Quorum sensing inhibitory drugs as next generation antimicrobials: Worth the effort?

Bacterial resistance poses a major challenge to the development of new antimicrobial agents. Conventional antibiotics have an inherent obsolescence because they select for development of resistance. Bacterial infections have again become a serious threat in developed countries. Particularly, elderly, immunocompromised, and hospitalized patients are susceptible to infections caused by bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, and Staphylococcus epidermidis. These bacteria form chronic, biofilm-based infections, which are challenging because bacterial cells living as biofilms are more tolerant to antibiotics than their planktonic counterparts. Therefore, research should identify new antimicrobial agents and their corresponding targets to decrease the biofilm-forming capability or persistence of the infectious bacteria. Here, we review one such drug target: bacterial cell-to-cell communication systems, or quorum sensing.     

Bacterial biofilm in upper respiratory tract infections

The upper respiratory tract is of easy access to pathogens, and although it has evolved a number of defensive barriers to avoid invasion, acute and chronic infections of the ears, nose, and throat are common and present a huge challenge to the health-care system. Though most infections are viral, mild, and self-limiting, bacterial infection is responsible for considerable morbidity and has potential for life-threatening sequelae. Biofilms form when free-floating planktonic organisms adhere to a surface. Within a polymicrobial, biofilm organisms interact, exchanging metabolites, enzymes, and genetic material. The colony is protected, allowing bacteria to thrive in otherwise unfavorable conditions. A role for the biofilm in upper respiratory tract infections has been proposed because infections often run a persistent, remitting course, samples are sometimes difficult to culture, and resistance to medical management is common. This review presents recently published evidence of bacterial biofilms in established upper respiratory tract infections.     

The diabetic foot: The importance of biofilms and wound bed preparation

Biofilms are ubiquitous and medically important complex structures consisting of microbial-associated cells embedded in self-produced extracellular matrix of hydrated extrapolymeric substances, which are irreversibly attached to a biological or nonbiological surface. Bacteria that reside as biofilms are resistant to traditional therapy. This alternative community in which microbes exist has recently attracted interest as a potential reason why chronic wounds do not heal. This may be especially important for diabetic foot ulcers, which are often characterized by their refractory nature, their predisposition to have associated underlying infection, and their improvement with de’bridement. Animal and in vitro models have been developed to better study biofilms, which will allow a venue for therapeutic intervention. Potential opportunities exist that include prevention of bacterial attachment, prevention of biofilm formation, disruption of the biofilm to allow penetration of topical antimicrobial agents, interference with quorum sensing, and enhancement of bacteria dispersion from biofilms to a more easily destroyed planktonic state.     

Diagnóstico microbiológico de las infecciones relacionadas con la formación de biopelículas

Biofilm-related infections represent a serious health problem, accounting for 65- 80% of all infections. The infections are generally chronic and characterized by the persistence of the microorganism, due to the increased resistance of biofilms to both the immune system and antimicrobials. Biofilms can be located to almost every human body tissue and on exogenous devices such as catheters, pacemakers, prosthetic material, implants, urinary catheters, etc.Traditional antimicrobial susceptibility studies in clinical microbiology laboratories have lied on the study of planktonic form of microorganisms. However, this approach might lead to miss the biofilm characteristics and to a treatment failure. Microbiological diagnosis and antimicrobial susceptibility studies of biofilm-related infections are complex and, nowadays, represent a challenge that clinicians and microbiologists have to address as a team in the absence of consensus or standardized protocols.     

生物膜铜绿假单胞菌耐药机制

生物膜与浮游菌相比,表现出更高的抗生素耐药性,其耐药机制复杂,是多因素综合作用的结果,单一机制不能全面解释生物膜耐药。本文就近期研究结果中与生物膜铜绿假单胞菌耐药有关的机制进行综述。     

How much do Pseudomonas biofilms contribute to symptoms of pulmonary exacerbation in cystic fibrosis?

Recent studies suggest that chronic Pseudomonas aeruginosa lung infections in cystic fibrosis (CF) involve anaerobic biofilms, and that these biofilms are the reason chronic infections are rarely eradicated by antibiotic therapy, regardless of the in vitro susceptibility of infecting bacteria. These observations led to the development of an in vitro method for testing antibiotic susceptibility of CF clinical isolates in biofilms (Moskowitz et al., J Clin Microbiol 2004;42:1915-1922) and unearthed an apparent paradox. Antibiotics that remain cornerstones of clinical management of CF pulmonary exacerbations (e.g., beta-lactam antibiotics) appear to have little to no activity at clinically achievable concentrations when tested in vitro against clinical P. aeruginosa isolates growing in biofilms. The proven clinical efficacy of beta-lactam antibiotics in treating exacerbations, and the selection for beta-lactam resistance in vivo, suggest that planktonic bacteria play a significant role in pulmonary exacerbations of CF. A model of infection architecture is proposed in which biofilm and planktonic compartments each play a role in infection persistence and pulmonary exacerbation, respectively. Infection architecture may partially account for the observed lack of correlation between in vitro antibiotic susceptibility testing and clinical response to antibiotic therapy.     

Efficacy of Surgical/Wound Washes against Bacteria: Effect of Different In Vitro Models

Topical antiseptics are often used to treat chronic wounds with biofilm infections and during salvage of biofilm contaminated implants, but their antibacterial efficacy is frequently only tested against non-aggregated planktonic or free-swimming organisms. This study evaluated the antibacterial and antibiofilm efficacy of four commercial surgical washes Bactisure, TorrenTX, minimally invasive lavage (MIS), and Betadine against six bacterial species: Staphylococcus epidermidis, Staphylococcus aureus, Streptococcus pyogenes, Acinetobacter baumannii, Pseudomonas aeruginosa, and Escherichia coli, which are commonly isolated from surgical site infections and chronic wound infections using different in vitro models. We determined minimum planktonic inhibitory and eradication concentration and minimum 1-day-old biofilm inhibition and eradication concentration of antiseptics in 96-well plates format with 24 h contact time. We also tested the efficacy of antiseptics at in-use concentration and contact time in the presence of biological soil against 3-day-old biofilm grown on coupons with shear in a bioreactor, such that the results are more applicable to the clinical biofilm situations. In the 96-well plate model, the minimum concentration required to inhibit or kill planktonic and biofilm bacteria was lower for Bactisure and TorrenTX than for MIS and Betadine. However, Betadine and Bactisure showed better antibiofilm efficacy than TorrenTX and MIS in the 3-day-old biofilm bioreactor model at in-use concentration. The minimal concentration of surgical washes required to inhibit or kill planktonic bacterial cells and biofilms varies, suggesting the need for the development and use of biofilm-based assays to assess antimicrobial therapies, such as topical antiseptics and their effective concentrations. The antibiofilm efficacy of surgical washes against different bacterial species also varies, highlighting the importance of testing against various bacterial species to achieve a thorough understanding of their efficacy.     

Approach to resistant gram-negative bacterial pulmonary infections in patients with cystic fibrosis

PURPOSE OF THE REVIEW: Patients with cystic fibrosis are living longer with chronic pulmonary bacterial infections. One consequence of antibiotic treatment of these chronic infections has been the increasing prevalence of antibiotic resistance seen in bacterial isolates recovered from patients with cystic fibrosis. RECENT FINDINGS: Bacteria such as Pseudomonas aeruginosa and Burkholderia cepacia are able to acquire antibiotic resistance by either spontaneous mutation or gene transfer via plasmids or integrins. In addition, bacteria survive by forming antibiotic-resistant biofilms within the airways of patients with cystic fibrosis. Therapeutic approaches to dealing with antibiotic-resistant bacterial pulmonary infections include the use of in vitro synergy testing to determine optimal double antibiotic combinations or multiple-combination bactericidal testing to determine bactericidal double and triple antibiotic combinations to use against the bacteria in the clinical setting of acute exacerbations. SUMMARY: Therapy for antibiotic-resistant bacterial infections in cystic fibrosis involves the use of new laboratory methods (synergy testing or multiple-combination bactericidal testing) to optimize antibiotic treatment strategies. Clinical trials are required to address whether treatment guided by susceptibility testing improves clinical outcomes. Future novel approaches will likely include drugs that can disrupt bacterial biofilm formation and the use of cationic peptide antimicrobial compounds.     

Specific antibody to Cryptococcus neoformans glucurunoxylomannan antagonizes antifungal drug action against cryptococcal biofilms in vitro

The fungus Cryptococcus neoformans possesses a polysaccharide capsule and can form biofilms on medical devices. We investigated the efficacy that the combination of a specific antibody to the capsular polysaccharide and antifungal therapy has against cryptococcal biofilms. The antibody enhanced the susceptibility of planktonic cells to antifungal agents, but an antagonistic effect was observed for combination therapy against C. neoformans biofilms. Our findings suggest that antibody therapies for infectious diseases that involve biofilms may antagonize certain antimicrobial therapies, and they also imply that products of the immune response may contribute to drug resistance of biofilms formed in vivo.     

Biofilms: A clinical perspective

Biofilms play an increasingly recognized role in many aspects of human disease. Most of our understanding of infections is based on research that has examined free-living organisms. The results do not necessarily apply to biofilm organisms, since metabolic and synthetic characteristics of free-living organisms can change when they assume the biofilm mode of growth. Biofilms reduce our ability to eradicate infections, causing relapses after seemingly appropriate therapy. Awareness of biofilms, prevention of contamination of implanted or invasive devices, and use of appropriate antimicrobial dosing and treatment durations can limit the negative impact of biofilms while we strive for new technological solutions.     

Medicinal plant products targeting quorum sensing for combating bacterial infections

Traditional treatment of infectious diseases is based on compounds that aim to kill or inhibit bacterial growth. The bacterial resistance against antibiotics is a serious issue for public health. Today, new therapeutic targets other than the bacterial wall were deciphered. Quorum sensing or bacterial pheromones are molecules called auto-inducer secreted by bacteria to regulate some functions such as antibiotic resistance and biofilms formation. This therapeutic target is well-studied worldwide, nevertheless the scientific data are not updated and only recent researches started to look into its potential as a target to fight against infectious diseases. A major concern with this approach is the frequently observed development of resistance to antimicrobial compounds. Therefore, this paper aims to provide a current overview of the quorum sensing system in bacteria by revealing their implication in biofilms formation and the development of antibiotic resistance, and an update on their importance as a potential target for natural substances.     

Biofilm streamers cause catastrophic disruption of flow with consequences for environmental and medical systems

Biofilms are antibiotic-resistant, sessile bacterial communities that occupy most moist surfaces on Earth and cause chronic and medical device-associated infections. Despite their importance, basic information about biofilm dynamics in common ecological environments is lacking. Here, we demonstrate that flow through soil-like porous materials, industrial filters, and medical stents dramatically modifies the morphology of Pseudomonas aeruginosa biofilms to form 3D streamers, which, over time, bridge the spaces between obstacles and corners in nonuniform environments. We discovered that accumulation of surface-attached biofilm has little effect on flow through such environments, whereas biofilm streamers cause sudden and rapid clogging. We demonstrate that flow-induced shedding of extracellular matrix from surface-attached biofilms generates a sieve-like network that captures cells and other biomass, which add to the existing network, causing exponentially fast clogging independent of growth. These results suggest that biofilm streamers are ubiquitous in nature and strongly affect flow through porous materials in environmental, industrial, and medical systems.     

Antibiofilm Efficacy of the Pseudomonas aeruginosa Pbunavirus vB_PaeM-SMS29 Loaded onto Dissolving Polyvinyl Alcohol Microneedles

Resistant bacteria prevail in most chronic skin wounds and other biofilm-related topical skin infections. Bacteriophages (phages) have proven their antimicrobial effectiveness for treating different antibiotic-resistant and multidrug-resistant bacterial infections, but not all phages are effective against biofilms. Phages possessing depolymerases can reach different biofilm layers; however, those that do not have depolymerase activity struggle to penetrate and navigate in the intricate 3D biofilm structure and mainly infect bacteria lodged in the outer biofilm layers. To address this, Pseudomonas aeruginosa phage vB_PaeM-SMS29, a phage with poor antibiofilm properties, was incorporated into polyvinyl alcohol (PVA, Mowiol 4:88) supplemented with 0.1% (v/v) of glycerol, and cast onto two different microneedle arrays varying in geometry. The dissolving microneedles were thoroughly characterized by microscopy, force-displacement, swelling, phage release and stability. Furthermore, 48 h-old biofilms were formed using the colony biofilm procedure (absence of broth), and the antibiofilm efficacy of the phage-loaded microneedles was evaluated by viable cell counts and microscopy and compared to free phages. The phages in microneedles were fairly stable for six months when stored at 4 °C, with minor decreases in phage titers observed. The geometry of the microneedles influenced the penetration and force-displacement characteristics but not the antimicrobial efficacy against biofilms. The two PVA microneedles loaded with phages reduced P. aeruginosa PAO1 biofilms by 2.44 to 2.76 log₁₀ CFU·cm⁻² at 24 h. These values are significantly higher than the result obtained after the treatment with the free phage (1.09 log₁₀ CFU·cm⁻²). Overall, this study shows that the distribution of phages caused by the mechanical disruption of biofilms using dissolving microneedles can be an effective delivery method against topical biofilm-related skin infections.     

Novel approaches to the diagnosis, prevention, and treatment of medical device-associated infections

The pathogenesis of device-associated infections is related to biofilm bacteria that exhibit distinct characteristics with respect to growth rate, structural features, and protection from host immune mechanisms and antimicrobial agents when compared with planktonic counterparts. Biofilm-associated infections are prevented, diagnosed, and treated differently from infections not associated with biofilms. This article reviews innovative concepts for the prevention of biofilm formation, and novel treatment approaches. Specific approaches for the diagnosis and prevention of catheter-associated urinary tract and bloodstream infections, as well as infections associated with orthopedic implants and cardiovascular implantable electronic devices, are also discussed.     

Spatial vulnerability: bacterial arrangements, microcolonies, and biofilms as responses to low rather than high phage densities

The ability of bacteria to survive and propagate can be dramatically reduced upon exposure to lytic bacteriophages. Study of this impact, from a bacterium’s perspective, tends to focus on phage-bacterial interactions that are governed by mass action, such as can be observed within continuous flow or similarly planktonic ecosystems. Alternatively, bacterial molecular properties can be examined, such as specific phage‑resistance adaptations. In this study I address instead how limitations on bacterial movement, resulting in the formation of cellular arrangements, microcolonies, or biofilms, could increase the vulnerability of bacteria to phages. Principally: (1) Physically associated clonal groupings of bacteria can represent larger targets for phage adsorption than individual bacteria; and (2), due to a combination of proximity and similar phage susceptibility, individual bacteria should be especially vulnerable to phages infecting within the same clonal, bacterial grouping. Consistent with particle transport theory—the physics of movement within fluids—these considerations are suggestive that formation into arrangements, microcolonies, or biofilms could be either less profitable to bacteria when phage predation pressure is high or require more effective phage-resistance mechanisms than seen among bacteria not living within clonal clusters. I consider these ideas of bacterial ‘spatial vulnerability’ in part within a phage therapy context.     

Tigecycline: the answer to beta-lactam and fluoroquinolone resistance?

Patients with serious bacterial infections such as intra-abdominal infections and complicated skin and soft tissue infections are often treated empirically because a delay in appropriate initial antimicrobial therapy has been shown to significantly increase morbidity and mortality. Furthermore, pathogens that have developed resistance to mainstay therapeutic options are increasing in prevalence making these infections a challenge for physicians. Treatment guidelines for surgical and intra-abdominal infections recommend selection of an agent or a combination of agents with activity to cover both Gram-positive, Gram-negative organisms and anaerobes. Recommended agents include second-generation cephalosporins with anaerobic coverage, beta-lactam/beta-lactamase inhibitor agents, fluoroquinolone/metronidazole combinations and carbapenems. However, the effectiveness of these agents has come into question as once susceptible organisms are now showing signs of resistance to such antimicrobial therapies. Alternative agents specifically designed to overcome mechanisms of microbial resistance have been sought. The result of that search has been the development of a new class of antimicrobials termed glycylcyclines. The first of these novel antibacterials is tigecycline, with a broad spectrum of activity that includes coverage against vancomycin-resistant enterococci, methicillin-resistant S. aureus, and many species of multidrug-resistant Gram-negative bacteria. Tigecycline also has activity against most penicillin-susceptible and resistant Gram-positive organisms. Clinical trial experience with tigecycline has shown it to be at least as effective as current recommended regimens for the treatment of intra-abdominal infections and complicated skin and soft tissue infections. This new agent thus holds promise as an alternative to the beta-lactams and fluoroquinolones for the initial empiric treatment of serious bacterial infections.     

Biofilm formation: a clinically relevant microbiological process.

Microorganisms universally attach to surfaces and produce extracellular polysaccharides, resulting in the formation of a biofilm. Biofilms pose a serious problem for public health because of the increased resistance of biofilm-associated organisms to antimicrobial agents and the potential for these organisms to cause infections in patients with indwelling medical devices. An appreciation of the role of biofilms in infection should enhance the clinical decision-making process.     

Platelets enhance biofilm formation and resistance of endocarditis-inducing streptococci on the injured heart valve

Infective endocarditis is a typical biofilm-associated infectious disease frequently caused by commensal streptococci, but the contribution of host factors in biofilm formation is unclear. We found that platelets are essential for in vitro biofilm formation by Streptococcus mutans or Streptococcus gordonii grown in human plasma. The biofilms were composed of bacterial floes embedded with platelet aggregates in layers, and a similar architecture was also detected in situ on the injured valves of a rat model of experimental endocarditis. Similar to planktonic cells, the streptococci in biofilms were also able to induce platelet aggregation, which facilitates multilayer biofilm formation. Entrapping of platelets directly enhances the resistance of streptococcal biofilms to clindamycin. Prophylactic antibiotics or aspirin can reduce but not prevent or abolish biofilm formation on injured heart valves. Therefore, the platelet is a host factor for commensal streptococci in the circulation to consolidate biofilm formation and protect bacteria against antibiotics.     

Phage–Bacteria Interactions in Potential Applications of Bacteriophage vB_EfaS-271 against Enterococcus faecalis

Phage therapy is one of main alternative option for antibiotic treatment of bacterial infections, particularly in the era of appearance of pathogenic strains revealing resistance to most or even all known antibiotics. Enterococcus faecalis is one of such pathogens causing serious human infections. In the light of high level of biodiversity of bacteriophages and specificity of phages to bacterial species or even strains, development of effective phage therapy depend, between others, on identification and characterization of a large collection of these viruses, including understanding of their interactions with host bacterial cells. Recently, isolation of molecular characterization of bacteriophage vB_EfaS-271, infecting E. faecalis strains have been reported. In this report, phage–host interactions are reported, including ability of vB_EfaS-271 to infect bacteria forming biofilms, efficiency of eliminating bacterial cells from cultures depending on multiplicity of infection (m.o.i.), toxicity of purified phage particles to mammalian cells, and efficiency of appearance of phage-resistant bacteria. The presented results indicate that vB_EfaS-271 can significantly decrease number of viable E. faecalis cells in biofilms and in liquid cultures and reveals no considerable toxicity to mammalian cells. Efficiency of formation of phage-resistant bacteria was dependent on m.o.i. and was higher when the virion-cell ratio was as high as 10 than at low (between 0.01 and 0.0001) m.o.i. values. We conclude that vB_EfaS-271 may be considered as a candidate for its further use in phage therapy.