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Immunogenicity of a pentavalent recombinant Escherichia coli bacterin against enterotoxemia and botulism in sheep 五价重组大肠杆菌菌素对绵羊肠毒血症和肉毒中毒的免疫原性。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.anaerobe.2024.102895
Jaqueline Freitas Motta , Marcos Roberto A. Ferreira , Stefanie Bressan Waller , Rafael Rodrigues Rodrigues , Rafael Amaral Donassolo , Clóvis Moreira Júnior , Mariliana Luiza Ferreira Alves , Fernanda Dornelles Feijó , Fabricio Rochedo Conceição

Introduction

Producing commercial bacterins/toxoids against Clostridium spp. is laborious and hazardous. Conversely, developing prototype vaccines using purified recombinant toxoids, though safe and effective, is both laborious and costly for application in production animals.

Objective

Considering that inactivated recombinant Escherichia coli (bacterin) is a simple, cost-effective, and to be safe solution, we evaluated, for the first time, a pentavalent formulation of recombinant bacterins containing the alpha, beta, and epsilon toxins of Clostridium perfringens and C and D neurotoxins of Clostridium botulinum in sheep.

Methods

Subcutaneously, 18 Texel sheep received two doses (200 μg of each antigen) of recombinant bacterin (n = 7) or purified recombinant antigens (n = 6) on days 0 and 28, while the control group (n = 5) did not receive an immunization. Sera samples from days 0 (before the 1st dose), 28 (before the 2nd dose), and 56, 84, and 112 were used for measuring IgG (indirect ELISA) and neutralizing antibodies (mouse serum neutralization).

Results

Both formulations induced significant levels of IgG against all five toxins (p < 0.05) up to day 112, with peaks at days 28 and 56 post-immunization. The expected booster effect occurred only for the botulinum toxins. The neutralizing antibody titers were satisfactory against ETX (≥2 IU/ml for both formulations) and BoNT-D [5 IU/ml (bacterin) and 10 IU/ml (purified)].

Conclusion

While adjustments are required, the recombinant bacterin platform holds great potential for polyvalent vaccines due to its straightforward, safe, and cost-effective production, establishing it as a user-friendly technology for the veterinary immunobiological industry.

简介:生产针对梭状芽孢杆菌的商业细菌/类毒素既费力又危险。相反,使用纯化的重组类毒素开发疫苗原型虽然安全有效,但在生产动物中应用既费力又昂贵:考虑到灭活重组大肠杆菌(细菌素)是一种简单、经济、安全的解决方案,我们首次在绵羊身上评估了含有产气荚膜梭菌的α、β和ε毒素以及肉毒梭菌的C和D神经毒素的重组细菌素五价制剂:18只德克塞尔绵羊分别在第0天和第28天皮下注射两剂(每种抗原200微克)重组菌素(7只)或纯化重组抗原(6只),对照组(5只)不进行免疫接种。第 0 天(第 1 剂前)、第 28 天(第 2 剂前)、第 56、84 和 112 天的血清样本用于测量 IgG(间接 ELISA)和中和抗体(小鼠血清中和法):结果:两种制剂都能诱导出针对所有五种毒素的大量 IgG(pConclusion):虽然还需要进行调整,但重组菌素平台因其生产简单、安全、成本效益高,在多价疫苗方面具有巨大潜力,使其成为兽医免疫生物学行业的一项用户友好型技术。
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引用次数: 0
Changes in the solid-associated bacterial and fungal communities following ruminal in vitro fermentation of winery by-products: aspects of the bioactive compounds and feed safety 酿酒厂副产品经瘤胃体外发酵后固体相关细菌和真菌群落的变化:生物活性化合物和饲料安全问题。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.anaerobe.2024.102893
Ratchaneewan Khiaosa-ard , Cátia Pacífico , Mubarik Mahmood , Elsayed Mickdam , Julia Meixner , Laura-Sophie Traintinger , Qendrim Zebeli

Objectives

Feeding winery by-products (WBP) could affect the bovine microbiome because of their phenol compounds and a transfer of WBP-associated microbiota. This work examined changes in the underexplored solid-associated rumen microbiome following the inclusion of WBP.

Methods

Using the rumen simulation technique, fermenters were inoculated with the inoculum of donor cows and were fed one of six dietary treatments including a control diet of 70 % hay +30 % concentrate (CON), control diet + 3.7 % commercial grapeseed extract (EXT), 65 % hay + 25 % concentrate + 10 % grape pomace (GP-low), 56 % hay + 24 % concentrate + 20 % grape pomace (GP-high), 70 % hay + 25 % concentrate + 5 % grapeseed meal (GS-low), and 65 % hay + 25 % concentrate + 10 % grapeseed meal (GS-high) (dry matter basis). The compositional changes of bacteria, archaea and fungi in the solid fractions were based on 16S and ITS2 rRNA sequencing.

Results

The alpha- and beta-diversity of the microbiota were unaffected. However, treatment modified the bacterial composition at low taxonomic levels. Butyrivibrio fibrisolvens, Treponema bryantii, and bacterium MC2010 decreased in EXT, while Treponema berlinense was increased in GP-high and GP-low compared to CON. Concerning fungi, GS-high increased Candida spp., Lachancea spp., Microdochium spp., Mucor spp., Pichia spp., Saturnispora spp., and Zygosaccharomyces spp. compared to CON. Many non-Saccharomyces yeasts were detected in WBP samples but absent in donor cows and CON samples. The genera affected by treatment were not the major contributors to the ruminal degradation of nutrients.

Conclusions

The results indicate a sensitivity of rumen solid bacteria to grape phenols when delivered as an extract and a transfer of WBP-associated microbiota into the rumen.

目的:饲喂酿酒厂副产品(WBP)可能会影响牛的微生物组,因为它们含有酚类化合物,而且会转移 WBP 相关微生物群。本研究考察了在添加 WBP 后,未被充分探索的固体相关瘤胃微生物群的变化:方法:使用瘤胃模拟技术,在发酵罐中接种供体奶牛的接种物,并饲喂六种日粮处理之一,包括 70% 干草 + 30% 精料的对照日粮(CON)、对照日粮 + 3.7%的商品葡萄籽提取物(EXT)、65%干草+25%精料+10%葡萄渣(GP-低)、56%干草+24%精料+20%葡萄渣(GP-高)、70%干草+25%精料+5%葡萄籽粕(GS-低)和65%干草+25%精料+10%葡萄籽粕(GS-高)(干物质基础)。根据 16S 和 ITS2 rRNA 测序分析了固体馏分中细菌、古菌和真菌的组成变化:结果:微生物群的α-和β-多样性未受影响。然而,处理改变了低分类水平的细菌组成。与CON相比,EXT中的布氏纤毛虫、白喉特雷波菌和MC2010细菌减少了,而GP-高和GP-低中的柏林特雷波菌增加了。在真菌方面,与对照组相比,GS-高增加了念珠菌属、Lachancea 菌属、Microdochium 菌属、Mucor 菌属、Pichia 菌属、Saturnispora 菌属和 Zygosaccharomyces 菌属。在 WBP 样品中检测到了许多非酵母菌,但在供体牛和 CON 样品中却没有检测到。受处理影响的菌属并非瘤胃营养物质降解的主要贡献者:结论:研究结果表明,瘤胃固态菌对以提取物形式输送的葡萄酚很敏感,WBP相关微生物群会转移到瘤胃中。
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引用次数: 0
The impact of 30-day antecedent antibiotic exposure on Clostridioides difficile ribotype patterns and the relationship with clinical outcomes: A single center study 30 天前抗生素暴露对艰难梭菌核糖型模式的影响以及与临床结果的关系:单中心研究。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.anaerobe.2024.102894
Majd Alsoubani , Jennifer K. Chow , Angie Mae Rodday , Laura A. McDermott , Seth T. Walk , David R. Snydman

Background

Antibiotic exposure is a known risk factor for Clostridioides difficile infection (CDI) and recurrence and can lead to infection with specific C. difficile strains. In this study, we sought to explore the relationship between antecedent antibiotic exposure and C. difficile antimicrobial resistance, and the impact of resistance on clinical outcomes.

Methods

This was a single center retrospective study evaluating patients with CDI between 2011 and 2021. A logistic regression model was used to evaluate the relationship between antecedent antibiotics in the 30 days prior to CDI and resistance among isolates. In addition, an exploratory analysis using a cause-specific Cox proportional hazards model evaluated the association between resistance and a composite outcome of clinical failure, relapse at 30 days or CDI-related death.

Results

we analyzed one isolate from 510 patients; resistance was noted in 339 (66.5 %) of the isolates. Exposure to fluoroquinolones and macrolides was associated with 2.4 (95 % CI 1.4–4.4) and 4.7 (95 % CI 1.1–20.5) increased odds of having resistance compared to other antibiotic class exposure, respectively. There were 58 (17.0 %) patients in the resistance group who developed the composite outcome and 24 (14.2 %) patients who lacked resistance who developed the composite outcome (HR 1.32, 95 % CI 0.81–2.14).

Conclusion

These findings suggest that fluoroquinolone and macrolide exposure were significantly associated with isolating a resistant strain, but we did not find significant differences in clinical outcomes based on the presence of antimicrobial resistance.

背景:抗生素暴露是难辨梭状芽孢杆菌感染(CDI)和复发的已知风险因素,可导致感染特定的难辨梭状芽孢杆菌(C. difficile)菌株。在这项研究中,我们试图探讨前抗生素暴露与艰难梭菌抗菌药耐药性之间的关系,以及耐药性对临床结果的影响:这是一项单中心回顾性研究,评估了 2011-2021 年间的 CDI 患者。采用逻辑回归模型评估了 CDI 发病前 30 天内使用抗生素与分离菌耐药性之间的关系。此外,一项探索性分析使用了特定病因的 Cox 比例危险度模型,评估了耐药性与临床失败、30 天内复发或 CDI 相关死亡等综合结果之间的关系。与接触其他抗生素类药物相比,接触氟喹诺酮类药物和大环内酯类药物产生耐药性的几率分别增加了 2.4 (95% CI 1.4-4.4) 和 4.7 (95% CI 1.1-20.5)。耐药组中有58例(17.0%)患者出现综合结果,24例(14.2%)无耐药患者出现综合结果(HR 1.32,95% CI 0.81-2.14):这些研究结果表明,接触氟喹诺酮类药物和大环内酯类药物与分离出耐药菌株有显著相关性,但我们并未发现因抗菌药耐药性的存在而导致临床结果出现显著差异。
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引用次数: 0
Development of two recombinant vaccines against Clostridioides difficile infection and immunogenicity in pregnant sows and neonatal piglets 针对艰难梭菌感染的两种重组疫苗的开发以及妊娠母猪和新生仔猪的免疫原性。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-08-08 DOI: 10.1016/j.anaerobe.2024.102896
Carolina P. Ramos , Williane F. Siqueira , Laila A. Viana , João L.R. Cunha , Ricardo T. Fujiwara , Victor S. Amarante , Thayanne G.V. Souza , Rodrigo O.S. Silva

Introduction

Clostridioides difficile is the main cause of antibiotic-associated diarrhea in humans and is a major enteropathogen in several animal species. In newborn piglets, colonic lesions caused by C. difficile A and B toxins (TcdA and TcdB, respectively) cause diarrhea and significant production losses.

Objective

The present study aimed to develop two recombinant vaccines from immunogenic C-terminal fragments of TcdA and TcdB and evaluate the immune response in rabbits and in breeding sows. Two vaccines were produced: bivalent (rAB), consisting of recombinant fragments of TcdA and TcdB, and chimeric (rQAB), corresponding to the synthesis of the same fragments in a single protein. Groups of rabbits were inoculated with 10 or 50 μg of proteins adjuvanted with aluminum or 0.85 % sterile saline in a final volume of 1 mL/dose. Anti-TcdA and anti-TcdB IgG antibodies were detected in rabbits and sows immunized with both rAB and rQAB vaccines by ELISA. The vaccinated sows were inoculated intramuscularly with 20 μg/dose using a prime-boost approach.

Results

Different antibody titers (p ≤ 0.05) were observed among the vaccinated groups of sows (rAB and rQAB) and control. Additionally, newborn piglets from vaccinated sows were also positive for anti-TcdA and anti-TcdB IgGs, in contrast to control piglets (p ≤ 0.05). Immunization of sows with the rQAB vaccine conferred higher anti-TcdA and anti-TcdB responses in piglets, suggesting the superiority of this compound over rAB.

Conclusion

The synthesized recombinant proteins were capable of inducing antibody titers against C. difficile toxins A and B in sows, and were passively transferred to piglets through colostrum.

导言:艰难梭菌是导致人类抗生素相关性腹泻的主要原因,也是多种动物的主要肠道病原体。在新生仔猪中,艰难梭菌 A 型和 B 型毒素(分别为 TcdA 和 TcdB)引起的结肠病变会导致腹泻和严重的生产损失:本研究旨在利用 TcdA 和 TcdB 的免疫原 C 端片段开发两种重组疫苗,并评估兔子和繁殖母猪的免疫反应。我们生产了两种疫苗:由 TcdA 和 TcdB 重组片段组成的双价疫苗(rAB)和嵌合疫苗(rQAB),前者相当于将相同片段合成为单一蛋白。每组兔子接种 10 或 50 μg 的铝或 0.85% 无菌生理盐水佐剂蛋白,最终剂量为 1 mL/次。用酶联免疫吸附试验检测使用 rAB 和 rQAB 疫苗免疫的兔子和母猪体内的抗 TcdA 和抗 TcdB IgG 抗体。接种后的母猪肌肉注射 20 μg/dose 疫苗,采用原代-加强免疫法:结果:接种母猪组(rAB 和 rQAB)与对照组的抗体滴度不同(p ≤ 0.05)。此外,接种疫苗母猪的新生仔猪抗 TcdA 和抗 TcdB IgG 也呈阳性,与对照组仔猪形成鲜明对比(p≤0.05)。用 rQAB 疫苗免疫母猪可使仔猪产生更高的抗 TcdA 和抗 TcdB 反应,表明该化合物优于 rAB:结论:合成的重组蛋白能诱导母猪产生抗艰难梭菌毒素 A 和 B 的抗体滴度,并能通过初乳被动转移给仔猪。
{"title":"Development of two recombinant vaccines against Clostridioides difficile infection and immunogenicity in pregnant sows and neonatal piglets","authors":"Carolina P. Ramos ,&nbsp;Williane F. Siqueira ,&nbsp;Laila A. Viana ,&nbsp;João L.R. Cunha ,&nbsp;Ricardo T. Fujiwara ,&nbsp;Victor S. Amarante ,&nbsp;Thayanne G.V. Souza ,&nbsp;Rodrigo O.S. Silva","doi":"10.1016/j.anaerobe.2024.102896","DOIUrl":"10.1016/j.anaerobe.2024.102896","url":null,"abstract":"<div><h3>Introduction</h3><p><em>Clostridioides difficile</em> is the main cause of antibiotic-associated diarrhea in humans and is a major enteropathogen in several animal species. In newborn piglets, colonic lesions caused by <em>C. difficile</em> A and B toxins (TcdA and TcdB, respectively) cause diarrhea and significant production losses.</p></div><div><h3>Objective</h3><p>The present study aimed to develop two recombinant vaccines from immunogenic C-terminal fragments of TcdA and TcdB and evaluate the immune response in rabbits and in breeding sows. Two vaccines were produced: bivalent (rAB), consisting of recombinant fragments of TcdA and TcdB, and chimeric (rQAB), corresponding to the synthesis of the same fragments in a single protein. Groups of rabbits were inoculated with 10 or 50 μg of proteins adjuvanted with aluminum or 0.85 % sterile saline in a final volume of 1 mL/dose. <em>Anti</em>-TcdA and <em>anti</em>-TcdB IgG antibodies were detected in rabbits and sows immunized with both rAB and rQAB vaccines by ELISA. The vaccinated sows were inoculated intramuscularly with 20 μg/dose using a prime-boost approach.</p></div><div><h3>Results</h3><p>Different antibody titers (p ≤ 0.05) were observed among the vaccinated groups of sows (rAB and rQAB) and control. Additionally, newborn piglets from vaccinated sows were also positive for <em>anti</em>-TcdA and <em>anti</em>-TcdB IgGs, in contrast to control piglets (p ≤ 0.05). Immunization of sows with the rQAB vaccine conferred higher <em>anti</em>-TcdA and <em>anti</em>-TcdB responses in piglets, suggesting the superiority of this compound over rAB.</p></div><div><h3>Conclusion</h3><p>The synthesized recombinant proteins were capable of inducing antibody titers against <em>C. difficile</em> toxins A and B in sows, and were passively transferred to piglets through colostrum.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102896"},"PeriodicalIF":2.5,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911502","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
The growth-stimulating factor of Treponema phagedenis from bovine digital dermatitis lesions 牛数字皮炎皮损中的噬菌体生长刺激因子
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-08-01 DOI: 10.1016/j.anaerobe.2024.102882
Rathanon Khemgaew , Kaoru Hori , Satomi Sasaki , Naoaki Misawa , Takako Taniguchi

Bovine digital dermatitis (BDD) is an infectious skin disease of the hoof characterized by painful ulcerations that cause lameness in dairy cattle. Cell-free supernatants (CFS) of Falsiporphyromonas endometrii predominantly isolated from BDD lesions had the highest growth-stimulating effect on Treponema phagedenis among BDD-associated bacteria. Butyric acid was detected at a concentration of 45.4 mM in CFS of F. endometrii, and the growth of T. phagedenis was significantly promoted by butyric acid supplementation.

牛数字皮炎(BDD)是一种蹄部传染性皮肤病,其特点是溃疡疼痛,导致奶牛跛行。在与 BDD 相关的细菌中,主要从 BDD 病变中分离出的子宫内膜法氏囊无细胞上清液(CFS)对噬菌体的生长刺激作用最大。在 F. endometrii 的 CFS 中检测到浓度为 45.4 mM 的丁酸,丁酸补充剂显著促进了噬菌体的生长。
{"title":"The growth-stimulating factor of Treponema phagedenis from bovine digital dermatitis lesions","authors":"Rathanon Khemgaew ,&nbsp;Kaoru Hori ,&nbsp;Satomi Sasaki ,&nbsp;Naoaki Misawa ,&nbsp;Takako Taniguchi","doi":"10.1016/j.anaerobe.2024.102882","DOIUrl":"10.1016/j.anaerobe.2024.102882","url":null,"abstract":"<div><p>Bovine digital dermatitis (BDD) is an infectious skin disease of the hoof characterized by painful ulcerations that cause lameness in dairy cattle. Cell-free supernatants (CFS) of <em>Falsiporphyromonas endometrii</em> predominantly isolated from BDD lesions had the highest growth-stimulating effect on <em>Treponema phagedenis</em> among BDD-associated bacteria. Butyric acid was detected at a concentration of 45.4 mM in CFS of <em>F. endometrii</em>, and the growth of <em>T. phagedenis</em> was significantly promoted by butyric acid supplementation.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"88 ","pages":"Article 102882"},"PeriodicalIF":2.5,"publicationDate":"2024-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141726804","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
Identification of linear B cell epitopes on the leukotoxin protein of Fusobacterium necrophorum 鉴定坏死分枝杆菌白细胞毒素蛋白上的线性 B 细胞表位。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-07-25 DOI: 10.1016/j.anaerobe.2024.102884
Jiawei Xiao , Siwen Yu , Kai Jiang , Xianjing He , Lan Bi , Pengyu Zhao , Tianshuo Wang , Ning Yang , Donghua Guo

Objective

Fusobacterium necrophorum can casuse Lemierre's syndrome in humans and a range of illnesses, including foot rot and liver abscesses, in animals. The main virulence factor released by F. necrophorum is leukotoxin, which has been shown to have a strong correlation with the severity of the disease. Leukotoxin is commonly employed as the key antigen in the formulation of subunit vaccines. Therefore, identification of the B-cell epitope of F. necrophorum leukotoxin is necessary.

Methods

In this research, we utilized lymphocyte hybridoma technology to develop a monoclonal antibody (mAb), 3D7, targeting the F. necrophorum leukotoxin protein. Identification of B-cell epitopes recognized by 3D7 mAb was achieved through Western blot, ELISA and dot blots using leukotoxin-truncated recombinant proteins and peptides, and through SWISS-MODEL homology modeling and PyMOL visualization.

Results

The 3D7 mAb was identified as belonging to the IgG1 subclass with a κ-chain light chain. It demonstrated reactivity with the natural leukotoxin. The results showed that the 3D7 mAb recognizes a B-cell epitope of the F. necrophorum leukotoxin protein, I2168SSFGVGV2175 (EP-3D7). Sequence comparison analysis showed that EP-3D7 was highly conserved in F. necrophorum strains, but less conserved in other bacteria, indicating the specificity of EP-3D7. EP-3D7 is present on the surface of leukotoxin proteins in a β-folded manner.

Conclusions

In summary, these results establish EP-3D7 as a conserved antigenic epitope of F. necrophorum leukotoxin. It could be valuable in the development of vaccines and diagnostic reagents for F. necrophorum epitopes.
目的:坏死杆菌可导致人类患上莱米埃尔综合征,动物患上一系列疾病,包括烂脚病和肝脓肿。坏死杆菌释放的主要毒力因子是白细胞毒素,已被证明与疾病的严重程度密切相关。白细胞毒素通常被用作亚单位疫苗的关键抗原。因此,有必要鉴定 F. necrophorum 白细胞毒素的 B 细胞表位:在这项研究中,我们利用淋巴细胞杂交瘤技术开发出了一种针对F. necrophorum白细胞毒素蛋白的单克隆抗体(mAb)3D7。利用白细胞毒素截短重组蛋白和肽,通过Western印迹、ELISA和点印迹,并通过SWISS-MODEL同源建模和PyMOL可视化,鉴定了3D7 mAb识别的B细胞表位:结果:经鉴定,3D7 mAb 属于具有 κ 链轻链的 IgG1 亚类。它与天然白细胞毒素具有反应性。结果表明,3D7 mAb 能识别 F. necrophorum 白细胞毒素蛋白的 B 细胞表位 I2168SSFGVGV2175(EP-3D7)。序列对比分析表明,EP-3D7 在 F. necrophorum 菌株中高度保守,但在其他细菌中保守程度较低,这表明了 EP-3D7 的特异性。EP-3D7以β折叠的方式存在于白细胞毒素蛋白的表面:总之,这些结果确定了 EP-3D7 是 F. necrophorum 白细胞毒素的保守抗原表位。结论:这些结果确立了 EP-3D7 作为 F. necrophorum 白细胞毒素的保守抗原表位,它对疫苗和 F. necrophorum 表位诊断试剂的开发可能很有价值。
{"title":"Identification of linear B cell epitopes on the leukotoxin protein of Fusobacterium necrophorum","authors":"Jiawei Xiao ,&nbsp;Siwen Yu ,&nbsp;Kai Jiang ,&nbsp;Xianjing He ,&nbsp;Lan Bi ,&nbsp;Pengyu Zhao ,&nbsp;Tianshuo Wang ,&nbsp;Ning Yang ,&nbsp;Donghua Guo","doi":"10.1016/j.anaerobe.2024.102884","DOIUrl":"10.1016/j.anaerobe.2024.102884","url":null,"abstract":"<div><h3>Objective</h3><div><em>Fusobacterium necrophorum</em> can casuse Lemierre's syndrome in humans and a range of illnesses<em>,</em> including foot rot and liver abscesses, in animals. The main virulence factor released by <em>F. necrophorum</em> is leukotoxin, which has been shown to have a strong correlation with the severity of the disease. Leukotoxin is commonly employed as the key antigen in the formulation of subunit vaccines. Therefore, identification of the B-cell epitope of <em>F. necrophorum</em> leukotoxin is necessary.</div></div><div><h3>Methods</h3><div>In this research, we utilized lymphocyte hybridoma technology to develop a monoclonal antibody (mAb), 3D7, targeting the <em>F. necrophorum</em> leukotoxin protein. Identification of B-cell epitopes recognized by 3D7 mAb was achieved through Western blot, ELISA and dot blots using leukotoxin-truncated recombinant proteins and peptides, and through SWISS-MODEL homology modeling and PyMOL visualization.</div></div><div><h3>Results</h3><div>The 3D7 mAb was identified as belonging to the IgG1 subclass with a κ-chain light chain. It demonstrated reactivity with the natural leukotoxin. The results showed that the 3D7 mAb recognizes a B-cell epitope of the <em>F. necrophorum</em> leukotoxin protein, I<sup>2168</sup>SSFGVGV<sup>2175</sup> (EP-3D7). Sequence comparison analysis showed that EP-3D7 was highly conserved in <em>F. necrophorum</em> strains, but less conserved in other bacteria, indicating the specificity of EP-3D7. EP-3D7 is present on the surface of leukotoxin proteins in a <em>β</em>-folded manner.</div></div><div><h3>Conclusions</h3><div>In summary, these results establish EP-3D7 as a conserved antigenic epitope of <em>F. necrophorum</em> leukotoxin. It could be valuable in the development of vaccines and diagnostic reagents for <em>F. necrophorum</em> epitopes.</div></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"90 ","pages":"Article 102884"},"PeriodicalIF":2.5,"publicationDate":"2024-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141765001","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
An outbreak of botulism on a pig farm due to the newly described Clostridium botulinum type C 一个养猪场爆发肉毒中毒事件,病原体是新发现的 C 型肉毒梭状芽孢杆菌。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-07-24 DOI: 10.1016/j.anaerobe.2024.102885
Caroline Le Maréchal , Martine Denis , Louis Vanlemmens , Léa Jambou , Gauthier Delvallez , Typhaine Poezevara , Mouna Abed-Zahar , Laure Diancourt , Marie-Agnès Baudouard , Eric Eveno , Marie-Hélène Bayon-Auboyer , Mustapha Fellag , Sandra Rouxel , Nadia Amenna , Christelle Mazuet , Jean-Marc Guéguen , Nicolas Rose

This study reports a botulism outbreak on a pig farm. Clostridium botulinum type C was detected using PCR. The gene encoding the toxin corresponds to a novel type C neurotoxin recently described in a human botulism outbreak, raising the question of its prevalence in pigs and the related risks to humans.

本研究报告了一个养猪场爆发的肉毒中毒事件。通过 PCR 检测到了 C 型肉毒梭菌。编码该毒素的基因与最近在一起人类肉毒中毒疫情中描述的新型 C 型神经毒素相对应,这就提出了该毒素在猪中的流行程度以及对人类的相关风险问题。
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引用次数: 0
Succinate-mediated symbiosis between Dialister hominis and an uncharacterized Segatella-like pectinophile 琥珀酸介导的 Dialister hominis 与一种未定性的 Segatella 类嗜果胶菌之间的共生。
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-07-20 DOI: 10.1016/j.anaerobe.2024.102883
Tomoki Makiura , Minenosuke Matsutani , Hou-Chia Tseng , Naoshi Fujimoto , Akihiro Ohnishi

Objectives

Syntrophy has been documented between pectinophiles and methanol-utilizing bacteria, along with instances of cross-feeding between pectinophiles and methanogens. However, studies on the ecology of pectinophiles in anaerobic digestion (AD) are lacking. Therefore, in this study, we aimed to elucidate the ecology of pectinophiles by isolating novel pectinophile forms and conducting a comprehensive analysis of their physiology and ecology.

Methods

Complex microbial communities from AD systems were enriched in a pectin-containing medium; subsequently, specific strains were isolated using a pectinophile isolation method. The carbon source assimilation and growth ability of the isolates, along with their symbiotic relationships, were evaluated using batch tests.

Results

Strain LPYR103-Pre exhibited 16S rRNA gene sequence similarity and average nucleotide identity values of 94.3 % and 77.9 %, respectively, compared to its closest related species, Segatella cerevisiae. Strain LPYR103-Pre demonstrated attenuated growth in the presence of eight common sugars but exhibited remarkably high growth in the presence of pectin, d-galacturonate, and d-glucuronate, with succinate being identified as a primary metabolite. Accumulation of succinate inhibited the growth of strain LPYR103-Pre. However, this growth impediment was alleviated by Dialister hominis LPYG114-Dih, whose growth required succinate.

Conclusions

Our results elucidate the specific carbon source requirements of the Segatella-like strain LPYR103-Pre and succinate-mediated symbiosis involving D. hominis. These findings provide new insights into the degradation of pectin and its degradation products during AD, contributing to the identification of unknown pectinophiles.

目的:嗜果胶细菌与甲醇利用细菌之间的合成作用以及嗜果胶细菌与甲烷菌之间的交叉觅食均有文献记载。然而,有关厌氧消化(AD)中嗜果胶菌生态学的研究却很缺乏。因此,在本研究中,我们旨在通过分离新型嗜果胶菌并对其生理和生态学进行全面分析,来阐明嗜果胶菌的生态学:方法:在含果胶的培养基中富集厌氧消化(AD)系统中的复杂微生物群落,然后使用嗜果胶菌分离方法分离特定菌株。通过批量试验评估了分离菌株的碳源同化和生长能力及其共生关系:结果:菌株 LPYR103-Pre 与最亲缘的物种 Segatella cerevisiae 的 16S rRNA 基因序列相似度和平均核苷酸同一性分别为 94.3% 和 77.9%。菌株 LPYR103-Pre 在八种常见糖类的存在下表现出生长减弱,但在果胶、D-半乳糖醛酸和 D-葡萄糖醛酸的存在下表现出显著的高生长,其中琥珀酸被确定为主要代谢产物。琥珀酸的积累抑制了菌株 LPYR103-Pre 的生长。然而,Dialister hominis LPYG114-Dih 的生长需要琥珀酸,因此这种生长障碍得到了缓解:我们的研究结果阐明了类Segatella菌株LPYR103-Pre和琥珀酸介导的D. hominis共生对碳源的特殊要求。这些发现为了解AD过程中果胶的降解及其降解产物提供了新的视角,有助于识别未知的嗜果胶菌。
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引用次数: 0
Association between Bacteroides fragilis and Fusobacterium nucleatum infection and colorectal cancer in Vietnamese patients 越南患者中脆弱拟杆菌和核酸镰刀菌感染与结直肠癌之间的关系
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-06-26 DOI: 10.1016/j.anaerobe.2024.102880
Truong Nguyen Duy , Hoang Le Huy , Quyen Đao Thanh , Hoai Ngo Thi , Hanh Ngo Thi Minh , Manh Nguyen Dang , Song Le Huu , Trung Ngo Tat

Background

Colorectal cancer (CRC) is a significant global health concern, and understanding the role of specific bacterial infections in its development and progression is of increasing interest. This cross-sectional study investigated the associations between Bacteroides fragilis (B. fragilis) and Fusobacterium nucleatum (F. nucleatum) infections and Vietnamese CRC patients.

Methods

192 patients with either polyps or CRC at varying stages were recruited from May 2017 to December 2020. Real-time PCR assessed infection rates and bacterial loads in CRC tissues.

Results

B. fragilis infection was notably higher in CRC tissues (51.6 %) than polyps (9.4 %), with a fivefold higher relative load. Positive associations were found in stages II and III, indicating a fivefold increase in CRC progression risk. F. nucleatum infection rates were significantly higher in CRC tissues (55.2 %) than in polyps (10.5 %). In stage II, the infection rate exceeded that in adjacent tissues. The relative load of F. nucleatum was higher in stage III than in stages I and II. Positive F. nucleatum patients had a 3.2 times higher risk of CRC progression.

Conclusion

These findings suggest associations between loading of F. nucleatum or/and B. fragilis with the advanced stages of CRC.

背景:结肠直肠癌(CRC)是全球关注的重大健康问题,了解特定细菌感染在其发生和发展中的作用越来越受到关注。这项横断面研究调查了脆弱拟杆菌(B. fragilis)和核酸镰刀菌(F. nucleatum)感染与越南 CRC 患者之间的关联。方法:2017 年 5 月至 2020 年 12 月期间,招募了 192 名不同阶段的息肉或 CRC 患者。实时 PCR 评估了 CRC 组织中的感染率和细菌量:CRC组织中B. fragilis感染率(51.6%)明显高于息肉(9.4%),相对载量高出5倍。在 II 期和 III 期发现了正相关,表明 CRC 进展风险增加了五倍。CRC 组织中的 F. nucleatum 感染率(55.2%)明显高于息肉(10.5%)。在二期,感染率超过了邻近组织。在 III 期,F. nucleatum 的相对载量高于 I 期和 II 期。核酸桿菌陽性患者的 CRC 發病風險高出 3.2 倍:这些研究结果表明,F. nucleatum或/和B. fragilis的载量与CRC的晚期阶段有关。
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引用次数: 0
Description of Faecalibacterium wellingii sp. nov. and two Faecalibacterium taiwanense strains, aiding to the reclassification of Faecalibacterium species Faecalibacterium wellingii sp.
IF 2.5 3区 生物学 Q3 MICROBIOLOGY Pub Date : 2024-06-24 DOI: 10.1016/j.anaerobe.2024.102881

Objectives

The genus Faecalibacterium is one of the most important butyrate producers in the human intestinal tract and has been widely linked to health. Recently, several different species have been described, but still more phylogroups have been identified, suggesting that additional species may exist. Four strains HTF-FT, HTF-128, HTF-75H and HTF-76H, representing two different phylogenetic clusters, are evaluated in this study.

Methods

Phylogenomic analysis was performed using whole-genome sequences and 16S rRNA gene sequences. Chemotaxonomic analysis was done based on matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Physiological and phenotypical characteristics of these strains were also determined. All characteristics of these strains were compared with other validly published species within the genus Faecalibacterium.

Results

On a genomic level, the strains HTF-FT and HTF-128 shared an average nucleotide identity (ANI) of <95.0 % and digital DNA-DNA hybridization (dDDH) of <70.0 with other Faecalibacterium species, while between HTF-FT and HTF-128 the ANI-value was 97.18 % and the dDDH was 76.8 %. HTF-75H and HTF-76H had an ANI and dDDH value of 100 % (99.96 %) and 100 % (99.99 %) respectively. Both HTF-75H and HTF-76H were closely related to Faecalibacterium taiwanense HLW78T. 16S rRNA gene and chemotaxonomic analysis were in accordance with the genomic data, confirming that HTF-FT and HTF-128 represent a novel Faecalibacterium species and HTF-75H and HTF-76H belong to F. taiwanense.

Conclusions

Faecalibacterium strains HTF-FT (=DSM 117771T = NCIMB 15531T) and HTF-128 represent a novel species. The name Faecalibacterium wellingii with HTF-FT as type strain is proposed. Two novel isolates HTF-75H (=DSM 17770 = NCIMB 15530) and HTF-76H are described in this study and belong to the recently described Faecalibacterium taiwanense.

目的:粪杆菌属是人类肠道中最重要的丁酸生产者之一,与健康有着广泛的联系。最近,描述了几个不同的物种,但仍发现了更多的系统群,这表明可能存在更多的物种。本研究对代表两个不同系统发生群的 HTF-FT、HTF-128、HTF-75HT 和 HTF-76H 四个菌株进行了评估:方法:利用全基因组序列和 16S rRNA 基因序列进行系统发生组分析。基质辅助激光解吸/电离飞行时间质谱法进行了化学分类分析。还测定了这些菌株的生理和表型特征。将这些菌株的所有特征与粪便杆菌属内其他已发表的有效菌种进行了比较:在基因组水平上,四株菌株的平均核苷酸同一性(ANI)为 T,HTF-128 的 ANI 值为 97.18%,dDDH 为 76.8%。HTF-75HT 和 HTF-76H 的 ANI 值和 dDDH 值分别为 100%(99.96%)和 100%(99.99%)。16S rRNA 基因和化学分类学分析与基因组数据一致,证实这四株菌株代表了两个不同的粪杆菌属:结论:粪杆菌菌株 HTF-FT(=DSM 117771T =NCIMB 15531T)、HTF-128、HTF-75HT(=DSM 17770T =NCIMB 15530T)和 HTF-76H 代表两个新菌种。以 HTF-FT 为模式菌株的 Faecalibacterium wellingii 和以 HTF-75HT 为模式菌株的 Faecalibacterium langellae 被命名为 Faecalibacterium wellingii。
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引用次数: 0
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