Biology Commons^™

De Novo Sequencing, Assembly, And Annotation Of The Transcriptome For The Free-Living Testate Amoeba Arcella Intermedia, Giulia M. Ribeiro, Alfredo L. Porfírio-Sousa, Xyrus X. Maurer-Alcalá, Laura A. Katz, Daniel J.G. Lahr

Biological Sciences: Faculty Publications

Arcella, a diverse understudied genus of testate amoebae is a member of Tubulinea in Amoebozoa group. Transcriptomes are a powerful tool for charac- terization of these organisms as they are an efficient way of characterizing the protein-coding potential of the genome. In this work, we employed both sin- gle-cell and clonal populations transcriptomics to create a reference transcrip- tome for Arcella. We compared our results with annotations of Dictyostelium discoideum, a model Amoebozoan. We assembled a pool of 38 Arcella inter- media transcriptomes, which after filtering are composed of a total of 14,712 translated proteins. There are GO categories enriched …

Top-Down And Bottom-Up Controls On Microeukaryotic Diversity (I.E., Amplicon Analyses Of Sar Lineages) And Function (I.E., Metatranscriptome Analyses) Assessed In Microcosm Experiments, Jean David Grattepanche, Laura A. Katz

Biological Sciences: Faculty Publications

The availability of high-throughput sequencing (HTS) has transformed our understanding of the diversity of microbial eukaryotes (i.e., protists) across diverse habitats. Yet relating this biodiversity to function remains a challenge, particularly in the context of microbial food webs. Here we perform a set of microcosm experiments to evaluate the impact of changing predator and prey concentrations on a marine protist community, focusing on SAR (Stramenopila, Alveolata, and Rhizaria) lineages. We combine an estimate of taxonomic diversity through analysis of SSU-rDNA amplicons with metatranscriptomics, a proxy for function. We assess changes in a community sampled from New England waters with varying …

What Does Soil-Transmitted Helminth Elimination Look Like? Results From A Targeted Molecular Detection Survey In Japan, Mitsuko Hasegawa, Nils Pilotte, Mihoko Kikuchi, Arianna R. Means, Marina Papaiakovou, Andrew M. Gonzalez, Jacqueline R.M.A. Maasch, Hiroshi Ikuno, Toshihiko Sunahara, Kristjana H. Ásbjörnsdóttir, Judd L. Walson, Steven A. Williams, Shinjiro Hamano

Biological Sciences: Faculty Publications

Background: Japan is one of the few countries believed to have eliminated soil-transmitted helminths (STHs). In 1949, the national prevalence of Ascaris lumbricoides was 62.9%, which decreased to 0.6% in 1973 due to improvements in infrastructure, socioeconomic status, and the implementation of national STH control measures. The Parasitosis Prevention Law ended in 1994 and population-level screening ceased in Japan; therefore, current transmission status of STH in Japan is not well characterized. Sporadic cases of STH infections continue to be reported, raising the possibility of a larger-scale recrudescence of STH infections. Given that traditional microscopic detection methods are not sensitive to …

What Does Soil-Transmitted Helminth Elimination Look Like? Results From A Targeted Molecular Detection Survey In Japan, Mitsuko Hasegawa, Nils Pilotte, Mihoko Kikuchi, Arianna R. Means, Marina Papaiakovou, Andrew M. Gonzalez, Jacqueline R.M.A. Maasch, Hiroshi Ikuno, Toshihiko Sunahara, Kristjana H. Ásbjörnsdóttir, Judd L. Walson, Steven A. Williams, Shinjiro Hamano

Biological Sciences: Faculty Publications

Background: Japan is one of the few countries believed to have eliminated soil-transmitted helminths (STHs). In 1949, the national prevalence of Ascaris lumbricoides was 62.9%, which decreased to 0.6% in 1973 due to improvements in infrastructure, socioeconomic status, and the implementation of national STH control measures. The Parasitosis Prevention Law ended in 1994 and population-level screening ceased in Japan; therefore, current transmission status of STH in Japan is not well characterized. Sporadic cases of STH infections continue to be reported, raising the possibility of a larger-scale recrudescence of STH infections. Given that traditional microscopic detection methods are not sensitive to …

Phylogenomics Of The Epigenetic Toolkit Reveals Punctate Retention Of Genes Across Eukaryotes, Agnes K.M. Weiner, Mario A. Cerón-Romero, Ying Yan, Laura A. Katz

Biological Sciences: Faculty Publications

Epigenetic processes in eukaryotes play important roles through regulation of gene expression, chromatin structure, and genome rearrangements. The roles of chromatin modification (e.g., DNA methylation and histone modification) and non-protein-coding RNAs have been well studied in animals and plants. With the exception of a few model organisms (e.g., Saccharomyces and Plasmodium), much less is known about epigenetic toolkits across the remainder of the eukaryotic tree of life. Even with limited data, previous work suggested the existence of an ancient epigenetic toolkit in the last eukaryotic common ancestor. We use PhyloToL, our taxon-rich phylogenomic pipeline, to detect homologs of epigenetic genes …

Dietary Fish Oil Supplement Induces Age-Specific Contractile And Proteomic Responses In Muscles Of Male Rats, David W. Russ, Kalina Dimova, Emily Morris, Marguerite Pacheco, Sean M. Garvey, Stylianos P. Scordilis

Biological Sciences: Faculty Publications

Background: Dietary fish oil (DFO) has been identified as a micronutrient supplement with the potential to improve musculoskeletal health in old age. Few data are available for effects of DFO on muscle contractility, despite the significant negative impact of muscle weakness on age-related health outcomes. Accordingly, the effects of a DFO intervention on the contractile function and proteomic profile of adult and aged in an animal model of aging were investigated.

Methods: This preliminary study evaluated 14 adult (8 months) and 12 aged (22 months) male, Sprague-Dawley rats consuming a DFO-supplemented diet or a control diet for 8 weeks (7 …

Comparative Studies On The Polymorphism And Copy Number Variation Of Mtssu Rdna In Ciliates (Protista, Ciliophora): Implications For Phylogenetic, Environmental, And Ecological Research, Yurui Wang, Yaohan Jiang, Yongqiang Liu, Yuan Li, Laura A. Katz, Feng Gao, Ying Yan

Biological Sciences: Faculty Publications

No abstract provided.

Molecular Detection Of Intestinal Helminths And Protozoa Among Young Children In Dosso Region, Niger, Kristen Aiemjoy, Ahmed M. Arzika, Catherine Cook, Elodie Lebas, Nils Pilotte, Jessica R. Grant, Steven A. Williams, Thomas M. Lietman, Jeremy D. Keenan

Biological Sciences: Faculty Publications

Eukaryotic parasites are significant contributors to childhood illness in Niger. While helminthiases have received national attention through mass deworming efforts, the epidemiology of intestinal protozoa in Niger remains underexamined. This study employed real-time PCR diagnostics to describe the prevalence of two schistosomes, four soil-transmitted helminths, and one protozoan parasite in Boboye Department, Dosso Region. Prevalence was assessed using bulk stool specimens collected from a population-based sample of 86 children residing in 9 communities. Anthropometric measurements were used to calculate child growth z-scores and stool consistency was graded. Helminths were absent from the study population, with the exception of a single …

Misconceptions About Conception And Other Fallacies: Historical Bias In Reproductive Biology, Virginia Hayssen

Biological Sciences: Faculty Publications

Natural selection (differential reproduction) is a major tenet of evolutionary theory. In mammals the success of reproduction is primarily controlled by females who provide the majority of offspring care via gestation and lactation. In some species, maternal care also extends post-weaning. This primacy of female reproduction in evolution has not quite crept into our understanding of organismal adaptations in anatomy, physiology, and behavior. This cultural legacy has left its mark and led to misconceptions in our understanding of reproductive biology that are especially prominent in the understanding of reproduction in the general public. Here, I give examples of such misconceptions. …

The Wisdom Of A Crowd Of Near-Best Fits: Drug-Resistant Tuberculosis In The United States, Ellie Mainou, Gwen Spencer, Dylan Shepardson, Robert Dorit

Biological Sciences: Faculty Publications

Antibiotic-resistant tuberculosis (TB) strains pose a major challenge to TB eradication. Existing US epidemiological models have not fully incorporated the impact of antibiotic-resistance. To develop a more realistic model of US TB dynamics, we formulated a compartmental model integrating single-and multi-drug resistance. We fit twenty-seven parameters to twenty-two years of historical data using a genetic algorithm to minimize a non-differentiable error function. Since counts for several compartments are not available, many parameter combinations achieve very low error. We demonstrate that a crowd of near-best fits can provide compelling new evidence about the ranges of key parameters. While available data is …

Eukaryotic Plankton Communities Across Reef Environments In Bocas Del Toro Archipelago, Panamá, Andrea M. Rodas, Rachel M. Wright, Logan K. Buie, Hannah E. Aichelman, Karl D. Castillo, Sarah W. Davies

Biological Sciences: Faculty Publications

Variation in light and temperature can influence the genetic diversity and structure of marine plankton communities. While open-ocean plankton communities receive much scientific attention, little is known about how environmental variation affects plankton communities on tropical coral reefs. Here, we characterize eukaryotic plankton communities on coral reefs across the Bocas del Toro Archipelago, Panama´. Temperature loggers were deployed, and midday light levels were measured to quantify environmental differences across reefs at four inshore and four offshore sites (Inshore = Punta Donato, Smithsonian Tropical Research Institute (STRI) Point, Cristobal, Punta Laurel and Offshore = Drago Mar, Bastimentos North, Bastimentos South, and …

Combined Genome And Transcriptome Analyses Of The Ciliate Schmidingerella Arcuata (Spirotrichea) Reveal Patterns Of Dna Elimination, Scrambling, And Inversion, Susan A. Smith, Xyrus X. Maurer-Alcalá, Ying Yan, Laura A. Katz, Luciana F. Santoferrara, George B. Mcmanus

Biological Sciences: Faculty Publications

Schmidingerella arcuata is an ecologically important tintinnid ciliate that has long served as a model species in plankton trophic ecology. We present a partial micronuclear genome and macronuclear transcriptome resource for S. arcuata, acquired using single-cell techniques, and we report on pilot analyses including functional annotation and genome architecture. Our analysis shows major fragmentation, elimination, and scrambling in the micronuclear genome of S. arcuata. This work introduces a new nonmodel genome resource for the study of ciliate ecology and genomic biology and provides a detailed functional counterpart to ecological research on S. arcuata.

Complementary Metagenomic Approaches Improve Reconstruction Of Microbial Diversity In A Forest Soil, L. V. Alteio, F. Schulz, R. Seshadri, N. Varghese, W. Rodriguez-Reillo, E. Ryan, D. Goudeau, S. A. Eichorst, R. R. Malmstrom, R. M. Bowers, Laura A. Katz, J. L. Blanchard, T. Woyke

Biological Sciences: Faculty Publications

Soil ecosystems harbor diverse microorganisms and yet remain only partially characterized as neither single-cell sequencing nor whole-community sequencing offers a complete picture of these complex communities. Thus, the genetic and metabolic potential of this “uncultivated majority” remains underexplored. To address these challenges, we applied a pooled-cell-sorting-based mini-metagenomics approach and compared the results to bulk metagenomics. Informatic binning of these data produced 200 mini-metagenome assembled genomes (sorted-MAGs) and 29 bulk metagenome assembled genomes (MAGs). The sorted and bulk MAGs increased the known phylogenetic diversity of soil taxa by 7.2% with respect to the Joint Genome Institute IMG/M database and showed clade-specific …

Engaging New Audiences With Imaging And Microscopy, Michael J. Barresi, Enrico Coen, Elisabeth Kugler, Jamie Shuda, Derek C. Sung

Biological Sciences: Faculty Publications

In this Spotlight, we hear first-hand accounts from five scientists and educators who use microscopy and imaging to engage, entertain, educate and inspire new audiences with science and the field of developmental biology in particular. The 'voices' that follow each convey each authors' own personal take on why microscopy is such a powerful tool for capturing the minds, and the hearts, of scientists, students and the public alike. They discuss how microscopy and imaging can reveal new worlds, and improve our communication and understanding of developmental biology, as well as break down barriers and promote diversity for future generations of …

Δscope: A New Method To Quantify 3d Biological Structures And Identify Differences In Zebrafish Forebrain Development, Morgan S. Schwartz, Jake Schnabl, Mackenzie P.H. Litz, Benjamin Baumer, Michael Barresi

Biological Sciences: Faculty Publications

Research in the life sciences has traditionally relied on the analysis of clear morphological phenotypes, which are often revealed using increasingly powerful microscopy techniques analyzed as maximum intensity projections (MIPs). However, as biology turns towards the analysis of more subtle phenotypes, MIPs and qualitative approaches are failing to adequately describe these phenotypes. To address these limitations and quantitatively analyze the three-dimensional (3D) spatial relationships of biological structures, we developed the computational method and program called ∆SCOPE (Changes in Spatial Cylindrical Coordinate Orientation using PCA Examination). Our approach uses the fluorescent signal distribution within a 3D data set and reorients the …

The Concept Of The Hologenome, An Epigenetic Phenomenon, Challenges Aspects Of The Modern Evolutionary Synthesis, Adena Collens, Emma Kelley, Laura A. Katz

Biological Sciences: Faculty Publications

John Tyler Bonner's call to re-evaluate evolutionary theory in light of major transitions in life on Earth (e.g., from the first origins of microbial life to the evolution of sex, and the origins of multicellularity) resonate with recent discoveries on epigenetics and the concept of the hologenome. Current studies of genome evolution often mistakenly focus only on the inheritance of DNA between parent and offspring. These are in line with the widely accepted Neo-Darwinian framework that pairs Mendelian genetics with an emphasis on natural selection as explanations for the evolution of biodiversity on Earth. Increasing evidence for widespread symbioses complicates …

Gene Expression Of Endangered Coral (Orbicella Spp.) In Flower Garden Banks National Marine Sanctuary After Hurricane Harvey, Rachel M. Wright, Adrienne M.S. Correa, Lucinda A. Quigley, Lory Z. Santiago-Vázquez, Kathryn E.F. Shamberger, Sarah W. Davies

Biological Sciences: Faculty Publications

About 190 km south of the Texas–Louisiana border, the East and West Flower Garden Banks (FGB) have maintained > 50% coral cover with infrequent and minor incidents of disease or bleaching since monitoring began in the 1970s. However, a mortality event, affecting 5.6 ha (2.6% of the area) of the East FGB, occurred in late July 2016 and coincided with storm-generated freshwater runoff extending offshore and over the reef system. To capture the immediate effects of storm-driven freshwater runoff on coral and symbiont physiology, we leveraged the heavy rainfall associated with Hurricane Harvey in late August 2017 by sampling FGB corals …

Single-Cell Transcriptomics Reveal A Correlation Between Genome Architecture And Gene Family Evolution In Ciliates, Ying Yan, Xyrus X. Maurer-Alcalá, Rob Knight, Sergei L.Kosakovsky Pond, Laura A. Katz

Biological Sciences: Faculty Publications

Ciliates, a eukaryotic clade that is over 1 billion years old, are defined by division of genome function between transcriptionally inactive germline micronu-clei and functional somatic macronuclei. To date, most analyses of gene family evolution have been limited to cultivable model lineages (e.g., Tetrahymena, Paramecium, Oxytricha, and Stylonychia). Here, we focus on the uncultivable Karyorelictea and its understudied sister class Heterotrichea, which represent two extremes in genome architecture. Somatic macronuclei within the Karyorelictea are described as nearly diploid, while the Heterotrichea have hyperpolyploid somatic genomes. Previous analyses indicate that genome architecture impacts ciliate gene family evolution as the most diverse …

Ubiquity Or Not Ubiquity: That Is The Question, Robin S. Sleith, Laura A. Katz

Biological Sciences: Faculty Publications

The nature of population structure in eukaryotic microbes has been the subject of intense debate, but until recently the tools to test these hypotheses were either problematic (e.g., allozymes that cannot detect all genetic changes) or beyond financial and technological limits of most laboratories (e.g., high throughput sequencing). In a recent issue of Molecular Ecology, Craig et al. (2019) use a genomic approach to investigate the population structure of a model alga, the chlorophyte Chlamydomonas reinhardtii (Figure 1). Using high throughput sequencing, read mapping, and variant calling, they detected strong signals of differentiation at a continental scale, while local patterns …

Production Of Dynein And Kinesin Motor Ensembles On Dna Origami Nanostructures For Single Molecule Observation, Jingjie Hu, Nathan Derr

Biological Sciences: Faculty Publications

Summary

The goal of this protocol is to form ensembles of molecular motors on DNA origami nanostructures and observe the ensemble motility using total internal reflection fluorescence microscopy.

Abstract

Cytoskeletal motors are responsible for a wide variety of functions in eukaryotic cells, including mitosis, cargo transport, cellular motility, and others. Many of these functions require motors to operate in ensembles. Despite a wealth of knowledge about the mechanisms of individual cytoskeletal motors, comparatively less is known about the mechanisms and emergent behaviors of motor ensembles, examples of which include changes to ensemble processivity and velocity with changing motor number, location, …

Evolution And Unprecedented Variants Of The Mitochondrial Genetic Code In A Lineage Of Green Algae, David Zihala, Marek Eliáš, Laura A. Katz

Biological Sciences: Faculty Publications

Mitochondria of diverse eukaryotes have evolved various departures from the standard genetic code, but the breadth of possible modifications and their phylogenetic distribution are known only incompletely. Furthermore, it is possible that some codon reassignments in previously sequenced mitogenomes have been missed, resulting in inaccurate protein sequences in databases. Here we show, considering the distribution of codons at conserved amino acid positions in mitogenome-encoded proteins, that mitochondria of the green algal order Sphaeropleales exhibit a diversity of codon reassignments, including previously missed ones and some that are unprecedented in any translation system examined so far, necessitating redefinition of existing translation …

Positive Genetic Associations Among Fitness Traits Support Evolvability Of A Reef-Building Coral Under Multiple Stressors, Rachel M. Wright, Hanaka Mera, Carly D. Kenkel, Maria Nayfa, Line K. Bay, Mikhail V. Matz

Biological Sciences: Faculty Publications

Climate change threatens organisms in a variety of interactive ways that requires simultaneous adaptation of multiple traits. Predicting evolutionary responses requires an understanding of the potential for interactions among stressors and the genetic variance and covariance among fitness-related traits that may reinforce or constrain an adaptive response. Here we investigate the capacity of Acropora millepora, a reef-building coral, to adapt to multiple environmental stressors: rising sea surface temperature, ocean acidification, and increased prevalence of infectious diseases. We measured growth rates (weight gain), coral color (a proxy for Symbiodiniaceae density), and survival, in addition to nine physiological indicators of coral and …

A Case For Using Genomics And A Bioinformatics Pipeline To Develop Sensitive And Species-Specific Pcr-Based Diagnostics For Soil-Transmitted Helminths, Jessica R. Grant, Nils Pilotte, Steven A. Williams

Biological Sciences: Faculty Publications

The balance of expense and ease of use vs. specificity and sensitivity in diagnostic assays for helminth disease is an important consideration, with expense and ease often winning out in endemic areas where funds and sophisticated equipment may be scarce. In this review, we argue that molecular diagnostics, specifically new assays that have been developed with the aid of next-generation sequence data and robust bioinformatic tools, more than make up for their expense with the benefit of a clear and precise assessment of the situation on the ground. Elimination efforts associated with the London Declaration and the World Health Organization …

Pooling As A Strategy For The Timely Diagnosis Of Soil-Transmitted Helminths In Stool: Value And Reproducibility, Marina Papaiakovou, James Wright, Nils Pilotte, Darren Chooneea, Fabian Schär, James E. Truscott, Julia C. Dunn, Iain Gardiner, Judd L. Walson, Steven A. Williams, D. Timothy J. Littlewood

Biological Sciences: Faculty Publications

Background: The strategy of pooling stool specimens has been extensively used in the field of parasitology in order to facilitate the screening of large numbers of samples whilst minimizing the prohibitive cost of single sample analysis. The aim of this study was to develop a standardized reproducible pooling protocol for stool samples, validated between two different laboratories, without jeopardizing the sensitivity of the quantitative polymerase chain reaction (qPCR) assays employed for the detection of soil-transmitted helminths (STHs). Two distinct experimental phases were recruited. First, the sensitivity and specificity of the established protocol was assessed by real-time PCR for each one …

Genome Analyses Of The New Model Protist Euplotes Vannus Focusing On Genome Rearrangement And Resistance To Environmental Stressors, Xiao Chen, Yaohan Jiang, Feng Gao, Weibo Zheng, Timothy J. Krock, Naomi A. Stover, Chao Lu, Laura A. Katz, Weibo Song

Biological Sciences: Faculty Publications

As a model organism for studies of cell and environmental biology, the free-living and cosmopolitan ciliate Euplotes vannus shows intriguing features like dual genome architecture (i.e., separate germline and somatic nuclei in each cell/organism), “gene-sized” chromosomes, stop codon reassignment, programmed ribosomal frameshifting (PRF) and strong resistance to environmental stressors. However, the molecular mechanisms that account for these remarkable traits remain largely unknown. Here we report a combined analysis of de novo assembled high-quality macronuclear (MAC; i.e., somatic) and partial micronuclear (MIC; i.e., germline) genome sequences for E. vannus, and transcriptome profiling data under varying conditions. The results demonstrate that: (a) …

Causative Agent Of Canine Heartworm (Dirofilaria Immitis) Detected In Wild Lemurs, Sarah Zohdy, Kim Valenta, Bernadette Rabaoarivola, Weam I. Zaky, Nils Pilotte, Steven A. Williams, Colin A. Chapman, Zach J. Farris

Biological Sciences: Faculty Publications

The lemurs of Madagascar are threatened by human activities. We present the first molecular detection of canine heartworm (Dirofilaria immitis) in a wild non-human primate, the mouse lemur (Microcebus rufus). Zoonotic D. immitis infection has been associated with clinical pathology that includes serious and often fatal cardiac and pulmonary reactions. With human encroachment and associated increases in free-roaming dog populations in Madagascar, we examined lemurs for zoonotic canid pathogens. D. immitis presents a new potential conservation threat to lemurs. We highlight the need for wide-ranging and effective interventions, particularly near protected areas, to address this growing …

Phylotol: A Taxon/Gene-Rich Phylogenomic Pipeline To Explore Genome Evolution Of Diverse Eukaryotes, Mario A. Cerón-Romero, Xyrus X. Maurer-Alcalá, Jean David Grattepanche, Ying Yan, Miguel M. Fonseca, Laura A. Katz, Fabia Ursula Battistuzz

Biological Sciences: Faculty Publications

Estimating multiple sequence alignments (MSAs) and inferring phylogenies are essential for many aspects of comparative biology. Yet, many bioinformatics tools for such analyses have focused on specific clades, with greatest attention paid to plants, animals, and fungi. The rapid increase in high-throughput sequencing (HTS) data from diverse lineages now provides opportunities to estimate evolutionary relationships and gene family evolution across the eukaryotic tree of life. At the same time, these types of data are known to be error-prone (e.g., substitutions, contamination). To address these opportunities and challenges, we have refined a phylogenomic pipeline, now named PhyloToL, to allow easy incorporation …

Hybrid Genome Assembly Of A Neotropical Mutualistic Ant, Juliane Hartke, Tilman Schell, Evelien Jongepier, Hanno Schmidt, Philipp P. Sprenger, Juraj Paule, Erich Bornberg-Bauer, Thomas Schmitt, Florian Menzel, Markus Pfenninger, Barbara Feldmeyer, Laura A. Katz

Biological Sciences: Faculty Publications

The success of social insects is largely intertwined with their highly advanced chemical communication system that facilitates recognition and discrimination of species and nest-mates, recruitment, and division of labor. Hydrocarbons, which cover the cuticle of insects, not only serve as waterproofing agents but also constitute a major component of this communication system. Two cryptic Crematogaster species, which share their nest with Camponotus ants, show striking diversity in their cuticular hydrocarbon (CHC) profile. This mutualistic system therefore offers a great opportunity to study the genetic basis of CHC divergence between sister species. As a basis for further genome-wide studies high-quality genomes …

Frankia-Enriched Metagenomes From The Earliest Diverging Symbiotic Frankia Cluster: They Come In Teams, Thanh Van Nguyen, Daniel Wibberg, Theoden Vigil-Stenman, Fede Berckx, Kai Battenberg, Kirill N. Demchenko, Jochen Blom, Maria P. Fernandez, Takashi Yamanaka, Alison M. Berry, Jörn Kalinowski, Andreas Brachmann, Katharina Pawlowski, Laura A. Katz

Biological Sciences: Faculty Publications

Frankia strains induce the formation of nitrogen-fixing nodules on roots of actinorhizal plants. Phylogenetically, Frankia strains can be grouped in four clusters. The earliest divergent cluster, cluster-2, has a particularly wide host range. The analysis of cluster-2 strains has been hampered by the fact that with two exceptions, they could never be cultured. In this study, 12 Frankia-enriched metagenomes of Frankia cluster-2 strains or strain assemblages were sequenced based on seven inoculum sources. Sequences obtained via DNA isolated from whole nodules were compared with those of DNA isolated from fractionated preparations enhanced in the Frankia symbiotic structures. The results show …

Targeting A Highly Repeated Germline Dna Sequence For Improved Real-Time Pcr-Based Detection Of Ascaris Infection In Human Stool, Nils Pilotte, Jacqueline R.M.A. Maasch, Alice V. Easton, Eric Dahlstrom, Thomas B. Nutman, Steven A. Williams

Biological Sciences: Faculty Publications

Background: With the expansion of soil transmitted helminth (STH) intervention efforts and the corresponding decline in infection prevalence, there is an increased need for sensitive and specific STH diagnostic assays. Previously, through next generation sequencing (NGS)-based identification and targeting of non-coding, high copy-number repetitive DNA sequences, we described the development of a panel of improved quantitative real-time PCR (qPCR)-based assays for the detection of Necator americanus, Ancylostoma duodenale, Ancylostoma ceylanicum, Trichuris trichiura, and Strongyloides stercoralis. However, due to the phenomenon of chromosome diminution, a similar assay based on high copy-number repetitive DNA was not developed for the detection of Ascaris …