Tailocins are nano-scale phage tail-like protein complexes that can mediate antagonistic interactions between closely related bacterial species. While the capacity to produce R-type tailocin was found widely across Gammaproteobacteria, the production of F-type tailocins seems comparatively rare. In this study, we examined the freshwater isolate, Pragia fontium 24613, which can produce both R- and F-type tailocins. We investigated their inhibition spectrum, focusing on clinically relevant enterobacteria, and identified the associated tailocin gene cluster. Transmission electron microscopy confirmed that inactivation of the tape measure protein within the tailocin cluster disrupted R-tailocin production. Comparative analysis of Budviciaceae gene clusters showed high conservation of R-type tailocin genes, whereas F-type tailocin genes were found in only a few species, with little conservation. Our findings indicate a high prevalence of bacteriocin production among underexplored Enterobacteriales species. Detected tailocins showed potential as antimicrobials targeting clinically significant pathogens.
In 2019, Pantoea piersonii was initially isolated from the interior surfaces of the International Space Station. This microorganism is a species within the genus Pantoea in the family Erwiniaceae, belonging to the order Enterobacterales. Recent literature has documented four cases of its isolation. Despite initial predictions suggesting the non-pathogenicity of P. piersonii strains, evidence from observed cases indicates potential pathogenicity. According to documented evidence in the literature, this microorganism is capable of causing severe and life-threatening conditions, including sepsis. Traditional tests, as well as automated systems, may fail to provide complete differentiation due to these similarities. While MALDI-TOF MS is a valuable tool for identification in clinical diagnostic microbiology, sequencing may be necessary for precise identification. To determine the antibiotic susceptibility profile, various methods can be utilized, including minimum inhibitory concentration determination, disk diffusion testing (Kirby-Bauer test), genotypic resistance assays (PCR and sequencing), and automated systems. The literature reports a limited number of cases associating P. piersonii with human infection. This study contributes to this body of knowledge by reporting a novel case in which P. piersonii was isolated from a tissue sample for the first time. In this case report, the patient achieved recovery following the administration of appropriate antibiotic treatment based on the diagnosis. It underscores the need for precise identification and understanding of its pathogenicity.
- MeSH
- antibakteriální látky * farmakologie terapeutické užití MeSH
- enterobakteriální infekce * mikrobiologie diagnóza farmakoterapie MeSH
- lidé MeSH
- mikrobiální testy citlivosti * MeSH
- Pantoea * izolace a purifikace genetika patogenita MeSH
- Check Tag
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- kazuistiky MeSH
Feruloyl esterases (FAEs) are a crucial component of the hemicellulose-degrading enzyme family that facilitates the degradation of lignocellulose while releasing hydroxycinnamic acids such as ferulic acid with high added value. Currently, the low enzyme yield of FAEs is one of the primary factors limiting its application. Therefore, in this paper, we optimized the fermentation conditions for the expression of FAE BpFaeT132C-D143C with excellent thermal stability in Escherichia coli by experimental design. Firstly, we explored the effects of 11 factors such as medium type, isopropyl-β-D-thiogalactopyranoside (IPTG) concentration, and inoculum size on BpFaeT132C-D143C activity separately by the single factor design. Then, the significance of the effects of seven factors, such as post-induction temperature, shaker rotational speed, and inoculum size on BpFaeT132C-D143C activity, was analyzed by Plackett-Burman design. We identified the main factors affecting the fermentation conditions of E. coli expressing BpFaeT132C-D143C as post-induction temperature, pre-induction period, and post-induction period. Finally, we used the steepest ascent path design and response surface method to optimize the levels of these three factors further. Under the optimal conditions, the activity of BpFaeT132C-D143C was 3.58 U/ml, which was a significant 6.6-fold increase compared to the pre-optimization (0.47 U/ml), demonstrating the effectiveness of this optimization process. Moreover, BpFaeT132C-D143C activity was 1.52 U/ml in a 3-l fermenter under the abovementioned optimal conditions. It was determined that the expression of BpFaeT132C-D143C in E. coli was predominantly intracellular in the cytoplasm. This study lays the foundation for further research on BpFaeT132C-D143C in degrading agricultural waste transformation applications.
- MeSH
- Escherichia coli * genetika metabolismus enzymologie MeSH
- fermentace * MeSH
- isopropylthiogalaktosid metabolismus MeSH
- karboxylesterhydrolasy * genetika metabolismus chemie biosyntéza MeSH
- kultivační média chemie MeSH
- kyseliny kumarové metabolismus MeSH
- lignin MeSH
- rekombinantní proteiny genetika metabolismus biosyntéza chemie MeSH
- stabilita enzymů MeSH
- teplota MeSH
- Publikační typ
- časopisecké články MeSH
Biopriming seeds with beneficial bacteria has potential to enhance seed germination. Therefore, in this investigation, five sulphur-oxidizing bacterial cultures, viz., Pantoea dispersa SOB2, Bacillus velezensis SN06, Bacillus cereus SN20, Bacillus tropicus SN16, and Bacillus megaterium SN11, were evaluated for different plant growth-promoting traits and their impact on Vigna radiata L. (mung bean) and Brassica juncea L. (mustard) seed germination. Among these, three bacterial cultures Pantoea dispersa SOB2, Bacillus velezensis SN06, and Bacillus megaterium SN11 evinced potential for mineral solubilization on solid medium where Pantoea dispersa SOB2 had the maximum solubilization indices-3.06, 5.14, and 2.48 for phosphate, zinc, and potassium respectively. The culture also displayed higher indole acetic acid (113.12 μg/mL), gibberellic acid (162.66 μg/mL), ammonia (5.23 μg/mL), and siderophore (69.53%) production than other bacterial cultures whereas Bacillus cereus SN20 showed maximum exopolysaccharide production (9.26 g/L). Bacterial culture Pantoea dispersa SOB2 significantly ameliorated the germination rate (3.73 no./day) and relative seed germination (208%) of Brassica juncea L., while Bacillus velezensis SN06 and Bacillus cereus SN20 followed with germination rate and relative seed germination of 2.86 no./day and 207%, respectively. Pantoea dispersa SOB2 displayed lowest mean germination time 2.91 days followed by Bacillus megaterium SN11 with 3.19 days. Biopriming with sulphur-oxidizing bacterial cultures, germination parameters of Vigna radiata L. were also markedly improved. Pantoea dispersa SOB2 demonstrated the highest germination rate (6.72 no./day), relative seed germination (115.56%), and minimum mean generation time (1.73 days). Bacillus velezensis SN06 inoculation had a beneficial effect on the seedling growth of Vigna radiata L., whereas Pantoea dispersa SOB2 greatly aided the seedling growth of Brassica juncea L. Results corroborated a prominent positive correlation between seed germination and plant growth-promoting traits. This is the first study on Pantoea dispersa as sulphur oxidizer, displaying plant growth promoting traits and seed germination potential. The potent sulphur-oxidizing bacterial cultures possessing plant growth promoting activities enhanced seed germination under in vitro conditions that could be further explored in field as biofertilizers to enhance the growth and yield of Brassica juncea L. and Vigna radiata L. crop.
- MeSH
- Bacillus * metabolismus MeSH
- Bacteria * metabolismus MeSH
- hořčice rodu Brassica * růst a vývoj mikrobiologie MeSH
- klíčení * MeSH
- kyseliny indoloctové metabolismus MeSH
- oxidace-redukce MeSH
- Pantoea metabolismus MeSH
- regulátory růstu rostlin metabolismus MeSH
- semena rostlinná * mikrobiologie růst a vývoj MeSH
- síra * metabolismus MeSH
- vigna * růst a vývoj mikrobiologie MeSH
- Publikační typ
- časopisecké články MeSH
Foodborne diseases triggered by various infectious micro-organisms are contributing significantly to the global disease burden as well as to increasing mortality rates. Salmonella enterica belongs to the most prevalent form of bacteria accountable for significant burden of foodborne illness across the globe. The conventional therapeutic approach to cater to Salmonella enterica-based infections relies on antibiotic therapy, but the rapid emergence of the antibiotic resistance strains of Salmonella sp. necessitates the development of alternative treatment and prevention strategies. In light of this growing concern, the scientific community is rigorously exploring novel phytochemicals harnessed from medicinally important plants as a promising approach to curb Salmonella enterica infections. A variety of phytochemicals belonging to alkaloids, phenols, flavonoid, and terpene classes are reported to exhibit their inhibitory activity against bacterial cell communication, membrane proteins, efflux pumps, and biofilm formation among drug resistant Salmonella strains. The present review article delves to discuss the emergence of antibiotic resistance among Salmonella enterica strains, various plant sources, identification of phytochemicals, and the current state of research on the use of phytochemicals as antimicrobial agents against Salmonella enterica, shedding light on the promising potential of phytochemicals in the fight against this pathogen.
- MeSH
- antibakteriální látky * farmakologie MeSH
- bakteriální léková rezistence MeSH
- fytonutrienty * farmakologie chemie MeSH
- lidé MeSH
- nemoci přenášené potravou mikrobiologie prevence a kontrola farmakoterapie MeSH
- Salmonella enterica * účinky léků MeSH
- salmonelóza * mikrobiologie farmakoterapie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Úvod: Shigelóza je vysoce nakažlivé průjmové onemocnění s potenciálně velmi závažným průběhem. I s ohledem na třetinový nárůst případů v roce 2023 ve srovnání s rokem předchozím jsme si stanovili za cíl podat přehled aktuálních informací o onemocnění a analyzovat data nahlášených případů shigelózy v České republice (ČR). Metody: Zpracovali jsme narativní rešerši odborné literatury v českém a anglickém jazyce, zejména cílenou na evropské studie od roku 2018. Dále jsme provedli analýzu dat hlášených pod kódem diagnózy A03 v národním systému pro hlášení infekčních nemocí (ISIN) v letech 2018–2023. Soustředili jsme se na hlavní epidemiologické ukazatele, zejména pohlaví, věk, geografickou distribuci, sezonnost a hospitalizace. Použity byly programy Excel (verze 2016), STATA (verze 17) a Datawrapper GmbH. Výsledky: Celkem bylo nahlášeno 681 případů onemocnění shigelózou s průměrnou roční incidencí 1/100 000 obyvatel: do roku 2021 byla incidence mírně vyšší u žen, od roku 2022 evidujeme trend opačný. V pandemických letech byl zaznamenán významný pokles případů. V letech 2022 a 2023 byl počet případů mírně vyšší než v období před pandemií. Nejvíce případů evidujeme v ČR každoročně v měsících srpen až prosinec. Ze všech sérotypů shigel byla nejčastěji detekována S. sonnei (80 %), následovaná S. flexneri (15 %). Incidence na 100 000 obyvatel byla nejvyšší u osob ve věku 5–9 let: 2,6 (chlapci 2,4 a dívky 2,8), dále 1–4 roky: 2,4 (chlapci 2,2, dívky 2,6) a osob ve věku 25–34 let: 1,8 (muži 1,8 a ženy 1,7). Podle krajů byla průměrná roční specifická incidence nejvyšší v krajích Moravskoslezském, Olomouckém a v hlavním městě Praze. Hospitalizováno bylo 27 % případů, nejvíce ve věkových skupinách 25–34 a 5–9 let (shodně 17,9 %). Proporce hospitalizovaných případů v rámci jednotlivých věkových skupin byla nejvyšší ve věkové skupině 75+ let (69 %), dále věkových skupinách 1–4 roky, 5–9 let a 65–74let (32–37 %). V souvislosti s onemocněním bylo vykázáno jedno úmrtí muže ve věku 52 let. V rámci epidemického výskytu bylo nahlášeno 11 % případů. Importováno bylo 39 % nahlášených případů. Závěr: V ČR je shigelóza spíše málo zastoupeným gastrointestinálním onemocněním, přičemž téměř 40 % případů tvoří importované nákazy. V současnosti je hrozbou pro veřejné zdraví především globální šíření multirezistentních kmenů podpořené narůstajícím cestovním ruchem a volnými sexuálními praktikami. Rizikovými skupinami zůstávají děti, imunokompromitované osoby (včetně seniorů) a muži mající sex s muži. Očkování není v Evropě dostupné. Stěžejním je nadále dodržování základních hygienických pravidel, zejména v kolektivech a při práci s potravinami. Důraz by měl být dále kladen na zdravotní edukaci osob, včetně poučení před vycestováním do zahraničí. Důkladná anamnéza, včasné trasování, dohled a racionální volba eventuální antibiotické terapie jsou zásadní. V ČR musí být všechny suspektní kmeny zaslány do NRL ke konfirmaci. Celogenomovou sekvenaci a testy citlivosti na antibiotika je vhodné provádět u všech izolátů.
Introduction: Shigellosis is a highly contagious diarrheal disease, which could potentially be very serious. Considering the onethird increase in cases in 2023 compared to the previous year, we aimed to provide an update on the disease and to analyse data on reported cases of shigellosis in the Czech Republic (CZ). Methods: We conducted a narrative search of the literature in Czech and English, particularly targeting European studies from 2018 onwards. We also analysed data reported under the diagnosis code A03 to the National Infectious Disease Reporting System (ISIN) in 2018–2023. We focused on the main epidemiological indicators, i.e. gender, age, geographical distribution, seasonality, and hospitalizations. Excel (version 2016), STATA (version 17), and Datawrapper GmbH were used. Results: A total of 681 shigellosis cases were reported with an average annual incidence of 1/100,000 population: until 2021, the incidence was slightly higher in women, while from 2022 onwards, the trend was reversed. A significant decrease in cases was recorded in the pandemic years. In 2022 and 2023, the number of cases was slightly higher than in the pre-pandemic period. Most cases were detected in CZ in August and December each year. Of all shigella serotypes, S. sonnei was the most frequently detected (80%), followed by S. flexneri (15%). The incidence per 100.000 population was highest among children aged 5–9 years: 2.6 (boys 2.4 and girls 2.8), followed by 1–4-year-olds: 2.4 (2.2 and 2.6, respectively) and persons aged 25–34 years: 1.8 (males 1.8 and females 1.7). Within individual age group, the average annual specific incidence rates were highest in the Moravian-Silesian and Olomouc regions and the capital city Prague. Hospitalizations accounted for 27% of cases, with the highest numbers in the 25–34 and 5–9 age groups (both 17.9%). The proportion of hospitalized cases was highest in the age groups 75+ (69%), 1–4, 5–9, and 65–74 (32–37%). A 52-year-old man was reported to have die in relation to the disease. Eleven percent of cases were reported in outbreak settings. Thirty-nine percent of reported cases were imported. Conclusions: In CZ, shigellosis is a relatively rare gastrointestinal disease, with nearly 40% of cases being imported. At present, the threat to public health is posed mainly by the global spread of multi-resistant strains linked to increasing tourism and free sexual practices. Children, immunocompromised persons (including the elderly), and men who have sex with men remain risk groups. Vaccination is not available in Europe. Compliance with basic hygiene rules, especially in collectives and when working with food, is still a key concern. Emphasis should also be placed on the health education, including instructions before traveling abroad. A thorough medical history, early tracing, surveillance, and rational choice of antibiotic therapy if appropriate are essential. In CZ, all suspected strains shall be sent to the NRL for confirmation. Whole genome sequencing and antibiotic susceptibility testing should be performed on all isolates.
Super- and low-shedding phenomena have been observed in genetically homogeneous hosts infected by a single bacterial strain. To decipher the mechanisms underlying these phenotypes, we conducted an experiment with chicks infected with Salmonella Enteritidis in a non-sterile isolator, which prevents bacterial transmission between animals while allowing the development of the gut microbiota. We investigated the impact of four commensal bacteria called Mix4, inoculated at hatching, on chicken systemic immune response and intestinal microbiota composition and functions, before and after Salmonella infection. Our results revealed that these phenotypes were not linked to changes in cell invasion capacity of bacteria during infection. Mix4 inoculation had both short- and long-term effects on immune response and microbiota and promoted the low-shedder phenotype. Kinetic analysis revealed that Mix4 activated immune response from day 4, which modified the microbiota on day 6. This change promotes a more fermentative microbiota, using the aromatic compounds degradation pathway, which inhibited Salmonella colonization by day 11 and beyond. In contrast, control animals exhibited a delayed TNF-driven pro-inflammatory response and developed a microbiota using anaerobic respiration, which facilitates Salmonella colonization and growth. This strategy offers promising opportunities to strengthen the barrier effect against Salmonella and possibly other pathogens.
- MeSH
- Bacteria * klasifikace genetika izolace a purifikace MeSH
- kur domácí * mikrobiologie imunologie MeSH
- nemoci drůbeže * mikrobiologie imunologie prevence a kontrola MeSH
- Salmonella enteritidis * imunologie růst a vývoj MeSH
- salmonelová infekce u zvířat * mikrobiologie imunologie prevence a kontrola MeSH
- střevní mikroflóra * MeSH
- symbióza MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Bacterial β sliding clamp (β-clamp) is an emerging drug target currently lacking small-molecule inhibitors with good in vivo activity. Thus, there is a need for fast and simple screening methods for identifying inhibitor candidates. Here we demonstrate the use of nuclear magnetic resonance spectroscopy (NMR) for evaluating compound binding to the E. coli β-clamp. To identify suitable molecular probes, a series of tetrahydrocarbazoles were synthesized, some of which contain fluorine. Key challenges in the synthesis were formation of regioisomers during the Fischer indole reaction and reducing racemization at the stereogenic center. The tetrahydrocarbazoles were assayed against the E. coli β-clamp by saturation-transfer difference (STD) NMR, waterLOGSY and T1ρ. Analysis by isothermal titration calorimetry gave KD-values of 1.7-14 μM for three fluorinated probe candidates, and NMR chemical shift perturbation experiments confirmed these molecules to directly interact with the β-clamp binding pocket. Binding of the fluorinated molecules to β-clamp was easily observed with 19F-observed T2-based binding experiments, and proof of concept for a fluorine-based binding assay for E. coli β-clamp binders is provided.
- MeSH
- Escherichia coli * účinky léků MeSH
- halogenace MeSH
- karbazoly * chemie chemická syntéza farmakologie metabolismus MeSH
- magnetická rezonanční spektroskopie * MeSH
- molekulární sondy chemie chemická syntéza metabolismus MeSH
- molekulární struktura MeSH
- proteiny z Escherichia coli metabolismus antagonisté a inhibitory chemie MeSH
- vazba proteinů MeSH
- vazebná místa MeSH
- vztahy mezi strukturou a aktivitou MeSH
- Publikační typ
- časopisecké články MeSH
Urinary tract infections (UTIs) are one of the most common bacterial diseases both in communities and in hospitalized patients, and at the same, time they are one of the most common indications for the use of antibiotics. UTI guidelines are generally available nationally or internationally, but they do not address all aspects of UTI treatment for different patient cohorts, age, gender, or comorbidities. The aim of the study was to point out the importance of stratified cumulative antibiograms at the level of individual health care facilities and the significant differences between epidemiological data, not only at the national level, but also at the local level. Our study analyses data from 383 patients with UTIs from a hospital department, General University Hospital (GUH), and 272 patients from an outpatient medical facility, Urocentrum (UC). This analysis focuses on the most common UTI causative agent, Escherichia coli, its representation as the causative agent of UTI in patients with complicated acute cystitis (N30), and its representation in complicated acute cystitis in patients with prostate cancer (C61). In addition to the frequency of occurrence, a sub-analysis of the incidence of resistance of E. coli to commonly used antibiotics by age, gender, diagnosis, and medical facility was performed. Results: The most common causative agent of UTI was E. coli. In patients with N30, it was 70% in GUH and 54% in UC, but in oncological patients with UTI, it was only 39% and 35%, respectively. In patients with UTI in C61, there was a significant difference in susceptibility of E. coli between individual health care facilities. Lower resistance was found in UC opposite to GUH isolates in ampicillin, with 29.8% vs. 65%, p = 0.001; amoxicillin/clavulanic acid, with 8.5% vs. 30%, p = 0.01; with 2.1% vs. 17.5% in pivmecillinam, p = 0.01; with 10.6% vs. 37.5% in co-trimoxazole, p = 0.003; and ciprofloxacin, with 10.6% vs. 30%, p = 0.04. The study shows significant differences in the sensitivity of urinary E. coli isolates in patients in relation to age, gender, medical devices, and the presence of comorbidities.
- MeSH
- antibakteriální látky * terapeutické užití MeSH
- bakteriální léková rezistence MeSH
- dospělí MeSH
- Escherichia coli * účinky léků izolace a purifikace MeSH
- infekce močového ústrojí * farmakoterapie mikrobiologie epidemiologie MeSH
- infekce vyvolané Escherichia coli farmakoterapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- mikrobiální testy citlivosti * MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
The RNA chaperone Hfq plays crucial roles in bacterial gene expression and is a major facilitator of small regulatory RNA (sRNA) action. The toroidal architecture of the Hfq hexamer presents three well-characterized surfaces that allow it to bind sRNAs to stabilize them and engage target transcripts. Hfq-interacting sRNAs are categorized into two classes based on the surfaces they use to bind Hfq. By characterizing a systematic alanine mutant library of Hfq to identify amino acid residues that impact survival of Escherichia coli experiencing nitrogen (N) starvation, we corroborated the important role of the three RNA-binding surfaces for Hfq function. We uncovered two, previously uncharacterized, conserved residues, V22 and G34, in the hydrophobic core of Hfq, to have a profound impact on Hfq's RNA-binding activity in vivo. Transcriptome-scale analysis revealed that V22A and G34A Hfq mutants cause widespread destabilization of both sRNA classes, to the same extent as seen in bacteria devoid of Hfq. However, the alanine substitutions at these residues resulted in only modest alteration in stability and structure of Hfq. We propose that V22 and G34 have impact on Hfq function, especially critical under cellular conditions when there is an increased demand for Hfq, such as N starvation.
- MeSH
- bakteriální RNA * metabolismus genetika chemie MeSH
- dusík metabolismus MeSH
- Escherichia coli * genetika metabolismus MeSH
- hydrofobní a hydrofilní interakce * MeSH
- konzervovaná sekvence MeSH
- malá nekódující RNA * metabolismus genetika chemie MeSH
- mutace MeSH
- protein hostitelského faktoru 1 * metabolismus genetika chemie MeSH
- proteiny z Escherichia coli * metabolismus genetika chemie MeSH
- regulace genové exprese u bakterií MeSH
- stabilita RNA * genetika MeSH
- stanovení celkové genové exprese MeSH
- transkriptom genetika MeSH
- vazba proteinů MeSH
- Publikační typ
- časopisecké články MeSH
