EcoSal Plus

Members of the family |, such as |, are considered both serious and urgent public health threats. Biofilms formed by these health care-associated pathogens can lead to negative and costly health outcomes. The global spread of antibiotic resistance, coupled with increased tolerance to antimicrobial treatments in biofilm-associated bacteria, highlights the need for novel strategies to overcome treatment hurdles. Bacteriophages (phages), or viruses that infect bacteria, have reemerged as one such potential strategy. Virulent phages are capable of infecting and killing their bacterial hosts, in some cases producing depolymerases that are able to hydrolyze biofilms. Phage therapy does have its limitations, however, including potential narrow host ranges, development of bacterial resistance to infection, and the potential spread of phage-encoded virulence genes. That being said, advances in phage isolation, screening, and genome sequencing tools provide an upside in overcoming some of these limitations and open up the possibilities of using phages as effective biofilm control agents.CC999999/ImCDC/Intramural CDC HHSUnited States

A review of phage mediated antibacterial applications

Background: For over a decade, resistance to newly synthesized antibiotics has been observed worldwide. The challenge of antibiotic resistance has led to several pharmaceutical companies to abandon the synthesis of new drugs in fear of bacteria developing resistance in a short period hence limiting initial investment return. To this effect, alternative approaches such as the use of bacteriophages to treat bacterial infections are being explored. This review explores the recent advances in phage-mediated antibacterial applications and their limitations.Methods: We conducted a comprehensive literature search of PubMed, Lib Hub and Google Scholar databases from January 2019 to November 2019. The search key words used were the application of bacteriophages to inhibit bacterial growth and human phage therapy to extract full-text research articles and proceedings from International Conferences published only in English.Results: The search generated 709 articles of which 95 full-text research articles fulfilled the inclusion guidelines. Transmission Electron Microscopy morphological characterization conducted in 23 studies registered Myoviruses, Siphoviruses, Podoviruses, and Cytoviruses phage families while molecular characterization revealed that some phages were not safe to use as they harbored undesirable genes. All in vivo phage therapy studies in humans and model animals against multidrug-resistant (MDR) bacterial infection provided 100% protection. Ex vivo and in vitro phage therapy experiments exhibited overwhelming results as they registered high efficacies of up to 100% against MDR clinical isolates. Phage-mediated bio-preservation of foods and beverages and bio-sanitization of surfaces were highly successful with bacterial growth suppression of up to 100%. Phage endolysins revealed efficacies statistically comparable to those of phages and restored normal ethanol production by completely eradicating lactic acid bacteria in ethanol fermenters. Furthermore, the average multiplicity of infection was highest in ex vivo phage therapy (557,291.8) followed by in vivo (155,612.4) and in vitro (434.5)

AIMS Microbiol

The p-traps of hospital handwashing sinks represent a potential reservoir for antimicrobial-resistant organisms of major public health concern, such as carbapenemase-producing KPC+ | (CPKP). Bacteriophages have reemerged as potential biocontrol agents, particularly against biofilm-associated, drug-resistant microorganisms. The primary objective of our study was to formulate a phage cocktail capable of targeting a CPKP strain (CAV1016) at different stages of colonization within polymicrobial drinking water biofilms using a CDC biofilm reactor (CBR) p-trap model. A cocktail of four CAV1016 phages, all exhibiting depolymerase activity, were isolated from untreated wastewater using standard methods. Biofilms containing |, |, |, |, |, and | were established in the CBR p-trap model for a period of 28 d. Subsequently, CAV1016 was inoculated into the p-trap model and monitored over a period of 21 d. Biofilms were treated for 2 h at either 25 \ub0C or 37 \ub0C with the phage cocktail (10| PFU/ml) at 7, 14, and 21 d post-inoculation. The effect of phage treatment on the viability of biofilm-associated CAV1016 was determined by plate count on m-Endo LES agar. Biofilm heterotrophic plate counts (HPC) were determined using R2A agar. Phage titers were determined by plaque assay. Phage treatment reduced biofilm-associated CAV1016 viability by 1 log| CFU/cm| (p 0.05) at any time point or temperature. Supplementation with a non-ionic surfactant appears to enhance phage association within biofilms. The results of this study suggest the potential of phages to control CPKP and other carbapenemase-producing organisms associated with microbial biofilms in the healthcare environment.32226914PMC7099197822

Applications of Personalised Phage Therapy highlighting the importance of Bacteriophage Banks against Emerging Antimicrobial Resistance

Emerging antibiotic resistance is one of the most important microbiological issues of the 21st century. This poses a query regarding the future use of antibiotics and availability of other promising therapeutic alternatives. The awareness about antibiotic misuse has improved insufficiently and is evident by the increased incidences of multidrug resistant infections globally. Amongst different antibacterial therapeutic approaches phage therapy has created a niche of its own due to continuous use for treatment of human infections in Eastern Europe. Synergistic compounds along with phages have also been proposed as a better alternative compared to antibiotics or phage alone for treatment of chronic cases and seriously debilitating diseases. As such, why not allow custom made phage therapy for treatment of chronic infections? However, the success of phage therapy will depend upon instant availability of characterised bacteriophages from bacteriophage banks which may serve as the major catalyst in bringing Phage Therapy to main stream treatment alternatives or in combination therapy at least. In the current article we present a glimpse of comprehensive approach about utility of bacteriophage banks and further present personalised phage therapy in a synergistic role with antibiotics to overcome emerging antimicrobial resistance

Implications of bacterial viruses on pathogenic bacteria : from natural microbial communities to therapeutic applications

Bacterial viruses (i.e. phages) are ubiquitous intracellular parasites of bacteria, that along with protist grazers account for majority of bacterial mortality in nature. Phages impose strong selection for bacterial phage-resistance, which is often coupled with fitness costs on bacterial traits such as growth ability, virulence or motility. Traditionally phage-host interactions have been studied with two species systems in the laboratory, neglecting the complex web of interactions present in natural communities. The ability of phages to selectively kill bacteria has ignited an interest on phages as alternative antibacterials. However, in order to develop phage therapy, understanding of phage-host interactions in the eco-evolutionary context is essential. In this thesis I studied the implications of lytic phages on opportunistic pathogenic bacteria, as opportunists often have the ability reproduce and reside in outside-host environments, where they are predisposed to a variety of selection pressures. The role of phages in top-down control of bacterial biomass and the evolution of bacterial phage-resistance were studied in the presence of protist predators with differing feeding modes, in low-resource systems mimicking natural pond environment. Hypothesis of coincidental evolution suggests that virulence is a by-product of selection for traits that maximize bacterial fitness in environmental reservoirs. Yet, disease outbreaks by opportunists are relatively rare, suggesting that something constrains the selection for virulence. To assess the role of lytic phages on the evolution of virulence, bacteria were cultured in low-resource environment, accompanied with changes in temperature regime or changes in composition of the community of interacting bacterial enemy species, and the virulence of bacteria was measured in vivo. To study whether the potential phage-resistant bacteria surviving phage therapy would be coupled with lowered virulence, due to costs associated with phage-resistance, a clinical bacterial isolate was exposed to phage cocktails and the virulence of the phage-resistant bacteria was measured in vivo. given the strong selection for phage- resistance, the prospects of phage therapy depend a great deal on whether new phages infecting pathogenic bacteria can be readily isolated from environment. To address this, an attempt was made to isolate phages against clinical bacterial isolates harboring resistance genes to multiple antibiotics. A single lytic phage was shown to be a non- efficient top-down regulator of bacterial biomass. Rapidly emerging phage-resistant bacteria took over the bacterial populations after initial lysis by phages and protist grazers accounted for most of the long-term negative trophic effects on bacterial biomass. The presence of protist predators selected for bacteria that were less susceptible to infection by lytic phages, which suggests an overlap in the bacterial defense against a parasite and predatory protists. In general, the presence of lytic phages selected for lowered virulence in bacteria. High temperature selected for more virulent and more motile bacteria, but this was constrained by the presence of a lytic phage. In the multispecies communities the presence of all bacterial enemies led to decreased virulence in vivo. Altogether, these results contrast the hypothesis of coincidental evolution, and suggest that the presence of phages in natural reservoirs constrains the evolution of virulence, most likely through fitness costs associated with phage-resistance. Exposure to phage cocktails was also shown to be associated with decreased bacterial virulence in the phage-resistant bacteria. However, exposure to some individual phages resulted in more virulent bacteria, suggesting that the outcome of therapy could depend on the identity of the phage cocktail. Finally, a phage cocktail lysing a wide range of clinical strains was isolated from sewage. This, along with geographical patterns of phage infections suggest that new phages are available in environmental reservoirs for therapy, and the emergence of phage-resistance should not hinder the prospects of phage therapy in the global perspective.Bakteerivirukset (faagit) ovat bakteerien solunsisäisiä loisia, jotka yhdessä alkueliösaalistajien kanssa aiheuttavat suurimman osan bakteerien kuolleisuudesta luonnossa. Faagit luovat voimakkaan valintapaineen faagivastustuskyvylle ja vastustuskyvystä seuraa usein kustannus esimerkiksi bakteerin kasvu-, taudinaiheuttamis- tai liikkumiskyvylle. Perinteisesti faagin ja isännän välisiä vuorovaikutuksia on tutkittu kahden lajin kokeissa laboratoriossa, mutta lähestymistapa jättää huomiotta luonnollisissa yhteisöissä vallitsevien vuorovaikutusten kirjon. Koska faagit tappavat kohdennetusti tiettyjä bakteereita, on ehdotettu, että faageja voitaisiin käyttää antibiootteina. Faagiterapian kehittäminen vaatii kuitenkin faagi-isäntä vuorovaikutusten tuntemusta niin ekologisesta kuin evolutiivisestakin näkökulmasta. Tässä väitöskirjassa olen tutkinut lyyttisten bakteerivirusten vaikutuksia opportunistisiin taudinaiheuttajabakteereihin. Osa opportunistisista bakteereista kykenee elämään ja lisääntymään isäntänsä ulkopuolisissa ympäristöissä, missä ne altistuvat lukuisille erilaisille valintapaineille. Tutkin faagien merkitystä bakteeribiomassan vähentäjinä, sekä bakteerien faagivastustuskyvyn evoluutiota matalaravinteisissa lampea muistuttavassa ympäristössä, joissa oli faagien lisäksi läsnä ravinnonhankintatavoiltaan toisistaan poikkeavia alkueliöitä. Sattumanvaraisen evoluution hypoteesin (coincidental evolution hypothesis) mukaan taudinaiheuttamiskyky on seurausta sellaisiin elinkiertopiirteisiin kohdistuvasta valinnasta, jotka parantavat kelpoisuutta isännän ulkopuolisissa ympäristöissä. Opportunistien aiheuttamat tautiepidemiat ovat kuitenkin verraten harvinaisia, mikä viittaa siihen että jokin tekijä ympäristössä toimii vastavoimana taudinaiheuttamiskykyyn kohdistuvalle valinnalle. Faagien vaikutuksia opportunistibakteerien taudinaiheuttamiskyvyn evoluutioon tutkittiin niinikään matalaravinteisessa ympäristössä, joissa vaihtelevina ympäristötekijöinä olivat joko lämpötila tai bakteereja ravinnokseen käyttävien vihollisten eri yhdistelmät. Kokeiden päätteeksi taudinaiheuttamiskyky mitattiin hyönteisissä. Kliinistä taudinaiheuttajabakteerikantaa altistettiin faagi-koktaileille (phage cocktails) kokeessa, jonka tarkoituksena oli selvittää onko faageille vastustuskykyisten bakteerin taudinaiheuttamiskyky alentunut faagipuolustuksen aiheuttamien kustannusten seurauksena, ja mahdolliset muutokset taudinaiheuttamiskyvyssä mitattiin hyönteisissä. Faagivastustuskykyyn kohdistuvan voimakkaan valintapaineen huomioonottaen faagiterapian onnistumisen kannalta on merkittävää, voidaanko uusia faageja tarvittaessa eristää ympäristöstä. Tavoitteeksi asetettiin eristää uusia faageja, jotka estävät kliinisten antibiooteille vastustuskykyisten bakteereiden kasvua. Yksittäinen faagityyppi osoittautui tehottomaksi säätelemään bakteeribiomassan määrää pitkällä aikavälillä ja faageille vastustuskykyiset bakteerit palauttivat populaatiot nopeasti lähes kontrollitasolle. Alkueläimet näin ollen vastasivat pitkällä aikavälillä käytännössä kaikesta bakteeribiomassan vähentymisestä. Alkueliöt vaikuttivat myös bakteerien faagivastustuskyvyn evoluutioon: alkueliöille altistuneet bakteerit olivat vähemmän alttiita faagi-infektioille. Korkea lämpötila johti taudinaiheuttamiskykyvyn nousuun bakteereissa, mutta ilmiö kumoutui faagien vaikutuksesta. Monilajisissa yhteisöissä kaikkien bakteeripetojen läsnäolo taas alensi bakteerien taudinaiheuttamiskykyä. Nämä tulokset puhuvat sattumanvaraisen evoluution hypoteesia vastaan ja faagit näyttäisivät sen sijaan luovan valintapaineen vähemmän taudinaiheuttamiskykyisille bakteereille. Myös faagikoktailit aiheuttivat taudinaiheuttamiskyvyn laskua bakteereissa. Jotkut yksittäiset virukset näyttivät kuitenkin nostavan bakteerien taudinaiheuttamiskykyä vastustuskykyisissä bakteereissa, minkä vuoksi tuleekin noudattaa erityistä varovaisuutta valittaessa faageja koktaileihin ja faagiterapiaan. Jätevedestä onnistuttiin eristämään kokoelma faageja, jotka estivät kasvua laajassa joukossa kliinisiä antibiooteille vastustuskykyisiä bakteerikantoja. Tämä tulos yhdessä faagi-infektioiden maantiedettä koskevien havaintojen kanssa antaa olettaa, että uusia faageja on eristettävissä ympäristövarannoista ja että laaja-alaisen faagivastustuskyvyn syntyminen ei ole este faagiterapian kehittämiselle tulevaisuudessa

CAUTI’s next top model : model dependent Klebsiella biofilm inhibition by bacteriophages and antimicrobials

Klebsiella infections, including catheter associated urinary tract infections, are a considerable burden on health care systems. This is due to their difficulty to treat, caused by antimicrobial resistance and their ability to form biofilms. In this study, we investigated the use of a Klebsiella phage cocktail to reduce biofilm viability. We used two methodologies to investigate this, a standard 96-well plate assay and a more complicated Foley catheter-based model. The phage cocktail was used alone and in combination with clinically relevant antibiotic treatments. Viability was measured by both a resazurin based stain and colony forming unit counts, of cells sloughed off from the biofilm. We showed that phage infection dynamics and host survival vary significantly in different standard laboratory media, presumably due to the expression of different surface receptors and capsule composition by the bacteria effecting phage binding. This underscores the importance of a realistic model for developing phage therapy. We demonstrate that bacteriophage-based treatments are a viable option for preventing Klebsiella colonisation and biofilm formation on urinary catheters. Phage cocktails were able to significantly reduce the amount of biofilm that formed when they were present during early biofilm formation. The phages used in this study were unable to significantly reduce a pre-formed mature biofilm, despite encoding depolymerases. Phages applied together with antimicrobial treatments, showed synergistic interactions, in some cases the combined treatment was much more effective than antimicrobial treatments alone. We show that phage cocktails have the potential to prevent Klebsiella biofilms in catheters, if used early or as a preventative treatment and will work well alongside standard antibiotics in the treatment of catheter-associated urinary tract infections (CAUTI)

Phage therapy as an alternative or complementary strategy to prevent and control biofilm-related infections

The complex heterogeneous structure of biofilms confers to bacteria an important survival strategy. Biofilms are frequently involved in many chronic infections in consequence of their low susceptibility to antibiotics as well as resistance to host defences. The increasing need of novel and effective treatments to target these complex structures has led to a growing interest on bacteriophages (phages) as a strategy for biofilm control and prevention. Phages can be used alone, as a cocktail to broaden the spectra of activity, or in combination with other antimicrobials to improve their efficacy. Here, we summarize the studies involving the use of phages for the treatment or prevention of bacterial biofilms, highlighting the biofilm features that can be tackled with phages or combined therapy approaches.This work was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the project PTDC/BBB-BSS/6471/2014, the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684). This work was also supported by BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 – Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Innovative Strategies for the Control of Biofilm Formation in Clinical Settings

Biofilm formation in clinical settings is an increasingly important issue particularly due to the emergence of multidrug-resistant strains, as it resulted in increased mortality, which poses a considerable financial burden on healthcare systems. The bacterial biofilms are quite resistant to the routine antimicrobial-based therapies; therefore, the novel strategies are desired in addition to the conventional antibiotics for the effective control of infections caused by biofilm-forming microbes. So far, the approaches being proposed to control the biofilm formation in clinical practice settings include the use of biofilm inhibitors and the use of modified biomaterials for the development of medical devices to thwart the formation of biofilms. In this chapter, we have focused on the latest developments in the anti-biofilm strategies through the interruption of the quorum-sensing system, which is crucial for biofilm formation and have summarized the various classes of antibacterial compounds for the control of biofilm formation. This agrees with the recent approaches suggested by the National Institute of Health (NIH) that advocates the use of combinational therapies based on the conventional methods and complementary treatment to explore the potential utility and safety concerns of the natural products. The studies regarding these emerging strategies could possibly lead to the establishment of better therapeutic alternates compared to conventional treatments