Friend or Foe: Interbacterial Competition in the Nasal Cavity.

Like other microbes that live on or in the human body, the bacteria that inhabit the upper respiratory tract, in particular the nasal cavity, have evolved to survive in an environment that presents a number of physical and chemical challenges; these microbes are constantly bombarded with nutritional fluctuations, changes in humidity, the presence of inhaled particulate matter (odorants and allergens), and competition with other microbes. Indeed, only a specialized set of species is able to colonize this niche and successfully contend with the host's immune system and the constant threat from competitors. To this end, bacteria that live in the nasal cavity have evolved a variety of approaches to outcompete contenders for the limited nutrients and space; broadly speaking, these strategies may be considered a type of "bacterial warfare." A greater molecular understanding of bacterial warfare has the potential to reveal new approaches or molecules that can be developed as novel therapeutics. As such, there are many studies within the last decade that have sought to understand the complex polymicrobial interactions that occur in various environments. Here, we review what is currently known about the age-dependent structure and interbacterial relationships within the nasal microbiota and summarize the molecular mechanisms that are predicted to dictate bacterial warfare in this niche. Although the currently described interactions are complex, in reality, we have likely only scratched the surface in terms of a true understanding of the types of interbacterial competition and cooperation that are thought to take place in and on the human body.

Microbial Dysbiosis During Simian Immunodeficiency Virus Infection is Partially Reverted with Combination Anti-retroviral Therapy.

Human immunodeficiency virus (HIV) infection is characterized by a massive loss of CD4 T cells in the gastrointestinal tract (GIT) that is accompanied by changes in the gut microbiome and microbial translocation that contribute to inflammation and chronic immune activation. Though highly active antiretroviral therapy (HAART) has led to better long-term outcomes in HIV infected patients, it has not been as effective at reverting pathogenesis in the GIT. Using the simian immunodeficiency virus (SIV) infection model, we show that combination antiretroviral therapy (c-ART) partially reverted microbial dysbiosis observed during SIV infection. Though the relative abundance of bacteria, their richness or diversity did not significantly differ between infected and treated animals, microbial dysbiosis was evident via multiple beta diversity metrics: Jaccard similarity coefficient, Bray-Curtis similarity coefficient, and Yue & Clayton theta similarity coefficient. Principal coordinates analysis (PCoA) clustered SIV-infected untreated animals away from healthy and treated animals that were clustered closely, indicating that c-ART partially reversed the gut dysbiosis associated with SIV infection. Metastats analysis identified specific operational taxonomic units (OTUs) falling within the Streptococcus, Prevotella, Acinetobacter, Treponema, and Lactobacillus genera that were differentially represented across the three groups. Our results suggest that complete viral suppression with c-ART could potentially revert microbial dysbiosis observed during SIV and HIV infections.

Corynebacterium pseudodiphtheriticum Exploits Staphylococcus aureus Virulence Components in a Novel Polymicrobial Defense Strategy.

Commensal bacteria in the human nasal cavity are known to suppress opportunistic pathogen colonization by competing for limited space and nutrients. It has become increasingly apparent that some commensal bacteria also produce toxic compounds that directly inhibit or kill incoming competitors. Numerous studies suggest that microbial species-specific interactions can affect human nasal colonization by the opportunistic pathogen Staphylococcus aureus However, the complex and dynamic molecular interactions that mediate these effects on S. aureus nasal colonization are often difficult to study and remain poorly understood. Here, we show that Corynebacterium pseudodiphtheriticum, a common member of the normal nasal microbiota, mediates contact-independent bactericidal activity against S. aureus, including methicillin-resistant S. aureus (MRSA). Bacterial interaction assays revealed that S. aureus isolates that were spontaneously resistant to C. pseudodiphtheriticum killing could be recovered at a low frequency. To better understand the pathways associated with killing and resistance, a S. aureus transposon mutant library was utilized to select for resistant mutant strains. We found that insertional inactivation of agrC, which codes for the sensor kinase of the Agr quorum sensing (Agr QS) system that regulates expression of many virulence factors in S. aureus, conferred resistance to killing. Analysis of the spontaneously resistant S. aureus isolates revealed that each showed decreased expression of the Agr QS components. Targeted analysis of pathways regulated by Agr QS revealed that loss of the phenol-soluble modulins (PSMs), which are effectors of Agr QS, also conferred resistance to bactericidal activity. Transmission electron microscopy analysis revealed that C. pseudodiphtheriticum induced dramatic changes to S. aureus cell surface morphology that likely resulted in cell lysis. Taken together, these data suggest that C. pseudodiphtheriticum-mediated killing of S. aureus requires S. aureus virulence components. While S. aureus can overcome targeted killing, this occurs at the cost of attenuated virulence; loss of Agr QS activity would phenotypically resemble a S. aureus commensal state that would be unlikely to be associated with disease. Commensal competition resulting in dampened virulence of the competitor may represent an exciting and unexplored possibility for development of novel antimicrobial compounds.IMPORTANCE While some individuals are nasally colonized with S. aureus, the underlying factors that determine colonization are not understood. There is increasing evidence that indicates that resident bacteria play a role; some commensal species can eradicate S. aureus from the nasal cavity. Among these, Corynebacterium pseudodiphtheriticum can eliminate S. aureus from the human nose. We sought to understand this phenomenon at a molecular level and found that C. pseudodiphtheriticum produces a factor(s) that specifically kills S. aureus While resistant S. aureus isolates were recovered at a low frequency, resistance came at the cost of attenuated virulence in these strains. Molecular dissection of the specific strategies used by C. pseudodiphtheriticum to kill S. aureus could lead to the development of novel treatments or therapies. Furthermore, commensal competition that requires virulence components of the competitor may represent an exciting and unexplored possibility for development of novel antimicrobial compounds.

Antimicrobial Activity of Clinically Isolated Bacterial Species Against Staphylococcus aureus.

Bacteria often exist in polymicrobial communities where they compete for limited resources. Intrinsic to this competition is the ability of some species to inhibit or kill their competitors. This phenomenon is pervasive throughout the human body where commensal bacteria block the colonization of incoming microorganisms. In this regard, molecular epidemiological and microbiota-based studies suggest that species-specific interactions play a critical role in the prevention of nasal colonization of the opportunistic pathogen Staphylococcus aureus. Despite this, S. aureus exists as part of the microbiota of ∼25% of the population, suggesting that the interplay between S. aureus and commensals can be complex. Microbiota studies indicate that several bacterial genera are negatively correlated with S. aureus colonization. While these studies paint a broad overview of bacterial presence, they often fail to identify individual species-specific interactions; a greater insight in this area could aid the development of novel antimicrobials. As a proof of concept study designed to identify individual bacterial species that possess anti-S. aureus activity, we screened a small collection of clinical isolates from the Walter Reed National Military Medical Center for the ability to inhibit multiple S. aureus strains. We found that the majority of the isolates (82%) inhibited at least one S. aureus strain; 23% inhibited all S. aureus strains tested. In total, seven isolates mediated inhibitory activity that was independent of physical contact with S. aureus, and seven isolates mediated bactericidal activity. 16S rRNA based-sequencing revealed that the inhibitory isolates belonged to the Acinetobacter, Agromyces, Corynebacterium, Microbacteria, Mycobacterium, and Staphylococcus genera. Unexpectedly, these included seven distinct Acinetobacter baumannii isolates, all of which showed heterogeneous degrees of anti-S. aureus activity. Defined mechanistic studies on specific isolates revealed that the inhibitory activity was retained in conditioned cell free medium (CCFM) derived from the isolates. Furthermore, CCFM obtained from S. saprophyticus significantly decreased mortality of S. aureus-infected Galleria mellonella caterpillars. While future studies will seek to define the molecular mechanisms of the inhibitory activities, our current findings support the study of polymicrobial interactions as a strategy to understand bacterial competition and to identify novel therapeutics against S. aureus and other pathogens.

Investigation of the microbiota of the reproductive tract in women undergoing a total hysterectomy and bilateral salpingo-oopherectomy.

OBJECTIVE: To conduct a pilot study to investigate the possible presence of bacteria throughout the female reproductive tract and to make a preliminary assessment of whether there are differences in the composition of the microbial communities between these body sites and/or between patients. DESIGN: Prospective pilot study followed by 16S amplification and high-throughput sequencing. SETTING: Tertiary care military hospital. PATIENT(S): A total of 10 women underwent a total hysterectomy with bilateral salpingo-oopherectomy; tissue samples were collected from the vagina, resected cervix, uterus, fallopian tubes, and ovaries. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Microbial composition of samples within patients and between body sites. RESULT(S): The microbial composition of each sample was characterized by amplification and sequencing of the V1-V3 region of the 16S rRNA gene. Bacteria were identified in 95% of the samples; the remaining 5% of samples showed no evidence of bacterial 16S rRNA. The microbial communities present at each anatomical location were highly related across the samples and across the patients. The Firmicutes phylum was highly abundant as was the Lactobacillus genus. CONCLUSION(S): This study is the first global evaluation of the distribution of bacteria throughout the female reproductive tract in its entirety. Bacteria were detected by 16S sequencing from anatomical sites including the fallopian tubes and ovaries. The microbial profiles were closely related regardless of which body site or patient the samples originated from. The results of this trial will serve as the basis for future work correlating the colonization of the female reproductive tract with both obstetric and gynecologic conditions.