Exacerbation of virulence of multi-drug resistant Escherichia coli O104:H4 by subinhibitory concentrations of ampicillin
Little is known about how subinhibitory concentrations of antibiotics to which bacteria are resistant affect bacterial virulence. In this study, the effect of subinhibitory concentrations of ampicillin on the virulence of E. coli O104:H4 was analyzed. Bacteria were pre-exposed to 0.1, 0.3, or 0.5 mg/mL of ampicillin in LB media and incubated for 4 h at 37 °C. Transformation capacity (using plasmids and PCR-amplified DNA sequences), swarming motility, biofilm production, curli formation, and virulence gene expression were determined. Ampicillin increased the transformation of E. coli O104:H4, with the highest number of transformants (>10 CFU/ng DNA; p ≤ 0.05) detected after exposure to DNA sequences of spectinomycin. In addition, bacteria pre-treated with 0.5 mg/mL of ampicillin exhibited higher swarming motility (7.6 cm, vs 6.0 cm for control; p ≤ 0.05) and biofilm production (up to 1.9-fold; p ≤ 0.05) when subsequently exposed to 0.1 and 0.3 mg/mL of antibiotic compared with the control. Also, significant overexpression of the virulence-related genes flhC (≤16.1-fold), fliA (≤22.1-fold), csgA (≤3.6-fold), csgD (≤9.1-fold), stx2a (≤32.2-fold), and the antibiotic resistance gene blaTEM-1 (≤5.5-fold) was observed. In conclusion, ampicillin-resistant E. coli O104:H4 increased the expression of its virulence factors when exposed to most subinhibitory concentrations of ampicillin analyzed in this study.
Staphylococcus aureus: Dynamics of pathogenicity and antimicrobial-resistance in hospital and community environments - Comprehensive overview
This study reviews Staphylococcus aureus, a significant pathogen in both hospital and community-acquired infections, addressing its epidemiology, pathogenesis, and antimicrobial resistance. It highlights virulence mechanisms, such as adhesion factors, toxins, enzymes, and biofilms, which contribute to survival and immune evasion. The spread of resistance occurs through the transfer of mobile genetic elements like SCCmec and genetic mutations. The analysis also compares hospital and community strains, including multidrug-resistant lineages like MRSA, VISA, and VRSA. The study concludes that S. aureus presents a major public health challenge, requiring new therapeutic approaches and preventive strategies.
Actinomycetes studies in Tunisia
Tunisia, located in North Africa, has a diverse topography along the Mediterranean Sea to the Sahara Desert. These environments encompass oases, rhizosphere soils, desert deposits, saline wetlands, offshore oilrigs, and ancient monument rocks. The country's varied environments have led to the isolation of a multitude of actinomycetes. A phylogenetic analysis based on the 16S rRNA sequences of one hundred isolated actinomycetes strains revealed that the majority belong to the genus Streptomyces. Secondary metabolite studies from these actinomycetes yielded 33 natural products. Notably, compound 12, 3-O-methylviridicatin, exhibited antitumor activity and suppressed HIV expression. This showcases Tunisia's potential for natural product research.
The roots of the Institut Pasteur's "Grand Cours"
The article presents an analysis of the history of the microbiology course delivered during the inaugural operational year of the Institut Pasteur in Paris. The year 1889 is examined through the lens of three hitherto unknown volumes that bring together the microbiology lectures delivered at the end of the 19th century. The course was entirely independent from the teaching provided by the universities or faculties of medicine and rapidly gained international recognition. Indeed, the course provided the students with the theoretical knowledge of Pasteurian theories regarding the completely new discipline of microbiology and the specific techniques used to cultivate, conserve, and observe microbes. The steady increase in the number of lectures between 1889 and 1914 reflects the expansion of microbiological knowledge during this period. The contributions of researchers such as Émile Roux (1853-1933), Élie Metchnikoff (1845-1916), and Amédée Borrel (1867-1936) illuminated the collaboration and the growing diversification of expertise at the heart of the Institut Pasteur (IP). Furthermore, this study highlights the international influence of the course, as evidenced by the participation of foreign students. It examines the history of the course as a powerful tool for disseminating knowledge of new microbiological techniques and the results of research carried out in Pasteur's laboratories. It also examines how the course served as a political instrument, asserting the authority of the Institut Pasteur in the field of microbiology in France and extending its influence worldwide.
Characterization and identification of Pseudomonas sp. AW4, an arsenic-resistant and plant growth-promoting bacteria isolated from the soybean (Glycine max L.) rhizosphere
Pseudomonas sp. AW4 is a highly arsenic (As) resistant bacterium with plant growth promoting properties, originally isolated from the soybean (Glycine max L.) rhizosphere. In order to safely use this isolate in diverse bioformulations, its characterization needs to be completed and a reliable identification must be provided. In the present work, we analyzed the morpho-physiological, biochemical and genomic characteristics of Pseudomonas sp. AW4. Identification of the isolate varied according to the parameters analyzed, mainly biochemical and physiological tests or individual genes and phylogenetic analyses. In this regard, we performed massive sequencing of its genome, in order to consistently complete its characterization and identification. Pseudomonas sp. AW4 formed a monophyletic clade with P. urmiensis SWRI10, presenting 3.08 % of unique genes against this reference isolate. More than 70 % of AW4 genes were also shared with P. oryziphila strain 1257 NZ and with P. reidholzensis strain CCOS 865. The search for genes related to As resistance evidenced the presence of the operon arsHRBC. Taken together, results of the present work allow identification of this bacterium as Pseudomonas urmiensis AW4 and open up a number of opportunities to study this strain and understand the mechanisms of arsenic resistance and plant growth promotion.
Novel insights into Bacillus thuringiensis: Beyond its role as a bioinsecticide
This review explores the diverse applications of Bacillus thuringiensis (Bt) beyond its traditional role as a bioinsecticide. Bt produces a variety of compounds with distinct chemical structures and biological activities. These include antimicrobial agents effective against plant pathogens and bioactive compounds that promote plant growth through the production of siderophores, hormones, and enzymes. Additionally, Bt's industrial potential is highlighted, encompassing biofuel production, bioplastics, nanoparticle synthesis, food preservation, anticancer therapies, and heavy metal bioremediation. This critical analysis emphasizes recent advancements and applications, providing insights into Bt's role in sustainable agriculture, biotechnology, and environmental management.
Cutibacterium acnes and its complex host interaction in prosthetic joint infection: Current insights and future directions
Cutibacterium acnes is a commensal Gram-positive anaerobic bacterium that can also act as an opportunistic pathogen in various diseases, particularly in prosthetic joint infections (PJI). Throughout this review, we delve into the current understanding of the intricate interactions between C. acnes and host cells and discuss bacterial persistence in the host. C. acnes colonization and subsequent PJI set-up represent complex processes involving bacterial adhesion, immune recognition, and host response mechanisms. We highlight existing knowledge and gaps in specific host-pathogen interactions and stress the importance of acquiring additional information to develop targeted strategies for preventing and treating C. acnes-related PIJ.
Influence of 5'-UTR nucleotide composition on translation efficiency in Escherichiacoli
Translation initiation for 5'-UTR contributes primarily to the efficient protein expression in Escherichia coli. Many studies have focused on constructing random 5'-UTR libraries to investigate the impact of mRNA features on protein translation efficiency. However, the study on the effect of the absence of specific types of nucleotides in the entire 5'-UTR region on translation efficiency has not yet been reported. Here, we constructed four reporter plasmid libraries encoding the sfGFP fluorescent protein, each preceded by 5'-UTRs that lack one specific nucleotide (25B, 25D, 25H, 25V). Each library was transformed into E. coli cells, and the fluorescence distribution among the different libraries was analyzed by flow cytometer. Additionally, we quantified the activity of 256 unique 5'-UTR sequences and analyzed the impact of the corresponding mRNA sequence features on translation efficiency. We found that the 25D library, which lacks the C nucleotide, exhibited the highest overall translation efficiency compared to the other three libraries. Moreover, the minimum free energy and 16S rRNA hybridization energy of the 5'-UTR sequence could work coordinately to influence translation efficiency. The 5'-UTR sequences lacking the C nucleotide also achieve efficient protein translation. These findings may provide several guiding principles for precisely tuning protein expression.
Characterization of tailocins of Pragia fontium 24613 and the tailocin loci within the family Budviciaceae
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.
Navigating dual-species fungal biofilms: The competitive and cooperative dynamics of Candidaalbicans
Research on microbial biofilms has primarily concentrated on bacterial-bacterial and bacterial-fungal interactions, leaving fungal-fungal dynamics underexplored. The present study examines interactions within dual-species fungal biofilms, with a particular emphasis on Candida albicans. The behavior and pathogenicity of this yeast are significantly influenced by its interactions with other fungal species in biofilms, where its ability to shift between yeast and hyphal forms contributes significantly to biofilm formation. These fungal species biofilms exhibit a complex interplay of synergistic cooperation and antagonistic competition, depending on the environmental context and resource availability. Understanding these interactions is essential for advancing our knowledge of fungal biofilm.
Targeting MurG enzyme in Klebsiella pneumoniae: An in silico approach to novel antimicrobial discovery
Antibiotic resistance poses a global crisis fuelled by widespread antibiotic use, particularly against Gram-negative bacteria like Klebsiella pneumoniae, a leading cause of hospital-acquired infections with high mortality rates. Urgent identification of effective drug targets is imperative, with a focus on metabolic pathways to inhibit bacterial growth. Targeting the crucial metabolic pathways of K. pneumoniae would be a more efficient way to prevent its growth and the diseases that it causes. The present study focused on inhibiting the UDP-N-acetylglucosamine--N-acetylmuramyl-(pentapeptide)pyrophosphoryl-undecaprenol N-acetylglucosamine transferase (MurG) enzyme, which is a key enzyme in peptidoglycan biosynthesis pathway. A high throughput virtual screening was used to find possible lead molecules from Enamine -High-Throughput Screening Center library. The resulting high binding affinity ligands were further assessed for their drug-likeness and other pharmacokinetic properties. Based on these analyses, the three ligands Z95813755_1, Z324718246_1 and Z324718246_2 were selected for further molecular dynamic simulation studies. The molecular dynamic simulation results and MM/PBSA analysis predicted that both Z95813755_1 and Z324718246_2, molecules show higher binding affinity towards MurG. For the first time we are reporting potential candidate inhibitors against MurG from K. pneumoniae, providing new insights in management of multi drug resistant K. pneumoniae infections.
Metabolic complexities and heterogeneity in quorum sensing signaling molecules in bacteria isolated from black band disease in a Caribbean coral
Coral diseases contribute to the worldwide loss of coral reefs, with the Black Band Disease (BBD) being a prominent example. BBD is an infectious condition with lesions with a pigmented mat composed of cyanobacteria, sulphate-reducing, sulphide-oxidizing, and heterotrophic bacteria. We compared the heterotrophic bacterial communities of healthy and BBD-affected colonies of the Caribbean coral Orbicella faveolata using culture-dependent and -independent techniques. Twenty and 23 bacterial isolates were identified from healthy and diseased tissues, respectively, which differed in their capacities to metabolize carbohydrates and citrate, either anaerobically or aerobically. They also differed in their quorum-sensing (QS) activity, as QS signaling molecules were found exclusively, and QS-inhibition was found primarily, in isolates from diseased tissues. Screening of bacterial diversity by 16SrDNA metabarcoding showed that members of the bacterial genera Muricauda and Maritimimonas were dominant in healthy tissues whereas members of the cyanobacterial genus Roseofilum were dominant in diseased tissues. These results suggest that bacterial dysbiosis can be linked with altered bacterial communication, likely leading to diachrony and imbalance that may participate in the progression of BBD. Investigating physiological traits and QS-based communication offers insights into the onset and progression of coral infections, paving the way for novel strategies to mitigate their impact.
Both GacS-regulated lipopeptides and the type three secretion system contribute to Pseudomonas cichorii induced necrosis in lettuce and chicory
Pseudomonas cichorii SF1-54, the causal agent of lettuce midrib rot disease, produces lipopeptides cichofactins and cichopeptins which are important virulence factors. The GacS/GacA two-component system is well known to regulate production of lipopeptides in pseudomonads. Additionally, the functions of the type three secretion system (T3SS) in P. cichorii-plant interactions are not clarified. In this study, we investigated the role of the GacS-regulated lipopeptides and the T3SS in pathogenicity of P. cichorii SF1-54 on two host plants, chicory and lettuce, by constructing mutants in hrpL, which encodes the key sigma factor to control T3SS expression, and gacS. Compared with the wildtype, the hrpL mutant produced lipopeptides at a similar level but the gacS mutant was strongly impaired in lipopeptide production. The mutant deficient in hrpL did not significantly differ from the wildtype in virulence on chicory and lettuce. The gacS mutant exhibited significantly less symptoms on both host plants compared to the wildtype and the hrpL mutant. Intriguingly, the gacS hrpL-double mutant no longer produced lipopeptides, lost virulence and showed impaired colonization on chicory, but was still weakly virulent on lettuce. Thus, contribution of both the GacS-regulated lipopeptides and T3SS to virulence of P. cichorii SF1-54 is host plant dependent.
Characterisation of a new virulent phage isolated from Hainan Island with potential against multidrug-resistant Pseudomonas aeruginosa infections
Multidrug-resistant (MDR) Pseudomonas aeruginosa is a serious life-threatening pathogen. The rise in P. aeruginosa resistance rates has renewed interest in phages as an alternative therapeutic approach for treating bacterial infections. In this study, we investigated the characteristics of the first Pseudomonas phage, vB_PaP_HN01, isolated from Hainan, the only tropical island in China. The lytic rate of this phage against P. aeruginosa reached 64.3 % (27/42). Under the optimal multiplicity of infection (MOI) of 0.1, more than 90 % of phage particles absorb onto the host cell within 10 min, with an eclipse period of around 15 min, and a high titer phage production (10 PFU/ml) within 90 min was demonstrated. vB_PaP_HN01 maintains a robust titer after 1 h exposure to pH values and temperatures (up to 50 °C). Genome annotation revealed that vB_PaP_HN01 did not contain drug-resistance or lysogeny-associated genes. It can effectively inhibit the formation of biofilms of MDR P. aeruginosa and eliminated aggressive biofilms (removal rate about 70 %). In the in vivo infection models, it was demonstrated that the survival rate and lifespan of Galleria mellonella larvae were increased alongside the injection of vB_PaP_HN01. These data revealed the potential of vB_PaP_HN01 against P. aeruginosa in clinic.
Evolutionary trends indicate a coherent organization of sap operons
Human hosts possess a complex network of immune responses against microbial pathogens. The production of antimicrobial peptides (AMPs), which target the pathogen cell membranes and inhibit them from inhabiting the hosts, is one such mechanism. However, pathogens have evolved systems that encounter these host-produced AMPs. The Sap (sensitivity to antimicrobial peptides) transporter uptakes AMPs inside the microbial cell and proteolytically degrades them. The Sap transporters comprise five subunits encoded by genes in an operon. Despite its ubiquitous nature, its subunits are not found to be in tandem with many organisms. In this study, a total of 421 Sap transporters were analyzed for their operonic arrangement. Out of 421, a total of 352 operons were found to be in consensus arrangement, while the remaining 69 show a varying arrangement of genes. The analysis of the intergenic distance between the subunits of the sap operon suggests a signature pattern with sapAB (-4), sapBC (-14), sapCD (-1), and sapDF (-4 to 1). An evolutionary analysis of these operons favors the consensus arrangement of the Sap transporter systems, substantiating its prevalence in most of the Gram-negative pathogens. Overall, this study provides insight into bacterial evolution, favoring the maintenance of the genetic organization of essential pathogenicity factors.
Endophyte mediated biocontrol mechanisms of phytopathogens in agriculture
The global human population is growing and demand for food is increasing. Global agriculture faces numerous challenges, including excessive application of synthetic pesticides, emergence of herbicide-and pesticide-resistant pathogenic microbes, and more frequent natural disasters associated with global warming. Searches for valuable endophytes have increased, with the aim of making agriculture more sustainable and environmentally friendly. Endophytic microbes are known to have a variety of beneficial effects on plants. They can effectively transfer nutrients from the soil into plants, promote plant growth and development, increase disease resistance, increase stress tolerance, prevent herbivore feeding, reduce the virulence of pathogens, and inhibit the growth of rival plant species. Endophytic microbes can considerably minimize the need for agrochemicals, such as fertilizers, fungicides, bactericides, insecticides, and herbicides in the cultivation of crop plants. This review summarizes current knowledge on the roles of endophytes focusing on their mechanisms of disease control against phytopathogens through the secretion of antimicrobial substances and volatile organic compounds, and the induction of systemic resistance in plants. Additionally, the beneficial roles of these endophytes and their metabolites in the control of postharvest diseases in plants have been summarized.
Effect of gamma irradiation on the proteogenome of cold-acclimated Kocuria rhizophila PT10
The effects of ionizing radiation (IR) on the protein dynamics of cold-stressed cells of a radioresistant actinobacterium, Kocuria rhizophila PT10, isolated from the rhizosphere of the desert plant Panicum turgidum were investigated using a shotgun methodology based on nanoflow liquid chromatography coupled to tandem mass spectrometry. Overall, 1487 proteins were certified, and their abundances were compared between the irradiated condition and control. IR of cold-acclimated PT10 triggered the over-abundance of proteins involved in (1) a strong transcriptional regulation, (2) amidation of peptidoglycan and preservation of cell envelope integrity, (3) detoxification of reactive electrophiles and regulation of the redox status of proteins, (4) base excision repair and prevention of mutagenesis and (5) the tricarboxylic acid (TCA) cycle and production of fatty acids. Also, one of the more significant findings to emerge from this study is the SOS response of stressed PT10. Moreover, a comparison of top hits radio-modulated proteins of cold-acclimated PT10 with proteomics data from gamma-irradiated Deinococcus deserti showed that stressed PT10 has a specific response characterised by a high over-abundance of NemA, GatD, and UdgB.
Lipid lysination by MprF contributes to hemolytic pigment retention in group B Streptococcus
Group B Streptococcus (GBS) is the leading cause of neonatal sepsis and meningitis. A major virulence factor is a pigmented beta-haemolytic/cyto-lysin (β-h/c) toxin with an ornithine rhamnolipid structure. We initially observed that absence of MprF enzyme altered pigmentation and haemolytic activity in GBS. Next, we showed that MprF-dependent lipid lysination contributes to the retention of the ornithine rhamnolipid within GBS membrane. Furthermore, cationic lipidation by MprF altered membrane properties contributing to resistance to the cyclic lipopeptide daptomycin and to acidic pH. This study highlights the importance of cationic lipids in cell envelope homeostasis and in modulating β-h/c activity.
BSC2 modulates AmB resistance via the maintenance of intracellular sodium/potassium ion homeostasis in Saccharomyces cerevisiae
Previous studies on BSC2 have shown that it enhances yeast cell resistance to AmB via antioxidation and induces multidrug resistance by contributing to biofilm formation. Herein, we found that BSC2 overexpression could reverse the sensitivity of pmp3Δ to AmB and help the tested strains restore the intracellular sodium/potassium balance under exposure to AmB. Meanwhile, overexpression of the chitin gene CHS2 could simulate BSC2 to reverse the sensitivity of pmp3Δ and nha1Δ to high salt or AmB. However, BSC2 overexpression in flo11Δ failed to induce AmB resistance, form biofilms, and affect cell wall biogenesis, while CHS2 overexpression compensated the resistance of flo11Δ to AmB. Additionally, BSC2 levels were positively correlated with maintaining cell membrane integrity under exposure to AmB, CAS, or a combination of both. BSC2 overexpression in nha1Δ exhibited a similar function of CHS2, which can compensate for the sensitivity of the mutant to high salt. Altogether, the results demonstrate for the first time that BSC2 may promote ion equilibrium by strengthening cell walls and inhibiting membrane damage in a FLO path-dependent manner, thus enhancing the resistance of yeast cells to AmB. This study also reveals the possible mechanism of antifungal drugs CAS and AmB combined to inhibit fungi.
Construction of core genome multi-locus sequence typing schemes for population structure analyses of Nocardia species
Nocardia, a member of the Actinobacteria phylum, populates diverse habitats globally, with certain species being the cause of various clinical infections in humans. There is paucity of data regarding the population structure of this genus and of established genomic-based phylogenetic methods. We examined the whole genome sequences of 193 isolates spanning five major pathogenic Nocardia species sourced from public databases, encompassing diverse geographic regions. Using the chewBBACA pipeline, a species-specific core genome multilocus sequence typing (cgMLST) schema was created for N. cyriacigeorgica, N. farcinica, N. brasiliensis, N. wallacei, and N. abscessus. Additional genomic features that were examined included virulence factor (VF) profile, total length and open-reading frame count, the core genome length and core gene count, and GC content. Our findings indicated that: (i) N. brasiliensis diverges significantly from the other four species, underscoring its distinct evolutionary trajectory; (ii) the population structures of all species were polyclonal, with phylogenetic clustering occurring in the minority of isolates; (iii) clonal complexes were largely restricted to specific geographical locations, rather than demonstrating a global distribution, and (iv) initial evidence suggests no direct common-source transmission amongst the studied strains. Our study establishes a comprehensive genome-based phylogenetic methodology for population structure of Nocardia species.
Biosynthetic capabilities of Antarctic yeast Sporobolomyces roseus AL: Temperature influence on intracellular metabolites and characterization of the exopolysaccharide
The purpose of the study was to investigate the biosynthetic properties of the Antarctic yeast strain Sporobolomyces roseus AL in response to temperature changes, to perform intracellular metabolic profiling, and to reveal the chemical and functional characteristics of the synthesized exopolysaccharide (EPS). The results show that the yeast strain needed a shorter time to reach a stationary phase at 22 °C contrary to that of 5 °C. An NMR analysis revealed differences in metabolic profiles of amino acids, glucose, trehalose, glycerol, uridine, etc. EPS (2.9 g/L) was characterized by high-molecular-weight with total carbohydrate, uronic acids, and protein content of 66 %, 10.5 %, and 2.5 %, respectively. Mannose (74 mol%) and galactose (19 mol%) were the major constituents. The FT-IR data suggested the presence of β-(1-4)-mannan. DSC thermogram, WVTR, mechanical properties, and moisture sorption of the EPS film showed thermal stability up to 220 °C and hydrophilic behavior. The newly obtained polymer film was studied for the first time and the data showed possibilities for its successful application as a film-forming material in the preparation of packaging materials. In conclusion, the temperature influenced the metabolic profile of the Antarctic yeast producer. The biotechnological process could be directed to obtain the target intracellular or extracellular metabolites.