In the past 30 years, Salmonella bloodstream infections have become a significant health problem in sub-Saharan Africa and are responsible for the deaths of an estimated 390,000 people each year. The disease is predominantly caused by a recently described sequence type of Salmonella Typhimurium: ST313, which has a distinctive set of prophage sequences. We have thoroughly characterized the ST313-associated prophages both genetically and experimentally. ST313 representative strain D23580 contains 5 full-length prophages: BTP1, Gifsy-2D23580, ST64BD23580, Gifsy- 1 D23580 and BTP5. We show that common S. Typhimurium prophages Gifsy-2, Gifsy-1 and ST64B are inactivated in ST313 by mutations. Prophage BTP1 was found to be a functional novel phage, and the first isolate of the proposed new species “Salmonella virus BTP1”, belonging to the P22virus genus. Surprisingly, ~10^9 BTP1 virus particles per ml were detected in the supernatant of non-induced, stationary-phase cultures of strain D23580, representing the highest spontaneously-induced phage titer so far reported for a bacterial prophage. High spontaneous induction is shown to be an intrinsic property of prophage BTP1, and indicates the phage-mediated lysis of around 0.2% of the lysogenic population. The fact that BTP1 is highly conserved in ST313 poses interesting questions about the potential fitness costs and benefits of novel prophages in epidemic S. Typhimurium ST313.

We discovered a novel interaction between phage P22 and its host Salmonella Typhimurium LT2 that is characterized by a phage mediated and targeted derepression of the host dgo operon. Upon further investigation, this interaction was found to be instigated by an ORFan gene (designated pid for phage P22 encoded instigator of dgo expression) located on a previously unannotated moron locus in the late region of the P22 genome, and encoding an 86 amino acid protein of 9.3 kDa. Surprisingly, the Pid/dgo interaction was not observed during strict lytic or lysogenic proliferation of P22, and expression of pid was instead found to arise in cells that upon infection stably maintained an unintegrated phage chromosome that segregated asymmetrically upon subsequent cell divisions. Interestingly, among the emerging siblings, the feature of pid expression remained tightly linked to the cell inheriting this phage carrier state and became quenched in the other. As such, this study is the first to reveal molecular and genetic markers authenticating pseudolysogenic development, thereby exposing a novel mechanism, timing, and populational distribution in the realm of phage–host interactions. Viruses of bacteria, also referred to as (bacterio)phages, are the most abundant biological entity on earth and have a tremendous impact on the ecology of their hosts. It has traditionally been recognized that upon infection by a temperate phage the host cell is forced either to produce and release new virions during lytic development or to replicate and segregate the phage chromosome together with its own genetic material during lysogenic development. These developmental paths are orchestrated by a dedicated set of phage–host interactions that are able to sense and redirect host cell physiology. In addition to this classical bifurcation of temperate phage development, many studies on phage biology in natural ecosystems hypothesize the existence and significance of stable phage carrier cells that are not engaged in either lytic or lysogenic proliferation. Using Salmonella Typhimurium and phage P22 as a model system, we provide substantial evidence authenticating the existence of the phage carrier state and demonstrate that this state (i) is asymmetrically inherited among carrier cell siblings and (ii) enables the execution of a novel phage–host interaction that is not encountered during lytic or lysogenic proliferation.

The Developing World Urgently Needs Phages to Combat Pathogenic Bacteria

Although the study of phage infection has a long history and catalyzed much of our current understanding in bacterial genetics, molecular biology, evolution and ecology, it seems that microbiologis...

Monitoring the complex transmission dynamics of a bacterial virus (temperate phage P22) throughout a population of its host (Salmonella Typhimurium) at single cell resolution revealed the unexpected existence of a transiently immune subpopulation of host cells that emerged from peculiarities preceding the process of lysogenization. More specifically, an infection event ultimately leading to a lysogen first yielded a phage carrier cell harboring a polarly tethered P22 episome. Upon subsequent division, the daughter cell inheriting this episome became lysogenized by an integration event yielding a prophage, while the other daughter cell became P22-free. However, since the phage carrier cell was shown to overproduce immunity factors that are cytoplasmically inherited by the P22-free daughter cell and further passed down to its siblings, a transiently resistant subpopulation was generated that upon dilution of these immunity factors again became susceptible to P22 infection. The iterative emergence and infection of transiently resistant subpopulations suggests a new bet-hedging strategy by which viruses could manage to sustain both vertical and horizontal transmission routes throughout an infected population without compromising a stable co-existence with their host. Extensive co-evolution with their host has shaped bacterial viruses into the most abundant and sophisticated pathogens known to date. However, how these important viral pathogens manage to safely exploit their host without jeopardizing stable co-existence remains a central question, since horizontal (lytic) transmission can reduce the number of susceptible host cells and cause pathogen extinction, while vertical (lysogenic) transmission impairs pathogen abundance. Scrutinizing transmission of temperate phage P22 throughout a bacterial population at single cell resolution now revealed that this phage is able to disseminate immunity factors that allow the emergence of transiently resistant subpopulations of host cells. The continued fostering and consumption of such subpopulations points to an entirely new strategy by which viruses could manage to sustain an active infection with their host.

One of the world’s fastest-growing human populations is in Sub-Saharan Africa (SSA), accounting for more than 950 million people, which is approximately 13% of the global population. Livestock farming is vital to SSA as a source of food supply, employment, and income. With this population increase, meeting this demand and the choice for a greater income and dietary options come at a cost and lead to the spread of zoonotic diseases to humans. To control these diseases, farmers have opted to rely heavily on antibiotics more often to prevent disease than for treatment. The constant use of antibiotics causes a selective pressure to build resistant bacteria resulting in the emergence and spread of multi-drug resistant (MDR) organisms in the environment. This necessitates the use of alternatives such as bacteriophages in curbing zoonotic pathogens. This review covers the underlying problems of antibiotic use and resistance associated with livestock farming in SSA, bacteriophages as a suitable alternative, what attributes contribute to making bacteriophages potentially valuable for SSA and recent research on bacteriophages in Africa. Furthermore, other topics discussed include the creation of phage biobanks and the challenges facing this kind of advancement, and the regulatory aspects of phage development in SSA with a focus on Kenya.