Escherichia albertii

艾氏大肠杆菌
  • 文章类型: Journal Article
    白僵菌是一种引起腹泻的新兴食源性病原体。艾伯蒂大肠杆菌已从各种食物中分离出来,包括猪肉和鸡肉,和环境水域,比如河水。尽管已经报道了许多食物中毒病例,对食物和环境水中细菌种群行为的分析不足。在这项研究中,我们在25克猪肉中接种了2-5个日志CFU。鸡肉,日本岩石牡蛎,太平洋牡蛎,和300毫升4℃的井水和海水,10°C,20°C,30°C,并分析了细菌在食品和环境水体中的种群行为。在4°C下3天后,食品中的艾伯蒂菌菌株EA21和EA24的种群保持约4logCFU/25g。在10°C下3天后,猪肉和牡蛎中的艾伯蒂菌菌株数量保持约4logCFU/25g,并且在鸡肉中增加到大约5-6logCFU/25g。在20°C下2天后,E.albertii菌株在猪肉和鸡肉中生长到大约6-7logCFU/25g,在日本牡蛎中,艾伯蒂尼菌株EA21而不是EA24增长到4.5logCFU/25g,在太平洋牡蛎中,阿伯蒂大肠杆菌菌株EA21而不是EA24略有增长,达到3.1logCFU/25g。在30°C下1天后,E.albertii菌株在鸡肉和猪肉中生长到大约7-8logCFU/25g,在日本牡蛎中增长到大约4-6logCFU/25g,太平洋牡蛎和6-7logCFU/25g。这些结果表明,在4°C以下,阿贝尔蒂大肠杆菌可以存活而不生长,在20°C和30°C的食物中生长迅速,尤其是肉类。艾氏大肠杆菌菌株在4°C的井水和海水中没有生长,10°C,20°C,和30°C。井水和海水中的艾伯蒂菌菌株在30℃时比在4℃时下降得更快,10°C,20°C,这表明阿伯蒂大肠杆菌在30°C的环境水中的生存力较低。
    Escherichia albertii is an emerging foodborne pathogen that causes diarrhea. E. albertii has been isolated from various foods, including pork and chicken meat, and environmental waters, such as river water. Although many food poisoning cases have been reported, there have been insufficient analyses of bacterial population behaviors in food and environmental water. In this study, we inoculated 2-5 log CFU of E. albertii into 25 g of pork, chicken meat, Japanese rock oyster, Pacific oyster, and 300 mL of well water and seawater at 4°C, 10°C, 20°C, and 30°C, and analyzed the bacterial population behavior in food and environmental water. After 3 days at 4°C, the population of E. albertii strain EA21 and EA24 in foods maintained approximately 4 log CFU/25 g. After 3 days at 10°C, the population of E. albertii strains in pork and oysters maintained approximately 4 log CFU/25 g, and that in chicken meat increased to approximately 5-6 log CFU/25 g. After 2 days at 20°C, E. albertii strains grew to approximately 6-7 log CFU/25 g in pork and chicken meat, and E. albertii strain EA21 but not EA24 grew to 4.5 log CFU/25 g in Japanese rock oyster, E. albertii strain EA21 but not EA24 slightly grew to 3.1 log CFU/25 g in Pacific oyster. After 1 day at 30°C, E. albertii strains grew to approximately 7-8 log CFU/25 g in chicken meat and pork, grew to approximately 4-6 log CFU/25 g in Japanese rock oyster, and 6-7 log CFU/25 g in Pacific oyster. These results suggest that E. albertii survives without growth below 4°C and grew rapidly at 20°C and 30°C in foods, especially in meat. E. albertii strains did not grow in well water and seawater at 4°C, 10°C, 20°C, and 30°C. The population of E. albertii strains in well water and seawater decreased faster at 30°C than at 4°C, 10°C, and 20°C, suggesting that E. albertii has low viability at 30°C in environmental water.
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  • 文章类型: Journal Article
    埃希氏菌(E.albertii)是一种新兴的腹泻病原体,与散发性感染和人类胃肠道暴发有关。eae基因,在肠上皮细胞脱落(LEE)操纵子的位点编码内膜蛋白,有助于附着和去除(A/E)病变的建立。从各种来源收集越来越多的艾氏大肠杆菌菌株导致eae亚型的数量迅速增加。这项研究系统地调查了从人类分离的E.albertii菌株中eae的患病率和遗传多样性。动物,和食物。eae基因存在于452/459(98.5%)株中,鉴定出23种亚型,包括两个新亚型,命名为eae-α11和η3。eae-σ亚型是人类中最主要的亚型,动物,和食物来源的菌株,而eae-γ3,τ,和α11在人源菌株中是独特的。此外,LEE岛也进行了基因组分析,转录,通过基因组分析和功能水平,定量逆转录PCR,和HEp-2细胞粘附试验,分别。eae转录水平是可变的,并与eae亚型相关。三种不同的坚持模式,包括局部粘附样(LAL),弥漫性粘附(DA),和分离(DE),在艾伯特菌株中观察到。这项研究证明了从人类中分离出的E.albertii菌株中功能内膜蛋白的高度多样性,动物,和食物。需要进一步的体内和体外研究以更好地阐明内膜蛋白或LEE在不同遗传背景下的作用。
    Escherichia albertii (E. albertii) is an emerging diarrheagenic pathogen associated with sporadic infections and human gastroenteric outbreaks. The eae gene, which encodes intimin in the locus of enterocyte effacement (LEE) operon, contributes to the establishment of the attaching and effacing (A/E) lesion. Increasing collection of E. albertii strains from various sources has resulted in a rapid increase in the number of eae subtypes. This study systematically investigated the prevalence and genetic diversity of eae among E. albertii strains isolated from humans, animals, and food. The eae gene was present in 452/459 (98.5%) strains and 23 subtypes were identified including two novel subtypes, named eae-α11 and η3. The eae-σ subtype was the most predominant among humans, animals, and food-derived strains, while eae-γ3, τ, and α11 were unique in human-derived strains. Additionally, the LEE island was also analyzed at genomic, transcriptional, and functional levels through genomic analysis, quantitative reverse transcription PCR, and HEp-2 cell adherence assays, respectively. The eae transcript levels were variable and associated with eae subtypes. Three different adherence patterns, including localized adherence-like (LAL), diffuse adherence (DA), and detachment (DE), were observed among E. albertii strains. This study demonstrated a high diversity of functional intimin in E. albertii strains isolated from humans, animals, and food. Further in vivo and in vitro studies are warranted to better elucidate the role of intimin or LEE in different genetic backgrounds.
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  • 文章类型: Journal Article
    艾氏大肠杆菌越来越被认为是一种重要的新兴人畜共患肠病原体。浣熊被证明是这种病原体最重要的储库之一。在大阪的1,606只野生浣熊中,有993只(62%)被发现。日本从2017年到2020年通过Eacdt-PCR。5月至12月(冬季)艾贝检出率升高,1月至4月(春季)检出率逐渐降低。此外,我们可以从30%(196/664)的Eacdt-PCR阳性样本中分离出艾氏大肠杆菌,每月的分离率似乎与其检出率相关。这些数据表明,野生浣熊的艾伯蒂菌患病率有季节性,冬季较高,春季较低。
    Escherichia albertii has increasingly been recognized as an important emerging zoonotic enteropathogen. Raccoon is shown to be one of the most vital reservoirs of this pathogen. E. albertii has been detected in 993 (62%) out of 1,606 wild raccoons in Osaka, Japan from 2017 to 2020 by Eacdt-PCR. The detection rate of E. albertii was increased from May to December (winter) and gradually decreased from January to April (spring). Furthermore, we could isolate E. albertii from 30% (196/664) of Eacdt-PCR positive samples and the monthly isolation rate seems to correlate with its detection rate. These data indicate that there is a seasonality regarding the prevalence of E. albertii in wild raccoon being higher in winter and lower in spring.
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  • 文章类型: Case Reports
    一例罕见的由艾伯蒂菌引起的菌血症,据报道,一名50岁男性患有肝硬化。清除,无色,在37℃需氧培养24小时后,在血琼脂上回收环状菌落。使用MALDI-TOF/MS将分离株鉴定为阿氏大肠杆菌,并通过诊断性三重PCR靶向clpX进行确认,lysP,和MDH基因。哌拉西林/他唑巴坦静脉给药(每8小时4.5g)3天是有效的。这项研究表明,阿伯替尼的特定菌株与人类肠道外感染有关,与肠外致病性大肠杆菌(ExPEC)相似。然而,对潜在致病机制的全面了解需要进一步探索。
    A rare case of bacteremia caused by Escherichia albertii, in a 50-year-old male with liver cirrhosis was reported. Clear, colorless, and circular colonies were recovered on blood agar after 24 h of aerobic incubation at 37 °C. The isolate was identified as E. albertii using MALDI-TOF/MS and confirmed by the diagnostic triplex-PCR targeting clpX, lysP, and mdh genes. The administration of piperacillin/tazobactam intravenously (4.5g every 8 hours) for 3 days was effective. This study suggested that specific strains of E. albertii have been implicated in causing extraintestinal infections in humans, similar to extraintestinal pathogenic E. coli (ExPEC). However, a comprehensive understanding of the underlying pathogenic mechanisms requires further exploration.
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  • 文章类型: Journal Article
    白僵菌是一种新兴的食源性致病菌。为了更好地了解这种病原体的发病机理和健康风险,比较基因组学和表型表征被用于评估从加利福尼亚州一个主要农业区的野生鸟类中分离的阿伯氏菌菌株的致病性潜力。志贺毒素基因stx2f存在于所有禽类菌株中。对20个完整基因组的Pangenome分析揭示了总共11,249个基因,其中近80%是辅助基因。基于核心基因的系统发育和基于辅助基因的相关性分析一致地将三个stx2f阳性临床菌株与五个携带ST7971的禽类菌株分组。在确定的三个Stx2f转换的预言整合站点中,ssrA是最常见的一种。除了肠上皮细胞流出和三种分泌系统的位置,高致病性岛,鉴定了OI-122和六型分泌系统。毒力基因库的实质性菌株变异,志贺毒素生产,和细胞毒性被揭示。6株禽类菌株的细胞毒性明显高于stx2f阳性大肠杆菌,其中三个表现出与肠出血性大肠杆菌爆发菌株相当的细胞毒性水平,这表明野生鸟类可以作为艾伯蒂菌菌株的储库,有可能在人类中引起严重疾病。
    Escherichia albertii is an emerging foodborne pathogen. To better understand the pathogenesis and health risk of this pathogen, comparative genomics and phenotypic characterization were applied to assess the pathogenicity potential of E. albertii strains isolated from wild birds in a major agricultural region in California. Shiga toxin genes stx2f were present in all avian strains. Pangenome analyses of 20 complete genomes revealed a total of 11,249 genes, of which nearly 80% were accessory genes. Both core gene-based phylogenetic and accessory gene-based relatedness analyses consistently grouped the three stx2f-positive clinical strains with the five avian strains carrying ST7971. Among the three Stx2f-converting prophage integration sites identified, ssrA was the most common one. Besides the locus of enterocyte effacement and type three secretion system, the high pathogenicity island, OI-122, and type six secretion systems were identified. Substantial strain variation in virulence gene repertoire, Shiga toxin production, and cytotoxicity were revealed. Six avian strains exhibited significantly higher cytotoxicity than that of stx2f-positive E. coli, and three of them exhibited a comparable level of cytotoxicity with that of enterohemorrhagic E. coli outbreak strains, suggesting that wild birds could serve as a reservoir of E. albertii strains with great potential to cause severe diseases in humans.
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  • 文章类型: Journal Article
    艾氏大肠杆菌是一种新的人类和动物肠病原体。该研究的目的是使用流行病学和基因组研究评估在波兰东北部分离的阿伯氏大肠杆菌菌株的患病率和致病性。2015-2018年,来自儿童和成人的粪便样本共1154份,497鸟粪,212份食物样本,92个水样,和500个乳糖阴性大肠杆菌菌株被测试。共分离出42株艾氏大肠杆菌。该方法适用于快速鉴定。总的来说,33.3%的艾氏大肠杆菌对一种抗生素耐药,和16.7%对二。分离株对头孢吡肟敏感,亚胺培南,左氧氟沙星,庆大霉素,甲氧苄啶/磺胺甲恶唑,并且不产生ESBLβ-内酰胺酶。已经证明了艾贝氏杆菌的高遗传变异性。在PFGE方法中,90.5%的菌株具有明显的脉冲型。在MLST打字中,85.7%的菌株被分配了不同的序列类型(STs),其中64%为新型ST型。在100%的艾氏大肠杆菌分离物中发现了细胞致死膨胀毒素(CDT)和Paa毒素基因。编码毒素的基因,伊贝,CdtB类型2,Tsh和Shiga(Stx2f),在26.2%中发现,9.7%,1.7%,和0.4%的艾伯蒂菌分离株,分别。测试菌株的染色体大小范围为4,573,338至5,141,010bp(平均4,784,003bp),并且至少一个质粒存在于所有菌株中。该研究有助于更准确地评估阿伯蒂大肠杆菌的遗传多样性及其对公共卫生的潜在威胁。
    Escherichia albertii is a new enteropathogen of humans and animals. The aim of the study was to assess the prevalence and pathogenicity of E. albertii strains isolated in northeastern Poland using epidemiological and genomic studies. In 2015-2018, a total of 1154 fecal samples from children and adults, 497 bird droppings, 212 food samples, 92 water samples, and 500 lactose-negative E. coli strains were tested. A total of 42 E. albertii strains were isolated. The PCR method was suitable for their rapid identification. In total, 33.3% of E. albertii isolates were resistant to one antibiotic, and 16.7% to two. Isolates were sensitive to cefepime, imipenem, levofloxacin, gentamicin, trimethoprim/sulfamethoxazole, and did not produce ESBL β-lactamases. High genetic variability of E. albertii has been demonstrated. In the PFGE method, 90.5% of the strains had distinct pulsotypes. In MLST typing, 85.7% of strains were assigned distinct sequence types (STs), of which 64% were novel ST types. Cytolethal distending toxin (CDT) and Paa toxin genes were found in 100% of E. albertii isolates. Genes encoding toxins, IbeA, CdtB type 2, Tsh and Shiga (Stx2f), were found in 26.2%, 9.7%, 1.7%, and 0.4% of E. albertii isolates, respectively. The chromosome size of the tested strains ranged from 4,573,338 to 5,141,010 bp (average 4,784,003 bp), and at least one plasmid was present in all strains. The study contributes to a more accurate assessment of the genetic diversity of E. albertii and the potential threat it poses to public health.
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  • 文章类型: Journal Article
    BACKGROUND: Escherichia albertii is an emerging zoonotic foodborne pathogen. Its prevalence, distribution, and reservoirs are not yet clearly defined. In this study, we evaluated the occurrence and genomic characteristics of E. albertii in livestock from Switzerland. A total of 515 caecal samples from sheep, cattle, calves, and fattening swine were collected between May 2022 and August 2022 at abattoir level. Using an E. albertii-specific PCR targeting the Eacdt-gene, 23,7 % (51/215) of swine from 24 different farms were positive. One (1 %) out of 100 calves showed a positive PCR result, while all samples from sheep and cattle were PCR negative. Eight E. albertii isolates could be recovered from swine samples and were analysed using whole-genome sequencing. All eight isolates belonged to ST2087 or a ST4619 group subclade, as did most genomes of the 11 available global swine isolates from public databases. These two clusters shared the presence of a virulence plasmid harboring the sitABCD and iuc genes. In summary, we demonstrate that fattening swine constitute an E. albertii reservoir in Switzerland and describe specific swine-associated lineages.
    BACKGROUND: Escherichia albertii ist ein neu auftretender zoonotischer lebensmittelassoziierter Krankheitserreger. Prävalenz, Verbreitung und mögliche Reservoire sind noch nicht eindeutig definiert. In dieser Studie haben wir das Vorkommen und die genomischen Eigenschaften von E. albertii bei Nutztieren in der Schweiz evaluiert. Zwischen Mai und August 2022 wurden insgesamt 515 Blinddarmproben von Schafen, Rindern, Kälbern und Mastschweinen während des Schlachtvorganges entnommen. Unter Verwendung einer E. albertii-spezifischen PCR, die auf das Eacdt-Gen abzielte, waren 23,7 % (51/215) der Schweine aus 24 verschiedenen Betrieben positiv. Eines (1 %) von 100 Kälbern zeigte ein positives PCR-Ergebnis, während alle Proben von Schafen und Rindern PCR-negativ waren. Acht E. albertii-Isolate konnten aus Schweineproben gewonnen und mittels Ganzgenomsequenzierung analysiert werden. Alle acht Isolate gehörten zu den MLST Sequenztypen ST2087 oder ST4619, ebenso wie die meisten Genome der 11 verfügbaren globalen Schweineisolate aus öffentlichen Datenbanken. Diese beiden Cluster teilten das Vorhandensein eines Virulenzplasmids, das die sitABCD- und iuc-Gene beherbergte. Zusammenfassend zeigen wir, dass Mastschweine ein E. albertii-Reservoir in der Schweiz darstellen und beschreiben spezifische Schweine-assoziierte Bakteriencluster.
    BACKGROUND: Escherichia albertii est un pathogène alimentaire zoonotique émergent. Sa prévalence, sa distribution et ses réservoirs ne sont pas encore clairement définis. Dans cette étude, nous avons évalué l’occurrence et les caractéristiques génomiques d’E. albertii chez le bétail en Suisse. Au total, 515 échantillons cæcaux d’ovins, de bovins, de veaux et de porcs d’engraissement ont été prélevés entre mai 2022 et août 2022 au niveau de l’abattoir. En utilisant une PCR spécifique à E. albertii ciblant le gène Eacdt, 23,7 % (51/215) des porcs provenant de 24 exploitations différentes étaient positifs. Un (1 %) veau sur 100 a présenté un résultat positif à la PCR, tandis que tous les échantillons d’ovins et de bovins étaient négatifs à la PCR. Huit isolats d’E. albertii ont pu être récupérés à partir d’échantillons provenant de porcs et ont été analysés par séquençage du génome entier. Les huit isolats appartenaient au groupe ST2087 ou à un sous-clade du groupe ST4619, comme la plupart des génomes des 11 isolats porcins mondiaux disponibles dans les bases de données publiques. Ces deux groupes partageaient la présence d’un plasmide de virulence hébergeant les gènes sitABCD et iuc. En résumé, nous démontrons que les porcs d’engraissement constituent un réservoir d’E. albertii en Suisse et décrivons des lignées spécifiques associées aux porcs.
    BACKGROUND: La Escherichia albertii è un patogeno zoonotico emergente negli alimenti. La sua prevalenza, distribuzione e i serbatoi non sono stati ancora chiaramente identificati. In questo studio abbiamo valutato la presenza e le caratteristiche genomiche di E. albertii negli animali da allevamento in Svizzera. Tra il maggio e l’agosto 2022, un totale di 515 campioni cecali sono stati raccolti da ovini, bovini, vitelli e suini da ingrasso durante il processo di macellazione. Utilizzando una PCR specifica per E. albertii mirata al gene Eacdt, il 23,7 % (51/215) dei suini provenienti da 24 aziende diverse è risultato positivo. Un vitello (1 %) su 100 ha mostrato un risultato positivo alla PCR, mentre tutti i campioni di ovini e bovini sono risultati negativi alla PCR. È stato possibile ottenere otto isolati di E. albertii da campioni di suini e sono stati analizzati mediante il sequenziamento dell’intero genoma. Tutti gli otto isolati appartenevano al gruppo ST2087 o a un sottoclade del gruppo ST4619, così come la maggior parte dei genomi degli 11 isolati globali di suini disponibili nelle banche dati pubbliche. Questi due cluster condividono la presenza di un plasmide di virulenza che ospita i geni sitABCD e iuc. In conclusione, abbiamo dimostrato che i suini da ingrasso rappresentano un serbatoio di E. albertii in Svizzera e abbiamo descritto specifici lignaggi associati ai suini.
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  • 文章类型: Journal Article
    艾氏大肠杆菌是一种新兴的,肠道病原体的意义。它于2003年首次从孟加拉国的小儿腹泻样本中分离出来。在这项研究中,遵循全面的计算机战略,首先从核心列出抗生素抗性基因,艾伯蒂大肠杆菌95个可用基因组的辅助和独特基因组部分。然后,从核心必需基因组中鉴定出56个药物靶标。最后,ZipA,一种必需的细胞分裂蛋白,通过交联FtsZ原丝来稳定它们,并作为Z环的细胞质膜锚,被选择用于进一步的下游处理。它是使用线程方法计算建模的,然后对两个植物化学库进行虚拟筛选,阿育吠陀(n=2103个化合物)和中药(n=36,043个化合物)。ADMET分析,随后在中央车身舱进行PBPK建模,在250名没有患病的人群中,尝试了250名肝硬化患者和250名肾脏受损患者。中药库的ZINC85624912显示出最高的生物利用度和血浆保留率。这是首次尝试通过PBPK模拟体内天然产物的命运。还对来自两个文库的前三种化合物进行20ns的动力学模拟以验证化合物的稳定性。从当前研究中获得的信息可以帮助湿实验室科学家研究从自然资源中筛选出的药物样化合物的支架,并且可以用于我们寻求对抗生素耐药的E.albertii的治疗。
    Escherichia albertii is an emerging, enteric pathogen of significance. It was first isolated in 2003 from a pediatric diarrheal sample from Bangladesh. In this study, a comprehensive in silico strategy was followed to first list out antibiotic-resistant genes from core, accessory and unique genome fractions of 95 available genomes of E. albertii. Then, 56 drug targets were identified from the core essential genome. Finally, ZipA, an essential cell division protein that stabilizes the FtsZ protofilaments by cross-linking them and serves as a cytoplasmic membrane anchor for the Z ring, was selected for further downstream processing. It was computationally modeled using a threading approach, followed by virtual screening of two phytochemical libraries, Ayurvedic (n = 2103 compounds) and Traditional Chinese Medicine (n = 36,043 compounds). ADMET profiling, followed by PBPK modeling in the central body compartment, in a population of 250 non-diseased, 250 cirrhotic and 250 renally impaired people was attempted. ZINC85624912 from Chinese medicinal library showed the highest bioavailability and plasma retention. This is the first attempt to simulate the fate of natural products in the body through PBPK. Dynamics simulation of 20 ns for the top three compounds from both libraries was also performed to validate the stability of the compounds. The obtained information from the current study could aid wet-lab scientists to work on the scaffold of screened drug-like compounds from natural resources and could be useful in our quest for therapy against antibiotic-resistant E. albertii.
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  • 文章类型: Journal Article
    艾氏大肠杆菌是一种新兴的人畜共患食源性肠病原体,可导致人类胃肠炎暴发。尽管从被认为是这种细菌的潜在宿主的鸟类中分离出了艾伯蒂菌,它在候鸟中的流行很少被描述。在这项研究中,使用全基因组测序对鄱阳湖候鸟中的艾伯蒂菌进行了调查和表征。收集了来自9种候鸟的81个粪便样品,并对24/81(29.6%)的艾贝特异性基因进行了PCR阳性测试。从24个PCR阳性样品中的18个中回收了47个分离株。所有分离株都携带eae和cdtB基因。将这些分离株分为8种阿伯替尼O基因型(EAOgs)(包括3种新型EAOgs)和3种阿伯替尼H基因型(EAHgs)。全基因组系统发育将候鸟分离株分为不同的谱系,本研究中的一些分离株在系统发育上与家禽或患者来源的菌株紧密分类.我们的发现表明,候鸟可能是异质艾伯蒂埃的重要水库,从而作为阿尔伯蒂大肠杆菌向人类的潜在传播媒介。
    Escherichia albertii is an emerging zoonotic foodborne enteropathogen leading to human gastroenteritis outbreaks. Although E. albertii has been isolated from birds which have been considered as the potential reservoirs of this bacterium, its prevalence in migratory birds has rarely been described. In this study, E. albertii in migratory birds from Poyang Lake was investigated and characterized using whole genome sequencing. Eighty-one fecal samples from nine species of migratory birds were collected and 24/81 (29.6%) tested PCR-positive for E. albertii-specific genes. A total of 47 isolates was recovered from 18 out of 24 PCR-positive samples. All isolates carried eae and cdtB genes. These isolates were classified into eight E. albertii O-genotypes (EAOgs) (including three novel EAOgs) and three E. albertii H-genotypes (EAHgs). Whole genome phylogeny separated migratory bird-derived isolates into different lineages, some isolates in this study were phylogenetically closely grouped with poultry-derived or patient-derived strains. Our findings showed that migratory birds may serve as an important reservoir for heterogeneous E. albertii, thereby acting as potential transmission vehicles of E. albertii to humans.
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  • 文章类型: Journal Article
    艾氏大肠杆菌是一种新兴的肠病原体。日本已经报道了几起食源性的阿伯蒂伊病毒暴发;然而,与大多数疫情相关的食物仍未被确认。因此,需要特异性和灵敏地检测阿氏大肠杆菌的聚合酶链反应(PCR)测定法。设计了用于靶向阿氏大肠杆菌特异性基因(EA-rtPCR)的实时PCR测定的引物和探针。74株,包括43株艾伯蒂埃及其近亲,EA-rtPCR特异性扩增阿氏大肠杆菌;因此,然后评估EA-rtPCR的敏感性。艾氏大肠杆菌培养物和接种病原体的富集鸡培养物的检测限为2.8和2.0-3.2对数菌落形成单位(CFU)/mL,分别。此外,通过EA-rtPCR从接种有0.1logCFU的病原体的25g鸡肉中检测到艾氏大肠杆菌。通过与巢式PCR方法的比较,还评估了EA-rtPCR检测鸡肉中的白葡萄酒。并测试了28个零售鸡肉和来自21个鸡肉尸体的193个解剖身体部位。1个和3个鸡肉在巢式PCR和EA-rtPCR中呈阳性,分别。14具尸体至少有一个身体部位对EA-rtPCR呈阳性,36和48个样本的巢式PCR和EA-rtPCR检测呈阳性,分别。从从6只鸡car体中获得的7个PCR阳性样品中分离出37株艾氏大肠杆菌。所有艾氏大肠杆菌分离株都带有eae基因,并通过EAO基因分型分类为艾氏O基因型(EAOg)3或EAOg4。本研究中开发的EA-rtPCR有可能改善食品中的艾贝氏杆菌检测并促进艾贝氏杆菌感染的研究。
    Escherichia albertii is an emerging enteropathogen. Several foodborne outbreaks of E. albertii have been reported in Japan; however, foods associated with most outbreaks remain unidentified. Therefore, polymerase chain reaction (PCR) assays detecting E. albertii specifically and sensitively are required. Primers and probe for real-time PCR assays targeting E. albertii-specific gene (EA-rtPCR) was designed. With 74 strains, including 43 E. albertii strains and several of its close relatives, EA-rtPCR specifically amplified E. albertii; therefore, the sensitivity of EA-rtPCR was then evaluated. The detection limits were 2.8 and 2.0-3.2 log colony-forming unit (CFU)/mL for E. albertii culture and enriched chicken culture inoculated with the pathogen, respectively. In addition, E. albertii was detected from 25 g of chicken meat inoculated with 0.1 log CFU of the pathogen by EA-rtPCR. The detection of E. albertii from chicken meat by EA-rtPCR was also evaluated by comparing with the nested-PCR assay, and 28 retail chicken meat and 193 dissected body parts from 21 chicken carcass were tested. One and three chicken meat were positive in the nested-PCR assay and EA-rtPCR, respectively. Fourteen carcasses had at least one body part that was positive for EA-rtPCR, and 36 and 48 samples were positive for the nested-PCR assay and EA-rtPCR, respectively. A total of 37 strains of E. albertii were isolated from seven PCR-positive samples obtained from six chicken carcass. All E. albertii isolates harbored eae gene, and were classified as E. albertii O-genotype (EAOg)3 or EAOg4 by EAO-genotyping. The EA-rtPCR developed in this study has potential to improve E. albertii detection in food and advance research on E. albertii infection.
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