Life Sciences Commons^™

A Newly Discovered Bordetella Species Carries A Transcriptionally Active Crispr-Cas With A Small Cas9 Endonuclease, Yury V. Ivanov, Nikki Shariat, Karen B. Register, Bodo Linz, Israel Rivera, Kai Hu, Edward G. Dudley, Eric T. Harvill

Biology Faculty Publications

Background

Clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated genes (cas) are widely distributed among bacteria. These systems provide adaptive immunity against mobile genetic elements specified by the spacer sequences stored within the CRISPR.

Methods

The CRISPR-Cas system has been identified using Basic Local Alignment Search Tool (BLAST) against other sequenced and annotated genomes and confirmed via CRISPRfinder program. Using Polymerase Chain Reactions (PCR) and Sanger DNA sequencing, we discovered CRISPRs in additional bacterial isolates of the same species of Bordetella. Transcriptional activity and processing of the CRISPR have been assessed via RT-PCR.

Results

Here we …

Application Of Β-Lactamase Reporter Fusions As An Indicator Of Effector Protein Secretion During Infections With The Obligate Intracellular Pathogen Chlamydia Trachomatis , Konrad E. Mueller, Kenneth A. Fields

Microbiology, Immunology, and Molecular Genetics Faculty Publications

Chlamydia spp. utilize multiple secretion systems, including the type III secretion system (T3SS), to deploy host-interactive effector proteins into infected host cells. Elucidation of secreted proteins has traditionally required ectopic expression in a surrogate T3SS followed by immunolocalization of endogenous candidate effectors to confirm secretion by chlamydiae. The ability to transform Chlamydia and achieve stable expression of recombinant gene products has enabled a more direct assessment of secretion. We adapted TEM-1 β-lactamase as a reporter system for assessment of chlamydial protein secretion. We provide evidence that this system facilitates visualization of secretion in the context of infection. Specifically, our findings …

The Ocean Sampling Day Consortium, Anna Kopf, Mesude Bicak, Renzo Kottman, Julia Schnetzer, Ivaylo Kostadinov, Katja Lehmann, Antonio Fernandez-Guerra, Christian Jeanthon, Eyal Rahav, Matthias Ulrich, Antje Wichels, Gunnar Gerdts, Paraskevi Polymenakou, Giorgos Kotoulas, Rania Siam, Rehab Z. Abdallah, Eva C. Sonnenschein, Thierry Cariou, Fergal O'Gara, Stephen Jackson, Sandi Orlic, Michael Steinke, Julia Busch, Bernardo Duarte, Isabel Caçador, João Canning-Clode, Oleksandra Bobrova, Viggo Marteinsson, Eyjolfur Reynisson, Clara Magalhães Loureiro, Gian Marco Luna, Grazia Marina Quero, Carolin R. Löscher, Anke Kremp, Marie E. Delorenzo, Lise Øvreås, Jennifer Tolman, Julie Laroche, Antonella Penna, Marc Frischer, Timothy Davis, Barker Katherine, Christopher P. Meyer, Sandra Ramos, Catarina Magalhães, Florence Jude-Lemeilleur, Ma Leopoldina Aguirre-Macedo, Shiao Y. Wang, Nicole Poulton, Scott Jones, Rachel Collin, Jed A. Fuhrman, Pascal Conan, Cecilia Alonso, Noga Stambler, Kelly Goodwin, Michael M. Yakimov, Federico Baltar, Levente Bodrossy, Jodie Van De Kamp, Dion Mf Frampton, Martin Ostrowski, Paul Van Ruth, Paul Malthouse, Simon Claus, Klaas Deneudt, Jonas Mortelmans, Sophie Pitois, David Wallom, Ian Salter, Rodrigo Costa, Declan C. Schroeder, Mahrous M. Kandil, Valentina Amaral, Florencia Biancalana, Rafael Santana, Maria Luiza Pedrotti, Takashi Yoshida, Hiroyuki Ogata, Tim Ingleton, Kate Munnik, Naiara Rodriguez-Ezpeleta, Veronique Berteaux-Lecellier, Patricia Wecker, Ibon Cancio, Daniel Valout, Christina Bienhold, Hassan Ghazal, Bouchra Chaouni, Soumya Essayeh, Sara Ettamimi, El Houcine Zaid, Noureddine Boukhatem, Abderrahim Bouali, Rajaa Chahboune, Said Barrijal, Mohammed Timinouni, Fatima El Otmani, Mohamed Bennani, Marianna Mea, Nadezhda Todorova, Ventzislav Karamfilov, Petra Ten Hoopten, Guy Cochrane, Stephanie L'Haridon, Kemal Can Bizel, Alessandro Vezzi, Federico M. Lauro, Patrick Martin, Rachelle M. Jensen, Jamie Hinks, Susan Gebbels, Riccardo Rosselli, Fabio De Pascale, Riccardo Schiavon, Antonina Dos Santos, Emilie Villar, Stéphane Pesant, Bruno Cataletto, Francesca Malfatti, Ranjith Edirisinghe, Jorge A. Herrera Silveira, Michele Barbier, Valentina Turk, Tinkara Tinta, Wayne J. Fuller, Ilkay Salihoglu, Nedime Serakinci, Mahmut Cerkez Ergoren, Eileen Bresnan, Juan Iriberri, Paul Anders Fronth Nyhus, Edvardsen Bente, Hans Erik Karlsen, Peter N. Golyshin, Josep M. Gasol, Snejana Moncheva, Nina Dzhembekova, Zackary Johnson, Christopher Daivd Sinigalliano, Maribeth Louise Gidley, Adriana Zingone, Roberto Danovaro, George Tsiamis, Melody S. Clark, Ana Cristina Costa, Monia El Bour, Ana M. Martins, R. Eric Collins, Anne-Lise Ducluzeau, Jonathan Martinez, Mark J. Costello, Linda A. Amaral-Zettler, Jack A. Gilbert, Neil Davies, Dawn Field, Frank Oliver Glöckner

Faculty Publications

Ocean Sampling Day was initiated by the EU-funded Micro B3 (Marine Microbial Biodiversity, Bioinformatics, Biotechnology) project to obtain a snapshot of the marine microbial biodiversity and function of the world’s oceans. It is a simultaneous global mega-sequencing campaign aiming to generate the largest standardized microbial data set in a single day. This will be achievable only through the coordinated efforts of an Ocean Sampling Day Consortium, supportive partnerships and networks between sites. This commentary outlines the establishment, function and aims of the Consortium and describes our vision for a sustainable study of marine microbial communities and their embedded functional traits.

Antibacterial Activity Of Native California Medicinal Plant Extracts Isolated From Rhamnus Californica And Umbellularia Californica., Maria G. Carranza, Mary B. Sevigny, Debashree Banerjee, Lacie Fox-Cubley

Natural Sciences and Mathematics | Faculty Scholarship

BACKGROUND: Antimicrobial resistance (AMR) is a major threat to global public health. Medicinal plants have long been used as remedies for infectious diseases by native cultures around the world and have the potential for providing effective treatments for antibiotic-resistant infections. Rhamnus californica (Rhamnaceae) and Umbellularia californica (Lauraceae) are two indigenous California plant species historically used by Native Americans to treat skin, respiratory and gastrointestinal infections. This study aimed to assess the in vitro antimicrobial activity of methanolic extracts of leaves and bark of R. and U. californica against methicillin-resistant Staphylococcus aureus (MRSA) and other Gram-positive and Gram-negative bacteria.

METHODS: Methanolic …

Anaerobic Detoxification Of Acetic Acid In A Thermophilic Ethanologen, A Joe Shaw, Bethany B. Miller, Stephen R. Rogers, William Robert Kenealy, Alex Meola, Ashwini Bhandiwad, W Ryan Sillers, Indraneel Shikhare, David Hogsett, Christopher Herring

Dartmouth Scholarship

The liberation of acetate from hemicellulose negatively impacts fermentations of cellulosic biomass, limiting the concentrations of substrate that can be effectively processed. Solvent-producing bacteria have the capacity to convert acetate to the less toxic product acetone, but to the best of our knowledge, this trait has not been transferred to an organism that produces ethanol at high yield. We have engineered a five-step metabolic pathway to convert acetic acid to acetone in the thermophilic anaerobe Thermoanaerobacterium saccharolyticum.

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Is There Specificity In A Defensive Mutualism Against Soil Versus Lab Nematodes, Dictyostelium Discoideum Farmers And Their Bacteria?, Boahemaa Adu-Oppong,, David C. Queller, Joan E. Strassmann

Biology Faculty Publications & Presentations

Background: The social amoeba Dictyostelium discoideum is a soil-dwelling microbe, which lives most of its life cycle in the vegetative stage as a predator of bacteria and as prey for nematodes. When bacteria are sparse, amoebae aggregate into a multicellular fruiting body. Some clones of D. discoideum have agriculture (Brock et al., 2011). They carry bacteria through the social stage, eat them prudently, and use some bacteria as defence against non-farming D. discoideum competitors. Caenorhabditis elegans preys on D. discoideum in the laboratory but does not encounter it in nature because C. elegans lives on rotten fruit. The nematode …

Examining Phage Infection Utilizing Mycobacterium Smegmatis, Tanya L. Riddick

Undergraduate Research

Bacteriophages, also known as phages, are viruses that are ubiquitous and survive and replicate within the host of the bacterial cell, Mycobacterium smegmatis. They are considered one of the most abundant organisms on earth (10³¹). Structurally, they are 100-200nm in size and consist of a protein encapsulated head that contains DNA or RNA, a tail sheath and tail fibers. This research consisted of examining phage infection, by re-isolating a novel phage, Tango. Tango was originally isolated genetically in 2013 by a previous ISBT student, Anna Maccarrone. The phage was sent to genetic sequencing but two phages were discovered, …

Microbial Communities Of The Providence River, Jacqueline Kratch

Honors Projects in Science and Technology

The Providence River has been industrialized for over one-hundred years. Industries such as oil storage and metal recycling facilities have left high levels of pollutant metals, including lead (Pb), in the soil and water. The elevated Pb concentrations in these environments influence the selection of Pb resistance mechanisms in the bacterial community1. One mechanism of heavy metal resistance is the Pb efflux pump, consisting of proteins in the cell membrane that aid in the transport of Pb out of the cell2. In this study we investigated the co-occurrence of Pb efflux pumps and antibiotic efflux pumps in bacteria from Pb …

A Response Regulator From A Soil Metagenome Enhances Resistance To The Β-Lactam Antibiotic Carbenicillin In Escherichia Coli, Heather K. Allen, Ran An, Jo Handelsman, Luke A. Moe

Plant and Soil Sciences Faculty Publications

Functional metagenomic analysis of soil metagenomes is a method for uncovering as-yet unidentified mechanisms for antibiotic resistance. Here we report an unconventional mode by which a response regulator derived from a soil metagenome confers resistance to the β-lactam antibiotic carbenicillin in Escherichia coli. A recombinant clone (βlr16) harboring a 5,169 bp DNA insert was selected from a metagenomic library previously constructed from a remote Alaskan soil. The βlr16 clone conferred specific resistance to carbenicillin, with limited increases in resistance to other tested antibiotics, including other β-lactams (penicillins and cephalosporins), rifampin, ciprofloxacin, erythromycin, chloramphenicol, nalidixic acid, fusidic acid, and gentamicin. Resistance …

Mechanisms Of Antimicrobial Resistant Salmonella Enterica Transmission Associated With Starling–Livestock Interactions, James C. Carlson, Doreene R. Hyatt, Jeremy W. Ellis, David R. Pipkin, Anna M. Mangan, Michael Russell, Denise S. Bolte, Richard M. Engeman, Thomas J. Deliberto, George M. Linz

USDA Wildlife Services: Staff Publications

Bird–livestock interactions have been implicated as potential sources for bacteria within concentrated animal feeding operations (CAFO). European starlings (Sturnus vulgaris) in particular are known to contaminate cattle feed and water with Salmonella enterica through their fecal waste.We propose that fecal waste is not the only mechanisms through which starlings introduce S. enterica to CAFO. The goal of this study was to assess if starlings can mechanically move S. enterica. We define mechanical movement as the transportation of media containing S. enterica, on the exterior of starlings within CAFO.We collected 100 starlings and obtained external wash and gastrointestinal tract (GI) samples. …

Molecular Characterization Of Salmonella Enterica Isolates Associated With Starling–Livestock Interactions, James C. Carlson, Doreene R. Hyatt, Kevin Bentler, Anna M. Mangan, Michael Russell, Antoinette J. Piaggio, George M. Linz

USDA Wildlife Services: Staff Publications

Bird–livestock interactions have been implicated as potential sources for bacteria within concentrated animal feeding operations (CAFO). In this study we characterized XbaIdigested genomic DNA from Salmonella enterica using pulsed-field gel electrophoresis (PFGE). The PFGE analysis was conducted using 182 S. enterica isolates collected from a single CAFO between 2009 and 2012. Samples collected in 2012 were subjected to antimicrobial susceptibility testing. The analysis was limited to S. enterica serotypes, with at least 10 isolates, known to occur in both European starlings (Sturnus vulgaris) and cattle (Bos taurus) within this CAFO. A total of five different serotypes were screened; S. Anatum, …

Assessment Of A Metaviromic Dataset Generated From Nearshore Lake Michigan, Siobhan C. Watkins, Neil Kuehnle, C Anthony Ruggeri, Kema Malki, Katherine Bruder, Jinan Elayyan, Kristina Damisch, Naushin Vahora, Paul O'Malley, Brianne Ruggles-Sage, Zachary Romer, Catherine Putonti

Bioinformatics Faculty Publications

Bacteriophages are powerful ecosystem engineers. They drive bacterial mortality rates and genetic diversity, and affect microbially mediated biogeochemical processes on a global scale. This has been demonstrated in marine environments; however, phage communities have been less studied in freshwaters, despite representing a potentially more diverse environment. Lake Michigan is one of the largest bodies of freshwater on the planet, yet to date the diversity of its phages has yet to be examined. Here, we present a composite survey of viral ecology in the nearshore waters of Lake Michigan. Sequence analysis was performed using a web server previously used to analyse …

Growth And Gas Production Of A Novel Obligatory Heterofermentative Cheddar Cheese Nonstarter Lactobacilli Species On Ribose And Galactose, Western Dairy Center, Utah State University, Megan Jane Ostler, Jeff Broadbent, Craig J. Oberg, Donald J. Mcmahon

Nutrition, Dietetics, and Food Sciences Faculty Publications

An obligatory heterofermentative lactic acid bacterium, Lactobacillus wasatchii sp. nov., isolated from gassy Cheddar cheese was studied for growth, gas formation, salt tolerance, and survival against pasteurization treatments at 63°C and 72°C. Initially, Lb. wasatchii was thought to use only ribose as a sugar source and we were interested in whether it could also utilize galactose. We conducted experiments to determine the rate and extent of growth and gas production in carbohydraterestricted (CR) de Man, Rogosa, and Sharpe (MRS) medium under anaerobic conditions with various combinations of ribose and galactose at 12, 23, and 37°C, with 23°C being the optimum …

New Tracer To Estimate Community Predation Rates Of Phagotrophic Protists, Alexander B. Bochdansky, Melissa A. Clouse

OES Faculty Publications

Predation of eukaryotic microbes on prokaryotes is one of the most important trophic interactions on Earth, representing a major mortality term and shaping morphology and composition of prokaryotic communities. Here we introduce and validate a new tracer to determine predation rates on prokaryotes. Minicells of Escherichia coli marked with a bright green fluorescent protein (GFP) vector have many operational advantages over previously used prey analogs such as fluorescently labeled bacteria. GFP-minicells are similar in size to naturally occurring bacteria from a variety of environments including the oligotrophic open ocean and the deep sea. They are relatively stable against microbial and …