Understanding the efficacy of wastewater surveillance for SARS-CoV-2 in two diverse communities.

During the COVID-19 pandemic, wastewater-based surveillance has been shown to be a useful tool for monitoring the spread of disease in communities and the emergence of new viral variants of concern. As the pandemic enters its fourth year and clinical testing has declined, wastewater offers a consistent non-intrusive way to monitor community health in the long term. This study sought to understand how accurately wastewater monitoring represented the actual burden of disease between communities.

Validity assessment of Michigan's proposed qPCR threshold value for rapid water-quality monitoring of E. coli contamination.

Michigan's water-quality standards specify that E. coli concentrations at bathing beaches must not exceed 300 E. coli per 100 mL, as determined by the geometric mean of culture-based concentrations in three or more representative samples from a given beach on a given day.

RNase E Promotes Expression of Type III Secretion System Genes in Pseudomonas aeruginosa.

is an important opportunistic pathogen that employs a type III secretion system (T3SS) to inject effector proteins into host cells. Using a protein depletion system, we show that the endoribonuclease RNase E positively regulates expression of the T3SS genes. We also present evidence that RNase E antagonizes the expression of genes of the type VI secretion system and limits biofilm production in Thus, RNase E, which is thought to be the principal endoribonuclease involved in the initiation of RNA degradation in , plays a key role in controlling the production of factors involved in both acute and chronic stages of infection.

Modeling Pollen-Mediated Virus Spread in Bee Colonies as a Classroom Activity.

Using a hands-on approach, this activity introduces students to the concept of viral spread and honey bee pathogenesis by illustrating pathogen transmission throughout the hive. This viral transmission activity, designed for introductory biology, virology, or microbiology classes, can be used in laboratory or lecture settings. Students are provided with information on viral transmission and hive structure.

Efficacy of a nitric oxide dressing in decreasing bacterial counts on human skin.

Objective: Inhibiting the proliferation of skin bacteria, using nitric oxide (NO), is a potential strategy to prevent infections. This study evaluated the efficacy of using a new NO releasing film dressing to decrease resident human bacterial skin flora compared with the measured microbial activity underneath control sterile Tegaderm transparent dressings.

Methods: A within-subjects design using a sample of convenience compared the bacterial counts under the skin of experimental dressings to those under control dressings.

Cellular Light Scattering for the Identification of Bacteria and Its Application to the Identification of .

Rapid identification of bacteria is critical in clinical and food safety applications. This paper describes a novel instrument and data analysis method for identifying bacteria based on the measurement of laser light scattering as the beam interacts with bacterial cells suspended in water. A description of the technology is followed by an identification performance study for a set of strains from the genus Staphylococcus (the inclusive target organisms) and a set of non-Staphylococcus strains (the exclusive organisms).

Regulation of acetyl-CoA synthetase transcription by the CrbS/R two-component system is conserved in genetically diverse environmental pathogens.

The CrbS/R two-component signal transduction system is a conserved regulatory mechanism through which specific Gram-negative bacteria control acetate flux into primary metabolic pathways. CrbS/R governs expression of acetyl-CoA synthase (acsA), an enzyme that converts acetate to acetyl-CoA, a metabolite at the nexus of the cell's most important energy-harvesting and biosynthetic reactions. During infection, bacteria can utilize this system to hijack host acetate metabolism and alter the course of colonization and pathogenesis.

Growth phase-dependent control of transcription start site selection and gene expression by nanoRNAs.

Prokaryotic and eukaryotic RNA polymerases can use 2- to ∼4-nt RNAs, "nanoRNAs," to prime transcription initiation in vitro. It has been proposed that nanoRNA-mediated priming of transcription can likewise occur under physiological conditions in vivo and influence transcription start site selection and gene expression. However, no direct evidence of such regulation has been presented.

NanoRNAs prime transcription initiation in vivo.

It is often presumed that, in vivo, the initiation of RNA synthesis by DNA-dependent RNA polymerases occurs using NTPs alone. Here, using the model Gram-negative bacterium Pseudomonas aeruginosa, we demonstrate that depletion of the small-RNA-specific exonuclease, Oligoribonuclease, causes the accumulation of oligoribonucleotides 2 to ∼4 nt in length, "nanoRNAs," which serve as primers for transcription initiation at a significant fraction of promoters. Widespread use of nanoRNAs to prime transcription initiation is coupled with global alterations in gene expression.

Bacillus subtilis RNase J1 endonuclease and 5' exonuclease activities in the turnover of DeltaermC mRNA.

RNase J1, a ribonuclease with 5' exonuclease and endonuclease activities, is an important factor in Bacillus subtilis mRNA decay. A model for RNase J1 endonuclease activity in mRNA turnover has RNase J1 binding to the 5' end and tracking to a target site downstream, where it makes a decay-initiating cleavage. The upstream fragment from this cleavage is degraded by 3' exonucleases; the downstream fragment is degraded by RNase J1 5' exonuclease activity.

Rsd family proteins make simultaneous interactions with regions 2 and 4 of the primary sigma factor.

Bacterial anti-sigma factors typically regulate sigma factor function by restricting the access of their cognate sigma factors to the RNA polymerase (RNAP) core enzyme. The Escherichia coli Rsd protein forms a complex with the primary sigma factor, sigma(70), inhibits sigma(70)-dependent transcription in vitro, and has been proposed to function as a sigma(70)-specific anti-sigma factor, thereby facilitating the utilization of alternative sigma factors. In prior work, Rsd has been shown to interact with conserved region 4 of sigma(70), but it is not known whether this interaction suffices to account for the regulatory functions of Rsd.

Local and global regulators linking anaerobiosis to cupA fimbrial gene expression in Pseudomonas aeruginosa.

The cupA gene cluster of Pseudomonas aeruginosa encodes components and assembly factors of a putative fimbrial structure that enable this opportunistic pathogen to form biofilms on abiotic surfaces. In P. aeruginosa the control of cupA gene expression is complex, with the H-NS-like MvaT protein functioning to repress phase-variable (on/off) expression of the operon.

Effect of 5'-proximal elements on decay of a model mRNA in Bacillus subtilis.

Previous work showed that a 42-nucleotide sequence from an SP82 bacteriophage early RNA functions as a 5' mRNA stabilizer in Bacillus subtilis. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of decay of a model mRNA with alterations at the 5'-end was used to elucidate the mechanism of SP82-mediated stability. A predicted 5'-terminal stem-loop structure was essential for stabilization.

TetL tetracycline efflux protein from Bacillus subtilis is a dimer in the membrane and in detergent solution.

The TetL antiporter from the Bacillus subtilis inner membrane is a tetracycline-divalent cation efflux protein that is energized by the electrochemical proton gradient across the membrane. In this study, we expressed tetL in Escherichia coli and investigated the oligomeric state of TetL in the membrane and in detergent solution. Evidence for an oligomeric state of TetL emerged from SDS-PAGE and Western blot analysis of membrane samples as well as purified protein samples from cells that expressed two differently tagged TetL species.

Effect of translational signals on mRNA decay in Bacillus subtilis.

A 254-nucleotide model mRNA, designated deltaermC mRNA, was used to study the effects of translational signals and ribosome transit on mRNA decay in Bacillus subtilis. DeltaermC mRNA features a strong ribosome-binding site (RBS) and a 62-amino-acid-encoding open reading frame, followed by a transcription terminator structure. Inactivation of the RBS or the start codon resulted in a fourfold decrease in the mRNA half-life, demonstrating the importance of ternary complex formation for mRNA stability.

Endonuclease cleavage of messenger RNA in Bacillus subtilis.

A deletion derivative of the ermC gene was constructed that expresses a 254-nucleotide mRNA. The small size of this mRNA facilitated the detection of processing products that did not differ greatly in size from the full-length transcript. In the presence of erythromycin, which induces ribosome stalling near the 5' end of ermC mRNA, the 254-nucleotide mRNA was cleaved endonucleolytically at the site of ribosome stalling.