Pub Date : 2024-08-28DOI: 10.1016/j.anaerobe.2024.102901
Nicholas Geremia , Gianfranco Sanson , Luigi Principe , Roberta Maria Antonello , Verena Zerbato , Roberto Luzzati
Clostridium perfingens bloodstream infections (BSIs) can be associated with high mortality rates. We performed a subanalysis of all C. perfringens BSIs enrolled during a multicentric retrospective observational study (ITANAEROBY). Data were collected from January 2016 to December 2020. C. perfringens BSIs were 134 (134/1960, 6.8 %). The highest resistance rate was observed for clindamycin (26/120, 21.6 %), penicillin (11/71, 15.4 %) and metronidazole (14/131, 10.7 %). In conclusion, C. perfringens reduced susceptibility phenotype to first-line therapy.
{"title":"A subanalysis of Clostridium perfringens bloodstream infections from a 5-year retrospective nationwide survey (ITANAEROBY)","authors":"Nicholas Geremia , Gianfranco Sanson , Luigi Principe , Roberta Maria Antonello , Verena Zerbato , Roberto Luzzati","doi":"10.1016/j.anaerobe.2024.102901","DOIUrl":"10.1016/j.anaerobe.2024.102901","url":null,"abstract":"<div><p><em>Clostridium perfingens</em> bloodstream infections (BSIs) can be associated with high mortality rates. We performed a subanalysis of all <em>C. perfringens</em> BSIs enrolled during a multicentric retrospective observational study (ITANAEROBY). Data were collected from January 2016 to December 2020. <em>C. perfringens</em> BSIs were 134 (134/1960, 6.8 %). The highest resistance rate was observed for clindamycin (26/120, 21.6 %), penicillin (11/71, 15.4 %) and metronidazole (14/131, 10.7 %). In conclusion, <em>C. perfringens</em> reduced susceptibility phenotype to first-line therapy.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"90 ","pages":"Article 102901"},"PeriodicalIF":2.5,"publicationDate":"2024-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142103748","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}
Pub Date : 2024-08-24DOI: 10.1016/j.anaerobe.2024.102902
Stefanie Bressan Waller , Cleideanny Cancela Galvão , Rafael Rodrigues Rodrigues , Cleiderson de Lima Aguirres , Pedro Henrique Dala Nora Quatrin , Mariliana Luiza Ferreira Alves , Marcos Roberto Alves Ferreira , Fabricio Rochedo Conceição
Introduction
Chickens with Necrotic Enteritis (NE), caused by Clostridium perfringens, exhibit acute and chronic symptoms that are difficult to diagnose, leading to significant economic losses. Vaccination is the best method for controlling and preventing NE. However, only two vaccines based on the CPA and NetB toxins have been commercialized, offering partial protection, highlighting the urgent need for more effective vaccines.
Objective
This review aimed to identify promising antigens for NE vaccine formulation and discuss factors affecting their effectiveness.
Methods
A systematic review using five scientific databases identified 30 eligible studies through the Rayyan tool, which were included for quality review.
Results
We identified 25 promising antigens, including CPA, NetB, FBA, ZMP, CnaA, FimA, and FimB, categorized by their role in disease pathogenesis. This review discusses the biochemical, physiological, and genetic traits of recombinant antigens used in vaccine prototypes, their expression systems, and immunization potential in chickens challenged with virulent C. perfringens strains. Market supply challenges, immunogenic potential, vaccine platforms, adjuvants, and factors related to vaccination schedules—such as administration routes, dosing intervals, and age at immunization—are also addressed. Additionally, the study notes that vaccine formulations tested under mild challenges may not offer adequate field-level protection due to issues replicating aggressive conditions, strain virulence loss, and varied methodologies.
Conclusions
An ideal NE vaccine should incorporate multiple antigens, molecular adjuvants, and delivery systems via in ovo and oral routes. The review underscores the challenges in developing and validating NE vaccines and the urgent need for a standardized protocol to replicate aggressive challenges accurately.
导言:由产气荚膜梭菌(Clostridium perfringens)引起的鸡坏死性肠炎(Necrotic Enteritis,NE)表现出急性和慢性症状,难以诊断,导致重大经济损失。接种疫苗是控制和预防 NE 的最佳方法。然而,目前仅有两种基于 CPA 和 NetB 毒素的疫苗实现了商业化,只能提供部分保护,因此迫切需要更有效的疫苗:本综述旨在确定有望用于配制 NE 疫苗的抗原,并讨论影响其有效性的因素:方法:利用五个科学数据库进行系统性综述,通过Rayyan工具确定了30项符合条件的研究,并将其纳入质量审查:结果:我们确定了 25 种有前景的抗原,包括 CPA、NetB、FBA、ZMP、CnaA、fimA 和 fimB,并根据它们在疾病发病机制中的作用进行了分类。报告讨论了疫苗原型中使用的重组抗原的生化、生理和遗传特征、表达系统,以及在鸡受到毒性产气荚膜杆菌菌株挑战时的免疫潜力。报告还探讨了市场供应挑战、免疫原潜力、疫苗平台、佐剂以及与疫苗接种计划相关的因素,如给药途径、给药间隔和免疫年龄。此外,报告还指出,在温和挑战下测试的疫苗配方可能无法提供足够的实地保护,原因是存在复制侵袭性条件、菌株毒力下降和方法不同等问题:结论:理想的近亲繁殖疫苗应包含多种抗原、分子佐剂以及通过胎内和口服途径给药的给药系统。综述强调了开发和验证近亲疫苗所面临的挑战,以及准确复制侵袭性挑战的标准化方案的迫切需要。
{"title":"Clostridium perfringens antigens and challenges for development of vaccines against necrotic enteritis in poultry","authors":"Stefanie Bressan Waller , Cleideanny Cancela Galvão , Rafael Rodrigues Rodrigues , Cleiderson de Lima Aguirres , Pedro Henrique Dala Nora Quatrin , Mariliana Luiza Ferreira Alves , Marcos Roberto Alves Ferreira , Fabricio Rochedo Conceição","doi":"10.1016/j.anaerobe.2024.102902","DOIUrl":"10.1016/j.anaerobe.2024.102902","url":null,"abstract":"<div><h3>Introduction</h3><p>Chickens with Necrotic Enteritis (NE), caused by <em>Clostridium perfringens</em>, exhibit acute and chronic symptoms that are difficult to diagnose, leading to significant economic losses. Vaccination is the best method for controlling and preventing NE. However, only two vaccines based on the CPA and NetB toxins have been commercialized, offering partial protection, highlighting the urgent need for more effective vaccines.</p></div><div><h3>Objective</h3><p>This review aimed to identify promising antigens for NE vaccine formulation and discuss factors affecting their effectiveness.</p></div><div><h3>Methods</h3><p>A systematic review using five scientific databases identified 30 eligible studies through the Rayyan tool, which were included for quality review.</p></div><div><h3>Results</h3><p>We identified 25 promising antigens, including CPA, NetB, FBA, ZMP, CnaA, FimA, and FimB, categorized by their role in disease pathogenesis. This review discusses the biochemical, physiological, and genetic traits of recombinant antigens used in vaccine prototypes, their expression systems, and immunization potential in chickens challenged with virulent <em>C. perfringens</em> strains. Market supply challenges, immunogenic potential, vaccine platforms, adjuvants, and factors related to vaccination schedules—such as administration routes, dosing intervals, and age at immunization—are also addressed. Additionally, the study notes that vaccine formulations tested under mild challenges may not offer adequate field-level protection due to issues replicating aggressive conditions, strain virulence loss, and varied methodologies.</p></div><div><h3>Conclusions</h3><p>An ideal NE vaccine should incorporate multiple antigens, molecular adjuvants, and delivery systems via <em>in ovo</em> and oral routes. The review underscores the challenges in developing and validating NE vaccines and the urgent need for a standardized protocol to replicate aggressive challenges accurately.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102902"},"PeriodicalIF":2.5,"publicationDate":"2024-08-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142071803","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}
Pub Date : 2024-08-21DOI: 10.1016/j.anaerobe.2024.102900
Farrukh Raza Amin , Habiba Khalid , Jingjing Wang , Yaxiang Li , Longxue Ma , Wuxi Chen , Yu Duan , Yida Zhai , Demao Li
Objectives
This study investigated the codigestion of corn straw (CS) with cow manure (CM), cow digestion solution (CD), and a strain consortium (SC) for enhanced volatile fatty acid (VFA) production. The aims of this study were to develop a sustainable technique to increase VFA yields, examine how combining microbial reagents with CS affects VFA production by functional microorganisms, and assess the feasibility of improving microbial diversity through codigestion.
Methods
Batch experiments evaluated VFA production dynamics and microbial community changes with different combinations of CS substrates with CM, CD, and SC. Analytical methods included measuring VFAs by GC, ammonia and chemical oxygen demand (COD) by standard methods and microbial community analysis by 16S rRNA gene sequencing.
Results
Codigesting CS with the strain consortium yielded initial VFA concentrations ranging from 0.6 to 1.0 g/L, which were greater than those of the other combinations (0.05–0.3 g/L). Including CM, and CD further increased VFA production to 1.0–2.0 g/L, with the highest value of 2.0 g/L occurring when all four substrates were codigested. Significant ammonium reduction (194–241 mg/L to 29–37 mg/L) and COD reduction (3310–5250 mg/L to 730–1210 mg/L) were observed. Codigestion with CM and CD had greater Shannon diversity indices (3.19–3.24) than did codigestion with the other consortia (2.26). Bacillota dominated (96.5–99.6 %), with Clostridiales playing key roles in organic matter breakdown.
Conclusions
This study demonstrated the feasibility of improving VFA yields and harnessing microbial diversity through anaerobic codigestion of lignocellulosic and animal waste streams. Codigestion substantially enhanced VFA production, which was dominated by butyrate, reduced ammonium and COD, and enriched fiber-degrading and fermentative bacteria. These findings can help optimize codigestion for sustainable waste management and high-value chemical production.
{"title":"High value-added chemical production through anaerobic codigestion of corn straw with a microbial consortium, cow manure and cow digestion solution","authors":"Farrukh Raza Amin , Habiba Khalid , Jingjing Wang , Yaxiang Li , Longxue Ma , Wuxi Chen , Yu Duan , Yida Zhai , Demao Li","doi":"10.1016/j.anaerobe.2024.102900","DOIUrl":"10.1016/j.anaerobe.2024.102900","url":null,"abstract":"<div><h3>Objectives</h3><p>This study investigated the codigestion of corn straw (CS) with cow manure (CM), cow digestion solution (CD), and a strain consortium (SC) for enhanced volatile fatty acid (VFA) production. The aims of this study were to develop a sustainable technique to increase VFA yields, examine how combining microbial reagents with CS affects VFA production by functional microorganisms, and assess the feasibility of improving microbial diversity through codigestion.</p></div><div><h3>Methods</h3><p>Batch experiments evaluated VFA production dynamics and microbial community changes with different combinations of CS substrates with CM, CD, and SC. Analytical methods included measuring VFAs by GC, ammonia and chemical oxygen demand (COD) by standard methods and microbial community analysis by 16S rRNA gene sequencing.</p></div><div><h3>Results</h3><p>Codigesting CS with the strain consortium yielded initial VFA concentrations ranging from 0.6 to 1.0 g/L, which were greater than those of the other combinations (0.05–0.3 g/L). Including CM, and CD further increased VFA production to 1.0–2.0 g/L, with the highest value of 2.0 g/L occurring when all four substrates were codigested. Significant ammonium reduction (194–241 mg/L to 29–37 mg/L) and COD reduction (3310–5250 mg/L to 730–1210 mg/L) were observed. Codigestion with CM and CD had greater Shannon diversity indices (3.19–3.24) than did codigestion with the other consortia (2.26). <em>Bacillota</em> dominated (96.5–99.6 %), with <em>Clostridiales</em> playing key roles in organic matter breakdown.</p></div><div><h3>Conclusions</h3><p>This study demonstrated the feasibility of improving VFA yields and harnessing microbial diversity through anaerobic codigestion of lignocellulosic and animal waste streams. Codigestion substantially enhanced VFA production, which was dominated by butyrate, reduced ammonium and COD, and enriched fiber-degrading and fermentative bacteria. These findings can help optimize codigestion for sustainable waste management and high-value chemical production.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102900"},"PeriodicalIF":2.5,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141999256","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}
Oxygen tolerance of anaerobes is a virulence factor, but can also be a beneficial property. Many species have evolved to tolerate or take advantage of the presence of low, especially nanaerobic (≤0.14 %) oxygen concentrations. Oxygen tolerance is genus-, species- and strain-dependent according to their protective mechanisms. It was better expressed in some pathogenic species such as Bacteroides fragilis, Clostridioides difficile, and Clostridium perfringens, as well as in Akkermansia muciniphila than in other potential probiotics such as Alistipes, Blautia and Roseburia spp. Different degrees of oxygen sensitivity were found between the strains of Anaerostipes, Faecalibacterium, and Bifidobacterium spp. Importantly, clostridial spores and anaerobes in biofilms are protected from oxidation. Rubrerythrins and flavodiiron proteins and two regulators (sigma factor B and PerR) contribute to C. difficile protection from reactive oxygen species (ROS). The frequent pathogen, B. fragilis, has numerous protective factors such as enzymes (catalase, superoxide dismutase, alkyl hydroperoxidase, thioredoxin peroxidase, and aerobic-type NrdAB ribonucleotide reductase), and nanaerobic respiration. Seven proteins confer strain-specific oxygen adaptation of Faecalibacterium prausnitzii. Oxygen tolerance protects anaerobes from ROS, shields their DNA and modulates gene expression. Furthermore, oxygen can induce mutations leading to antibiotic resistance as shown in Prevotella melaninogenica. Some Faecalibacterium, Anaerostipes, Bifidobacterium, and Akkermansia strains from the intestinal microbiota exhibiting oxygen tolerance may become next-generation probiotic candidates. Further studies are needed to reveal oxygen effects on more anaerobic species and strains, and the influence of oxygen on antibiotic resistance. More studies on oxygen-tolerant probiotic strains can be useful to optimize biotechnological methods.
{"title":"Oxygen tolerance in anaerobes as a virulence factor and a health-beneficial property","authors":"Lyudmila Boyanova , Liliya Boyanova , Petyo Hadzhiyski , Raina Gergova , Rumyana Markovska","doi":"10.1016/j.anaerobe.2024.102897","DOIUrl":"10.1016/j.anaerobe.2024.102897","url":null,"abstract":"<div><p>Oxygen tolerance of anaerobes is a virulence factor, but can also be a beneficial property. Many species have evolved to tolerate or take advantage of the presence of low, especially nanaerobic (≤0.14 %) oxygen concentrations. Oxygen tolerance is genus-, species- and strain-dependent according to their protective mechanisms. It was better expressed in some pathogenic species such as <em>Bacteroides fragilis, Clostridioides difficile</em>, and <em>Clostridium perfringens,</em> as well as in <em>Akkermansia muciniphila</em> than in other potential probiotics such as <em>Alistipes, Blautia</em> and <em>Roseburia</em> spp. Different degrees of oxygen sensitivity were found between the strains of <em>Anaerostipes, Faecalibacterium,</em> and <em>Bifidobacterium</em> spp. Importantly, clostridial spores and anaerobes in biofilms are protected from oxidation. Rubrerythrins and flavodiiron proteins and two regulators (sigma factor B and PerR) contribute to <em>C. difficile</em> protection from reactive oxygen species (ROS). The frequent pathogen, <em>B. fragilis,</em> has numerous protective factors such as enzymes (catalase, superoxide dismutase, alkyl hydroperoxidase, thioredoxin peroxidase, and aerobic-type NrdAB ribonucleotide reductase), and nanaerobic respiration. Seven proteins confer strain-specific oxygen adaptation of <em>Faecalibacterium prausnitzii</em>. Oxygen tolerance protects anaerobes from ROS, shields their DNA and modulates gene expression. Furthermore, oxygen can induce mutations leading to antibiotic resistance as shown in <em>Prevotella melaninogenica</em>. Some <em>Faecalibacterium, Anaerostipes, Bifidobacterium,</em> and <em>Akkermansia</em> strains from the intestinal microbiota exhibiting oxygen tolerance may become next-generation probiotic candidates. Further studies are needed to reveal oxygen effects on more anaerobic species and strains, and the influence of oxygen on antibiotic resistance. More studies on oxygen-tolerant probiotic strains can be useful to optimize biotechnological methods.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102897"},"PeriodicalIF":2.5,"publicationDate":"2024-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141999257","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}
Pub Date : 2024-08-13DOI: 10.1016/j.anaerobe.2024.102898
Kasper K. Mortensen , Hans Linde Nielsen , Kirstine K. Søgaard
Objectives
Bacteremia with anaerobic bacteria is generally a marker of severe prognosis. However, population-based data is lacking. Our aim was to describe the epidemiology and the 30-day mortality rate of anaerobic bacteremia in a Danish population-based setting.
Methods
In this population-based cohort study, all first-time episodes of anaerobic bacteremia from the North Denmark Bacteremia Research Database during 1994–2019 were identified. Information on comorbidities, discharge diagnoses, and mortality was retrieved. 30-day mortality rates were calculated and a multivariate logistic regression analysis to identify risk factors for death was performed.
Results
1750 episodes with anaerobic bacteremia were identified, corresponding to an incidence rate of 12.5 per 100,000 inhabitants (increasing from 11.2 in 1994–2014 to 17.7 in 2015–2019). Of these episodes, a third were polymicrobial, and the majority (70 %) of patients had one or more comorbid conditions. Abdominal infection was the source of bacteremia in 61 % of patients, while it was unknown for 15 %. The most frequently isolated genera were Bacteroides (45 %), Clostridium (20 %) and Fusobacterium (6 %). The overall crude 30-day mortality rate was 27 %, but rates were even higher for patients of high age, with liver disease, and solid tumors. The odds ratio (OR) for 30-day mortality was 1.32 for Clostridium species, and 1.27 for polymicrobial bacteremia with aerobic bacteria.
Conclusions
The incidence rate of anaerobic bacteremia increased, and the 30-day mortality rate remained high during the study period. Multiple factors influence 30-day mortality rates, including high age, liver disease, solid tumor, polymicrobial bacteremia, and bacteremia with Clostridium species.
{"title":"Clinical and microbiological characteristics of anaerobic bacteremia during 1994–2019: A Danish population-based cohort study","authors":"Kasper K. Mortensen , Hans Linde Nielsen , Kirstine K. Søgaard","doi":"10.1016/j.anaerobe.2024.102898","DOIUrl":"10.1016/j.anaerobe.2024.102898","url":null,"abstract":"<div><h3>Objectives</h3><p>Bacteremia with anaerobic bacteria is generally a marker of severe prognosis. However, population-based data is lacking. Our aim was to describe the epidemiology and the 30-day mortality rate of anaerobic bacteremia in a Danish population-based setting.</p></div><div><h3>Methods</h3><p>In this population-based cohort study, all first-time episodes of anaerobic bacteremia from the North Denmark Bacteremia Research Database during 1994–2019 were identified. Information on comorbidities, discharge diagnoses, and mortality was retrieved. 30-day mortality rates were calculated and a multivariate logistic regression analysis to identify risk factors for death was performed.</p></div><div><h3>Results</h3><p>1750 episodes with anaerobic bacteremia were identified, corresponding to an incidence rate of 12.5 per 100,000 inhabitants (increasing from 11.2 in 1994–2014 to 17.7 in 2015–2019). Of these episodes, a third were polymicrobial, and the majority (70 %) of patients had one or more comorbid conditions. Abdominal infection was the source of bacteremia in 61 % of patients, while it was unknown for 15 %. The most frequently isolated genera were <em>Bacteroides</em> (45 %), <em>Clostridium</em> (20 %) and <em>Fusobacterium</em> (6 %). The overall crude 30-day mortality rate was 27 %, but rates were even higher for patients of high age, with liver disease, and solid tumors. The odds ratio (OR) for 30-day mortality was 1.32 for <em>Clostridium</em> species, and 1.27 for polymicrobial bacteremia with aerobic bacteria.</p></div><div><h3>Conclusions</h3><p>The incidence rate of anaerobic bacteremia increased, and the 30-day mortality rate remained high during the study period. Multiple factors influence 30-day mortality rates, including high age, liver disease, solid tumor, polymicrobial bacteremia, and bacteremia with <em>Clostridium</em> species.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102898"},"PeriodicalIF":2.5,"publicationDate":"2024-08-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141987268","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}
Flagellin protein, an integral component of flagella, provides motility to several bacterial species and also acts as a candidate antigen in diagnostics and subunit vaccines. The bulk production of flagellin with retention of all conformational epitopes using recombinant protein technology is of paramount importance in the development of pathogen-specific immuno-assays and vaccines. We describe the production of highly soluble and immuno-reactive rFliA(C) protein of Clostridium chauvoei, a causative agent of blackleg or black quarter (BQ) affecting cattle and small ruminants worldwide. The bacterium is known to possess peritrichous flagella that provide motility and also act as a virulence factor with high protective antigenicity.
Methods
Upon sequence and structural analysis, a partial fliA(C) gene from Clostridium chauvoei was cloned and the recombinant mature protein with N- and C- terminal truncation was over-expressed as a His-tagged fusion protein (∼25 kDa) in Escherichia coli. Subsequently, rFliA(C) protein was purified by single-step affinity chromatography and characterized for its immuno-reactivity in laboratory animals, Western blot, and indirect-ELISA format.
Results
rFliA(C) was highly soluble and was purified in high quantity and quality. rFliA(C) elicited antigen-specific conformational polyclonal antibodies in rabbit and guinea pig models, as well as anti-Clostridium chauvoei-specific antibodies being specifically detected in BQ-vaccinated and convalescent sera of bovines in Western blot and in indirect-ELISA format. Further, no cross reactivity was noted with antibodies against major bovine diseases (e.g., foot-and-mouth disease, IBR, LSDV, hemorrhagic septicaemia, brucellosis, and leptospirosis).
Conclusion
The study indicated the production of conformational recombinant flagellin—rFliA(C)—antigen and its potential utility in development of diagnostics for detection of Clostridium chauvoei-specific antibodies in BQ-recovered and/or vaccinated animals.
目的:鞭毛蛋白是鞭毛的一个组成部分,它为多种细菌提供运动能力,也是诊断和亚单位疫苗的候选抗原。利用重组蛋白技术批量生产能保留所有构象表位的鞭毛蛋白,对于开发病原体特异性免疫测定和疫苗至关重要。我们描述了高可溶性和免疫反应性 rFliA(C)梭菌蛋白的生产过程,梭菌是影响全球牛和小反刍动物的黑腿病(BQ)的致病菌。众所周知,该细菌具有富周鞭毛,可提供运动能力,同时也是一种具有高度保护性抗原性的毒力因子:方法:通过序列和结构分析,克隆了来自 Chauvoei梭菌的部分 fliA(C) 基因,并在大肠杆菌中过度表达了 N 端和 C 端截断的重组成熟蛋白,即 His 标记的融合蛋白(25 kDa)。结果:rFliA(C)具有很高的可溶性,纯化的数量和质量都很高。rFliA(C)能在家兔和豚鼠模型中激发抗原特异性构象多克隆抗体,并能在接种过 BQ 疫苗的牛和恢复期牛的血清中通过 Western 印迹和间接-ELISA 方法特异性地检测到抗梭状芽孢杆菌特异性抗体。此外,与主要牛病(如口蹄疫、IBR、LSDV、出血性败血症、布鲁氏菌病和钩端螺旋体病)的抗体没有交叉反应:该研究表明,重组鞭毛蛋白-rFliA(C)-抗原的制备及其在开发用于检测BQ恢复和/或疫苗接种动物体内梭状芽孢杆菌特异性抗体的诊断中的潜在用途。
{"title":"Production of highly soluble and immuno-reactive recombinant flagellin protein of Clostridium chauvoei","authors":"Awadhesh Prajapati , Roopa Anandamurthy Hemanth , Mandira Ramakrishna Namrutha , Suresh Bindu , Revanaiah Yogisharadhya , Nihar Nalini Mohanty , Mohammed Mudassar Chanda , Sathish Bhadravati Shivachandra","doi":"10.1016/j.anaerobe.2024.102899","DOIUrl":"10.1016/j.anaerobe.2024.102899","url":null,"abstract":"<div><h3>Objective</h3><p>Flagellin protein, an integral component of flagella, provides motility to several bacterial species and also acts as a candidate antigen in diagnostics and subunit vaccines. The bulk production of flagellin with retention of all conformational epitopes using recombinant protein technology is of paramount importance in the development of pathogen-specific immuno-assays and vaccines. We describe the production of highly soluble and immuno-reactive rFliA(C) protein of <em>Clostridium chauvoei,</em> a causative agent of blackleg or black quarter (BQ) affecting cattle and small ruminants worldwide. The bacterium is known to possess peritrichous flagella that provide motility and also act as a virulence factor with high protective antigenicity.</p></div><div><h3>Methods</h3><p>Upon sequence and structural analysis, a partial <em>fliA(C)</em> gene from <em>Clostridium chauvoei</em> was cloned and the recombinant mature protein with N- and C- terminal truncation was over-expressed as a His-tagged fusion protein (∼25 kDa) in <em>Escherichia coli</em>. Subsequently, rFliA(C) protein was purified by single-step affinity chromatography and characterized for its immuno-reactivity in laboratory animals, Western blot, and indirect-ELISA format.</p></div><div><h3>Results</h3><p>rFliA(C) was highly soluble and was purified in high quantity and quality. rFliA(C) elicited antigen-specific conformational polyclonal antibodies in rabbit and guinea pig models, as well as anti-<em>Clostridium chauvoei-</em>specific antibodies being specifically detected in BQ-vaccinated and convalescent sera of bovines in Western blot and in indirect-ELISA format. Further, no cross reactivity was noted with antibodies against major bovine diseases (e.g., foot-and-mouth disease, IBR, LSDV, hemorrhagic septicaemia, brucellosis, and leptospirosis).</p></div><div><h3>Conclusion</h3><p>The study indicated the production of conformational recombinant flagellin—rFliA(C)—antigen and its potential utility in development of diagnostics for detection of <em>Clostridium chauvoei-</em>specific antibodies in BQ-recovered and/or vaccinated animals.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102899"},"PeriodicalIF":2.5,"publicationDate":"2024-08-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141981547","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}
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 Escherichiacoli (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 Clostridiumperfringens and C and D neurotoxins of Clostridiumbotulinum 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.
{"title":"Immunogenicity of a pentavalent recombinant Escherichia coli bacterin against enterotoxemia and botulism in sheep","authors":"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","doi":"10.1016/j.anaerobe.2024.102895","DOIUrl":"10.1016/j.anaerobe.2024.102895","url":null,"abstract":"<div><h3>Introduction</h3><p>Producing commercial bacterins/toxoids against <em>Clostridium</em> 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.</p></div><div><h3>Objective</h3><p>Considering that inactivated recombinant <em>Escherichia</em> <em>coli</em> (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 <em>Clostridium</em> <em>perfringens</em> and C and D neurotoxins of <em>Clostridium</em> <em>botulinum</em> in sheep.</p></div><div><h3>Methods</h3><p>Subcutaneously, 18 Texel sheep received two doses (200 μg of each antigen) of recombinant bacterin (<em>n</em> = 7) or purified recombinant antigens (<em>n</em> = 6) on days 0 and 28, while the control group (<em>n</em> = 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).</p></div><div><h3>Results</h3><p>Both formulations induced significant levels of IgG against all five toxins (<em>p</em> < 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)].</p></div><div><h3>Conclusion</h3><p>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.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102895"},"PeriodicalIF":2.5,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911503","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}
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.
{"title":"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","authors":"Ratchaneewan Khiaosa-ard , Cátia Pacífico , Mubarik Mahmood , Elsayed Mickdam , Julia Meixner , Laura-Sophie Traintinger , Qendrim Zebeli","doi":"10.1016/j.anaerobe.2024.102893","DOIUrl":"10.1016/j.anaerobe.2024.102893","url":null,"abstract":"<div><h3>Objectives</h3><p>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.</p></div><div><h3>Methods</h3><p>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.</p></div><div><h3>Results</h3><p>The alpha- and beta-diversity of the microbiota were unaffected. However, treatment modified the bacterial composition at low taxonomic levels. <em>Butyrivibrio fibrisolvens</em>, <em>Treponema bryantii</em>, and bacterium MC2010 decreased in EXT, while <em>Treponema berlinense</em> was increased in GP-high and GP-low compared to CON. Concerning fungi, GS-high increased <em>Candida</em> spp.<em>, Lachancea</em> spp.<em>, Microdochium</em> spp.<em>, Mucor</em> spp.<em>, Pichia</em> spp.<em>, Saturnispora</em> spp.<em>,</em> and <em>Zygosaccharomyces</em> spp. compared to CON. Many non-<em>Saccharomyces</em> 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.</p></div><div><h3>Conclusions</h3><p>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.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102893"},"PeriodicalIF":2.5,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1075996424000763/pdfft?md5=f26adcc727df8f717fb35c58701764e2&pid=1-s2.0-S1075996424000763-main.pdf","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2024-08-08DOI: 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。
{"title":"The impact of 30-day antecedent antibiotic exposure on Clostridioides difficile ribotype patterns and the relationship with clinical outcomes: A single center study","authors":"Majd Alsoubani , Jennifer K. Chow , Angie Mae Rodday , Laura A. McDermott , Seth T. Walk , David R. Snydman","doi":"10.1016/j.anaerobe.2024.102894","DOIUrl":"10.1016/j.anaerobe.2024.102894","url":null,"abstract":"<div><h3>Background</h3><p>Antibiotic exposure is a known risk factor for <em>Clostridioides difficile</em> infection (CDI) and recurrence and can lead to infection with specific <em>C. difficile</em> strains. In this study, we sought to explore the relationship between antecedent antibiotic exposure and <em>C. difficile</em> antimicrobial resistance, and the impact of resistance on clinical outcomes.</p></div><div><h3>Methods</h3><p>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.</p></div><div><h3>Results</h3><p>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).</p></div><div><h3>Conclusion</h3><p>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.</p></div>","PeriodicalId":8050,"journal":{"name":"Anaerobe","volume":"89 ","pages":"Article 102894"},"PeriodicalIF":2.5,"publicationDate":"2024-08-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141911471","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}