DOCSLIB.ORG

The Development of a Dual Target Mycoplasma Bovis Taqman Real-Time PCR System for the Rapid Analysis of Bovine Semen THESIS Pres

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
The Development of a Dual Target Mycoplasma Bovis Taqman Real-Time PCR System for the Rapid Analysis of Bovine Semen THESIS Pres The Development of a Dual Target Mycoplasma bovis TaqMan real-time PCR System for the Rapid Analysis of Bovine Semen THESIS Presented in Partial Fulfillment of the Requirements for the Degree Master of Science in the Graduate School of The Ohio State University By Kristina Marie McDonald, B.A. Graduate Program in Veterinary Preventive Medicine The Ohio State University 2012 Master's Examination Committee: Dr. Gustavo Schuenemann, Advisor Dr. Ziv Raviv Dr. Päivi Rajala-Schultz Copyrighted by Kristina Marie McDonald 2012 Abstract Mycoplasma bovis is a major bovine pathogen causing respiratory disease, arthritis, otitis media, mastitis, and genital disorders in cattle worldwide. M. bovis infections tend to persist in affected herds and are often resistant to antibiotics. Spread by close contact of infected animals or a heavily contaminated environment, M. bovis can colonize mucosal surfaces and disseminate to other organ sites. Rapid identification and isolation of diseased animals is critical to prevent the spread of infection. Artificial insemination with M. bovis contaminated semen is a common source of infection of the female bovine genital tract. In this study, we report the development and validation of a dual target TaqMan real-time PCR system for the rapid detection of M. bovis in bovine semen. The assays target the housekeeping genes fusA and oppD/F. Both assays exclusively amplified M. bovis when tested against a panel 15 bovine mycoplasma species including 59 field and laboratory strains of M. bovis. Quantification was determined by testing serial dilutions of plasmids containing the target sequences; both the fusA and oppD/F assays demonstrated a detection limit of 1 plasmid copy/uL and efficiencies of 1.975 and 1.977 respectively. ii Bovine semen was spiked with serial dilutions of M. bovis in order to compare the detection limits of the real-time PCR assays and culture in clinical samples. Culture demonstrated a detection limit of 310 organisms/µl, while the fusA and oppD/F assays detected 3.1 organisms/µl. Fresh semen ejaculates from 26 commercial bulls were obtained from an artificial insemination center and evaluated for M. bovis by real-time PCR and culture. All samples were negative for the organism by both real-time PCR and culture. Although real-time PCR systems have been described for the detection of M. bovis in other bovine fluids and tissues, no PCR assays have been developed to address the specific evaluation of M. bovis in bovine semen. In this study, we present two highly specific and sensitive TaqMan real-time PCRs for the rapid detection and quantification of M. bovis in bovine semen. iii This thesis is dedicated to my cherished husband Rick, who supported me unconditionally throughout this journey. iv Acknowledgments I would like to thank my advisor, Dr. Ziv Raviv, for his guidance and support throughout my time in the Veterinary Preventive Medicine Department. His enthusiasm and dedication to my program ensured that I gained exceptional knowledge, not only about mycoplasmas, but about many molecular techniques as well. I would also like to recognize the members of my master’s examination committee, Dr. Päivi Rajala-Shultz and Dr. Gustavo Schuenemann, for their support and advice, which made this document possible. Finally, I would like to thank Select Sires, Inc. for funding my education and research. Throughout my career they have continued to support all of my ambitions and goals. v Vita 1999 - Rutherford B. Hayes High School, Delaware, OH 2007 - B.A. Microbiology, Ohio Wesleyan University 2007 to present - Select Sires, Inc., Plain City, OH 2010 to present - Graduate Student, The Ohio State University Publications McDonald, K., Wetzel, A., Raviv, Z. The Development of Dual Target Mycoplasma bovis TaqMan Real-Time PCRs for Rapid Analysis of Bull Semen. Publication Pending Wetzel, A., Lefevre, K., Raviv, Z. Revised Mycoplasma synoviae vlhA PCRs. Avian Diseases 2010; 54:1292-1297. Fields of Study Major Field: Veterinary Preventive Medicine Studies in Mycoplasma vi Table of Contents Abstract ............................................................................................................. ii Dedication ......................................................................................................... iv Acknowledgments ............................................................................................. x Vita .................................................................................................................... vi List of Tables ..................................................................................................... ix List of Figures .................................................................................................... x Chapter 1 Introduction ....................................................................................... 1 1.1 List of References .................................................................................. 4 Chapter 2 Literature review ............................................................................... 8 2.1 The Class Mollicutes.............................................................................. 8 2.2 The Genus Mycoplasma ........................................................................ 9 2.3 Bovine Mycoplasmas ............................................................................. 11 2.4 Mycoplasma bovis ................................................................................. 13 2.4.1 The distribution and economic impact of M. bovis ........................ 13 2.4.2 Pathogenesis of M. bovis .............................................................. 15 2.4.3 Transmission of M. bovis .............................................................. 16 2.4.4 M. bovis in the bovine genital tract................................................ 18 2.4.5 M. bovis Detection ........................................................................ 20 2.4.6 M. bovis treatment, prevention, and control .................................. 23 2.5 Conclusion ............................................................................................. 26 2.6 List of References .................................................................................. 30 vii Chapter 3 The development of a dual target Mycoplasma bovis TaqMan real-time PCR system for rapid the rapid analysis of bovine semen ................................ 36 3.1 Abstract ................................................................................................. 36 3.2 Introduction ............................................................................................ 37 3.3 Materials and Methods .......................................................................... 38 3.4 Results ................................................................................................... 46 3.5 Discussion ............................................................................................. 48 3.6 List of References .................................................................................. 69 Chapter 4 Conclusions and further directions .................................................... 74 4.1 List of References .................................................................................. 78 Bibliography ..................................................................................................... 81 viii List of Tables Table 2.1 Gross properties of bacterial genomes ........................................... 28 Table 2.2 Properties distinguishing Mollicutes from other Eubacteria ............ 29 Table 3.1 Mycoplasma isolates utilized in this study ...................................... 54 Table 3.2 Primer and probe sequences used for PCR systems ..................... 58 Table 3.3 Detection limits and reproducibility of the FusA real-time PCR ....... 59 Table 3.4 Detection limits and reproducibility of the OppD/F real-time PCR .. 60 Table 3.5 Summary of M. bovis spiked semen results ................................... 61 ix List of Figures Figure 3.1 Protocol for the isolation of mycoplasmas from semen ................. 62 Figure 3.2 Alignment of FusA sequences ....................................................... 63 Figure 3.3 Alignment of OppD/F sequences ................................................... 65 Figure 3.4 FusA standard curve ..................................................................... 67 Figure 3.5 OppD/F standard curve ................................................................. 68 x Chapter 1 Introduction The genus Mycoplasma, belonging to the class Mollicutes, contains small prokaryotic organisms which lack a cell wall and are bound by a plasma membrane. Mycoplasmas’ genome is the smallest known among free living and self-replicating organisms and the G/C content of their genome is low (23-40%). Because of their restricted genomic potential, mycoplasmas have limited functional metabolic pathways, and live as saprophytes or parasites in their hosts. Among the over 200 known species, several mycoplasmas are recognized as important human and animal pathogens [1]. Mycoplasma bovis (Mb) is the most economically significant and pathogenic bovine mycoplasma of intensely farmed cattle in countries free of contagious bovine pleuropneumonia [2-7]. M. bovis colonizes mucosal surfaces producing several diseases including pneumonia, arthritis, otitis media, and meningitis, in calves, and mastitis and genital infections in adult bovines [8,
Load more

Recommended publications
  • Investigating Prevalence and Geographical Distribution of Mycoplasma Sp. in the Gut of Atlantic Salmon (Salmo Salar L.)
    Master’s Thesis 2020 60 ECTS Faculty of Chemistry, Biotechnology and Food Science Investigating prevalence and geographical distribution of Mycoplasma sp. in the gut of Atlantic Salmon (Salmo salar L.) Mari Raudstein MSc Biotechnology ACKNOWLEDGEMENTS This master project was performed at the Faculty of Chemistry, Biotechnology and Food Science, at the Norwegian University of Life Sciences (NMBU), with Professor Knut Rudi as primary supervisor and associate professor Lars-Gustav Snipen as secondary supervisor. To begin with, I want to express my gratitude to my supervisor Knut Rudi for giving me the opportunity to include fish, one of my main interests, in my thesis. Professor Rudi has helped me execute this project to my best ability by giving me new ideas and solutions to problems that would occur, as well as answering any questions I would have. Further, my secondary supervisor associate professor Snipen has helped me in acquiring and interpreting shotgun data for this thesis. A big thank you to both of you. I would also like to thank laboratory engineer Inga Leena Angell for all the guidance in the laboratory, and for her patience when answering questions. Also, thank you to Ida Ormaasen, Morten Nilsen, and the rest of the Microbial Diversity group at NMBU for always being positive, friendly, and helpful – it has been a pleasure working at the MiDiv lab the past year! I am very grateful to the salmon farmers providing me with the material necessary to study salmon gut microbiota. Without the generosity of Lerøy Sjøtroll in Bømlo, Lerøy Aurora in Skjervøy and Mowi in Chile, I would not be able to do this work.
    [Show full text]
  • Bovine Mycoplasmosis
    Bovine mycoplasmosis The microorganisms described within the genus Mycoplasma spp., family Mycoplasmataceae, class Mollicutes, are characterized by the lack of a cell wall and by the little size (0.2-0.5 µm). They are defined as fastidious microorganisms in in vitro cultivation, as they require specific and selective media for the growth, which appears slower if compared to other common bacteria. Mycoplasmas can be detected in several hosts (mammals, avian, reptiles, plants) where they can act as opportunistic agents or as pathogens sensu stricto. Several Mycoplasma species have been described in the bovine sector, which have been detected from different tissues and have been associated to variable kind of gross-pathology lesions. Moreover, likewise in other zoo-technical areas, Mycoplasma spp. infection is related to high morbidity, low mortality and to the chronicization of the disease. Bovine Mycoplasmosis can affect meat and dairy animals of different ages, leading to great economic losses due to the need of prophylactic/metaphylactic measures, therapy and to decreased production. The new holistic approach to multifactorial or chronic diseases of human and veterinary interest has led to a greater attention also to Mycoplasma spp., which despite their elementary characteristics are considered fast-evolving microorganisms. These kind of bacteria, historically considered of secondary importance apart for some species, are assuming nowadays a greater importance in the various zoo-technical fields with increased diagnostic request in veterinary medicine. Also Acholeplasma spp. and Ureaplasma spp. are described as commensal or pathogen in the bovine sector. Because of the phylogenetic similarity and the common metabolic activities, they can grow in the same selective media or can be detected through biomolecular techniques targeting Mycoplasma spp.
    [Show full text]
  • The Mysterious Orphans of Mycoplasmataceae
    The mysterious orphans of Mycoplasmataceae Tatiana V. Tatarinova1,2*, Inna Lysnyansky3, Yuri V. Nikolsky4,5,6, and Alexander Bolshoy7* 1 Children’s Hospital Los Angeles, Keck School of Medicine, University of Southern California, Los Angeles, 90027, California, USA 2 Spatial Science Institute, University of Southern California, Los Angeles, 90089, California, USA 3 Mycoplasma Unit, Division of Avian and Aquatic Diseases, Kimron Veterinary Institute, POB 12, Beit Dagan, 50250, Israel 4 School of Systems Biology, George Mason University, 10900 University Blvd, MSN 5B3, Manassas, VA 20110, USA 5 Biomedical Cluster, Skolkovo Foundation, 4 Lugovaya str., Skolkovo Innovation Centre, Mozhajskij region, Moscow, 143026, Russian Federation 6 Vavilov Institute of General Genetics, Moscow, Russian Federation 7 Department of Evolutionary and Environmental Biology and Institute of Evolution, University of Haifa, Israel 1,2 [email protected] 3 [email protected] 4-6 [email protected] 7 [email protected] 1 Abstract Background: The length of a protein sequence is largely determined by its function, i.e. each functional group is associated with an optimal size. However, comparative genomics revealed that proteins’ length may be affected by additional factors. In 2002 it was shown that in bacterium Escherichia coli and the archaeon Archaeoglobus fulgidus, protein sequences with no homologs are, on average, shorter than those with homologs [1]. Most experts now agree that the length distributions are distinctly different between protein sequences with and without homologs in bacterial and archaeal genomes. In this study, we examine this postulate by a comprehensive analysis of all annotated prokaryotic genomes and focusing on certain exceptions.
    [Show full text]
  • Role of Protein Phosphorylation in Mycoplasma Pneumoniae
    Pathogenicity of a minimal organism: Role of protein phosphorylation in Mycoplasma pneumoniae Dissertation zur Erlangung des mathematisch-naturwissenschaftlichen Doktorgrades „Doctor rerum naturalium“ der Georg-August-Universität Göttingen vorgelegt von Sebastian Schmidl aus Bad Hersfeld Göttingen 2010 Mitglieder des Betreuungsausschusses: Referent: Prof. Dr. Jörg Stülke Koreferent: PD Dr. Michael Hoppert Tag der mündlichen Prüfung: 02.11.2010 “Everything should be made as simple as possible, but not simpler.” (Albert Einstein) Danksagung Zunächst möchte ich mich bei Prof. Dr. Jörg Stülke für die Ermöglichung dieser Doktorarbeit bedanken. Nicht zuletzt durch seine freundliche und engagierte Betreuung hat mir die Zeit viel Freude bereitet. Des Weiteren hat er mir alle Freiheiten zur Verwirklichung meiner eigenen Ideen gelassen, was ich sehr zu schätzen weiß. Für die Übernahme des Korreferates danke ich PD Dr. Michael Hoppert sowie Prof. Dr. Heinz Neumann, PD Dr. Boris Görke, PD Dr. Rolf Daniel und Prof. Dr. Botho Bowien für das Mitwirken im Thesis-Komitee. Der Studienstiftung des deutschen Volkes gilt ein besonderer Dank für die finanzielle Unterstützung dieser Arbeit, durch die es mir unter anderem auch möglich war, an Tagungen in fernen Ländern teilzunehmen. Prof. Dr. Michael Hecker und der Gruppe von Dr. Dörte Becher (Universität Greifswald) danke ich für die freundliche Zusammenarbeit bei der Durchführung von zahlreichen Proteomics-Experimenten. Ein ganz besonderer Dank geht dabei an Katrin Gronau, die mich in die Feinheiten der 2D-Gelelektrophorese eingeführt hat. Außerdem möchte ich mich bei Andreas Otto für die zahlreichen Proteinidentifikationen in den letzten Monaten bedanken. Nicht zu vergessen ist auch meine zweite Außenstelle an der Universität in Barcelona. Dr. Maria Lluch-Senar und Dr.
    [Show full text]
  • Mycoplasma Agalactiae MEMBRANE PROTEOME
    UNIVERSITÀ DEGLI STUDI DI SASSARI SCUOLA DI DOTTORATO IN SCIENZE BIOMOLECOLARI E BIOTECNOLOGICHE INDIRIZZO MICROBIOLOGIA MOLECOLARE E CLINICA XXIII Ciclo CHARACTERIZATION OF Mycoplasma agalactiae MEMBRANE PROTEOME Direttore: Prof. Bruno Masala Tutor: Dr. Alberto Alberti Tesi di dottorato della Dott.ssa Carla Cacciotto ANNO ACCADEMICO 2009-2010 TABLE OF CONTENTS 1. Abstract 2. Introduction 2.1 Mycoplasmas: taxonomy and main biological features 2.2 Metabolism 2.3 In vitro cultivation 2.4 Mycoplasma lipoproteins 2.5 Invasivity and pathogenicity 2.6 Diagnosis of mycoplasmosis 2.7 Mycoplasma agalactiae and Contagious Agalactia 3. Research objectives 4. Materials and methods 4.1 Media and buffers 4.2 Bacterial strains and culture conditions 4.3 Total DNA extraction and PCR 4.4 Total proteins extraction 4.5 Triton X-114 fractionation 4.6 SDS-PAGE 4.7 Western immunoblotting 4.8 2-D PAGE 4.9 2D DIGE 4.10 Spot picking and in situ tryptic digestion 4.11 GeLC-MS/MS 4.12 MALDI-MS 4.13 LC-MS/MS 4.14 Data analysis Dott.ssa Carla Cacciotto, Characterization of Mycoplasma agalactiae membrane proteome. Tesi di Dottorato in Scienze Biomolecolari e Biotecnologiche, Università degli Studi di Sassari. 5. Results 5.1 Species identification 5.2 Extraction of bacterial proteins and isolation of liposoluble proteins 5.3 2-D PAGE/MS of M. agalactiae PG2T liposoluble proteins 5.4 2D DIGE of liposoluble proteins among the type strain and two field isolates of M. agalactiae 5.5 GeLC-MS/MS of M. agalactiae PG2T liposoluble proteins 5.6 Data analysis and classification 6. Discussion 7.
    [Show full text]
  • Genomic Islands in Mycoplasmas
    G C A T T A C G G C A T genes Review Genomic Islands in Mycoplasmas Christine Citti * , Eric Baranowski * , Emilie Dordet-Frisoni, Marion Faucher and Laurent-Xavier Nouvel Interactions Hôtes-Agents Pathogènes (IHAP), Université de Toulouse, INRAE, ENVT, 31300 Toulouse, France; [email protected] (E.D.-F.); [email protected] (M.F.); [email protected] (L.-X.N.) * Correspondence: [email protected] (C.C.); [email protected] (E.B.) Received: 30 June 2020; Accepted: 20 July 2020; Published: 22 July 2020 Abstract: Bacteria of the Mycoplasma genus are characterized by the lack of a cell-wall, the use of UGA as tryptophan codon instead of a universal stop, and their simplified metabolic pathways. Most of these features are due to the small-size and limited-content of their genomes (580–1840 Kbp; 482–2050 CDS). Yet, the Mycoplasma genus encompasses over 200 species living in close contact with a wide range of animal hosts and man. These include pathogens, pathobionts, or commensals that have retained the full capacity to synthesize DNA, RNA, and all proteins required to sustain a parasitic life-style, with most being able to grow under laboratory conditions without host cells. Over the last 10 years, comparative genome analyses of multiple species and strains unveiled some of the dynamics of mycoplasma genomes. This review summarizes our current knowledge of genomic islands (GIs) found in mycoplasmas, with a focus on pathogenicity islands, integrative and conjugative elements (ICEs), and prophages. Here, we discuss how GIs contribute to the dynamics of mycoplasma genomes and how they participate in the evolution of these minimal organisms.
    [Show full text]
  • Impacts of Mycoplasma Agalactiae Restriction-Modification Systems on Pan-Epigenome Dynamics and Genome Plasticity
    bioRxiv preprint doi: https://doi.org/10.1101/2021.06.21.448925; this version posted June 21, 2021. The copyright holder for this preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission. Impacts of Mycoplasma agalactiae restriction-modification systems on pan-epigenome dynamics and genome plasticity Emilie Dordet-Frisoni1, Céline Vandecasteele3, Rachel Contarin 1, Eveline Sagné2, Eric Baranowski2, Christophe Klopp4, Laurent Xavier Nouvel2, and Christine Citti2* 1 INTHERES, Université de Toulouse, INRAE, ENVT, Toulouse, France 2 IHAP, Université de Toulouse, INRAE, ENVT, Toulouse, France 3 INRAE, US 1426, Get-PlaGe, GenoToul, Castanet-Tolosan, France 4 INRA, UR875 MIAT, Sigenae, BioInfo Genotoul, BioInfoMics, F-31326 Auzeville, France Abstract DNA methylation plays an important role in the biology of bacteria. Often associated with restriction modification (RM) systems, they also provide a defence against foreign DNA. Little is known regarding the methylome of the mycoplasma genus, which encompasses several pathogenic species with small genomes. Here, single molecule real-time (SMRT) and bisulphite sequencing combined with whole- genome analysis identified 19 methylated motifs associated with three orphan methyltransferases (MTases) and eight RM systems in Mycoplasma agalactiae, a ruminant pathogen and a model organism. All systems had a homolog in at least one phylogenetically distinct Mycoplasma spp. Our study also revealed that several superimposed genetic events may participate in the M. agalactiae dynamic epigenome landscape. These included (i) DNA shuffling and frameshift mutations that affect the MTase and restriction endonuclease content of a clonal population and (ii) gene duplication, erosion, and horizontal transfer that modulate MTase and RM repertoires of the species.
    [Show full text]
  • Large-Scale Analysis of the Mycoplasma Bovis Genome Identified Non-Essential, Adhesion- and Virulence-Related Genes
    fmicb-10-02085 September 13, 2019 Time: 16:34 # 1 ORIGINAL RESEARCH published: 13 September 2019 doi: 10.3389/fmicb.2019.02085 Large-Scale Analysis of the Mycoplasma bovis Genome Identified Non-essential, Adhesion- and Virulence-Related Genes Christoph Josi1,2, Sibylle Bürki1, Sara Vidal1, Emilie Dordet-Frisoni3, Christine Citti3, Laurent Falquet4† and Paola Pilo1*† 1 Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, Institute of Veterinary Bacteriology, University of Bern, Bern, Switzerland, 2 Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland, 3 UMR 1225, IHAP, Université de Toulouse, INRA, ENVT, Toulouse, France, 4 Department of Biology, Faculty of Science and Medicine, Swiss Institute of Bioinformatics, University of Fribourg, Fribourg, Switzerland Edited by: Mycoplasma bovis is an important pathogen of cattle causing bovine mycoplasmosis. Feng Gao, Tianjin University, China Clinical manifestations are numerous, but pneumonia, mastitis, and arthritis cases are Reviewed by: mainly reported. Currently, no efficient vaccine is available and antibiotic treatments Yong-Qiang He, are not always satisfactory. The design of new, efficient prophylactic and therapeutic Guangxi University, China Angelika Lehner, approaches requires a better understanding of the molecular mechanisms responsible University of Zurich, Switzerland for M. bovis pathogenicity. Random transposon mutagenesis has been widely used in *Correspondence: Mycoplasma species to identify potential gene functions. Such an approach can also be Paola Pilo used to screen genomes and search for essential and non-essential genes for growth. | downloaded: 29.4.2020 [email protected] Here, we generated a random transposon mutant library of M. bovis strain JF4278 †These authors have contributed equally to this work containing approximately 4000 independent insertion sites.
    [Show full text]
  • Phylogenomics of Expanding Uncultured Environmental Tenericutes
    bioRxiv preprint doi: https://doi.org/10.1101/2020.01.21.914887; this version posted January 23, 2020. The copyright holder for this preprint (which was not certified by peer review) is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under aCC-BY-NC-ND 4.0 International license. 1 Phylogenomics of expanding uncultured environmental Tenericutes 2 provides insights into their pathogenicity and evolutionary 3 relationship with Bacilli 4 Yong Wang1,*, Jiao-Mei Huang1,2, Ying-Li Zhou1,2, Alexandre Almeida3,4, Robert D. 5 Finn3, Antoine Danchin5,6, Li-Sheng He1 6 1Institute of Deep Sea Science and Engineering, Chinese Academy of Sciences, Sanya, 7 Hai Nan, China 8 2 University of Chinese Academy of Sciences, Beijing, China 9 3European Molecular Biology Laboratory, European Bioinformatics Institute 10 (EMBL-EBI), Wellcome Genome Campus, Hinxton, UK 11 4Wellcome Sanger Institute, Wellcome Genome Campus, Hinxton, UK. 12 5Department of Infection, Immunity and Inflammation, Institut Cochin INSERM 13 U1016 - CNRS UMR8104 - Université Paris Descartes, 24 rue du Faubourg 14 Saint-Jacques, 75014 Paris, France 15 6School of Biomedical Sciences, Li Kashing Faculty of Medicine, University of Hong 16 Kong, 21 Sassoon Road, SAR Hong Kong, China 17 18 *Corresponding author: 19 Yong Wang, PhD 20 Institute of Deep Sea Science and Engineering, Chinese Academy of Sciences 21 No. 28, Luhuitou Road, Sanya, Hai Nan, P.R. of China 22 Phone: 086-898-88381062 23 E-mail: [email protected] 24 Running title: Genomics of environmental Tenericutes 25 Keywords: Bacilli; autotrophy; pathogen; gut microbiome; environmental 26 Tenericutes 1 bioRxiv preprint doi: https://doi.org/10.1101/2020.01.21.914887; this version posted January 23, 2020.
    [Show full text]
  • Comparison of Culture and PCR for Detection of Field Isolates of Bovine
    Kafkas Univ Vet Fak Derg Kafkas Universitesi Veteriner Fakultesi Dergisi ISSN: 1300-6045 e-ISSN: 1309-2251 26 (3): 337-342, 2020 Journal Home-Page: http://vetdergikafkas.org Research Article DOI: 10.9775/kvfd.2019.23106 Online Submission: http://submit.vetdergikafkas.org Comparison of Culture and PCR for Detection of Field Isolates of Bovine Milk Mollicutes Abd Al-Bar AL-FARHA 1,a Farhid HEMMATZADEH 2,b Rick TEARLE 3,c Razi JOZANI 4,d Andrew HOARE 5,e Kiro PETROVSKI 6,f 1 Department of Animal Production, Technical Agricultural College, Northern Technical University, Mosul, IRAQ 2 University of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIA 3 Davies Centre, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIA 4 Tabriz University, Department of Veterinary Clinical Sciences, Tabriz, IRAN 5 South East Vets, South Australia, AUSTRALIA 6 Australian Centre for Antimicrobial Resistance Ecology, The University of Adelaide, School of Animal and Veterinary Sciences, South Australia, AUSTRALIA a ORCID: 0000-0003-1742-6350; b ORCID: 0000-0002-4572-8869; c ORCID: 0000-0003-2243-5091; d ORCID: 0000-0003-2889-4686 e ORCID: 0000-0002-5848-6707; f ORCID: 0000-0003-4016-2576 Article ID: KVFD-2019-23106 Received: 25.07.2019 Accepted: 07.02.2020 Published Online: 07.02.2020 How to Cite This Article Al-Farha AAB, Hemmatzadeh F, Tearle R, Jozani R, Hoare A, PetrovskI K: Comparison of culture and PCR for detection of field isolates of bovine milk mollicutes. Kafkas Univ Vet Fak Derg, 26 (3): 337-342, 2020. DOI: 10.9775/kvfd.2019.23106 Abstract Mycoplasma mastitis raises significant concerns in the dairy industry worldwide.
    [Show full text]
  • Reconstructing the Functions of Endosymbiotic Mollicutes in Fungus
    RESEARCH ARTICLE Reconstructing the functions of endosymbiotic Mollicutes in fungus- growing ants Panagiotis Sapountzis*, Mariya Zhukova, Jonathan Z Shik, Morten Schiott, Jacobus J Boomsma* Centre for Social Evolution, Department of Biology, University of Copenhagen, Copenhagen, Denmark Abstract Mollicutes, a widespread class of bacteria associated with animals and plants, were recently identified as abundant abdominal endosymbionts in healthy workers of attine fungus- farming leaf-cutting ants. We obtained draft genomes of the two most common strains harbored by Panamanian fungus-growing ants. Reconstructions of their functional significance showed that they are independently acquired symbionts, most likely to decompose excess arginine consistent with the farmed fungal cultivars providing this nitrogen-rich amino-acid in variable quantities. Across the attine lineages, the relative abundances of the two Mollicutes strains are associated with the substrate types that foraging workers offer to fungus gardens. One of the symbionts is specific to the leaf-cutting ants and has special genomic machinery to catabolize citrate/glucose into acetate, which appears to deliver direct metabolic energy to the ant workers. Unlike other Mollicutes associated with insect hosts, both attine ant strains have complete phage-defense systems, underlining that they are actively maintained as mutualistic symbionts. DOI: https://doi.org/10.7554/eLife.39209.001 *For correspondence: [email protected] (PS); Introduction [email protected] (JJB) Bacterial endosymbionts, defined here as comprising both intra- and extra-cellular symbionts Competing interests: The (Bourtzis and Miller, 2006), occur in all eukaryotic lineages and range from parasites to mutualists authors declare that no (Bourtzis and Miller, 2006; Martin et al., 2017).
    [Show full text]
  • Large-Scale Analysis of the Mycoplasma Bovis Genome Identified Non-Essential, Adhesion- and Virulence-Related Genes
    fmicb-10-02085 September 13, 2019 Time: 16:34 # 1 ORIGINAL RESEARCH published: 13 September 2019 doi: 10.3389/fmicb.2019.02085 Large-Scale Analysis of the Mycoplasma bovis Genome Identified Non-essential, Adhesion- and Virulence-Related Genes Christoph Josi1,2, Sibylle Bürki1, Sara Vidal1, Emilie Dordet-Frisoni3, Christine Citti3, Laurent Falquet4† and Paola Pilo1*† 1 Department of Infectious Diseases and Pathobiology, Vetsuisse Faculty, Institute of Veterinary Bacteriology, University of Bern, Bern, Switzerland, 2 Graduate School for Cellular and Biomedical Sciences, University of Bern, Bern, Switzerland, 3 UMR 1225, IHAP, Université de Toulouse, INRA, ENVT, Toulouse, France, 4 Department of Biology, Faculty of Science and Medicine, Swiss Institute of Bioinformatics, University of Fribourg, Fribourg, Switzerland Edited by: Mycoplasma bovis is an important pathogen of cattle causing bovine mycoplasmosis. Feng Gao, Tianjin University, China Clinical manifestations are numerous, but pneumonia, mastitis, and arthritis cases are Reviewed by: mainly reported. Currently, no efficient vaccine is available and antibiotic treatments Yong-Qiang He, are not always satisfactory. The design of new, efficient prophylactic and therapeutic Guangxi University, China Angelika Lehner, approaches requires a better understanding of the molecular mechanisms responsible University of Zurich, Switzerland for M. bovis pathogenicity. Random transposon mutagenesis has been widely used in *Correspondence: Mycoplasma species to identify potential gene functions. Such an approach can also be Paola Pilo used to screen genomes and search for essential and non-essential genes for growth. [email protected] Here, we generated a random transposon mutant library of M. bovis strain JF4278 †These authors have contributed equally to this work containing approximately 4000 independent insertion sites.
    [Show full text]