Type VI protein secretion system (T6SS) is important for bacterial competition through contact-dependent killing of competitors. T6SS delivers effectors to neighboring cells and corresponding antagonistic proteins confer immunity against that are delivered by sister cells. Although has been found in more than 100 gram-negative bacteria including many human pathogens, few T6SS-dependent effector have experimentally determined. Here we report a high-throughput approach using transposon...

One of the hallmarks opportunistic pathogens is their ability to adjust and respond a wide range environmental host-associated conditions. The human pathogen Pseudomonas aeruginosa has an thrive in variety hosts cause acute chronic infections individuals with impaired host defenses or cystic fibrosis. Here we report in-depth transcriptional profiling this organism when grown at host-related temperatures. Using RNA-seq samples from P. 28°C 37°C detected genes preferentially expressed body...

Determining how an organism responds to its environment by altering gene expression is key understanding ecology. Here, we used RNA-seq comprehensively and quantitatively assess the transcriptional response of bacterial opportunistic cystic fibrosis (CF) pathogen endemic soil dweller, Burkholderia cenocepacia, in conditions mimicking these 2 environments. By sequencing 762 million bases cDNA from closely related B. cenocepacia strains (one isolated a CF patient one soil), identified number...

High-throughput sequencing of transposon (Tn) libraries created within entire genomes identifies and quantifies the contribution individual genes operons to fitness organisms in different environments. We used insertion-sequencing (INSeq) analyze all non-essential chromosome Pseudomonas aeruginosa strain PA14 based on a library ∼300,000 Tn insertions. In vitro growth LB provided baseline for comparison with survival insertion strains following 6 days colonization murine gastrointestinal...

Significance It is thought antibiotic resistance carries a fitness cost and reduces microbial virulence. Using high-throughput sequencing analysis of transposon insertion bank in Pseudomonas aeruginosa , we found enhanced for vivo mucosal colonization systemic spread strains with insertions the oprD gene. This conferred to carbapenem antibiotics as well killing at acidic pH by normal human serum along increased cytotoxicity against murine macrophages. RNA-sequencing revealed that deficiency...

In the process of clone-based genome sequencing, initial assemblies frequently contain cloning gaps that can be resolved using cloning-independent methods, but reason for their occurrence is largely unknown. By analyzing 9,328,693 sequencing clones from 393 microbial genomes, we systematically mapped more than 15,000 genes residing in and experimentally showed expression products are toxic to Escherichia coli host. A subset these sequences was further evaluated through a series functional...

P. gingivalis is an important cause of serious periodontal diseases, and emerging as a pathogen in several systemic conditions including some forms cancer. Initial colonization by involves interaction with gingival epithelial cell the organism can also access host tissues spread haematogenously. To better understand mechanisms underlying these properties, we utilized highly saturated transposon insertion library assessed fitness mutants during survival murine abscess model high-throughput...

Respiratory melioidosis is a disease presentation of the biodefense pathogen, Burkholderia pseudomallei, which frequently associated with lethal septicemic spread bacteria. We have recently developed an improved respiratory model to study pathogenesis pseudomallei in lung (intubation-mediated intratracheal [IMIT] inoculation), more closely models descriptions human melioidosis, including prominent from and reduced involvement upper tract. previously demonstrated that Type 3 Secretion System...

ABSTRACT Staphylococcus aureus asymptomatically colonizes the nasal cavity and pharynx of up to 60% human population and, as an opportunistic pathogen, can breach its normal habitat, resulting in life-threatening infections. S. infections are additional concern for populations with impaired immune function such those cystic fibrosis (CF) or chronic granulomatous disease. Multi-drug resistance is increasingly common infections, creating urgent need new antimicrobials compounds that improve...

Our understanding of the Escherichia genus is heavily biased toward pathogenic or commensal isolates from human animal hosts. Recent studies have recovered that persist, and even grow, outside these Although environmental are typically phylogenetically distinct, they highly related to phenotypically indistinguishable their counterparts, including for coliform test. To gain insights into genomic diversity diverse habitats, freshwater, soil, animal, sources, we carried out comparative DNA-DNA...

Burkholderia dolosa is a member of the cepacia complex (BCC), which group bacteria that cause chronic lung infection in patients with cystic fibrosis (CF) and can be associated outbreaks carrying high morbidity mortality. While investigating genomic diversity B. strains collected from an outbreak among CF patients, we previously identified fixL as gene showing signs strong positive selection. This has homology to rhizobial FixL/FixJ two-component system. The goals this study were determine...

Burkholderia cenocepacia is an endemic soil dweller and emerging opportunistic pathogen in patients with cystic fibrosis (CF). The identification of virulence factors potential therapeutic targets has been hampered by the genomic diversity within species as many are not shared among pathogenic members species.In this study, global putative was performed analyzing transcriptome two related strains B. (one clinical, one environmental) under conditions mimicking sputum versus soil. Soil a...

Porphyromonas gingivalis is a Gram-negative anaerobe and keystone periodontal pathogen. A mariner transposon insertion mutant library has recently been used to define 463 genes as putatively essential for the in vitro growth of P. ATCC 33277 planktonic culture (Library 1). We have independently generated 2) same strain herein compare that are complex media, defined by both libraries. In all, 281 (61%) identified Library 1 were common 2. Many these involved fundamentally important metabolic...

Students of color, particularly women face substantial barriers in STEM disciplines higher education due to social isolation and interpersonal, technological, institutional biases. For example, online exam proctoring software often uses facial detection technology identify potential cheating behaviors. Undetected faces result flagging notifying instructors these as “suspicious” instances needing manual review. However, algorithms employed by may be biased against students with certain skin...

Exploration of interspecies interactions between microorganisms can have taxonomic, ecological, evolutionary, or medical applications. To better explore it is important to establish the ideal conditions that ensure survival all species involved. In this study, we sought identify biotic and abiotic factors would result in high co-culture viability two interkingdom species, Pseudomonas aeruginosa Acanthamoeba castellanii , soil dwelling microbes. There been limited studies showing long-term...

Pseudomonas aeruginosa is an opportunistic bacterial pathogen capable of causing a wide range disease manifestations, including severe pneumonia. Recently, clinics have reported rise in nosocomial infections with multidrug resistant (MDR) species, MDR strains P. aeruginosa. In order to quickly evaluate the efficacy new therapeutics for infections, highly reproducible and validated animal models need be developed pre-clinical testing. Here, we describe characterization two murine study...

The bacterium Myxococcus xanthus employs extracellular signals to coordinate aggregation and sporulation during multicellular development. Extracellular, contact-dependent signaling that involves the CsgA protein (called C-signaling) activates FruA, a putative response regulator governs branched pathway inside cells. One branch regulates cell movement, leading aggregation. other gene expression, sporulation. C-signaling is required for full expression of most genes induced after 6 h into...

ABSTRACT Porphyromonas gingivalis is considered a major etiologic agent in adult periodontitis. Gingipains are among its most important virulence factors, but their release unique strain HG66. We present the genome sequence of HG66 with single contig 2,441,680 bp and G+C content 48.1%.

In this paper we analyze an experiment for the use of low-cost gas sensors intended to detect bacteria in wounds using a non-intrusive technique. Seven different genera/species microbes tend be present most wound infections. Detection these usually requires sample and laboratory testing which is costly, inconvenient time-consuming. The validation processes with nineteen types (1 Candida, 2 Enterococcus, 6 Staphylococcus, 1 Aeromonas, Micrococcus, E. coli Pseudomonas) are presented here, four...

Abstract Exploration of interspecies interactions between microorganisms can have taxonomic, ecological, evolutionary, or medical applications. To better explore it is important to establish the ideal conditions that ensure survival all species involved. In this study, we sought identify biotic and abiotic factors would result in high co-culture viability two interkingdom species, Pseudomonas aeruginosa Acanthamoeba castellanii , soil dwelling microbes. Long-term these organisms has...

Closing of gaps in draft assemblies using Next Generation Sequencing (NGS) data is becoming increasingly important. In this paper we present CloG, a software pipeline that uses NGS to close assemblies. Firstly, CloG the VELVET assembler generate hybrid assembly from mixture reads: short reads and original (treated as long reads). It then closes between adjacent contigs by reconciling (i.e., "stitching") two By exploiting strengths both stitching reassembly, able outperform its contemporaries...

Many pathogenic microbial ecosystems are polymicrobial, and community function can be shaped by interbacterial interactions. Little is known, however, regarding the genetic determinants required for fitness in heterotypic environments.

ABSTRACT Myxococcus xanthus utilizes extracellular signals during development to coordinate cell movement, differentiation, and changes in gene expression. One of these signals, the C signal, regulates expression many genes, including Ω4400, a identified by an insertion Tn 5 lac into chromosome. Expression Ω4400 is reduced csgA mutant cells, which fail perform signaling, promoter region has several sequences similar found regulatory regions other C-signal-dependent genes. such gene, Ω4403,...

The bacterium Myxococcus xanthus undergoes multicellular development during times of nutritional stress and uses extracellular signals to coordinate cell behavior. C-signal affects gene expression late in development, including that Omega4499, an operon identified by insertion Tn5 lac into the M. chromosome. Omega4499 promoter region has several sequences common with those found previously be important for other C-signal-dependent promoters. To determine if these are activity, effects...