Bacteriophage therapy and infective endocarditis - is it realistic?

Infective endocarditis (IE) is a severe infection of the inner heart. Even with current standard treatment, the mean in-hospital mortality is as high as 15-20%, and 1-year mortality is up to 40% for left-sided IE. Importantly, IE mortality rates have not changed substantially over the past 30 years, and the incidence of IE is rising. The treatment is challenging due to the bacterial biofilm mode of growth inside the heart valve vegetations, resulting in antibiotic tolerance. Achieving sufficient antibiotic anti-biofilm concentrations in the biofilms of the heart valve vegetations is problematic, even with high-dose and long-term antibiotic therapy. The increasing prevalence of IE caused by antibiotic-resistant bacteria adds to the challenge. Therefore, adjunctive antibiotic-potentiating drug candidates and strategies are increasingly being investigated. Bacteriophage therapy is a reemerging antibacterial treatment strategy for difficult-to-treat infections, mainly biofilm-associated and caused by multidrug-resistant bacteria. However, significant knowledge gaps regarding the safety and efficacy of phage therapy impede more widespread implementation in clinical practice. Hopefully, future preclinical and clinical testing will reveal whether it is a viable treatment. The objective of the present review is to assess whether bacteriophage therapy is a realistic treatment for IE.

Phage SEP1 hijacks Staphylococcus epidermidis stationary cells' metabolism to replicate.

In nature, bacteria often survive in a stationary state with low metabolic activity. Phages use the metabolic machinery of the host cell to replicate, and, therefore, their efficacy against non-dividing cells is usually limited. Nevertheless, it was previously shown that the Staphylococcus epidermidis phage SEP1 has the remarkable capacity to actively replicate in stationary-phase cells, reducing their numbers. Here, we studied for the first time the transcriptomic profiles of both exponential and stationary cells infected with SEP1 phage using RNA-seq to gain a better understanding of this rare phenomenon. We showed that SEP1 successfully takes over the transcriptional apparatus of both exponential and stationary cells. Infection was, however, delayed in stationary cells, with genes within the gp142-gp154 module putatively implicated in host takeover. S. epidermidis responded to SEP1 infection by upregulating three genes involved in a DNA modification system, with this being observed already 5 min after infection in exponential cells and later in stationary cells. In stationary cells, a significant number of genes involved in translation and RNA metabolic and biosynthetic processes were upregulated after 15 and 30 min of SEP1 infection in comparison with the uninfected control, showing that SEP1 activates metabolic and biosynthetic pathways necessary to its successful replication.IMPORTANCEMost phage-host interaction studies are performed with exponentially growing cells. However, this cell state is not representative of what happens in natural environments. Additionally, most phages fail to replicate in stationary cells. The Staphylococcus epidermidis phage SEP1 is one of the few phages reported to date to be able to infect stationary cells. Here, we unveiled the interaction of SEP1 with its host in both exponential and stationary states of growth at the transcriptomic level. The findings of this study provide valuable insights for a better implementation of phage therapy since phages able to infect stationary cells could be more efficient in the treatment of recalcitrant infections.

Exploring the transcriptional landscape of phage-host interactions using novel high-throughput approaches.

In the last decade, powerful high-throughput sequencing approaches have emerged to analyse microbial transcriptomes at a global scale. However, to date, applications of these approaches to microbial viruses such as phages remain scarce. Tailoring these techniques to virus-infected bacteria promises to obtain a detailed picture of the underexplored RNA biology and molecular processes during infection. In addition, transcriptome study of stress and perturbations induced by phages in their infected bacterial hosts is likely to reveal new fundamental mechanisms of bacterial metabolism and gene regulation. Here, we provide references and blueprints to implement emerging transcriptomic approaches towards addressing transcriptome architecture, RNA-RNA and RNA-protein interactions, RNA modifications, structures and heterogeneity of transcription profiles in infected cells that will provide guides for future directions in phage-centric therapeutic applications and microbial synthetic biology.

M13 phage grafted with peptide motifs as a tool to detect amyloid-ß oligomers in brain tissue.

Oligomeric clusters of amyloid-ß (Aß) are one of the major biomarkers for Alzheimer's disease (AD). However, proficient methods to detect Aß-oligomers in brain tissue are lacking. Here we show that synthetic M13 bacteriophages displaying Aß-derived peptides on their surface preferentially interact with Aß-oligomers. When exposed to brain tissue isolated from APP/PS1-transgenic mice, these bacteriophages detect small-sized Aß-aggregates in hippocampus at an early age, prior to the occurrence of Aß-plaques. Similarly, the bacteriophages reveal the presence of such small Aß-aggregates in post-mortem hippocampus tissue of AD-patients. These results advocate bacteriophages displaying Aß-peptides as a convenient and low-cost tool to identify Aß-oligomers in post-mortem brain tissue of AD-model mice and AD-patients.

Bacteriophage Control of Infectious Biofilms.

Biofilm formation, a strategy of bacterial survival, is a significant concern in different areas, including health, where infectious biofilms are very difficult to combat with conventional antimicrobial therapies. Bacteriophages, the viruses that infect bacteria, are promising agents to prevent and control biofilm-related infections. This chapter describes a series of standard procedures that can be used to study the potential of bacteriophages for biofilm control, from biofilm formation to bacteriophage treatment and evaluation of its efficacy.

Synthetic Biology to Engineer Bacteriophage Genomes.

Recent advances in the synthetic biology field have enabled the development of new molecular biology techniques used to build specialized bacteriophages with new functionalities. Bacteriophages have been engineered toward a wide range of applications, including pathogen control and detection, targeted drug delivery, or even assembly of new materials.In this chapter, two strategies that have been successfully used to genetically engineer bacteriophage genomes will be addressed: the bacteriophage recombineering of electroporated DNA (BRED) and the yeast-based phage-engineering platform.

Farnesol in combination with N-acetylcysteine against Staphylococcus epidermidis planktonic and biofilm cells

Staphylococcus epidermidis is the most frequent cause of nosocomial sepsis and catheter-related infections, in which biofilm formation is considered to be the main virulence mechanism. In biofilm environment, microbes exhibit enhanced resistance to antimicrobial agents. This fact boosted the search of possible alternatives to antibiotics. Farnesol and N-acetylcysteine (NAC) are non-antibiotic drugs that have demonstrated antibacterial properties. In this study, the effect of farnesol and NAC isolated or in combination (farnesol+NAC) was evaluated. NAC at 10 × MIC caused a total cell death in planktonic cells. On the other hand, S. epidermidis biofilms exhibited 4 log reduction in viable cell number after a 24h treatment with NAC at the former concentration. Our results demonstrated that there was a higher CFU log reduction of S. epidermidis planktonic cells when farnesol was combined with NAC at 1 × MIC relatively to each agent alone. However, these results were not relevant because NAC alone at 10 × MIC was always the condition which gave the best results, having a very high killing effect on planktonic cells and a significant bactericidal effect on biofilm cells. This study demonstrated that no synergy was observed between farnesol and NAC. However, the pronounced antibacterial effect of NAC against S. epidermidis, on both lifestyles, indicates the use of NAC as a potential therapeutic agent in alternative to antibiotics.

Oral Candida carriage of patients attending a dental clinic in Braga, Portugal

Antecedentes. La capacidad que poseen las diferentes especies de Candida de colonizar las superficies, puede ser considerada como un factor de riesgo para la infección oral. Objetivos. Establecer la colonización oral por Candida en pacientes que asisten a una clínica dental en Braga, Portugal. Métodos. Un total de 97 pacientes fueron estudiados. Se recogieron muestras bucales con hisopo y fueron cultivadas directamente en CHROMagar Candida. Las levaduras seleccionadas se identificaron mediante reacción en cadena de la polimerasa. Resultados. De las muestras analizadas 54,6% (n=53) fueron positivas para Candida. Candida albicans fue la especie más frecuentemente aislada, representado el 79% de todas las especies identificadas. Las especies de Candida no-Candida albicans (CNCA) aisladas fueron Candida parapsilosis, Candida glabrata, Candida tropicalis y Candida guilliermondii. En la población estudiada no se observó asociación entre la presencia de C. albicans o CNCA con la edad, el sexo o el uso de prótesis. En el 17% de los casos (n=9) se encontró una colonización mixta con dos especies de Candida. Conclusiones. Este estudio muestra una alta incidencia de colonización por Candida en esta población; por lo tanto, se sugiere la necesidad de un diagnóstico preciso para la identificación de las especies de Candida(AU)

Background. The ability of the Candida species to colonize surfaces can be considered as a risk factor for oral infection. Aims. To establish oral Candida carriage in patients attending a dental clinic in Braga, Portugal. Methods. A total of 97 patients were analysed. Swab samples were collected and directly cultured onto CHROMagar Candida. Representative yeasts were identified by polymerase chain reaction. Results. From the samples analysed 54.6% (n=53) were Candida positive, and Candida albicans was the most frequently isolated species, accounting for 79% of all the species identified. Non-C. albicans Candida (NCAC) species recovered included Candida parapsilosis, Candida glabrata, Candida tropicalis, and Candida guilliermondii. There was a lack of association between the presence of C. albicans or NCAC species, and age, gender, or prostheses wearing in this population. In 17% of the cases (n=9), polymicrobial cultures, with two different Candida species, were identified. Conclusions. This study shows a high Candida carriage rate among this population, thus pointing to the relevance of an accurate diagnostic approach in Candida species identification(AU)

Adhesion of Salmonella Enteritidis to stainless steel surfaces

Adhesion of microorganisms to food processing surfaces and the problems it causes are a matter of strong concern to the food industry. Contaminated food processing surfaces may act as potential sources of transmission of pathogens in food industry, catering and in the domestic environments. Several studies have shown that adhesion of bacteria to surfaces partly depends upon the nature of the inert surfaces and partly upon the bacterial surface properties. The aim of this study was to compare the adhesion of four different strains of Salmonella Enteritidis to stainless steel 304 (SS 304). The effect of surface hydrophobicity and surface elemental composition on the adhesion process was also analysed. Hydrophobicity was evaluated through contact angle measurements using the sessile drop method. All the strains studied showed positive values of the degree of hydrophobicity (deltaGlwl) and so can be considered hydrophilic while stainless steel revealed a hydrophobic character. Bacterial cell surface composition was measured using X-ray photoelectron spectroscopy (XPS). The XPS results corroborated the similarity of the values of the degree of hydrophobicity obtained by contact angles. The different Salmonella strains showed similar elemental composition and cell surface physico-chemical properties. Nevertheless, S. Enteritidis MUSC presented higher adhesion ability to SS 304 (p<0.05). It can be concluded that the physico-chemical properties of the strain does not explain the ability of adhesion to stainless steel. Other factors like the production of polysaccharides must be considered.

A adesão de microrganismos a superfícies de processamento de alimentos e os problemas que daí resultam são matéria de grande preocupação para a indústria alimentar. Superfícies de processamento de alimentos contaminadas podem actuar como uma potencial fonte de transmissão de patogénicos na indústria alimentar, restauração e em ambientes domésticos. Diversos estudos têm demonstrado que a adesão de bactérias a superfícies depende, por um lado, da natureza das superfícies inertes e, por outro, das propriedades superficiais das bactérias. O objectivo deste trabalho consistiu na comparação da capacidade de adesão de 4 cepas diferentes de Salmonella Enteritidis ao aço inoxidável 304 (SS 304). Analisou-se também o efeito da hidrofobicidade e da composição elementar no processo de adesão. A hidrofobicidade foi determinada através da medição de ângulos de contacto usando o método da gota séssil. Todas as cepas apresentaram valores positivos do grau de hidrofobicidade (deltaGlwl) podendo, assim, ser consideradas hidrofílicas enquanto o aço inoxidável revelou um carácter hidrofóbico. A composição elementar da superfície das células bacterianas foi medida através de espectroscopia de fotoelectrões X (XPS). Os resultados do XPS corroboraram a similaridade de valores do grau de hidrofobicidade obtidos por ângulos de contacto. As diferentes cepas de Salmonella apresentaram uma composição elementar e propriedades físico-químicas semelhantes. No entanto, a Salmonella MUSC apresentou uma capacidade de adesão ao aço inoxidável mais elevada (p<0.05). Pode então concluir-se que as propriedades físico-químicas das cepas não explicam a capacidade de adesão ao aço inoxidável, devendo ser considerados outros factores tais como a produção de exopolissacáridos.