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The emergence of antibiotic resistance has been raised due to the inappropriate use of antibiotics that might be the urgent health crisis. Antibiotics in animals and food chain have been widely used as growth promoters, prophylaxis, and treatment of infectious diseases, which provides the selective pressure on foodborne pathogens. It is essential to develop novel therapeutic weapon for controlling the appearance of resistance and inhibiting imminent threat of pandemics possible by means of multidrug resistant (MDR) pathogens. Nowadays, the control of MDR pathogens has been improved with the development of bacteriophage (phage)-based approaches such as single phage therapy, synergistic approach, phage cocktail, engineered phage, and anti-clustered regularly interspaced short palindromic repeats (anti-CRISPR) system. This review overwhelmingly focuses on slash the emergence of food-borne pathogens and its bacterial resistances using phage components and also discusses on the impact of phage components to diminish the outbreak of viral infections.

Stan Falkow looked at the world with his eyes peering from the outer membrane of a gram-negative bacterium. It was a great vantage point from which to dream about the possible superpowers these organisms might have. He mused about these dreams with his trainees who sometimes found through rigorous scientific exploration that the superpowers really were there! Stan also realized that bacterial pathogenesis by definition was a two-way street, and that masterful understanding of bacterial virulence factors also required a masterful understanding of host cell processes against which the virulence factors were deployed. In my own scientific journey, I have sought to explore bacterial-host interactions that result in subtle alterations of the host's adaptive immune response. Here, as an example, I describe an interaction between Chlamydia trachomatis and host T cells that may contribute to the establishment of persistent infection.

Bacterial membrane vesicles (MVs) mediate bacterial virulence by enabling secretion and long distance delivery of bacterial effector molecules. Staphylococcus haemolyticus has now been demonstrated to produce membrane vesicles (MVs). The protein content of S. haemolyticus MVs was identified by Mass spectrometry and compared to proteins identified in the total secretome. This information is presented in this data article. Further background and interpretation of the data can be found in the article: Comparative exoproteome profiling of an invasive and a commensal S. haemolyticus isolate (Cavanagh et al., in press). Data are available via Proteome Xchange with identifier PXD010389.

SUMMARYInvestigation of plant-bacteria interactions requires quantification of in planta bacterial titers by means of colony counting assays. However, colony counting assays are cumbersome and time-consuming, and are unable to detect spatial patterns of bacterial colonization in plants. Here, to overcome these shortcomings, we devised a broadly applicable genetic engineering tool for bioluminescence-based quantitative and spatial detection of bacteria in plants. We developed plasmid vectors that have broad host ranges and enable Tn7 transposon-mediated integration of the luxCDABE luciferase operon into a specific genomic location ubiquitously found across bacterial phyla. These vectors allowed for generation of bioluminescent transformants of various plant pathogenic bacteria belonging to the genera Pseudomonas, Rhizobium (Agrobacterium), and Ralstonia. The bioluminescent transformant of Pseudomonas syringae pv. tomato DC3000 (Pto-lux) was as virulent in Arabidopsis thaliana as its parental strain. Direct luminescence measurements of Pto-lux-inoculated plant tissues reported bacterial titers in A. thaliana, Solanum lycopersicum, Nicotiana benthamiana, and Marchantia polymorpha as accurately as conventional colony counting assays. We further showed the utility of our vectors for converting the previously generated Pto derivatives to isogenic bioluminescent strains. Importantly, quantitative bioluminescence assays using these Pto-lux strains accurately reported the effects of plant immunity and bacterial effectors on bacterial growth with a dynamic range of 4 orders of magnitude. Moreover, macroscopic bioluminescence imaging illuminated spatial colonization patterns of the Pto-lux in/on inoculated plant tissues. Taken together, our vectors offer untapped opportunities for developing bioluminescence-based quantitative and spatial analysis of bacterial growth in a variety of plant-bacteria interactions.SIGNIFICANCE STATEMENTWe developed broad-host-range plasmid vectors that integrate the luciferase operon, luxCDABE, into a specific genomic location ubiquitously found across bacterial phyla. Using these vectors, we established a high-throughput method for bioluminescence-based quantitative assays of in planta bacterial growth with a dynamic range of 4 orders of magnitude and visualized spatiotemporal patterns of bacterial colonization in/on inoculated plant tissues.

New classes of antimicrobials that are effective therapies for infections with multi-drug resistant pathogens are urgently needed. The nematode Caenorhabditis elegans has been incorporated into small molecule screening platforms to identify anti-infective compounds that provide protection of a host during infection. The use of a live animal in these screening systems offers several advantages, including the ability to identify molecules that boost innate immune responses in a manner advantageous to host survival and compounds that disrupt bacterial virulence mechanisms. In addition, new classes of antimicrobials that target the pathogen have been uncovered, as well as interesting chemical probes that can be used to dissect new mechanisms of host-pathogen interactions.

The glyoxylate shunt bypasses the oxidative decarboxylation steps of the tricarboxylic acid (TCA) cycle, thereby conserving carbon skeletons for gluconeogenesis and biomass production. In Escherichia coli, carbon flux is redirected through the first enzyme of the glyoxylate shunt, isocitrate lyase (ICL), following phosphorylation and inactivation of the TCA cycle enzyme, isocitrate dehydrogenase (ICD), by the kinase/phosphatase, AceK. In contrast, mycobacterial species lack AceK and employ a phosphorylation-insensitive isocitrate dehydrogenase (IDH), which is allosterically activated by the product of ICL activity, glyoxylate. However, Pseudomonas aeruginosa expresses IDH, ICD, ICL, and AceK, raising the question of how these enzymes are regulated to ensure proper flux distribution between the competing pathways. Here, we present the structure, kinetics, and regulation of ICL, IDH, and ICD from P. aeruginosa. We found that flux partitioning is coordinated through reciprocal regulation of these enzymes, linking distribution of carbon flux to the availability of the key gluconeogenic precursors, oxaloacetate and pyruvate. Specifically, a greater abundance of these metabolites activated IDH and inhibited ICL, leading to increased TCA cycle flux. Regulation was also exerted through AceK-dependent phosphorylation of ICD; high levels of acetyl-CoA (which would be expected to accumulate when oxaloacetate is limiting) stimulated the kinase activity of AceK, whereas high levels of oxaloacetate stimulated its phosphatase activity. In summary, the TCA cycle–glyoxylate shunt branch point in P. aeruginosa has a complex enzymology that is profoundly different from those in other species characterized to date. Presumably, this reflects its predilection for consuming fatty acids, especially during infection scenarios.

Transition metals are required cofactors for many proteins that are critical for life, and their concentration within cells is carefully maintained to avoid both deficiency and toxicity. To defend against bacterial pathogens, vertebrate immune proteins sequester metals, in particular zinc, iron, and manganese, as a strategy to limit bacterial acquisition of these necessary nutrients in a process termed “nutritional immunity.” In response, bacteria have evolved elegant strategies to access metals and counteract this host defense. In mammals, metal abundance can drastically shift due to changes in dietary intake or absorption from the intestinal tract, disrupting the balance between host and pathogen in the fight for metals and altering susceptibility to disease. This review describes the current understanding of how dietary metals modulate host-microbe interactions and the subsequent impact on the outcome of disease.

The aim of the study was to determine the prevalence of virulence genes responsible for the adhesion (flaA, cadF and racR) and invasion (virB11, iam and pldA) in Campylobacter isolates from cattle and swine and determine their adherence and invasion abilities. The studies conducted revealed high prevalence rate of adherence and invasion associated genes irrespective of the isolates origin. All Campylobacter strains of swine and cattle origin adhered to HeLa cells at mean level 0.1099% ± SD 0.1341% and 0.0845% ± SD 0.1304% of starting viable inoculum, respectively. However swine isolates exhibited higher invasion abilities (0.0012% ± SD 0.0011%) compared to bovine isolates (0.00038% ± SD 0.00055%). The results obtained revealed significantly positive correlation between invasion and adherence abilities of swine origin isolates (R = 0.4867 in regard to C. jejuni and R = 0.4507 in regard to C. coli) and bovine origin isolates (R = 0.726 in regard to C. jejuni). Bacterial virulence is multifactorial and it is affected by the expression of virulence genes. Moreover the presence of virulence genes determines the ability of Campylobacter isolates to adhere and invade the cells.

Highlights • Bacterial Pro-Pro-endopeptidase (PPEP) is the latest member of the metalloendopeptidase class (E.C. 3.4.24.89). • PPEP homologs are found in two firmicutes orders, clostridiales and bacillales spread over 9 genera and more than 130 species. • Some PPEP homologs have acquired additional anchor domains that bind noncovalently to various elements of the bacterial peptidoglycan cell wall. • Prototype family members, PPEP-1 and PPEP-2, target bacterial surface adhesion proteins, but homologs could target other extracellular proteins., Pro-Pro-endopeptidases (PPEP, EC 3.4.24.89) are secreted, zinc metalloproteases that have the unusual capacity to cleave a peptide bond between two prolines, a bond that is generally less sensitive to proteolytic cleavage. Two well studied members of the family are PPEP-1 and PPEP-2, produced by Clostridioides difficile, a human pathogen, and Paenibacillus alvei, a bee secondary invader, respectively. Both proteases seem to be involved in mediating bacterial adhesion by cleaving cell surface anchor proteins on the bacterium itself. By using basic alignment and phylogenetic profiling analysis, this work shows that the complete family of proteins that contain a PPEP domain includes proteins from more than 130 species spread over 9 genera. These analyses also suggest that the PPEP domain spread through horizontal gene transfer events between species within the Firmicutes’ classes Bacilli and Clostridia. Bacterial species containing PPEP homologs are found in diverse habitats, varying from human pathogens and gut microbiota to free-living bacteria, which were isolated from various environments, including extreme conditions such as hot springs, desert soil and salt lakes. The phylogenetic tree reveals the relationships between family members and suggests that smaller subgroups could share cleavage specificity, substrates and functional similarity. Except for PPEP-1 and PPEP-2, no cleavage specificity, specific physiological target, or function has been assigned for any of the other PPEP-family members. Some PPEP proteins have acquired additional domains that recognize and bind noncovalently to various elements of the bacterial peptidoglycan cell-wall, anchoring these PPEPs. Secreted or anchored to the cell-wall surface PPEP proteins seem to perform various functions., Graphical abstract Image, graphical abstract

Methylation of outer membrane proteins (OMPs) has been implicated in bacterial virulence. Lysine methylation in rickettsial OmpB is correlated with rickettsial virulence, and N- and O-methylations are also observed in virulence-relevant OMPs from several pathogenic bacteria that cause typhus, leptospirosis, tuberculosis, and anaplasmosis. We summarize recent findings on the structure of methylated OmpB, biochemical characterization, and crystal structures of OmpB methyltransferases. Native rickettsial OmpB purified from highly virulent strains contains multiple clusters of trimethyllysine, in contrast with mostly monomethyllysine, and no trimethyllysine is found in an avirulent strain. Crystal structure of the methyltransferases reveals mechanistic insights for catalysis, and a working model is discussed for this unusual post-translational modification.

The present study has examined the virulence potential of Escherichia coli isolates harboring at least one virulence gene (associated with ExPEC or InPEC pathotype and belonging to different phylogenetic groups: A, B1, B2 or D), isolated from free ranging Tibetan yak feces. The E. coli isolates (n = 87) were characterized for different serogroups and a mouse model of subcutaneous–infection was used to envisage the virulence within these E. coli strains. Of the 87 E. coli isolates examined, 23% of the E. coli isolates caused lethal infections in a mouse model of subcutaneous infection and were classified as killer. Moreover, the majority of the killer strains belonged to phylogroup A (65%) and serogroup O60 or O101 (35%). Phylogroup B1, serogroups O60 and O101 were statistically associated with the killer status (P 1) were observed between the killer status isolates and all other bacterial virulence traits. This study comprises the first report on the virulence potential of E. coli strains isolated from free-ranging Tibetan yaks feces. Our findings suggest that pathogenic E. coli of free ranging yaks is highly worrisome, as these feces are used as manures by farmers and therewith pose a health risk to humans upon exposure.

The bacterial communication through signaling molecules termed quorum sensing has gained a lot of attention due to its applicability to different fields of study. Quorum sensing regulates several bacterial activities such as biofilm formation, antibiotic resistance, bioluminescence and bacterial virulence. These coordinated activities are only realized when the bacterial population reaches a high population density. The signaling molecules used in quorum sensing occur at very low concentrations which call for very sensitive methods for their identification and characterization. Many studies have been performed on the use of inhibitors in attempt to disrupt quorum sensing and consequently alleviate its negative effects. This article gives a description of the types of auto-inducers involved in quorum sensing and the fields of study they greatly affect especially the environmental one. It also covers types of analytical sample preparation and detection methods, and different types of inhibitors used for quorum quenching.

The continued rise of antibiotic-resistant bacterial infections has motivated alternative strategies for target discovery and treatment of infections. Antivirulence therapies function through inhibition of in vivo required virulence factors to disarm the pathogen instead of directly targeting viability or growth. This approach to treating bacteria-mediated diseases may have advantages over traditional antibiotics because it targets factors specific for pathogenesis, potentially reducing selection for resistance and limiting collateral damage to the resident microbiota. This review examines vulnerable molecular mechanisms used by bacteria to cause disease and the antivirulence compounds that sabotage these virulence pathways. By expanding the study of antimicrobial targets beyond those that are essential for growth, antivirulence strategies offer new and innovative opportunities to combat infectious diseases.

1. Department of Genetics, Ecology and Evolution (GEE), Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. 2. Institute of Health Sciences, Federal University of Bahia, Salvador, BA, Brazil. 3. Department of Microbiology, Immunology and Parasitology, Institute of Biological and Natural Sciences, Federal University of Triângulo Mineiro, Uberaba, MG, Brazil. 4. Laboratory of Clinical Pathology, Hospital das Clinicas, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. 5. Tiradentes University, Technology and Research Institute, Biomaterials Laboratory, Aracaju, Sergipe, Brazil. 6. Swedish University of Agricultural Sciences, Uppsala, Sweden. 7. Department of Computer Science, Virginia Commonwealth University, Richmond, Virginia, 23284, USA. 8. Institute of Integrative Omics and Applied Biotechnology (IIOAB), Nonakuri, Purba Medinipur, West Bengal 721137, India. 9. Molecular and Computational Biology of Fungi Laboratory, Department of Microbiology, Institute of Biological Sciences, Federal University of Minas Gerais, Belo Horizonte, Minas Gerais, Brazil. 10. AQUACEN – National Reference Laboratory of Aquatic Animal Disease, Ministry of Agriculture, Livestock and Food Supply, Veterinary School, Federal University of Minas Gerais, Belo Horizonte, MG, Brazil. 11. Institute of Veterinary Medicine, University of Gottingen, Burckhardtweg 2, Gottingen, Germany. 12. Goncalo Moniz Institute, Oswaldo Cruz Foundation, Salvador, Bahia, Brazil.

The looming antibiotic crisis has prompted the development of new strategies towards fighting infection. Traditional antibiotics target bacterial processes essential for viability, whereas proposed antivirulence approaches rely on the inhibition of factors that are required only for the initiation and propagation of infection within a host. Although antivirulence compounds have yet to prove their efficacy in the clinic, bacterial signal peptidase I (SPase) represents an attractive target in that SPase inhibitors exhibit broad-spectrum antibiotic activity, but even at sub-MIC doses also impair the secretion of essential virulence factors. The potential consequences of SPase inhibition on bacterial virulence have not been thoroughly examined, and are explored within this review. In addition, we review growing evidence that SPase has relevant biological functions outside of mediating secretion, and discuss how the inhibition of these functions may be clinically significant.

Inflammation, an old medical problem, is being recognized as an active, well orchestrated biological process. When dysregulated, chronic inflammation may ensue, leading to tissue-dependent diseases. Depending upon the ligand and cellular context, aryl hydrocarbon receptor (AHR) may accelerate or attenuate inflammation and subsequent resolution. Three examples are discussed in which AHR modulates inflammation by a mixture of genomic and non-genomic signaling pathways: (i) AHR-agonistic bacterial virulence factors are leading to both microbial defense and resolution of inflammatory responses. (ii) TCDD-mediated persistent AHR activation initially leads to inflammation by non-genomic signaling, and may potentially lead to chronic inflammation. (iii) AHR may modulate anti-inflammatory actions in obesity-mediated non-alcoholic fatty liver disease (NAFLD): Hepatic lipotoxicity triggers generation of danger-associated molecular patterns (DAMPs) that facilitate the development of hepatitis. AHR is mainly involved in the resolution phase by induction of lipoxin A4 and Il-22. Moderate AHR activation by phytochemicals and microbial AHR ligands may facilitate resolution. In control of inflammation, AHR appears to integrate environmental conditions with coordinated cellular functions.

International journal of medical microbiology : IJMM 310(2), 151392 (2020). doi:10.1016/j.ijmm.2020.151392, Published by Elsevier, München

Bacteriophages are types of viruses that infect bacteria. They are the most abundant and diverse entities in the biosphere, and influence the evolution of most bacterial species by promoting gene transfer, sometimes in unexpected ways. Although pac-type phages can randomly package and transfer bacterial DNA by a process called generalized transduction, some mobile genetic elements have developed elegant and sophisticated strategies to hijack the phage DNA-packaging machinery for their own transfer. Moreover, phage-like particles (gene transfer agents) have also evolved, that can package random pieces of the producing cell's genome. The purpose of this review is to give an overview of some of the various ways by which phages and phage-like particles can transfer bacterial genes, driving bacterial evolution and promoting the emergence of novel pathogens.

Neutrophil serine proteases (NSPs) are critical for the effective functioning of neutrophils and greatly contribute to immune protection against bacterial infections. Thanks to their broad substrate specificity, these chymotrypsin-like proteases trigger multiple reactions that are detrimental to bacterial survival such as direct bacterial killing, generation of antimicrobial peptides, inactivation of bacterial virulence factors and formation of neutrophil extracellular traps. Recently, the importance of NSPs in antibacterial defenses has been further underscored by discoveries of unique bacterial evasion strategies to combat these proteases. Bacteria can indirectly disarm NSPs by protecting bacterial substrates against NSP cleavage, but also produce inhibitory molecules that potently block NSPs. Here we review recent insights in the functional contribution of NSPs in host protection against bacterial infections and the elegant strategies that bacteria use to counteract these responses.

Single-nucleotide polymorphisms (SNPs) are the most common source of genetic variation within a species; however, few investigations demonstrate how naturally occurring SNPs may increase strain virulence. We recently used group A Streptococcus as a model pathogen to study bacteria strain genotype-patient disease phenotype relationships. Whole-genome sequencing of approximately 800 serotype M59 group A Streptococcus strains, recovered during an outbreak of severe invasive infections across North America, identified a disproportionate number of SNPs in the gene encoding multiple gene regulator of group A Streptococcus (mga). Herein, we report results of studies designed to test the hypothesis that the most commonly occurring SNP, encoding a replacement of arginine for histidine at codon 201 of Mga (H201R), significantly increases virulence. Whole transcriptome analysis revealed that the H201R replacement significantly increased expression of mga and 54 other genes, including many proven virulence factors. Compared to the wild-type strain, a H201R isogenic mutant strain caused significantly larger skin lesions in mice. Serial quantitative bacterial culture and noninvasive magnetic resonance imaging also demonstrated that the isogenic H201R strain was significantly more virulent in a nonhuman primate model of joint infection. These findings show that the H201R replacement in Mga increases the virulence of M59 group A Streptococcus and provide new insight to how a naturally occurring SNP in bacteria contributes to human disease phenotypes. Copyright © 2015 American Society for Investigative Pathology.

SummaryOtitis media (OM) is a public health problem in both developed and developing countries. It is the leading cause of hearing loss and represents a significant healthcare burden. In some cases, acute OM progresses to chronic suppurative OM (CSOM), characterized by effusion and discharge, despite antimicrobial therapy. The emergence of antibiotic resistance and potential ototoxicity of antibiotics has created an urgent need to design non-conventional therapeutic strategies against OM based on modern insights into its pathophysiology. In this article, we review the role of innate immunity as it pertains to OM and discuss recent advances in understanding the role of innate immune cells in protecting the middle ear. We also discuss the mechanisms utilized by pathogens to subvert innate immunity and thereby overcome defensive responses. A better knowledge about bacterial virulence and host resistance promises to reveal novel targets to design effective treatment strategies against OM. The identification and characterization of small natural compounds that can boost innate immunity may provide new avenues for the treatment of OM. There is also a need to design novel methods for targeted delivery of these compounds into the middle ear, allowing higher therapeutic doses and minimizing systemic side effects.

Equine infection with Corynebacterium pseudotuberculosis can manifest in several forms, including external or internal abscesses. The objective of this study was to phenotype clinical isolates of C. pseudotuberculosis and to investigate the relationship between lesion location and extent of lesions in the animals from which they were collected. One hundred and seventy-one C. pseudotuberculosis biovar equi isolates were collected from horses presenting to the University of California Veterinary Medical Teaching Hospital and two other sources in the period between September 1996 and December 2011. Bacterial isolates were grouped on the bases of biochemical characteristics and growth on brain heart infusion agar. Six phenotypes were identified: (1) large colonies that metabolized sucrose (n = 81); (2) large sucrose-negative colonies (n = 47); (3) medium sucrose-positive (n = 20); (4) medium sucrose-negative (n = 11); (5) small sucrose-positive (n = 7), and (6) small sucrose-negative (n = 5). Medical records corresponding to each isolate were accessed from the University's administrative computer system or from the submitting source in order to determine the anatomical site from which the isolate was collected (n = 171), as well as the extent of lesions (n = 164) in the patient. The relationship between phenotype, lesion location and extent of lesions was then investigated statistically. No significant relationship between strain and lesion location or extent of lesions was found. This suggests that phenotypic differences during in vitro culture does not account for external versus internal disease in horses. Further work to characterize strains genotypically and to identify determinants for bacterial virulence should be performed. Importantly, host and environmental factors should also be further investigated.

The spread of multidrug-resistant bacteria has accelerated sharply in the last decade. According to the World Health Organization they are responsible for an estimated 25 000 deaths in Europe each year. In addition, few new antibiotics are under development, raising the spectrum of a return to the "pre-antibiotic era". Non antibiotic antibacterial agents have recently attracted renewed interest. The most promising candidates are: i) phages (bacteria-infecting viruses) have been widely used in Eastern European countries since the 1930s but come up against logistic and regulatory obstacles due to the evolutionary nature of these biologic agents, while convincing clinical data are lacking; ii) bacteriocines are smallantibacterialpeptidesproducedby numerous bacteria; some have a rapid bactericidal effect, good tolerability, and a limited impact on the commensal flora; however, clinical use of bacteriocines is complicated by their fragility, poor penetration, and substantial risk of resistance selection ; iii) antisense oligonucleo tides act by inactivating genes through specific interaction with a complementary DNA or RNA fragment, potentially allowing specific inhibition of selected bacterial virulence factors. However, this therapeutic class may be more suitable for viral or genetic diseases than for multidrug-resistant bacterial infections, owing to the difficulty of delivering them inside bacteria.

Highlights • In vitro screens have very low success rate in follow up preclinical trials. • In vitro systems do not reflect the complexity of the host. • The main advantage of Drosophila is its significant similarity with humans. • Drug screens in Drosophila may increase the success of preclinical trials., Following an expansion in the antibiotic drug discovery in the previous century, we now face a bottleneck in the production of new anti-infective drugs. Traditionally, chemical libraries are screened either using in vitro culture systems or in silico to identify and chemically modify small molecules with antimicrobial properties. Nevertheless, almost all compounds passing through in vitro screening fail to pass preclinical trials. Drug screening in Drosophila offers to fill the gap between in vitro and mammalian model host testing by eliminating compounds that are toxic or have reduced bioavailability and by identifying others that may boost innate host defence or selectively reduce microbial virulence in a whole-organism setting. Such alternative screening methods in Drosophila, while low-throughput, may reduce the cost and increase the success rate of preclinical trials.

Shigella, the causative agent of bacillary dysentery, invades the colonic epithelium where it elicits an intense inflammation leading to tissular destruction. Key to bacterial virulence, type III effectors injected into host cells reorganize the actin cytoskeleton and regulate inflammatory responses. Much progress has been made recently in the characterization of these type III effectors. These findings have reshaped our view of Shigella invasion, suggesting a strategy to invade epithelial cells 'discretely' as an initial route of invasion, contrasting with the devastating inflammatory response associated with the disease's acute phase. The diverse roles of Shigella type III effectors highlight the complexity of an infection process where as little as a thousand bacteria are estimated sufficient to cause the disease in humans.

Introduction Recent studies have reported the color change and formation of precipitates containing para-chloroaniline (PCA) after a reaction of sodium hypochlorite (NaOCl) and chlorhexidine (CHX). Alexidine (ALX), a biguanide disinfectant similar to CHX, has greater affinity for bacterial virulence factors than CHX. This study determined by electrospray ionization mass spectrometry (ESI-MS) and scanning electron microscopy (SEM) whether the chemical interaction between ALX and NaOCl results in PCA or precipitates. Methods ESI-MS was performed on 4 different concentrations of ALX (1%, 0.5%, 0.25%, and 0.125%) with 4% NaOCl to detect the presence of PCA. As control groups, 1% ALX, 0.5% PCA, and a mixture of 2% CHX and 4% NaOCl were analyzed. The formation of precipitates on the dentinal surfaces of premolar root canals treated with the solutions of ALX and NaOCl (AN) or CHX and NaOCl (CN) was observed by SEM and the color change in the reaction solutions was also analyzed. Results ESI-MS showed that the peak (mass/charge ratio = 128.026) in the PCA spectrum was not detected in any of the 4 AN solutions, whereas the peak was found in the CN solution. SEM revealed precipitates covering dentinal surfaces in the CN solution. The AN solutions produced no precipitate. The AN solutions changed in color from light yellow to transparent with decreasing ALX concentration, whereas peach-brown discoloration was observed in the CN solution. Conclusions The interaction of ALX and NaOCl did not produce PCA or precipitates, and the color of the reacted solution changed transparent with decreasing ALX concentration.

Products like salt-cured fish contain approximately 15–21% NaCl and are rehydrated to 2–3% NaCl before preparation and consumption. These products are regarded as safe, but it has been shown that Listeria spp. is able to survive at extreme levels of salt and start to grow after rehydration. Thus, the ability of salt stressed Listeria monocytogenes to cause listeriosis, measured as its ability to invade Caco-2 cells was studied in this paper. Seven strains of L. monocytogenes and one Listeria innocua were cultivated in BHI to early and late stationary phase at 4 °C. At both phases, the strains were exposed to either no salt or to salt stress comparable to that applied in the production of rehydrated salt-cured cod, i.e. 21% NaCl followed by dilution to 2% NaCl. In addition, the eight strains were cultivated in BHI with 2% NaCl, which is similar to the salt content as in rehydrated salt-cured cod and other ready-to-eat (RTE) products as well. The ability of non salt stressed L. monocytogenes strains to enter Caco-2 cells was significant higher ( p > 0.05) compared to the corresponding strains exposed to 21% NaCl for 96 h, followed by 2% NaCl for 48 h. On the other hand, L. monocytogenes cultivated in BHI with 2% NaCl showed a higher invasiveness of Caco-2 cells than both the other sample categories. As the ability to invade Caco-2 cells correlates with bacterial virulence, the results suggests that L. monocytogenes represent a lower food safety risk when exposed to salt-curing with extreme NaCl concentrations than exposure of a constant and moderate level of salt commonly used in RTE products.

Although there has been a relentless increase in resistance to antimicrobial agents amongst important bacterial pathogens throughout the world, it is well known that the number of new antimicrobial agents being brought to the market has undergone a steady decline in the past several decades. There are a number of reasons for this, which are detailed in this article, but there is also a great deal of continuing research to find new effective antimicrobials, much of it now being carried out in academic centres and especially in small biotechnology companies, rather than by large pharma. Whilst classic screening methods and chemical modification of known antimicrobial agents continue to produce potential leads for new antimicrobial agents, a number of other approaches are being investigated. These include the search for potentiators of the activity of known antimicrobial agents and the development of hybrid agents, novel membrane-active drugs, and inhibitors of bacterial virulence and pathogenesis. A number of new bacterial targets are also being exploited, as are bacteriophages and their lytic enzymes. Given the amount of investigation presently underway, it is clear that although the antibiotic pipeline is not as promising as it was half a century ago, it is far from dry.

Background: Rheumatic fever (RF) classically occurs after group A Streptococcus (GAS) pharyngitis in children aged over 5 years in developing countries. The present report describes the bacterial and host determinants in non-related toddlers who developed RF diagnostic criteria after toxic shock syndrome (TSS). Methods and results: A 13-month-old boy and a 14-month-old girl presented GAS TSS. After several weeks, multiple subcutaneous nodules as well as migratory polyarthritis or monoarthritis developed in both children, fulfilling Jones criteria of RF. The relevance of the Jones criteria for very young children is, however, debatable and their use might lead to the unnecessary prescribing of secondary prophylaxis. A molecular analysis of both bacterial and host factors was carried out in an attempt to decipher the combination that could have led to such uncommon, but very similar presentations. The two GAS isolates belonged to the usual, although distinct, invasive emm-types 1 and 3. Both isolates carried a wide set of prophage-encoded virulence factors, with only the speG and speA superantigen-encoding genes in common. Both patients shared the HLA DQB1*0301 allele, which has been associated with susceptibility to GAS necrotizing fasciitis. Conclusions: Our study exemplifies the particularity of RF in young children and the complex role of superantigens and streptodornases in GAS-related pathologies. © 2009 International Society for Infectious Diseases., JOURNAL ARTICLE, SCOPUS: ar.j, info:eu-repo/semantics/published

Pyocyanin is produced by Ps. aeruginosa as a result of quorum sensing during wound colonisation increasing bacterial virulence and damaging host physiology, both of which contribute to an increased risk of infection. The use of carbon fibre tow as an electrochemical sensing matrix for assessing pyocyanin production is evaluated. Prototype sensor assemblies have been developed and response characteristics towards pyocyanin are detailed. The sensitive and linear quantification of pyocyanin is presented (r(2)=0.998) across the biomedically relevant concentration range (1-100 microM). Precise electrochemical measurements of pyocyanin by square wave voltammetry are established using carbon fibre assemblies (coefficient of variance=1.2 and 1.4% for 10 and 50 microM pyocyanin, respectively). Further testing of the sensors in bacterial cultures shows the ability to monitor pyocyanin production by Ps. aeruginosa in agreement with the chloroform-acid/photometric method and in the presence of other bacterially derived pigments and metabolites. The proposed small and inexpensive sensor assembly is suggested for use in monitoring Ps. aeruginosa growth.

Urinary tract infection (UTI) is a leading cause of serious bacterial infection in young children. Vesicoureteral reflux (VUR), a common pediatric urologic disorder, is believed to predispose to UTI, and both are associated with renal scarring. The complex interaction of bacterial virulence factors and host defense mechanisms influence renal damage. However, some renal parenchymal abnormalities associated with VUR are noninfectious in origin. Long-term, renal parenchymal injury may be associated with hypertension, pregnancy complications, proteinuria, and renal insufficiency. Optimal management of VUR and UTI is controversial because of the paucity of appropriate randomized controlled trials; there is a need for well-designed studies. The recently launched Randomized Intervention for children with VesicoUreteral Reflux (RIVUR) study hopefully will provide insight into the role of antimicrobial prophylaxis of UTI in children with VUR.

Many host and bacterial factors contribute to the development of different Escherichia coli extra-intestinal infections. The aim of this study was to evaluate the roles of host and bacterial factors in different extra-intestinal E. coli infections. A total of 221 E. coli isolates collected from urine, bile and peritoneal fluid were included in this retrospective study. Four main phylogenetic groups of E. coli, 14 genetic determinants, static biofilm formation and antimicrobial resistance data were assessed, as well as the immunological status of the hosts. Group B2 was the most common phylogenetic group (30%), especially in cases of asymptomatic bacteriuria (ABU), urinary tract infection (UTI), acute appendicitis/gastrointestinal perforation, and spontaneous bacterial peritonitis (SBP), and was associated with elevated prevalence of papG III, fimH, sfa, iha, hlyA, cnf1, ompT and usp. Phylogenetic group A was most common in the isolates from asymptomatic bacteriocholia, biliary tract infection, and peritoneal dialysis (PD)-related peritonitis. There was similarity with respect to both phylogenetic groups and virulence factors in strains from faeces and ABU, and in strains from faeces and SBP/PD-related peritonitis. Host characteristics were important in patients with ABU, UTI, and SBP/PD-related peritonitis. Immunocompetence of hosts was associated with a relatively high prevalence of papG II, afa and iha, and relatively low antimicrobial resistance to fluoroquinolones. This study demonstrates that, in most E. coli extra-intestinal infections, phylogenetic group B2 was predominant and was more virulent than the three other phylogenetic groups in the Taiwanese population studied. The diverse patterns of host and bacterial factors demonstrate that there were different host and bacterial factors dominating in different extra-intestinal E. coli infections.

Membrane disruption can efficiently alter cellular function; indeed, pore-forming toxins (PFTs) are well known as important bacterial virulence factors. However, recent data have revealed that structures similar to those found in PFTs are found in membrane active proteins across disparate phyla. Many similarities can be identified only at the 3D-structural level. Of note, domains found in membrane-attack complex proteins of complement and perforin (MACPF) resemble cholesterol-dependent cytolysins from Gram-positive bacteria, and the Bcl family of apoptosis regulators share similar architectures with Escherichia coli pore-forming colicins. These and other correlations provide considerable help in understanding the structural requirements for membrane binding and pore formation.

Here we discuss the application of the zebrafish as a relatively new model host for the study of mycobacterial pathogenesis. Recent advances in our understanding of host-mycobacteria interactions from the zebrafish include insights into the role of the innate immune system in both controlling and facilitating infection. Analysis in the zebrafish has revealed that innate macrophages restrict initial bacterial growth, but also convey infecting bacteria into the granuloma, which serves as a place for bacterial growth and spread. Bacterial virulence determinants interact with these processes at different steps in pathogenesis, which can be dissected in these living see-through hosts. As these studies uncover new facets of the bacteria-host interactions in tuberculosis they raise even more questions for future investigation.

Symptomatic urinary tract infections (UTI) are either uncomplicated or complicated. Uncomplicated infections occur in healthy women in the community and are usually caused by Escherichia coli . Complicated infections are associated with anatomical, functional, or metabolic abnormalities of the urinary tract that disable the natural innate host defences and lead to tissue injury. Patients with symptomatic infections will have >10 5 bacteria/ml and inflammatory cells in freshly voided urine. A third group is commonly seen whose symptoms may suggest UTI, but in whom there is no objective evidence for infection. Careful history, examination and investigation are important to avoid repeated and unnecessary courses of antibiotics. Infection is determined by bacterial virulence offset by a complex of innate host defences and some acquired immunity. Urine flow and regular and complete bladder emptying are the first priority; any cause of urine stagnation will promote infection. Investigation is, therefore, primarily aimed at ensuring there is no obstruction and that the bladder voids to completion. This is achieved with plain X-ray, ultrasound of kidneys and the bladder after voiding, and urine flow rate. Acute uncomplicated infection does not require more than 3 days antibiotics. Asymptomatic bacteriuria requires treatment only in infants, pregnancy and before urological intervention. For recurrent and complicated infections, it is mandatory to identify the organism and its sensitivity.

Despite advances in diagnosis and treatment, Mycobacterium tuberculosis causes active disease in about 8 million people worldwide annually. The study of the interplay between the host and the pathogen at the site of infection in human TB may contribute to elucidate the pathogenesis of the disease. In this work, using macroarray technology and real-time PCR, we analyzed the modulation of 847 genes encoding immune-inflammatory mediators in BALF samples of patients affected by active pulmonary TB (PTB) and control patients affected by non-TB diseases. The data show that the PTB milieu contains a complex network of gene activation. Different genes with adhesive properties and involved in tissue repair and fibrosis were modulated. In TB patients, we observed the up-regulation of cytokines, including IFN-gamma and IFN-gamma pathway genes, of several apoptotic genes, and of potent transcriptional activators. These findings can contribute to elucidate the mechanisms of MTB pathogenicity in humans.

In bacteria, two-component systems (TCS) are widely used signal transduction devices which are engaged in a multitude of gene regulatory systems that respond to changing growth conditions. Many pathogenic bacteria encounter different microenvironments during their infectious cycle and their ability to efficiently adapt to different niches inside and outside of their host organisms is frequently mediated by TCSs, which can, therefore, be considered as an essential prerequisite for their pathogenicity. Although significant progress has been made in the elucidation of basic principles of the signal transduction process itself, in many pathogens the contribution of TCS to bacterial virulence is insufficiently recognized.

Uncomplicated urinary tract infection presenting as cystitis or pyelonephritis is common and a significant cause of ill health, although there are generally few sequelae. Unfortunately, the vast body of research now accumulating on predisposing factors and bacterial virulence has had little therapeutic consequence. Nevertheless, standardisation of diagnostic criteria and formulation of guidelines on treatment will go some way to limit the anarchy of excessive investigation and over-treatment which has hitherto contributed to rising antibiotic resistance rates. Truly effective alternative treatments and prophylactic vaccines have yet to be developed.

The role of bacterial variation in the waxing and waning of epidemics was a subject of lively debate in late nineteenth and early twentieth-century bacteriology and epidemiology. The notion that changes in bacterial virulence were responsible for the rise and fall of epidemic diseases was an often-voiced, but little investigated hypothesis made by late nineteenth-century epidemiologists. It was one of the first hypotheses to be tested by scientists who attempted to study epidemiological questions using laboratory methods. This paper examines how two groups of experimental epidemiologists, the British group led by W. W. C. Topley and Major Greenwood, and an American group directed by Leslie T. Webster at the Rockefeller Institute, studied the role of variations in bacterial virulence in the course of laboratory epidemics of mouse typhoid. Relying on Ludwik Fleck’s concept of thought styles and thought collectives, the paper analyzes the fundamental conceptual differences between these two groups of researchers and analyzes the kinds of innovations they introduced as they attempted to integrate bacteriological and epidemiological approaches. The paper shows that the stylistic differences between the two groups can be understood better in the context of the institutional histories and disciplinary relations of epidemiology and bacteriology in the two countries.

There is now considerable evidence that suggests that the H. pylori organism isa human pathogen. The strong association between H. pylori and gastroduodenal disease is well documented. A number of hypotheses have been suggested for the pathogenic mechanisms of H. pylori-induced gastroduodenal disease, including the presence of bacterial virulence factors, the production of inflammatory mediators, disregulation of acid secretion, and the host immune response. At the present time, treatment with a combination of a proton pump inhibitor and antimicrobial agents continues to be recommended for the treatment of H. pylori-associated peptic ulcer disease.

RfaH is a bacterial virulence factor that belongs to the universally conserved NusG family of transcription factors and is commonly referred as the first example of a transformer protein. In contrast to other NusG proteins, the C-terminal domain (CTD) of RfaH folds into an α-helical hairpin that is stabilized by interactions with the N-terminal domain (NTD), which in turn masks an RNA polymerase (RNAp) binding site and thus constitutes an autoinhibited state.This autoinhibition is relieved when binding to a specific DNA element in the transcription elongation complex triggers domain dissociation, allowing the NTD to interact with RNAp to facilitate transcription while the CTD dramatically refolds into a NusG-like β-barrel that interacts with the ribosome to activate translation, thus changing both the structure and function of RfaH.Since this phenomenon is experimentally challenging, their structural details remain to be elucidated. Here, we explore the mechanism of the conformational switching of RfaH in the full-length protein using a dual-basin structure-based model. Our simulations capture several features described experimentally, such as the requirement of disruption of interdomain contacts to trigger the dramatic α-to-β transformation of RfaH, confirms the roles of previously indicated residues E48 and R138, and suggests a new important role for F130, in the stability of the interdomain interaction. These native basins are connected through an intermediate state that builds up upon binding to the NTD and shares features from both folds. We also examine the competitive binding between RfaH-CTD and RNAp for the NTD, showing that RNAp binding favors the β fold.Our study shows that native-biased models are appropriate for interrogating detailed mechanisms of the structural rearrangements during the native state interconversion of RfaH. FUNDING: Fondecyt 11140601.

In Louis Pasteur's scientific career it is striking to note the exponential character of the research he introduced in all the fields he opened up. He offered fabulous opportunities to stereochemistry. He is acknowledged as one of the founders of microbiology. He established the possibility of anaerobic life. He pointed the way to epidemiology, public health, and the bacteriologic fight. He struggled against the idea of spontaneous generation of life. He irrevocably substantiated the microbial theory of infectious diseases. He demonstrated that bacterial virulence could be attenuated, he evidenced immunity and generalised the vaccination principle. He also was an incomparable experimenter.

The EMBO-FEBS lecture course on the 'Molecular Basis of Bacterial Virulence and Survival Within Infected Hosts and in the Environment', organized by Pascale Cossart, Efstathios Gonos and Roberto Kolter, was held on the island of Spetsai, Greece, September 3-13, 2002.

Summary Staphylococcus aureus colonization contributes to skin inflammation in diseases such as atopic dermatitis, but the signaling pathways involved are unclear. Herein, epicutaneous S. aureus exposure to mouse skin promoted MyD88-dependent skin inflammation initiated by IL-36, but not IL-1α/β, IL-18, or IL-33. By contrast, an intradermal S. aureus challenge promoted MyD88-dependent host defense initiated by IL-1β rather than IL-36, suggesting that different IL-1 cytokines trigger MyD88 signaling depending on the anatomical depth of S. aureus cutaneous exposure. The bacterial virulence factor PSMα, but not α-toxin or δ-toxin, contributed to the skin inflammation, which was driven by IL-17-producing γδ and CD4 + T cells via direct IL-36R signaling in the T cells. Finally, adoptive transfer of IL-36R-expressing T cells to IL-36R-deficient mice was sufficient for mediating S. aureus -induced skin inflammation. Together, this study defines a previously unknown pathway by which S. aureus epicutaneous exposure promotes skin inflammation involving IL-36R/MyD88-dependent IL-17 T cell responses.

A wide variety of Gram-negative bacterial pathogens use a ‘type III' protein secretion system to deliver bacterial virulence factors into host cells. Recent results suggest that Gram-positive pathogens may employ similar methods to deliver virulence factors into host cells.

In the last decade there have been modest strides made in our understanding of the pathogenesis of urinary tract infections (UTIs). Matching the advances achieved in unraveling the complexities of bacterial virulence factors using molecular-genetic techniques has been a parallel increase in knowledge of host defense mechanisms. Host susceptibility to UTI in the past has been predominantly studied in patients with structural (obstruction) or functional abnormalities (diabetes mellitus); however, most patients with UTIs have no such demonstrable abnormalities, hence attention has now focused on host behavioral factors in addition to genetically determined cellular mechanisms that predispose to sporadic and recurrent UTI.