Evaluation of a Multiplex Real-Time PCR for Detection of Four Bacterial

Total Page:16

File Type:pdf, Size:1020Kb

Evaluation of a Multiplex Real-Time PCR for Detection of Four Bacterial Wisselink et al. BMC Veterinary Research (2017) 13:221 DOI 10.1186/s12917-017-1141-1 RESEARCHARTICLE Open Access Evaluation of a multiplex real-time PCR for detection of four bacterial agents commonly associated with bovine respiratory disease in bronchoalveolar lavage fluid Henk J. Wisselink* , Jan B.W.J. Cornelissen, Fimme J. van der Wal, Engbert A. Kooi, Miriam G. Koene, Alex Bossers, Bregtje Smid, Freddy M. de Bree and Adriaan F.G. Antonis Abstract Background: Pasteurella multocida, Mannheimia haemolytica, Histophilus somni and Trueperella pyogenes are four bacterial agents commonly associated with bovine respiratory disease (BRD). In this study a bacterial multiplex real-time PCR (the RespoCheck PCR) was evaluated for the detection in bronchoalveolar lavage fluid (BALF) of these four bacterial agents. Results: The analytical sensitivity of the multiplex real-time PCR assay determined on purified DNA and on bacterial cells of the four target pathogens was one to ten fg DNA/assay and 4 × 10−1 to 2 × 100 CFU/assay. The analytical specificity of the test was, as evaluated on a collection of 118 bacterial isolates, 98.3% for M. haemolytica and 100% for the other three target bacteria. A set of 160 BALF samples of calves originating from ten different herds with health problems related to BRD was examined with bacteriological methods and with the RespoCheck PCR. Using bacteriological examination as the gold standard, the diagnostic sensitivities and specificities of the four bacterial agents were respectively between 0.72 and 1.00 and between 0.70 and 0.99. Kappa values for agreement between results of bacteriological examination and PCRs were low for H. somni (0.17), moderate for P. multocida (0.52) and M. haemolytica (0.57), and good for T. pyogenes (0.79). The low and moderate kappa values seemed to be related to limitations of the bacteriological examination, this was especially the case for H. somni. Conclusion: It was concluded that the RespoCheck PCR assay is a valuable diagnostic tool for the simultaneous detection of the four bacterial agents in BALF of calves. Keywords: Bovine respiratory disease, PCR, Histophilus somni, Mannheimia haemolytica, Pasteurella multocida, Trueperella pyogenes * Correspondence: [email protected] Wageningen Bioveterinary Research, P.O. Box 65, 8200, AB, Lelystad, The Netherlands © The Author(s). 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. Wisselink et al. BMC Veterinary Research (2017) 13:221 Page 2 of 10 Background analysis of BRD associated organisms. To offer veteri- In veal production, bovine respiratory disease (BRD) is narians a fast and complete laboratory result, a set of the most common and economically important disease three multiplex real-time PCRs was developed for de- [1–3]. BRD is a multifactorial disease, involving multiple tection of i) viral, ii) bacterial and iii) mycoplasma potentially pathogenic microorganisms that causes eco- BRD-associated pathogens in bronchoalveolar lavage nomic losses due to morbidity, mortality, medication fluid (BALF) of calves, under the name RespoCheck costs, increased time on feeding and associated labour (WBVR, Lelystad, The Netherlands). In this study the costs [4]. In the Dutch veal industry, calves at an age RespoCheck bacterial multiplex real-time PCR (abbre- between ten days and five weeks are sold by dairy viated to RespoCheck PCR) was evaluated with com- farmers to traders who transport calves to the calf col- monly used culture methods as reference. This PCR lection centres, from where they are transported to fat- has been developed for the detection of four bacterial tening farms where they are mingled with other young pathogens commonly associated with BRD [9, 13], i.e. calves, reared and fattened until slaughter at an age of Pasteurella multocida, Mannheimia haemolytica, His- 6–7 months. The young age of the calves, their inad- tophilus somni and Trueperella pyogenes.Theresults equate immunity and several stress factors including showed that the RespoCheck PCR assay is a sensitive transportation and mingling with other young calves and valuable diagnostic tool suitable for the simultan- makes that the calves are vulnerable for infections, a eous detection of four BRD associated bacterial targets scenario which is recognised as resulting in the frequent in BALF of calves. occurrence of disease problems including BRD [5]. To protect calves, at the end of the twentieth century veal Methods farmers commonly administered antibiotics prophylac- Bacterial strains and growth conditions tically immediately after arrival on the farm. Growing A total of 118 bacterial isolates, representing 46 different concerns about the increase in antibiotic resistance species, were used for evaluation of specificity of the including methicillin resistant Staphylococcus aureus RespoCheck PCR. At least five representatives of each of (MRSA) and extended-spectrum βeta-lactamase produ- the four target species, as well as bacterial strains from cing Escherichia coli in livestock resulted in a policy non-target species, consisting of isolates of phylogenetic- aimed to substantially reduce the use of antibiotics in ally closely related non-target species, and isolates of The Netherlands [6]. Part of this policy was a ban of all species from cattle with diseases other than BRD were prophylactic use of antimicrobials in the Netherlands in included. Reference strains were obtained from the 2012 (Besluit Diergeneesmiddelen, 2 November 2012; American Type Culture Collection (ATCC) and the http://wetten.overheid.nl/BWBR0032386/2015-01-01). Culture Collection of the University of Gothenburg To implement and regulate this reduction in the use of (Sweden) (CCUB) and field strains were isolated from antibiotics in the Netherlands, in recent years a collab- lungs of calves in a Dutch field study on BRD. An over- oration was set up between stakeholders in livestock view of the strains is provided in Table 1. Bacteria other production and the national government [6, 7]. This than H. somni were grown overnight at 37 °C on heart lead to registration of all antibiotics administered to infusion agar (ACU 7269C, Acumedia Manufacturers livestock at farm level [8] and to mandatory treatment Inc. Lansing, MI) supplemented with 5% defibrinated and health plans to be developed for each farm to sheep blood. H. somni was grown overnight at 37 °C enforce that antimicrobial treatment is solely based on in air with 5% CO2 on chocolate blood agar, using 7% proper diagnosis [6]. However, the availability of rapid defibrinated horse blood and Columbia blood agar and reliable diagnostic tools is limited. (CM 331, Oxoid, Badhoevedorp, The Netherlands). In BRD, several viral and bacterial pathogens may be involved [9–11]. For detection of these potential patho- Field samples gens a variety of diagnostic tests have been described, In a Dutch field study on BRD from October 2013 till including culture and molecular methods, see review by March 2014, veal calf farms (n = 10) were selected. On Fulton and Confer [12]. These methods all have their each farm, the study period started shortly after arrival benefits and limitations in terms of speed of analysis, of the calves (D0) and ended 84 days (D84) later. At the sensitivity and specificity [12]. In case of BRD, interpret- start (D0) and end (D84) of the study period and during ation of results is sometimes challenging and diagnostic outbreaks of BRD, bronchoalveolar lavage (BAL) samples laboratories usually offer diagnostic tests for a limited were taken from calves without and with clinical problems number of potential pathogens which can be involved in of BRD (as defined by severe upper respiratory tract and/ BRD and which are sometimes also time-consuming. or lower respiratory tract disease). These are drawbacks that can hamper veterinarians and BAL samples were collected as previously described farmers for sending samples to diagnostic laboratory for [14]. Between 35 and 75 mL BAL was obtained from each Wisselink et al. BMC Veterinary Research (2017) 13:221 Page 3 of 10 Table 1 List of 118 bacterial isolates and results of the RespoCheck bacterial multiplex real-time PCR Identificationa CCUG identificationb Source RespoCheck bacterial multiplex real-time PCR(Ct values) (number of isolates tested) P. multocida M. haemolytica H. somni T. pyogenes Acidovorax spp. (n =3) NAc WBVR collectiond - g --- Actinomyces spp NA WBVR collectiond -- -- Aerococcus viridans NA WBVR collectione -- -- Biberstenia trehalosi NA WBVR collectiond -- -- Brucella abortus NA WBVR collectione -- -- Comamonas kerstersii NA WBVR collectiond -- -- Corynebacterium bovis (n =2) NA WBVR collectione -- -- Corynebacterium pseudotuberculosis NA WBVR collectione -- -- Escherichia coli NA WBVR collectione -- -- Gallibacterium anatis (n = 5) NA WBVR collectiond -- -- Hafnia alvei NA WBVR collectiond -- -- Histophilus somni NA ATCC 22132f - - 15.2 - Histophilus somni (n = 4) NA WBVR collectiond - - 12.3–14.1 - Klebsiella oxytoca NA WBVR
Recommended publications
  • Identification of Pasteurella Species and Morphologically Similar Organisms
    UK Standards for Microbiology Investigations Identification of Pasteurella species and Morphologically Similar Organisms Issued by the Standards Unit, Microbiology Services, PHE Bacteriology – Identification | ID 13 | Issue no: 3 | Issue date: 04.02.15 | Page: 1 of 28 © Crown copyright 2015 Identification of Pasteurella species and Morphologically Similar Organisms Acknowledgments UK Standards for Microbiology Investigations (SMIs) are developed under the auspices of Public Health England (PHE) working in partnership with the National Health Service (NHS), Public Health Wales and with the professional organisations whose logos are displayed below and listed on the website https://www.gov.uk/uk- standards-for-microbiology-investigations-smi-quality-and-consistency-in-clinical- laboratories. SMIs are developed, reviewed and revised by various working groups which are overseen by a steering committee (see https://www.gov.uk/government/groups/standards-for-microbiology-investigations- steering-committee). The contributions of many individuals in clinical, specialist and reference laboratories who have provided information and comments during the development of this document are acknowledged. We are grateful to the Medical Editors for editing the medical content. For further information please contact us at: Standards Unit Microbiology Services Public Health England 61 Colindale Avenue London NW9 5EQ E-mail: [email protected] Website: https://www.gov.uk/uk-standards-for-microbiology-investigations-smi-quality- and-consistency-in-clinical-laboratories UK Standards for Microbiology Investigations are produced in association with: Logos correct at time of publishing. Bacteriology – Identification | ID 13 | Issue no: 3 | Issue date: 04.02.15 | Page: 2 of 28 UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England Identification of Pasteurella species and Morphologically Similar Organisms Contents ACKNOWLEDGMENTS .........................................................................................................
    [Show full text]
  • Prevalence and Antibiotic Susceptibility of Mannheimia
    Veterinary World, EISSN: 2231-0916 RESEARCH ARTICLE Available at www.veterinaryworld.org/Vol.13/September-2020/28.pdf Open Access Prevalence and antibiotic susceptibility of Mannheimia haemolytica and Pasteurella multocida isolated from ovine respiratory infection: A study from Karnataka, Southern India Swati Sahay1,2 , Krithiga Natesan1, Awadhesh Prajapati1, Triveni Kalleshmurthy1 , Bibek Ranjan Shome1, Habibur Rahman3 and Rajeswari Shome1 1. Indian Council of Agricultural Research-National Institute of Veterinary Epidemiology and Disease Informatics, Bengaluru, Karnataka, India; 2. Department of Microbiology, Centre for Research in Pure and Applied Sciences, Jain University, Bengaluru, Karnataka, India; 3. International Livestock Research Institute, CG Centre, NASC Complex, DPS Marg, Pusa, New Delhi, India. Corresponding author: Rajeswari Shome, e-mail: [email protected] Co-authors: SS: [email protected], KN: [email protected], AP: [email protected], TK: [email protected], BRS: [email protected], HR: [email protected] Received: 25-04-2020, Accepted: 29-07-2020, Published online: 23-09-2020 doi: www.doi.org/10.14202/vetworld.2020.1947-1954 How to cite this article: Sahay S, Natesan K, Prajapati A, Kalleshmurthy T, Shome BR, Rahman H, Shome R (2020) Prevalence and antibiotic susceptibility of Mannheimia haemolytica and Pasteurella multocida isolated from ovine respiratory infection: A study from Karnataka, Southern India, Veterinary World, 13(9): 1947-1954. Abstract Background and Aim: Respiratory infection due to Mannheimia haemolytica and Pasteurella multocida are responsible for huge economic losses in livestock sector globally and it is poorly understood in ovine population. The study aimed to investigate and characterize M. haemolytica and P. multocida from infected and healthy sheep to rule out the involvement of these bacteria in the disease.
    [Show full text]
  • How Mannheimia Haemolytica Defeats Host Defence Through a Kiss of Death Mechanism Laurent Zecchinon, Thomas Fett, Daniel Desmecht
    How Mannheimia haemolytica defeats host defence through a kiss of death mechanism Laurent Zecchinon, Thomas Fett, Daniel Desmecht To cite this version: Laurent Zecchinon, Thomas Fett, Daniel Desmecht. How Mannheimia haemolytica defeats host de- fence through a kiss of death mechanism. Veterinary Research, BioMed Central, 2005, 36 (2), pp.133- 156. 10.1051/vetres:2004065. hal-00902968 HAL Id: hal-00902968 https://hal.archives-ouvertes.fr/hal-00902968 Submitted on 1 Jan 2005 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Vet. Res. 36 (2005) 133–156 133 © INRA, EDP Sciences, 2005 DOI: 10.1051/vetres:2004065 Review article How Mannheimia haemolytica defeats host defence through a kiss of death mechanism Laurent ZECCHINON, Thomas FETT, Daniel DESMECHT* Department of Pathology, Faculty of Veterinary Medicine, University of Liège, FMV Sart-Tilman B43, 4000 Liège, Belgium (Received 22 June 2004; accepted 6 October 2004) Abstract – Mannheimia haemolytica induced pneumonias are only observed in goats, sheep and cattle. The bacterium produces several virulence factors,whose principal ones are lipopolysaccharide and leukotoxin. The latter is cytotoxic only for ruminant leukocytes, a phenomenon that is correlated with its ability to bind and interact with the ruminant β2-integrin Lymphocyte Function-associated Antigen 1.
    [Show full text]
  • Review on the Potential Effects of Mannheimia Haemolytica and Its Immunogens on the Female Reproductive Physiology and Performance of Small Ruminants
    Journal of Animal Health and Production Review Article A Review on the Potential Effects of Mannheimia haemolytica and its Immunogens on the Female Reproductive Physiology and Performance of Small Ruminants 1,2 2 1,3 FAEZ FIRDAUS ABDULLAH JESSE *, MOHAMED ABDIRAHMAN BOOREI , ERIC LIM TEIK CHUNG , 2 4 2 2 FITRI WAN-NOR , MOHD AZMI MOHD LILA , MOHD JEFRI NORSIDIN , KAMARULRIZAL MAT ISA , 1 1 5 6 NUR AZHAR AMIRA , ARSALAN MAQBOOL , MOHAMMED NAJI ODHAH , YUSUF ABBA , ASINAMAI 7 6 6 2,6 ATHLIAMAI BITRUS , IDRIS UMAR HAMBALI , INNOCENT DAMUDU PETER , BURA THLAMA PAUL 1Institute of Tropical Agriculture and Food Security, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; 2Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; 3Department of Animal Science, Faculty of Agriculture, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia; 4Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Universiti Putra Malaysia, 43400 Serdang, Selangor, Malaysia; 5Department of Veterinary Clinical Studies, Faculty of Veterinary Medicine, Universiti Malaysia Kelantan, Pengakalan Chepa 16100, Kota Bharu, Kelantan, Malaysia; 6Faculty of Veterinary Medicine, University of Maiduguri, PMB 1069 Maiduguri, Borno Nigeria; 7Faculty of Veterinary Science, University of Jos, P.M.B 2084 Jos, Plateau Nigeria. Abstract | Mannheimia haemolytica causes pneumonic pasteurellosis (mannheimiosis) in ruminants which is the most economically significant infectious disease. Mannheimia belongs to the family Pasteurellaceae, are nonmotile, non- spore-forming, facultatively anaerobic, oxidase-positive and fermentative gram-negative rods or coccobacilli which are frequent respiratory and digestive tract commensals in both domestic and wild animals. They can produce infection in animals with compromised immune states.
    [Show full text]
  • CHARACTERIZATION of MANNHEIMIA HAEMOL JT/C4-SPECIFIC BACTERIOPHAGES YU-HUNG HSU Bachelor of Science, University of British Colum
    CHARACTERIZATION OF MANNHEIMIA HAEMOL JT/C4-SPECIFIC BACTERIOPHAGES YU-HUNG HSU Bachelor of Science, University of British Columbia, 2007 A Thesis Submitted to the School of Graduate Studies of the University of Lethbridge in Partial Fulfillment of the Requirements for the Degree MASTER OF SCIENCE (Molecular Biology/ Microbiology) Department of Biological Sciences University of Lethbridge LETHBRIDGE, ALBERTA, CANADA © Yu-Hung Hsu, 2011 Library and Archives Bibliotheque et Canada Archives Canada Published Heritage Direction du Branch Patrimoine de I'edition 395 Wellington Street 395, rue Wellington Ottawa ON K1A0N4 Ottawa ON K1A 0N4 Canada Canada Your file Votre reference ISBN: 978-0-494-88386-0 Our file Notre reference ISBN: 978-0-494-88386-0 NOTICE: AVIS: The author has granted a non­ L'auteur a accorde une licence non exclusive exclusive license allowing Library and permettant a la Bibliotheque et Archives Archives Canada to reproduce, Canada de reproduire, publier, archiver, publish, archive, preserve, conserve, sauvegarder, conserver, transmettre au public communicate to the public by par telecommunication ou par I'lnternet, preter, telecommunication or on the Internet, distribuer et vendre des theses partout dans le loan, distrbute and sell theses monde, a des fins commerciales ou autres, sur worldwide, for commercial or non­ support microforme, papier, electronique et/ou commercial purposes, in microform, autres formats. paper, electronic and/or any other formats. The author retains copyright L'auteur conserve la propriete du droit d'auteur ownership and moral rights in this et des droits moraux qui protege cette these. Ni thesis. Neither the thesis nor la these ni des extraits substantiels de celle-ci substantial extracts from it may be ne doivent etre imprimes ou autrement printed or otherwise reproduced reproduits sans son autorisation.
    [Show full text]
  • Dynamics and Stabilization of the Rumen Microbiome in Yearling
    www.nature.com/scientificreports OPEN Dynamics and stabilization of the rumen microbiome in yearling Tibetan sheep Lei Wang1,2,4, Ke Zhang3,4, Chenguang Zhang3, Yuzhe Feng1, Xiaowei Zhang1, Xiaolong Wang 3 & Guofang Wu1,2* The productivity of ruminants depends largely on rumen microbiota. However, there are few studies on the age-related succession of rumen microbial communities in grazing lambs. Here, we conducted 16 s rRNA gene sequencing for bacterial identifcation on rumen fuid samples from 27 Tibetan lambs at nine developmental stages (days (D) 0, 2, 7, 14, 28, 42, 56, 70, and 360, n = 3). We observed that Bacteroidetes and Proteobacteria populations were signifcantly changed during the growing lambs’ frst year of life. Bacteroidetes abundance increased from 18.9% on D0 to 53.9% on D360. On the other hand, Proteobacteria abundance decreased signifcantly from 40.8% on D0 to 5.9% on D360. Prevotella_1 established an absolute advantage in the rumen after 7 days of age. The co-occurrence network showed that the diferent microbial of the rumen presented a complex synergistic and cumbersome relationship. A phylogenetic tree was constructed, indicating that during the colonization process, may occur a phenomenon in which bacteria with close kinship are preferentially colonized. Overall, this study provides new insights into the colonization of bacterial communities in lambs that will beneft the development of management strategies to promote colonization of target communities to improve functional development. Rumen microbes are essential for ruminant health. Ruminants rely upon the action of microbial fermentation in the rumen to digest plant fbers and convert some nutrients that cannot be directly utilized into animal proteins for host utilization1.
    [Show full text]
  • MICRO-ORGANISMS and RUMINANT DIGESTION: STATE of KNOWLEDGE, TRENDS and FUTURE PROSPECTS Chris Mcsweeney1 and Rod Mackie2
    BACKGROUND STUDY PAPER NO. 61 September 2012 E Organización Food and Organisation des Продовольственная и cельскохозяйственная de las Agriculture Nations Unies Naciones Unidas Organization pour организация para la of the l'alimentation Объединенных Alimentación y la United Nations et l'agriculture Наций Agricultura COMMISSION ON GENETIC RESOURCES FOR FOOD AND AGRICULTURE MICRO-ORGANISMS AND RUMINANT DIGESTION: STATE OF KNOWLEDGE, TRENDS AND FUTURE PROSPECTS Chris McSweeney1 and Rod Mackie2 The content of this document is entirely the responsibility of the authors, and does not necessarily represent the views of the FAO or its Members. 1 Commonwealth Scientific and Industrial Research Organisation, Livestock Industries, 306 Carmody Road, St Lucia Qld 4067, Australia. 2 University of Illinois, Urbana, Illinois, United States of America. This document is printed in limited numbers to minimize the environmental impact of FAO's processes and contribute to climate neutrality. Delegates and observers are kindly requested to bring their copies to meetings and to avoid asking for additional copies. Most FAO meeting documents are available on the Internet at www.fao.org ME992 BACKGROUND STUDY PAPER NO.61 2 Table of Contents Pages I EXECUTIVE SUMMARY .............................................................................................. 5 II INTRODUCTION ............................................................................................................ 7 Scope of the Study ...........................................................................................................
    [Show full text]
  • Antimicrobial Resistance in Pasteurella and Mannheimia: Epidemiology and Genetic Basis
    Antimicrobial resistance in Pasteurella and Mannheimia: epidemiology and genetic basis Corinna Kehrenberg, Gundula Schulze-Tanzil, Jean-Louis Martel, Elisabeth Chaslus-Dancla, Stefan Schwarz To cite this version: Corinna Kehrenberg, Gundula Schulze-Tanzil, Jean-Louis Martel, Elisabeth Chaslus-Dancla, Stefan Schwarz. Antimicrobial resistance in Pasteurella and Mannheimia: epidemiology and genetic basis. Veterinary Research, BioMed Central, 2001, 32 (3-4), pp.323-339. 10.1051/vetres:2001128. hal- 00902701 HAL Id: hal-00902701 https://hal.archives-ouvertes.fr/hal-00902701 Submitted on 1 Jan 2001 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Vet. Res. 32 (2001) 323–339 323 © INRA, EDP Sciences, 2001 Review article Antimicrobial resistance in Pasteurella and Mannheimia: epidemiology and genetic basis Corinna KEHRENBERGa, Gundula SCHULZE-TANZILa, Jean-Louis MARTELc, Elisabeth CHASLUS-DANCLAb, Stefan SCHWARZa* a Institut für Tierzucht und Tierverhalten, Bundesforschungsanstalt für Landwirtschaft (FAL), 29223 Celle, Germany b Institut National de la Recherche Agronomique, Pathologie Aviaire et Parasitologie, 37380 Nouzilly, France c Agence Française de Sécurité Sanitaire des Aliments, Pathologie Bovine et Hygiène des viandes, 31 avenue Tony Garnier, 69364 Lyon Cedex 07, France (Received 23 November 2000; accepted 2 February 2001) Abstract – Isolates of the genera Pasteurella and Mannheimia cause a wide variety of diseases of great economic importance in poultry, pigs, cattle and rabbits.
    [Show full text]
  • Molecular Study on Pasteurella Multocida and Mannheimia Granulomatis from Kenyan Camels (Camelus Dromedarius) Ilona V
    Gluecks et al. BMC Veterinary Research (2017) 13:265 DOI 10.1186/s12917-017-1189-y RESEARCHARTICLE Open Access Molecular study on Pasteurella multocida and Mannheimia granulomatis from Kenyan Camels (Camelus dromedarius) Ilona V. Gluecks1†, Astrid Bethe2†, Mario Younan3† and Christa Ewers4* Abstract Background: Outbreaks of a Haemorrhagic Septicaemia (HS) like disease causing large mortalities in camels (Camelus dromedarius) in Asia and in Africa have been reported since 1890. Yet the aetiology of this condition remains elusive. This study is the first to apply state of the art molecular methods to shed light on the nasopharyngeal carrier state of Pasteurellaceae in camels. The study focused on HS causing Pasteurella multocida capsular types B and E. Other Pasteurellaceae, implicated in common respiratory infections of animals, were also investigated. Methods: In 2007 and 2008, 388 nasopharyngeal swabs were collected at 12 locations in North Kenya from 246 clinically healthy camels in 81 herds that had been affected by HS-like disease. Swabs were used to cultivate bacteria on blood agar and to extract DNA for subsequent PCR analysis targeting P. multocida and Mannheimia-specific gene sequences. Results: Forty-five samples were positive for P. multocida genes kmt and psl and for the P. multocida Haemorrhagic Septicaemia (HS) specific sequences KTSP61/KTT72 but lacked HS-associated capsular type B and E genes capB and capE. This indicates circulation of HS strains in camels that lack established capsular types. Sequence analysis of the partial 16S rRNA gene identified 17 nasal swab isolates as 99% identical with Mannheimia granulomatis,demonstrating a hitherto unrecognised active carrier state for M.
    [Show full text]
  • Genome Sequence and Description of Mannheimia Massilioguelmaensis Sp
    Genome sequence and description of Mannheimia massilioguelmaensis sp. nov. L. Hadjadj, Ahmed Aimen Bentorki, C. Michelle, K. Amoura, Abdelghani Djahoudi, Jean-Marc Rolain To cite this version: L. Hadjadj, Ahmed Aimen Bentorki, C. Michelle, K. Amoura, Abdelghani Djahoudi, et al.. Genome sequence and description of Mannheimia massilioguelmaensis sp. nov.. New Microbes and New Infec- tions, Wiley Online Library 2015, 8, pp.131-136. 10.1016/j.nmni.2015.10.005. hal-01772904 HAL Id: hal-01772904 https://hal-amu.archives-ouvertes.fr/hal-01772904 Submitted on 20 Apr 2018 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Distributed under a Creative Commons Attribution - NonCommercial - NoDerivatives| 4.0 International License TAXONOGENOMICS: GENOME OF A NEW ORGANISM Genome sequence and description of Mannheimia massilioguelmaensis sp. nov. L. Hadjadj1, A. A. Bentorki2, C. Michelle1, K. Amoura3, A. Djahoudi3 and J.-M. Rolain1 1) Unité de recherche sur les maladies infectieuses et tropicales émergentes (URMITE), UMR CNRS, IHU Méditerranée Infection, Faculté de Médecine et de Pharmacie, Aix-Marseille-Université, Marseille, France, 2) Laboratoire de Microbiologie, CHU Dorban and 3) Laboratoire de Microbiologie, Faculté de Médecine, Université Badji Mokhtar, Annaba, Algeria Abstract Strain MG13T sp.
    [Show full text]
  • Isolation and Identification of Mannheimia Haemolytica and Pasteurella Multocida Species from Ruminants in Six Different Regions in Morocco
    Journal of Agricultural Science and Technology A 8 (2018) 398-405 doi: 10.17265/2161-6256/2018.06.007 D DAVID PUBLISHING Isolation and Identification of Mannheimia haemolytica and Pasteurella multocida Species from Ruminants in Six Different Regions in Morocco Ghizlane Sebbar1, 2, Khalil Zro1, Faouzi Kichou3, Abdelkrim Fillali Maltouf2 and Bouchra Belkadi2 1. Society of Veterinary Pharmaceutical and Biological Productions (Biopharma), B.P. 4569, Rabat, Morocco 2. Laboratory of Microbiology and Molecular Biology, Faculty of Sciences, University Mohammed V, 4 Ibn Battouta Ave. BP 1014 RP Rabat, Morocco 3. Institute of Agronomy and Veterinary Medicine Hassan II, B.P. 6202, Rabat, Morocco Abstract: Pasteurella species is considered the principal pathogen of the respiratory tract. Mannheimia haemolytica and Pasteurella multocida were investigated and typed from nasal swabs and tissues taken from sheep, goat and cattle. Indeed, 41 lung and 121 nasal swabs samples were collected from animals with respiratory diseases during 2015 to 2017 in six different regions in Morocco. At first, a screening of Pasteurella species using the real time PCR (RT-PCR) was carried out, then all isolated strains on agar blood were confirmed by PCR gel based assay specific for M. haemolytica and P. multocida. Pathogenicity was evaluated in mice and histopathological examination was done on some of lung tissue. The results revealed that 34 samples of which 28 (55%) from nasal swabs and six (38%) from lungs were positive for M. haemolytica and nine samples of which seven (14%) from nasal swabs and two (13%) from lungs were positive on P. multocida serogroup A. Seventy-two percent (72%) isolates were highly pathogenic to mice, which is in accordance with the results obtained by histopathology examination.
    [Show full text]
  • Using Comparative Genomics to Identify Virulence Traits and Vaccine Candidates in Mannheimia Haemolytica
    Using comparative genomics to identify virulence traits and vaccine candidates in Mannheimia haemolytica A Thesis Submitted to the College of Graduate Studies and Research In Partial Fulfilment of the Requirements For the Degree of Doctor of Philosophy In the Department of Large Animal Clinical Sciences Western College of Veterinary Medicines University of Saskatchewan Saskatoon By Cassidy Klima Copyright Cassidy Klima, June 2015. All rights reserved Permission to Use In presenting this thesis/dissertation in partial fulfilment of the requirements for a Postgraduate degree from the University of Saskatchewan, I agree that the libraries of this University may make it freely available for inspection. I further agree that permission for copying of this thesis/dissertation in any manner, in whole or in part, for scholarly purposes may be granted by the professor or professors who supervised my thesis/dissertation work or, in their absence, by the Head of the Department or the Dean of the College in which my thesis work was done. It is understood that any copying or publication or use of this thesis/dissertation or parts thereof for financial gain shall not be allowed with-out my written permission. It is also understood that due recognition shall be given to me and to the University of Saskatchewan in any scholarly use which may be made of any material in my thesis/dissertation. Requests for permission to copy or to make other uses of materials in this thesis/dissertation in whole or part should be addressed to: Head of the Department of Large Animal Clinical Sciences Western College of Veterinary Medicine University of Saskatchewan Saskatoon, Saskatchewan, S7N 5B4 Canada OR Dean College of Graduate Studies and Research University of Saskatchewan 107 Administration Place Saskatoon, Saskatchewan, S7N 5A2 Canada i Abstract Bovine respiratory disease (BRD) is the principal cause of morbidity and mortality among feedlot cattle.
    [Show full text]