We compared the effectiveness of 4 sampling methods to recover Staphylococcus aureus, Klebsiella pneumoniae and Clostridioides difficile from contaminated environmental surfaces: cotton swabs, RODAC culture plates, sponge sticks with manual agitation, and sponge sticks with a stomacher. Organism type was the most important factor in bacterial recovery.

OBJECTIVES/GOALS: MiaA is a highly conserved prenyl transferase that catalyzes synthesis of the i6A37 tRNA modification in E. coli. While transcriptional regulation of MiaA is well characterized, there is no information on the MiaA post-transcriptional regulation. The aim of this study is to characterize the post-transcriptional regulation of the MiaA gene in E. coli. METHODS/STUDY POPULATION: To characterize the post-transcriptional regulation of miaA, we executed a targeted genetic screen of an E. coli small RNA library on a miaA-lacZ translational reporter fusion strain to identify small RNAs (sRNAs) that modulate MiaA translation or transcription termination. We also measured MiaA mRNA levels and miaA-lacZ activity in the absence or over-expression of candidate sRNA regulators of MiaA. We also measured MiaA mRNA levels in the absence of RNaseE and PNPase, two enzymes involved in mRNA turnover. Finally, we measured the ability of purified recombinant CsrA to bind to the MiaA mRNA transcript in vitro. RESULTS/ANTICIPATED RESULTS: We identified the carbon sensing sRNA CsrB and its cognate protein interaction partner CsrA, as potential post-transcriptional regulators of MiaA. Over-expression of CsrB fully repressed miaA-lacZ activity and MiaA mRNA levels. The absence of CsrA resulted in a defective miaA-lacZ activity and a 10-fold decrease in MiaA mRNA levels. We also identified an increase in the MiaA mRNA half-life particularly in the absence of RNaseE. Our results demonstrate an additional layer of regulation for the miaA operon by the CsrA/CsrB protein-sRNA system. DISCUSSION/SIGNIFICANCE: MiaA is a highly conserved bacterial protein. Our data may represent phenomena in an array of bacteria that could be targeted by novel antibiotics. The human MiaA homologue, TRIT1, plays a role in mitochondrial disorders. We anticipate that information garnered from MiaA studies will elucidate TRIT1 function and its role in mitochondrial disorders.

As practitioners of a historical science, paleontologists and geoscientists are well versed in the idea that the ability to understand and to anticipate the future relies upon our collective knowledge of the past. Despite this understanding, the fundamental role that the history of paleontology and the geosciences plays in shaping the structure and culture of our disciplines is seldom recognized and therefore not acted upon sufficiently. Here, we present a brief review of the history of paleontology and geology in Western countries, with a particular focus on North America since the 1800s. Western paleontology and geology are intertwined with systematic practices of exclusion, oppression, and erasure that arose from their direct participation in the extraction of geological and biological resources at the expense of Black, Indigenous, and People of Color (BIPOC). Our collective failure to acknowledge this history hinders our ability to address these issues meaningfully and systemically in present-day educational, academic, and professional settings. By discussing these issues and suggesting some ways forward, we intend to promote a deeper reflection upon our collective history and a broader conversation surrounding racism, colonialism, and exclusion within our scientific communities. Ultimately, it is necessary to listen to members of the communities most impacted by these issues to create actionable steps forward while holding ourselves accountable for the past.

ABSTRACT IMPACT: MiaA has a human homolog known as TRIT1. Mutations in TRIT1 have been associated with rare diseases such as MELAS and MERRF syndromes. These diseases are associated with mitochondrial disfunction.Understanding the mechanisms of bacterial sRNAs, and the miRNAs associated with these diseases could potentially afford the insight into effective cures. OBJECTIVES/GOALS: The aim is to investigate the regulation and function of tRNA isopentyladenine transferase enzyme in Escherichia coli. We aimed to execute screens for the identification of small RNA regulators of MiaA. The study will also investigate if i6A tRNA modification is necessary for the expression of major heat shock and mitochondrial proteins. METHODS/STUDY POPULATION: We constructed a chromosomal miaA-lacZ translational fusion driven by the arabinose responsive PBAD promoter and used it to screen against an Escherichia coli small RNA library. Using CsrB, one of our candidate sRNA regulators from our genetic screen, we measured the steady state levels of MiaA by Northern Blot in a PBAD-miaA2(P2HS)-lacZ translational fusion strain whereby pBR-pLac-csrB, pBR-pLac-csrA and the pBR-pLac vector are over-expressed, and under the control of an IPTG inducible promoter. Additionally, and in the same PBAD-miaA2(P2HS)-lacZ translational fusion strain background, we measured the steady state levels of MiaA in the wild type, csrA:zeo mutant strain, and csrA:zeo pBR-pLac-csrA complementation strain to determine if a combination of the pair would restore the wild-type genotype. RESULTS/ANTICIPATED RESULTS: Upon measuring the effect of small RNAs on miaA expression using quantitative b-galactosidase assays, we saw a 5-fold decrease in the expression of MiaA in the miaA-lacZ translational fusion containing sRNA CsrB, suggesting that this sRNA may play a role in the regulation of post-transcriptional expression of MiaA.From our northern blotting analysis, we observed a 6-fold decrease in MiaA expression in the absence of csrA, suggesting that csrA is essential for MiaA expression. DISCUSSION/SIGNIFICANCE OF FINDINGS: Identifying, mapping and characterizing how MiaA is regulated post-transcriptionally will give us an increased understanding in the maintenance and regulation of the normal function of E.coli to conserve homeostasis and translation fidelity.

The target site of glyphosate [N-(phosphonomethyl)glycine] inhibition in plants and bacteria is 5-enolpyruvylshikimate 3-phosphate (EPSP) synthase. Our strategy for developing glyphosate-resistant crops has been to genetically engineer plants with a gene that codes for EPSP synthase with low sensitivity in glyphosate. We cloned such a gene from the aroA locus of a glyphosate-resistant mutagenized strain of Salmonella typhimurium. The enzyme encoded by this gene has a single amino acid change resulting in lower affinity for glyphosate and higher affinity for substrates than either plant or wild-type bacterial counterpart. A chimaeric gene containing the mutant aroA gene behind the octopine synthase promoter was constructed and integrated into Agrobacterium T-DNA vectors. Analysis of gall tissue from Brassica campestris L. (turnip rape) infected with A. tumefaciens K12 containing this chimaera showed mRNA and protein expressed from the bacterial gene; 50% of the total EPSP synthase activity present had kinetic properties of the mutant bacterial enzyme. Tobacco (Nicotiana tabacum L. ‘Xanthi′) plants have been regenerated from cocultivation with A. rhizogenes containing the same construct; analysis indicates expression of the gene and enhanced tolerance to glyphosate.

Previous research with annual weed species indicates that critical timing of weed removal begins primarily after the two-leaf stage of onion, a time when postemergence (POST) herbicides can first be applied. Volunteer potato is difficult to manage and persists in onion fields of western United States. The purpose of this research was to quantify the duration of volunteer potato interference on yield and market grade of onion as well as potato tuber production. Volunteer potato interference caused a 5% yield loss before onions reached the two-leaf stage, at two of three locations. Relative to weed-free plots, onion bulb diameter was reduced as duration of interference increased, resulting in smaller proportions of marketable bulbs. Volunteer potato produced daughter tubers shortly after emergence, which explains, in part, weed persistence despite removal of shoots with contact herbicides, cultivation, and hand-weeding in onion. Significant losses in onion yield and bulb diameter are likely given current volunteer potato management systems.

Volunteer potato is highly competitive with onion and few control tactics are effective for removing this weed from an onion crop. Both volunteer potato density and duration of interference reduce onion yield, but the interaction of these factors is unknown. Field trials were conducted in 2003 in Idaho, Oregon, and Washington to determine the influence of volunteer potato density on the critical time of weed removal (CTWR) in onion. Yield losses of 2.5, 5.0, and 10% were estimated to occur at 534, 654, and 830 growing degree days (GDD) after onion emergence, respectively, with a volunteer potato density of 0.5 plants/m2. At 2.0 volunteer potato plants/m2, yield losses of 2.5, 5.0, and 10% were estimated to occur at 388, 481, and 598 GDD after onion emergence, respectively. Volunteer potato at 2.0 plants/m2 had to be removed at least one onion leaf stage sooner, compared to a weed density of 0.5 plants/m2, to avoid yield loss. Yield loss due to volunteer potato density or duration of interference was greatest among jumbo, colossal, and supercolossal market grades (P ≤ 0.1). Lowering potato tuber density in crops preceding onion will extend the critical time for weed removal and reduce the risk of crop loss.

Bulb onions are poor competitors and volunteer potato, commonly observed in western USA onion fields, is difficult to manage. To improve the understanding of onion and weed interactions, relationships were quantified among volunteer potato density, onion yield, and volunteer potato tuber production using hyperbolic or linear models. Onion yield losses because of volunteer potato interference occur at densities commonly observed in the field. A volunteer potato density as low as 0.067 plants m−2 resulted in a 10% reduction in crop yield. Asymptotic yield loss (A parameter) was 100% and achieved with 4 volunteer potato plants m−2. Volunteer potato competition limits onion bulb size, resulting in a lower quality and thus a less-valuable crop. Volunteer potato tuber density and biomass increased linearly with initial weed density as high as 8 volunteer potato m−2. Onion yield loss from volunteer potato competition occurs to a greater extent and at a lower weed density than demonstrated in previous research on small-seeded annual weed species.

The fossil record displays remarkable stasis in many species over long time periods, yet studies of extant populations often reveal rapid phenotypic evolution and genetic differentiation among populations. Recent advances in our understanding of the fossil record and in population genetics and evolutionary ecology point to the complex geographic structure of species being fundamental to resolution of how taxa can commonly exhibit both short-term evolutionary dynamics and long-term stasis.

Given their sheer number and evidence for long-term prehistoric occupation, atolls occupy a unique position in the peopling of the Pacific. However, they have frequently been overlooked in favor of larger islands due to a host of logistical and other issues. Once viewed as marginal environments, current research is now showing that small islands like these may have been more attractive to settlers than once thought. A new research program in Micronesia is dedicated to examining atolls to better develop baseline chronologies and investigate long-term human adaptations. As part of the initial stage of the project, we present the first radiocarbon dates (n=10) from Mwoakilloa (Mokil) atoll, which support a continuous occupation beginning between 1700–1560 cal BP (2σ). When compared to the settlement of other atoll groups in Micronesia such as the Marshall Islands—along with the nearby high volcanic islands of Pohnpei and Kosrae at approximately 2000–1800 yr ago—the dates from Mwoakilloa suggest a nearly contemporaneous or only slightly later occupation. The recovery of faunal material also demonstrates the translocation of at least two animals (Pacific rat and dog) to the island by humans that was coeval with early settlement. Additionally, there is evidence of landscape transformation in the form of a relatively large artificial mound created by debris and platform accumulation unseen elsewhere in central-eastern Micronesia, but common to atolls. These new dates reinforce the notion that Mwoakilloa and other atolls are integral to understanding prehistoric adaptations across the vast Pacific, though many questions still remain such as to the degree of interaction that occurred with nearby islands and whether settlement was continuous or intermittent through time.

Reports of bloodstream infections caused by methicillin-resistant Staphylococcus aureus among chronic hemodialysis patients to 2 Centers for Disease Control and Prevention surveillance systems (National Healthcare Safety Network Dialysis Event and Emerging Infections Program) were compared to evaluate completeness of reporting. Many methicillin-resistant S. aureus bloodstream infections identified in hospitals were not reported to National Healthcare Safety Network Dialysis Event.

Infect. Control Hosp. Epidemiol. 2016;37(2):205–207