Coliphages of the human urinary microbiota.

Due to its frequent association with urinary tract infections (UTIs), Escherichia coli is the best characterized constituent of the urinary microbiota (urobiome). However, uropathogenic E. coli is just one member of the urobiome. In addition to bacterial constituents, the urobiome of both healthy and symptomatic individuals is home to a diverse population of bacterial viruses (bacteriophages). A prior investigation found that most bacterial species in the urobiome are lysogens, harboring one or more phages integrated into their genome (prophages). Many of these prophages are temperate phages, capable of entering the lytic cycle and thus lysing their bacterial host. This transition from the lysogenic to lytic life cycle can impact the bacterial diversity of the urobiome. While many phages that infect E. coli (coliphages) have been studied for decades in the laboratory setting, the coliphages within the urobiome have yet to be cataloged. Here, we investigated the diversity of urinary coliphages by first identifying prophages in all publicly available urinary E. coli genomes. We detected 3,038 intact prophage sequences, representative of 1,542 unique phages. These phages include both novel species as well as species also found within the gut microbiota. Ten temperate phages were isolated from urinary E. coli strains included in our analysis, and we assessed their ability to infect and lyse urinary E. coli strains. We also included in these host range assays other urinary coliphages and laboratory coliphages. The temperate phages and other urinary coliphages were successful in lysing urinary E. coli strains. We also observed that coliphages from non-urinary sources were most efficient in killing urinary E. coli strains. The two phages, T2 and N4, were capable of lysing 83.5% (n = 86) of strains isolated from females with UTI symptoms. In conclusion, our study finds a diverse community of coliphages in the urobiome, many of which are predicted to be temperate phages, ten of which were confirmed here. Their ability to infect and lyse urinary E. coli strains suggests that urinary coliphages may play a role in modulating the E. coli strain diversity of the urobiome.

When Plaquing Is Not Possible: Computational Methods for Detecting Induced Phages.

High-throughput sequencing of microbial communities has uncovered a large, diverse population of phages. Frequently, phages found are integrated into their bacterial host genome. Distinguishing between phages in their integrated (lysogenic) and unintegrated (lytic) stage can provide insight into how phages shape bacterial communities. Here we present the Prophage Induction Estimator (PIE) to identify induced phages in genomic and metagenomic sequences. PIE takes raw sequencing reads and phage sequence predictions, performs read quality control, read assembly, and calculation of phage and non-phage sequence abundance and completeness. The distribution of abundances for non-phage sequences is used to predict induced phages with statistical confidence. In silico tests were conducted to benchmark this tool finding that PIE can detect induction events as well as phages with a relatively small burst size (10×). We then examined isolate genome sequencing data as well as a mock community and urinary metagenome data sets and found instances of induced phages in all three data sets. The flexibility of this software enables users to easily include phage predictions from their preferred tool of choice or phage sequences of interest. Thus, genomic and metagenomic sequencing now not only provides a means for discovering and identifying phage sequences but also the detection of induced prophages.

Examination of Staphylococcus aureus Prophages Circulating in Egypt.

Staphylococcus aureus infections are of growing concern given the increased incidence of antibiotic resistant strains. Egypt, like several other countries, has seen alarming increases in methicillin-resistant S. aureus (MRSA) infections. This species can rapidly acquire genes associated with resistance, as well as virulence factors, through mobile genetic elements, including phages. Recently, we sequenced 56 S. aureus genomes from Alexandria Main University Hospital in Alexandria, Egypt, complementing 17 S. aureus genomes publicly available from other sites in Egypt. In the current study, we found that the majority (73.6%) of these strains contain intact prophages, including Biseptimaviruses, Phietaviruses, and Triaviruses. Further investigation of these prophages revealed evidence of horizontal exchange of the integrase for two of the prophages. These Egyptian S. aureus prophages are predicted to encode numerous virulence factors, including genes associated with immune evasion and toxins, including the Panton-Valentine leukocidin (PVL)-associated genes lukF-PV/lukS-PV. Thus, prophages are likely to be a major contributor to the virulence of S. aureus strains in circulation in Egypt.

Genomic Survey of E. coli From the Bladders of Women With and Without Lower Urinary Tract Symptoms.

Urinary tract infections (UTIs) are one of the most common human bacterial infections. While UTIs are commonly associated with colonization by Escherichia coli, members of this species also have been found within the bladder of individuals with no lower urinary tract symptoms (no LUTS), also known as asymptomatic bacteriuria. Prior studies have found that both uropathogenic E. coli (UPEC) strains and E. coli isolates that are not associated with UTIs encode for virulence factors. Thus, the reason(s) why E. coli sometimes causes UTI-like symptoms remain(s) elusive. In this study, the genomes of 66 E. coli isolates from adult female bladders were sequenced. These isolates were collected from four cohorts, including women: (1) without lower urinary tract symptoms, (2) overactive bladder symptoms, (3) urgency urinary incontinence, and (4) a clinical diagnosis of UTI. Comparative genomic analyses were conducted, including core and accessory genome analyses, virulence and motility gene analyses, and antibiotic resistance prediction and testing. We found that the genomic content of these 66 E. coli isolates does not correspond with the participant's symptom status. We thus looked beyond the E. coli genomes to the composition of the entire urobiome and found that the presence of E. coli alone was not sufficient to distinguish between the urobiomes of individuals with UTI and those with no LUTS. Because E. coli presence, abundance, and genomic content appear to be weak predictors of UTI status, we hypothesize that UTI symptoms associated with detection of E. coli are more likely the result of urobiome composition.

Mimicking prophage induction in the body: induction in the lab with pH gradients.

The majority of bacteria within the human body are lysogens, often harboring multiple bacteriophage sequences (prophages) within their genomes. While several different types of environmental stresses can trigger or induce prophages to enter into the lytic cycle, they have yet to be fully explored and understood in the human microbiota. In the laboratory, the most common induction method is the DNA damaging chemical Mitomycin C. Although pH has been listed in the literature as an induction method, it is not widely used. Here, we detail a protocol for prophage induction by culture under different pH conditions. We explored the effects of pH on prophage induction in bacterial isolates from the bladder, where the pH is well documented to vary significantly between individuals as well as between healthy individuals and individuals with urinary tract symptoms or disease. Using this protocol, we successfully induced phages from seven bladder E. coli strains. Testing conditions and stressors appropriate to the environment from which a lysogen is isolated may provide insight into community dynamics of the human microbiota.

Draft Genome Sequence of Actinomyces neuii UMB1295, Isolated from the Female Urinary Tract.

Actinomyces neuii is an opportunistic pathogen. Within the urogenital tract, it has been associated with bacterial vaginosis and overactive bladder symptoms. Here, we investigate a draft genome sequence of A. neuii UMB1295, which was isolated from a catheterized urine sample from a woman with a urinary tract infection.

Draft Genome Sequence of Lactobacillus crispatus UMB1163, Isolated from the Female Urinary Tract.

Lactobacillus crispatus is a Gram-positive bacterium shown to protect against urinary and vaginal infections. Here, we report the draft genome sequence of L. crispatus UMB1163, isolated from the female urinary tract.

Draft Genome Sequence of Streptococcus anginosus UMB0839, Isolated from the Female Urinary Tract.

Here, we present the draft genome sequence of Streptococcus anginosus UMB0839, isolated from the female urinary tract. The total size of the genome is 2,104,786 bp assembled into 42 contigs with a GC content of 38.8% and 284× genome coverage.

Draft Genome Sequence of Escherichia coli UMB1353, Isolated from the Female Urinary Tract.

Here, we present the draft genome sequence of Escherichia coli UMB1353, isolated from a patient with a urinary tract infection. The sequence of this antibiotic-resistant E. coli strain contains one intact P2-like phage.

Draft Genome Sequence of Staphylococcus epidermidis UMB8493, Isolated from the Female Urinary Tract.

Staphylococcus epidermidis is a native member of the human microbiota. Here, we report the draft genome sequence of S. epidermidis UMB8493, an isolate from a catheterized urine sample from a female with overactive bladder symptoms. The 2.54-Mbp draft genome encodes genes associated with beta-lactam and fosfomycin resistance.

Draft Genome Sequence of Escherichia coli UMB9246, Isolated from the Bladder of a Woman with Recurrent Urinary Tract Infection.

Escherichia coli is a Gram-negative, motile, rod-shaped bacterium that causes the majority of uncomplicated urinary tract infections (UTIs). Here, we report the draft genome of E. coli strain UMB9246, an isolate from a woman with recurrent UTI.

Introducing Lu-1, a Novel Lactobacillus jensenii Phage Abundant in the Urogenital Tract.

Bacteriophages (phages) play a key role in shaping microbial communities, including those of the human body. Phages are abundant members of the urogenital tract, most often persisting through the lysogenic life cycle as prophages integrated within the genomes of their bacterial hosts. While numerous studies of the urogenital microbiota have focused on the most abundant bacterial member of this niche-Lactobacillus species-very little is known about Lactobacillus phages. Focusing on Lactobacillus jensenii strains from the urinary tract, we identified numerous prophages related to the previously characterized Lv-1 phage from a vaginal L. jensenii strain. Furthermore, we identified a new L. jensenii phage, Lu-1. Evidence suggests that both phages are abundant within the urogenital tract. CRISPR spacer sequences matching to Lv-1 and Lu-1 prophages were identified. While first detected in urinary isolates, the Lu-1 phage was also discovered in L. jensenii isolates from vaginal and perineal swabs, and both phages were found in metagenomic data sets. The prevalence of these phages in the isolates suggests that both phages are active members of the urogenital microbiota.

Draft Genome Sequence of Corynebacterium aurimucosum UMB7769, Isolated from the Female Urinary Tract.

Here, we present the draft genome sequence of Corynebacterium aurimucosum UMB7769, isolated from the female urinary tract. The size of the genome is 2,731,818 bp, assembled in 50 contigs, with an observed GC content of 60.9% and an N 50 score of 129,518 bp. Annotation revealed 31 antibiotic resistance genes.

Draft Genome Sequence of Enterococcus faecalis UMB7780, Isolated from the Female Urinary Tract.

Here, we present the draft genome sequence of Enterococcus faecalis UMB7780, isolated from the female urinary tract. The genome size is 3,005,901 bp, with a GC content of 37.36%, genome coverage of 179×, and an N 50 score of 169,627 bp. Genome analysis identified evidence of antibiotic resistance, as well as intact prophages.

Draft Genome Sequence of Enterococcus faecalis UMB1309, Isolated from Catheterized Urine.

A strain of Enterococcus faecalis was isolated from catheterized urine. Here, we present the draft genome sequence of this isolate, E. faecalis UMB1309. Analysis of the genome revealed multiple genes coding for virulence factors, as well as genes associated with antibiotic resistance.

Draft Genome Sequence of Proteus mirabilis UMB1310, Isolated from the Female Urinary Tract.

Proteus mirabilis is a Gram-negative bacterium that is linked to common complications within the urinary tract. Here, we present the draft genome for P. mirabilis UMB1310, which was isolated from the urine of a woman with a urinary tract infection.

Draft Genome Sequence of Lactobacillus jensenii Strain UMB7766, Isolated from the Female Bladder.

Lactobacillus jensenii is a beneficial and prominent community member within both the vaginal and female urinary microbiota. As more genomes for L. jensenii strains are made publicly available, we gain more knowledge about this beneficial community member. Here, we present the draft genome sequence of L. jensenii UMB7766, which was isolated from a urine specimen from a catheterized female patient with recurrent urinary tract infections.

Draft Genome Sequence of Corynebacterium coyleae UMB8490, Isolated from the Female Urinary Tract.

Here, we present the draft genome sequence for Corynebacterium coyleae UMB8490, isolated from the catheterized urine of a female with overactive bladder symptoms. The size of the genome is 2,483,223 bp assembled in 62 contigs with an observed GC content of 61.42%.

Draft Genome Sequence of Lactobacillus jensenii UMB0847, Isolated from the Female Urinary Tract.

Lactobacillus jensenii is a protective bacterium native to the female urinary tract. Here, we present the 1.6-Mbp draft genome for Lactobacillus jensenii UMB0847, isolated from a catheterized urine sample obtained from a pregnant female.

Draft Genome Sequence of Klebsiella pneumoniae UMB7779, Isolated from the Female Urinary Tract.

Klebsiella pneumoniae is a pathogenic bacterium commonly responsible for urinary tract infections (UTIs). Here, we report the draft genome sequence of K. pneumoniae strain UMB7779, isolated from catheterized urine of a woman with a recurrent UTI.