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Closing Keynote. Navigating Change: It's A Whitewater Adventure, Marjorie J. Kostelnik

Innovation in Pedagogy and Technology Symposium

Some people think of higher education as a sleepy backwater, where hardly anything changes too often. But for all of us living through it right now, higher education has become a whitewater adventure! Change is happening all around us in hundreds of different ways. Today, we’ll explore the who, why, how and what of change.

• Who is changing…who is promoting change…who has to live with the change

• Why are some people just better at change than others and what can you do to help your organization move forward?

• How can you manage change best … both as …

Wpa News 107 (2018), World Pheasant Association

Galliformes Specialist Group and Affiliated Societies: Newsletters

WPA News (Winter 2018), number 107

Published by the World Pheasant Association

Effects Of Red Clover Isoflavones On Tall Fescue Seed Fermentation And Microbial Populations In Vitro, Emily A. Melchior, Jason K. Smith, Liesel G. Schneider, Travis Mulliniks, Gary E. Bates, Zachary D. Mcfarlane, Michael D. Flythe, James L. Klotz, Jack P. Goodman, Huihua Ji, Phillip R. Meyer

West Central Research and Extension Center, North Platte

Negative impacts of endophyte-infected Lolium arundinaceum (Darbyshire) (tall fescue) are responsible for over $2 billion in losses to livestock producers annually. While the influence of endophyte-infected tall fescue has been studied for decades, mitigation methods have not been clearly elucidated. Isoflavones found in Trifolium pratense (red clover) have been the subject of recent research regarding tall fescue toxicosis mitigation. Therefore, the aim of this study was to determine the effect of ergovaline and red clover isoflavones on rumen microbial populations, fiber degradation, and volatile fatty acids (VFA) in an in vitro system. Using a dose of 1.10 mg …

Comparison Of Three Regionalization Techniques For Predicting Streamflow In Ungaged Watersheds In Nebraska, Usa Using Swat Model, Michael W. Van Liew, Aaron R. Mittelstet

Department of Agricultural and Biological Systems Engineering: Faculty Publications

This study compared three approaches, regional averaging, nearest neighbor, and donor techniques, to regionalize parameters in the Soil and Water Assessment Tool (SWAT) on eleven watersheds located in the Dissected Plains, Plains, and Rolling Hills Landforms in the eastern portion of the State of Nebraska, USA. Within the Rolling Hills Landform, three watersheds were randomly selected as calibration watersheds while two were randomly selected as validation watersheds. Two watersheds were randomly selected as calibration watersheds while one was randomly selected as a validation watershed within each of the Dissected Plains and Plains Landforms. The seven calibration watersheds were used to …

Influence Of Invasive Hybrid Cattails On Habitat Use By Common Loons, Spencer L. Wesche, Benjamin J. O'Neal, Steve K. Windels, Bryce T. Olson, Max Larreur, Adam A. Ahlers

United States National Park Service: Publications

An invasive hybrid cattail species, Typha x glauca (T. x glauca), is rapidly expanding across the United States and Canada. Dense clonal stands of T. x glauca outcompete native wetland plants, reduce open-water habitats, and negatively affect native wetland plant diversity; however, effects of hybrid cattail expansions on native wildlife are still unclear. We used multiple surveys and single-season occupancy models to examine how the relative coverage of T. x glauca affected habitat use by common loons (Gavia immer) at 71 wetland sites in Voyageurs National Park, Minnesota, USA, during summer 2016. Delineated wetland sites (2 …

Resistance To Deltamethrin In Prairie Dog (Cynomys Ludovicianus) Fleas In The Field And In The Laboratory, David A. Eads, Dean E. Biggins, Jonathan Bowser, Janet C. Mcallister, Randall L. Griebel, Eddie Childers, Travis M. Livieri, Cristi Painter, Lindsey Sterling Krank, Kristy Bly

United States National Park Service: Publications

Sylvatic plague poses a substantial risk to black-tailed prairie dogs (Cynomys ludovicianus) and their obligate predator, the black-footed ferret (Mustela nigripes). The effects of plague on prairie dogs and ferrets are mitigated using a deltamethrin pulicide dust that reduces the spread of plague by killing fleas, the vector for the plague bacterium. In portions of Conata Basin, Buffalo Gap National Grassland, and Badlands National Park, South Dakota, US, 0.05% deltamethrin has been infused into prairie dog burrows on an annual basis since 2005. We aimed to determine if fleas (Oropsylla hirsuta) in portions of …

The Utility Of Point Count Surveys To Predict Wildlife Interactions With Wind Energy Facilities: An Example Focused On Golden Eagles, Maitreyi Sur, James R. Belthoff, Emily R. Bjerre, Brian A. Millsap, Todd Katzner

United States Fish and Wildlife Service: Publications

Wind energy development is rapidly expanding in North America, often accompanied by requirements to survey potential facility locations for existing wildlife. Within the USA, golden eagles (Aquila chrysaetos) are among the most high-profile species of birds that are at risk from wind turbines. To minimize golden eagle fatalities in areas proposed for wind development, modified point count surveys are usually conducted to estimate use by these birds. However, it is not always clear what drives variation in the relationship between on-site point count data and actual use by eagles of a wind energy project footprint. We used existing …

A Rapid And Efficient Method For Enriching Mitochondrial Dna From Plants, Mackenzie M. Strehle, Emma Purfeerst, Alan C. Christensen

School of Biological Sciences: Faculty Publications

Current mitochondrial purification techniques are tedious and protracted due to their emphasis on recovering physiologically active mitochondria. However, for studies that are exclusively interested in isolating mitochondrial DNA (mtDNA) for applications such as PCR and sequencing, respiring mitochondria - and the complex procedures that stem from the need to retain their function - are unnecessary. Still, global DNA extraction methods have proven insufficient for mitochondrial DNA isolation because nuclear mitochondrial DNA segments (NUMTs) pose unique challenges to accurate mtDNA quantification and characterization. We present a rapid and simple extraction technique that maximizes recovery of mitochondrial DNA from plant cells, while …

Genome-Wide Association Study For The Relationship Between Temperature And Feed Intake In Beef Cattle, Robel Araya Ghebrewold

Department of Animal Science: Dissertations, Theses, and Student Research

Environmental conditions, such as changes in ambient temperature, can cause changes in animal behavior and performance. In general, it is believed that as ambient temperature increases, dry matter intake (DMI) of beef cattle decreases. However, our hypothesis was that the degree to which animals adjust their daily DMI due to changes in ambient temperature is partially controlled by genetic effects. Consequently, the objective of this study was to estimate the genetic component of the regression of DMI on ambient temperature using an admixed beef cattle population consisting of various crosses of Angus, Simmental, and Piedmontese (n = 239). Ambient …

Rumen Bacterial Community Structure Impacts Feed Efficiency In Beef Cattle, Henry A. Paz, Kristin E. Hales, James E. Wells, Larry A. Kuehn, Harvey C. Freetly, Elaine D. Berry, Michael D. Flythe, Matthew L. Spangler, Samodha C. Fernando

Department of Animal Science: Faculty Publications

The importance of the rumen microbiota on nutrient cycling to the animal is well recognized; however, our understanding of the influence of the rumen microbiome composition on feed efficiency is limited. The rumen microbiomes of two large animal cohorts (125 heifers and 122 steers) were characterized to identify specific bacterial members (operational taxonomic units [OTUs]) associated with feed efficiency traits (ADFI, ADG, and G:F) in beef cattle. The heifer and steer cohorts were fed a forage-based diet and a concentrate-based diet, respectively. A rumen sample was obtained from each animal via esophageal tubing and bacterial community composition was …

Using Single Molecule Mrna Fluorescent In Situ Hybridization (Rna-Fish) To Quantify Mrnas In Individual Murine Oocytes And Embryos, Fang Xie, Kelsey A. Timme, Jennifer R. Wood

Department of Animal Science: Faculty Publications

Changes in abundance of mRNAs during oocyte growth and maturation and during pre-implantation embryo development have been documented using quantitative real-time RT-PCR (qPCR), microarray analyses, and whole genome sequencing. However, these techniques require amplification of mRNAs, normalization using housekeeping genes, can be biased for abundant transcripts, and/or require large numbers of oocytes and embryos which can be difficult to acquire from mammalian species. We optimized a single molecule RNA fluorescence in situ hybridization (RNA-FISH) protocol, which amplifies fluorescence signal to detect candidate transcripts, for use with individual oocytes and embryos. Quantification using the software Localize showed patterns of Gdf9 and …

Improved Reference Genome For The Domestic Horse Increases Assembly Contiguity And Composition, Theodore S. Kalbfleisch, Edward S. Rice, Michael S. Depriest Jr., Brian P. Walenz, Matthew S. Hestand, Joris R. Vermeesch, Brendan L. O'Connell, Ian T. Fiddes, Alisa O. Vershinina, Nedda F. Saremi, Jessica L. Petersen, Carrie J. Finno, Rebecca R. Bellone, Molly E. Mccue, Samantha A. Brooks, Ernest Bailey, Ludovic Orlando, Richard E. Green, Donald C. Miller, Douglas F. Antczak, James N. Macleod

Department of Animal Science: Faculty Publications

Recent advances in genomic sequencing technology and computational assembly methods have allowed scientists to improve reference genome assemblies in terms of contiguity and composition. EquCab2, a reference genome for the domestic horse, was released in 2007. Although of equal or better quality compared to other first-generation Sanger assemblies, it had many of the shortcomings common to them. In 2014, the equine genomics research community began a project to improve the reference sequence for the horse, building upon the solid foundation of EquCab2 and incorporating new short-read data, long-read data, and proximity ligation data. Here, we present EquCab3. The count of …

Genome-Wide Association Study Of Stayability And Heifer Pregnancy In Red Angus Cattle, S. E. Speidel, B. A. Buckley, R. J. Boldt, R. M. Enns, J. Lee, Matthew L. Spangler, M. G. Thomas

Department of Animal Science: Faculty Publications

Reproductive performance is the most important component of cattle production from the standpoint of economic sustainability of commercial beef enterprises. Heifer Pregnancy (HPG) and Stayability (STAY) genetic predictions are 2 selection tools published by the Red Angus Association of America (RAAA) to assist with improvements in reproductive performance. Given the importance of HPG and STAY to the profitability of commercial beef enterprises, the objective of this study was to identify QTL associated with both HPG and STAY in Red Angus cattle. A genome-wide association study (GWAS) was performed using deregressed HPG and …

Genome-Wide Association Study For Feed Efficiency Traits Using Snp And Haplotype Models, Kashly Rene Schweer, Stephen D. Kachman, Larry A. Kuehn, Harvey Freetly, John E. Pollak, Matthew L. Spangler

Department of Animal Science: Faculty Publications

Feed costs comprise the majority of variable expenses in beef cattle systems making feed efficiency an important economic consideration within the beef industry. Due to the expense of recording individual feed-intake phenotypes, a genomic-enabled approach could be advantageous toward improving this economically relevant trait complex. A genome-wide association study (GWAS) was performed using 748 crossbred steers and heifers representing seven sire breeds with phenotypes for ADG and ADFI. Animals were genotyped with the BovineSNP50v2 BeadChip containing approximately 54,000 SNP. Both traits were analyzed using univariate SNPbased (BayesC) and haplotype-based (BayesIM) models and jointly using BayesIM to perform a bivariate GWAS. …

Shinyaim: Shiny‐Based Application Of Interactive Manhattan Plots For Longitudinal Genome‐Wide Association Studies, Waseem Hussain, Malachy T. Campbell, Harkamal Walia, Gota Morota

Department of Animal Science: Faculty Publications

Owning to advancements in sensor‐based, non‐destructive phenotyping platforms, researchers are increasingly collecting data with higher temporal resolution. These phenotypes collected over several time points are cataloged as longitudinal traits and used for genome‐wide association studies (GWAS). Longitudinal GWAS typically yield a large number of output files, posing a significant challenge to data interpretation and visualization. Efficient, dynamic, and integrative data visualization tools are essential for the interpretation of longitudinal GWAS results for biologists; however, these tools are not widely available to the community. We have developed a flexible and user‐friendly Shiny‐based online application, ShinyAIM, to dynamically view and interpret temporal …

Nutr 250: Human Nutrition And Metabolism-A Peer Review Of Teaching Project Benchmark Portfolio, Karsten Koehler

UNL Faculty Course Portfolios

The purpose of this portfolio was to purposefully introduce and evaluate activities that are aimed at improving student learning and performance in NUTR250 – Human Metabolism, a large lecture-based 200-level course. Activities were designed to improve class preparation and ascertain student learning beyond exams (objective 1), provide time to review class material (objective 2), balance material related to concepts and knowledge (objective 3), and assess the effectiveness of activities (objective 4). To address objective 1, students completed weekly prequizzes, which improved student reading and preparation prior to class in approximately 2 out of 3 students. On average, students spend 33±27 …

Rna-Seq Of Kaposi's Sarcoma Reveals Alterations In Glucose And Lipid Metabolism, For Yue Tso, Andrew V. Kossenkov, Salum J. Lidenge, Owen Ngalamika, John R. Ngowi, Julius Mwaiselage, Jayamanna Wickramasinghe, Eun Hee Kwon, John T. West, Paul M. Lieberman, Charles Wood

School of Biological Sciences: Faculty Publications

Kaposi's sarcoma-associated herpesvirus (KSHV) is the etiologic agent of Kaposi's sarcoma (KS). It is endemic in a number of sub-Saharan African countries with infection rate of >50%. The high prevalence of HIV-1 coupled with late presentation of advanced cancer staging make KS the leading cancer in the region with poor prognosis and high mortality. Disease markers and cellular functions associated with KS tumorigenesis remain ill-defined. Several studies have attempted to investigate changes of the gene profile with in vitro infection of monoculture models, which are not likely to reflect the cellular complexity of the in vivo lesion environment. Our approach …

Efficacy Of Oral Administration Of Sodium Iodide To Prevent Bovine Respiratory Disease Complex, B. M. Shoemake, B. L. Vander Ley, B. W. Newcomer, M. C. Heller

School of Veterinary and Biomedical Sciences: Faculty Publications

Background: The prevention of bovine respiratory disease complex (BRD) in beef cattle is important to maintaining health and productivity of calves in feeding operations.

Objective: Determine whether BRD bacterial and viral pathogens are susceptible to the lactoperoxidase/hydrogen peroxide/ iodide (LPO/H₂O₂/I^-) system in vitro and to determine whether the oral administration of sodium iodide (NaI) could achieve sufficient concentrations of iodine (I) in the respiratory secretions of weaned beef calves to inactivate these pathogens in vivo.

Animals: Sixteen weaned, apparently healthy, commercial beef calves from the University of Missouri, College of Veterinary Medicine teaching herd. …

Methods, Systems, And Devices Relating To Force Control Surgical Systems, Shane M. Farritor, Thomas Frederick, Kearney Lackas, Joe Bartels, Jacob Greenburg

Department of Mechanical and Materials Engineering: Faculty Publications

The various embodiments herein relate to robotic surgical systems and devices that use force and/or torque sensors to measure forces applied at various components of the system or device. Certain implementations include robotic surgical devices having one or more force/torque sensors that detect or measure one or more forces applied at or on one or more arms. Other embodiments relate to systems having a robotic surgical device that has one or more sensors and an external controller that has one or more motors such that the sensors transmit information that is used at the controller to actuate the motors to …

Characterization Of Neofabraea Actinidiae And N. Brasiliensis As Causal Agents Of Apple Bull’S-Eye Rot In Southern Brazil, Amauri Bogo, Carla C. Comparin, Rosa M. Valdebenito Sanhueza, Patricia Ritschel, Ricardo T. Casa, Fábio N. Silva, Sydney E. Everhart

Department of Plant Pathology: Faculty Publications

The causal agents of apple bull’s-eye rot in southern Brazil have recently been described as Neofabraea actinidiae and N. brasiliensis. Isolates of both species were evaluated for response of mycelial growth index (MGI) to different temperatures, enzyme production, mycelial growth inhibition and effective concentrations (EC₅₀ and EC₁₀₀) of the fungicides triflumizole, pyrimethanil and thiophanate methyl, as well as aggressiveness on fruits of ‘Fuji’ hybrid and ‘Pink Lady’. There was significantly lower mycelium growth in N. brasiliensis compared with N. actinidiae at all temperatures tested. Neither species grew at 3 and 32°C. There were minor differences in …

Elucidating The Role Of Highly Homologous Nicotiana Benthamiana Ubiquitin E2 Gene Family Members In Plant Immunity Through An Improved Virus‑Induced Gene Silencing Approach, Bangjun Zhou, Lirong Zeng

Department of Plant Pathology: Faculty Publications

Background: Virus-induced gene silencing (VIGS) has been used in many plant species as an attractive post transcriptional gene silencing (PTGS) method for studying gene function either individually or at large-scale in a high-throughput manner. However, the specificity and efficiency for knocking down members of a highly homologous gene family have remained to date a significant challenge in VIGS due to silencing of off-targets. Results: Here we present an improved method for the selection and evaluation of gene fragments used for VIGS to specifically and efficiently knock down members of a highly homologous gene family. Using this method, we knocked down …

The Tomato U-Box Type E3 Ligase Pub13 Acts With Group Iii Ubiquitin E2 Enzymes To Modulate Fls2-Mediated Immune Signaling, Bangjun Zhou, Lirong Zeng

Department of Plant Pathology: Faculty Publications

In Arabidopsis and rice, the ubiquitin ligase PUB13-mediated protein degradation plays a significant role in plant pattern-triggered immunity (PTI) and flowering time control. The Arabidopsis PUB13 has been shown to attenuate the pattern recognition receptor FLS2-mediated immune signaling by ubiquitinating FLS2 and consequently promoting its degradation by the 26S proteasome. Nevertheless, the cognate ubiquitin-conjugating enzymes (E2) with which PUB13 acts to modulate FLS2-mediated PTI are unknown. To address this question, we investigate here the tomato (Solanum lycopersicum) homolog of PUB13, SlPUB13 by utilizing the recently characterized complete set of tomato E2s. Of the 13 groups of tomato E2s, …

Proteomic Analysis Of Ubiquitinated Proteins In Rice (Oryza Sativa) After Treatment With Pathogen-Associated Molecular Pattern (Pamp) Elicitors, Xiao-Lin Chen, Xin Xie, Liye Wu, Caiyun Liu, Lirong Zeng, Xueping Zhou, Feng Luo, Guo-Liang Wang, Wende Liu

Department of Plant Pathology: Faculty Publications

Reversible protein ubiquitination plays essential roles in regulating cellular processes. Although many reports have described the functions of ubiquitination in plant defense responses, few have focused on global changes in the ubiquitome. To better understand the regulatory roles of ubiquitination in rice pattern-triggered immunity (PTI), we investigated the ubiquitome of rice seedlings after treatment with two pathogen-associated molecular patterns, the fungal-derived chitin or the bacterialderived flg22, using label-free quantitative proteomics. In chitin-treated samples, 144 and 167 lysine-ubiquitination sites in 121 and 162 proteins showed increased and decreased ubiquitination, respectively. In flg22-treated samples, 151 and 179 lysine-ubiquitination sites in 118 and …

Photosystem Ii Subunit S Overexpression Increases The Efficiency Of Water Use In A Field-Grown Crop, Katarzyna Glowacka, Johannes Kromdijk, Katherine Kucera, Jiayang Xie, Amanda P. Cavanagh, Lauriebeth Leonelli, Andrew D. B. Leakey, Donald R. Ort, Krishna K. Niyogi, Stephen P. Long

Department of Biochemistry: Faculty Publications

Insufficient water availability for crop production is a mounting barrier to achieving the 70% increase in food production that will be needed by 2050. One solution is to develop crops that require less water per unit mass of production. Water vapor transpires from leaves through stomata, which also facilitate the influx of CO2 during photosynthetic assimilation. Here, we hypothesize that Photosystem II Subunit S (PsbS) expression affects a chloroplastderived signal for stomatal opening in response to light, which can be used to improve wateruse efficiency. Transgenic tobacco plants with a range of PsbS expression, from undetectable to 3.7 …

Simulation Of Stimulation: Cytokine Dosage And Cell Cycle Crosstalk Driving Timing-Dependent T Cell Differentiation, Matteo Barberis, Tomáš Helikar, Paul Verbruggen

Department of Biochemistry: Faculty Publications

Triggering an appropriate protective response against invading agents is crucial to the effectiveness of human innate and adaptive immunity. Pathogen recognition and elimination requires integration of a myriad of signals from many different immune cells. For example, T cell functioning is not qualitatively, but quantitatively determined by cellular and humoral signals. Tipping the balance of signals, such that one of these is favored or gains advantage on another one, may impact the plasticity of T cells. This may lead to switching their phenotypes and, ultimately, modulating the balance between proliferating and memory T cells to sustain an appropriate immune response. …

Computational Systems Biology Approach For The Study Of Rheumatoid Arthritis: From A Molecular Map To A Dynamical Model, Vidisha Singh, Marek Ostaszewski, George D. Kalliolias, Gilles Chiocchia, Robert Olaso, Elisabeth Petit-Teixeira, Tomáš Helikar, Anna Niarakis

Department of Biochemistry: Faculty Publications

In this work we present a systematic effort to summarize current biological pathway knowledge concerning Rheumatoid Arthritis (RA). We are constructing a detailed molecular map based on exhaustive literature scanning, strict curation criteria, re-evaluation of previously published attempts and most importantly experts’ advice. The RA map will be web-published in the coming months in the form of an interactive map, using the MINERVA platform, allowing for easy access, navigation and search of all molecular pathways implicated in RA, serving thus, as an on line knowledgebase for the disease. Moreover the map could be used as a template for Omics data …

The Colomoto Interactive Notebook: Accessible And Reproducible Computational Analyses For Qualitative Biological Networks, Aurelien Naldi, Celine Hernandez, Nicolas Levy, Gautier Stoll, Pedro T. Monteiro, Claudine Chaouiya, Tomáš Helikar, Andrei Zinovyev, Laurence Calzone, Sarah Cohen-Boulakia, Denis Thieffry, Loic Pauleve

Department of Biochemistry: Faculty Publications

Analysing models of biological networks typically relies on workflows in which different software tools with sensitive parameters are chained together,many times with additional manual steps. The accessibility and reproducibility of such workflows is challenging, as publications often overlook analysis details, and because some of these tools may be difficult to install, and/or have a steep learning curve. The CoLoMoTo Interactive Notebook provides a unified environment to edit, execute, share, and reproduce analyses of qualitative models of biological networks. This framework combines the power of different technologies to ensure repeatability and to reduce users’ learning curve of these technologies. The framework …

A Mechanistic Computational Model Reveals That Plasticity Of Cd4+ T Cell Differentiation Is A Function Of Cytokine Composition And Dosage, Bhanwar Lal Puniya, Robert G. Todd, Akram Mohammed, Deborah M. Brown, Matteo Barberis, Tomáš Helikar

Department of Biochemistry: Faculty Publications

CD4+ T cells provide cell-mediated immunity in response to various antigens. During an immune response, naïve CD4+ T cells differentiate into specialized effector T helper (Th1, Th2, and Th17) cells and induced regulatory (iTreg) cells based on a cytokine milieu. In recent studies, complex phenotypes resembling more than one classical T cell lineage have been experimentally observed. Herein, we sought to characterize the capacity of T cell differentiation in response to the complex extracellular environment. We constructed a comprehensive mechanistic (logical) computational model of the signal transduction that regulates T cell differentiation. The model’s dynamics were characterized and analyzed under …

Genes For Membrane Transport Proteins: Not So Rare In Viruses, Timo Greiner, Anna Moroni, James L. Van Etten, Gerhard Thiel

James Van Etten Publications

Some viruses have genes encoding proteins with membrane transport functions. It is unknown if these types of proteins are rare or are common in viruses. In particular, the evolutionary origin of some of the viral genes is obscure, where other viral proteins have homologs in prokaryotic and eukaryotic organisms. We searched virus genomes in databases looking for transmembrane proteins with possible transport function. This effort led to the detection of 18 different types of putative membrane transport proteins indicating that they are not a rarity in viral genomes. The most abundant proteins are K+ channels. Their predicted structures vary between …

Size-Dependent Catalysis Of Chlorovirus Population Growth By A Messy Feeding Predator, John Delong, Zeina Al-Ameeli, Shelby Lyon, James L. Van Etten, David Dunigan

James Van Etten Publications

Many chloroviruses replicate in endosymbiotic zoochlorellae that are protected from infection by their symbiotic host. To reach the high virus concentrations that often occur in natural systems, a mechanism is needed to release zoochlorellae from their hosts. We demonstrate that the ciliate predator Didinium nasutum foraging on zoochlorellae-bearing Paramecium bursaria can release live zoochlorellae from the ruptured prey cell that can then be infected by chloroviruses. The catalysis process is very effective, yielding roughly 95% of the theoretical infectious virus yield as determined by sonication of P. bursaria. Chlorovirus activation is more effective with smaller Didinia, as larger …