者出现胃肠道问题(功能障碍、出血、腹腔内感染或炎症)。六例病例中观察到多种病原体,三例病例出现致命结果。有 8 个分离菌株的抗菌药敏感性数据,它们对青霉素(≤0.03 μg/mL)、氨苄西林-舒巴坦(≤0.结论 gnavus反刍球菌经常与腹腔内感染源有关,治疗策略应考虑多种病原体的可能性。
{"title":"Clinical and microbiological characteristics of Ruminococcus gnavus bacteremia and intra-abdominal infection","authors":"Naoki Watanabe ,&nbsp;Tomohisa Watari ,&nbsp;Yoshihito Otsuka ,&nbsp;Naoto Hosokawa ,&nbsp;Kazufumi Yamagata ,&nbsp;Miyuki Fujioka","doi":"10.1016/j.anaerobe.2024.102818","DOIUrl":"10.1016/j.anaerobe.2024.102818","url":null,"abstract":"<div><h3>Objectives</h3><p><span><em>Ruminococcus</em><em> gnavus</em></span><span> is a rare human pathogen, and clinical data on </span><em>R. gnavus</em> infection are insufficient. This retrospective study aimed to investigate the clinical characteristics of <em>R. gnavus</em> infections.</p></div><div><h3>Methods</h3><p><span>This study included 13 cases of bacteremia and three cases of non-bacteremia infections caused by </span><em>R. gnavus</em><span>. We evaluated the patient data, infection source, clinical outcomes, and antimicrobial susceptibility of </span><em>R. gnavus</em> isolates for these cases.</p></div><div><h3>Results</h3><p><span>The median age of patients was 75 years (range 47–95), and eight patients were female. Twelve cases were presumed to have an intra-abdominal infection source, and the remaining four cases had an unknown infection source. The most common underlying conditions were immunosuppression (seven cases), </span>solid tumors<span><span> (seven cases), and history of gastrointestinal surgery<span> (five cases). Thirteen patients exhibited gastrointestinal problems<span> (dysfunction, bleeding, intra-abdominal infection, or inflammation). Multiple pathogens<span> were observed in six cases, and fatal outcomes were recorded in three cases. Antimicrobial susceptibility data were available for eight isolates, all of which exhibited low minimum inhibitory concentrations to penicillin (≤0.03 μg/mL), ampicillin–sulbactam (≤0.5 μg/mL), piperacillin–tazobactam (≤4 μg/mL), and </span></span></span></span>metronidazole (≤0.5–1 μg/mL).</span></p></div><div><h3>Conclusion</h3><p><em>Ruminococcus gnavus</em><span> is frequently associated with an intra-abdominal infection source, and treatment strategies should consider the possibility of multiple pathogens.</span></p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"85 ","pages":"Article 102818"},"PeriodicalIF":2.3,"publicationDate":"2024-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139414985","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Production and purification of Clostridium perfringens type C beta-toxin and IgG and IgY antitoxins 生产和纯化 C 型产气荚膜梭菌 beta 型毒素以及 IgG 和 IgY 抗毒素
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2023-12-30 DOI: 10.1016/j.anaerobe.2023.102817
Guilherme Guerra Alves , Ronnie Antunes Assis , Victor Santos do Amarante , Carlos Augusto de Oliveira Júnior , Rodrigo Otávio Silveira Silva , Luiz Guilherme Dias Heneine , Francisco Carlos Faria Lobato

Objectives

This study aimed to produce and purify Clostridium perfringens type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken immunoglobulin Y (IgY).

Methods

Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a chelator and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY.

Results

All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %).

Conclusions

C. perfringens type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by C. perfringens type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.

目的 本研究旨在生产和纯化 C 型产气荚膜梭菌 beta-毒素、绵羊抗 beta-毒素免疫球蛋白 G (IgG) 和鸡免疫球蛋白 Y (IgY)。方法 采用两种方法纯化 beta-毒素:以锌为螯合剂的单步金属亲和层析法 (MAC) 和离子交换层析法 (IEX)。然后将纯化和灭活的 beta 毒素注射给羊和鸡,以产生 IgG 和 IgY。相比之下,MAC 在单步方案中每次运行可纯化超过 21 毫克的毒素。然后将纯化和灭活的 C 型毒素给绵羊和鸡注射,纯化出的 IgG 和 IgY 产率高,抗体滴度为 50 IU/mL,亲和力高(73.2%)。该方案可用于生产诊断和研究 C 型产气荚膜杆菌引起的坏死性肠炎的成分,也可用于评估现有疫苗和开发预防该疾病的新方法。
{"title":"Production and purification of Clostridium perfringens type C beta-toxin and IgG and IgY antitoxins","authors":"Guilherme Guerra Alves ,&nbsp;Ronnie Antunes Assis ,&nbsp;Victor Santos do Amarante ,&nbsp;Carlos Augusto de Oliveira Júnior ,&nbsp;Rodrigo Otávio Silveira Silva ,&nbsp;Luiz Guilherme Dias Heneine ,&nbsp;Francisco Carlos Faria Lobato","doi":"10.1016/j.anaerobe.2023.102817","DOIUrl":"10.1016/j.anaerobe.2023.102817","url":null,"abstract":"<div><h3>Objectives</h3><p>This study aimed to produce and purify <span><em>Clostridium perfringens</em></span><span><span> type C beta-toxin, sheep anti-beta toxin immunoglobulin G (IgG) and chicken </span>immunoglobulin Y (IgY).</span></p></div><div><h3>Methods</h3><p><span>Two methods were used for beta-toxin purification: single-step metal affinity chromatography (MAC) using zinc as a </span>chelator<span> and ion exchange chromatography (IEX). The purified and inactivated beta-toxoids were then administered to sheep and chickens in order to produce IgG and IgY.</span></p></div><div><h3>Results</h3><p>All assays using the IEX failed. In contrast, MAC purified more than 21 mg of toxin per run in a single-step protocol. The purified and inactivated beta-toxoids were then administered to sheep and chickens, and IgG and IgY were purified with a high yield, medium antibody titer of 50 IU/mL, and high avidity (73.2 %).</p></div><div><h3>Conclusions</h3><p><em>C. perfringens</em><span> type C beta-toxin and sheep or chicken anti-beta toxin IgG and IgY antibodies were successfully produced and purified using a simple protocol. This protocol can be used for the production of components used in the diagnosis and research of necrotic enteritis caused by </span><em>C. perfringens</em> type C, as well as for the evaluation of existing vaccines and the development of new preventive methods against this disease.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"85 ","pages":"Article 102817"},"PeriodicalIF":2.3,"publicationDate":"2023-12-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139063390","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Enhancing biogas generation from lignocellulosic biomass through biological pretreatment: Exploring the role of ruminant microbes and anaerobic fungi 通过生物预处理提高木质纤维素生物质的沼气产量:探索反刍微生物和厌氧真菌的作用
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2023-12-23 DOI: 10.1016/j.anaerobe.2023.102815
R. Tamilselvan, A. Immanuel Selwynraj

Objectives

Biogas production from Lignocellulosic Biomass (LB) via anaerobic digestion (AD) has gained attention for its potential in self-sustainability. However, the recalcitrance of LB cell walls pose a challenge to its degradability and biogas generation. Therefore, pretreatment of LB is necessary to enhance lignin removal and increase degradability. Among the different approaches, environmentally friendly biological pretreatment ispromising as it avoids the production of inhibitors.

Methods

The ruminal microbial community, including anaerobic fungi, bacteria, and protozoa, has shown an ability to effectively degrade LB through biomechanical and microbial penetration of refractory cell structures. In this review, we provide an overview of ruminant microbes dominating LB's AD, their degradation mechanism, and the bioaugmentation of the rumen. We also explore the potential cultivation of anaerobic fungi from the rumen, their enzyme potential, and their role in AD.

Results

The rumen ecosystem, comprising both bacteria and fungi, plays a crucial role in enhancing AD. This comprehensive review delves into the intricacies of ruminant microorganisms' adhesion to plant cells, elucidates degradation mechanisms, and explores integrated pretreatment approaches for the effective utilization of LB, minimizing the impact of inhibitors. The discussion underscores the considerable potential of ruminant microbes in pretreating LB, paving the way for sustainable biogas production.

Conclusions

Optimizing fungal colonization and ligninolytic enzyme production, such as manganese peroxidase and laccase, significantly enhances the efficiency of fungal pretreatment. Integrating anaerobic fungi through bioaugmentation during mainstream processing demonstrably increases methane production. This study opens promising avenues for further research and development of these microorganisms for bioenergy production.

目的 通过厌氧消化(AD)从木质纤维素生物质(LB)中生产沼气,因其潜在的自我可持续性而备受关注。然而,木质纤维素细胞壁的不稳定性对其降解性和沼气生产构成了挑战。因此,有必要对枸杞进行预处理,以提高木质素去除率和降解性。方法瘤胃微生物群落(包括厌氧真菌、细菌和原生动物)通过生物力学和微生物对难降解细胞结构的渗透,显示出有效降解枸杞的能力。在这篇综述中,我们概述了在枸橼酸反式脂肪酸中占主导地位的反刍微生物、它们的降解机制以及瘤胃的生物增量。我们还探讨了从瘤胃中培养厌氧真菌的可能性、它们的酶潜能以及它们在厌氧消化中的作用。结果由细菌和真菌组成的瘤胃生态系统在增强厌氧消化中起着至关重要的作用。本综述深入探讨了反刍微生物粘附植物细胞的复杂性,阐明了降解机制,并探讨了有效利用枸杞的综合预处理方法,最大限度地减少了抑制剂的影响。讨论强调了反刍微生物在枸杞预处理方面的巨大潜力,为可持续的沼气生产铺平了道路。结论优化真菌定殖和木质素分解酶(如锰过氧化物酶和漆酶等)的生产,可显著提高真菌预处理的效率。在主流处理过程中,通过生物增殖整合厌氧真菌可明显增加甲烷产量。这项研究为进一步研究和开发这些微生物用于生物能源生产开辟了前景广阔的道路。
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引用次数: 0
In vitro activity of delafloxacin against anaerobic bacteria compared with other antimicrobials 与其他抗菌药相比,德拉氧氟沙星对厌氧菌的体外活性
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2023-12-23 DOI: 10.1016/j.anaerobe.2023.102816
Fernando Cobo, Ana Franco-Acosta, Lina Martín-Hita, Javier Rodríguez-Granger, Antonio Sampedro-Martínez, Juan Antonio Reguera-Márquez, José María Navarro-Marí

The aim of this study was to describe the in vitro activity of delafloxacin against 230 anaerobic isolates and compare it with the activity of other antimicrobials used against infections caused by anaerobic microorganisms. Minimal inhibitory concentrations (MICs) were lower for delafloxacin than for all other antibiotics tested with the exception of piperacillin-tazobactam and meropenem against Gram-positive anaerobic cocci. Only two (0.8 %) isolates of Bacteroides spp. showed a MIC ≥4 μg/mL. With some exceptions, the present results show lower MICs for delafloxacin in comparison to the other antibiotics used against anaerobes.

本研究旨在描述德拉氧沙星对230株厌氧菌分离物的体外活性,并将其与其他用于治疗厌氧微生物感染的抗菌药的活性进行比较。除了哌拉西林-他唑巴坦和美罗培南之外,德拉氧沙星对革兰阳性厌氧球菌的最小抑菌浓度(MICs)低于所有其他受试抗生素。只有两例(0.8%)分离出的乳杆菌的 MIC 值≥4 μg/mL。除个别情况外,目前的结果显示,与其他用于厌氧菌的抗生素相比,德拉氧沙星的 MIC 值较低。
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引用次数: 0
TMT-based quantitative proteomics reveals the nutritional and stress resistance functions of anaerobic fungi in yak rumen during passage at different time intervals 基于tmt的定量蛋白质组学揭示了不同时间间隔牦牛瘤胃内厌氧真菌在传代过程中的营养和抗逆性功能。
IF 2.3 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2023-12-02 DOI: 10.1016/j.anaerobe.2023.102805
Jingrong Zhang , Yaqin Wei , Huizhen Qiu , Jiayi Han

Objectives

Anaerobic fungi are critical for nutrient digestion in the yak rumen. Although studies have reported the effects of passage at different time intervals on the community structure of yak rumen anaerobic fungi, it is unknown whether passage culture at different time intervals affects the microbial proteins of rumen anaerobic fungi and their functions.

Methods

Mycelium was obtained using the anaerobic continuous batch culture (CBC) of yak rumen fluid at intervals of 3 d, 5 d and 7 d. Quantitative analysis of fungal proteins and functional analysis was performed using tandem mass tagging (TMT) and bioinformatics.

Results

A total of 56 differential proteins (DPs) were found in 5 d vs. 3 d and 7 d vs. 3 d. Gene ontology (GO) enrichment indicated that the up-regulated proteins were mainly involved in biological regulation, cellular process, metabolic process, macromolecular complex, membrane, cell part, organelle, binding, catalytic activity and transporter activity. The downregulated proteins were mainly enriched in metabolic process, cell part, binding and catalytic activity. Furthermore, the downregulated proteins in 7 d vs. 3 d were related to membrane and organelle. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment results indicated that DPs were enriched in 14 pathways in 5 d vs. 3 d and 7 d vs. 3 d, mainly including terpenoid backbone biosynthesis, alaine, aspartate and glutamate metabolism, arginine biosynthesis, hypotaurine, cyanoamino acid, glutathione, β-alanine, pyrimidine, purine, galactose and propanate metabolism, steroid biosynthesis, ribosome biogenesis in eukaryotes and aminoacyl tRNA biosynthesis. The DPs were enriched in only 2 pathways in 5 d vs 3 d, lysine biosynthesis and cysteine and methionine metabolism. N-glycan biosynthesis and retinol metabolism are only found in the metabolism of DPs in 7 d vs 3 d.

Conclusions

Yak rumen anaerobic fungal proteins are involved in nutrition and stress tolerance during passage at different time intervals.

目的:厌氧真菌对牦牛瘤胃营养物质的消化具有重要作用。虽然已有研究报道了不同时间间隔传代对牦牛瘤胃厌氧真菌群落结构的影响,但不同时间间隔传代培养是否会影响瘤胃厌氧真菌的微生物蛋白及其功能尚不清楚。方法:采用厌氧连续间歇培养法(CBC)对牦牛瘤胃液进行菌丝体培养,培养间隔为3 d、5 d和7 d。采用串联质量标记(TMT)和生物信息学技术对真菌蛋白进行定量分析和功能分析。结果:在5个 d vs. 3d和7个 d vs. 3d共发现56个差异蛋白(dp)。基因本体(GO)富集表明,上调的蛋白主要参与生物调控、细胞过程、代谢过程、大分子复合物、膜、细胞部分、细胞器、结合、催化活性和转运蛋白活性。下调蛋白主要富集于代谢过程、细胞部分、结合和催化活性。此外,7 d和3 d中下调的蛋白与膜和细胞器有关。京都基因与基因组百科全书(KEGG)富集结果表明,DPs在5 d vs. 3 d和7 d vs. 3 d中富集了14条途径,主要包括萜类骨架生物合成、丙氨酸、天冬氨酸和谷氨酸代谢、精氨酸生物合成、次牛磺酸、氰氨基酸、谷胱甘肽、β-丙氨酸、嘧啶、嘌呤、半乳糖和丙氨酸代谢、类固醇生物合成、真核生物核糖体生物合成和氨基酰基tRNA生物合成。DPs仅在5 d vs 3 d、赖氨酸生物合成和半胱氨酸和蛋氨酸代谢两种途径中富集。n -聚糖的生物合成和视黄醇的代谢只在7 d vs 3 d的DPs代谢中发现。结论:牦牛瘤胃厌氧真菌蛋白在不同时间间隔传代过程中参与营养和胁迫耐受。
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