N-Linked Protein Glycosylation in Helicobacter Species

Total Page:16

File Type:pdf, Size:1020Kb

N-Linked Protein Glycosylation in Helicobacter Species N-linked Protein Glycosylation in Helicobacter Species A thesis submitted to the University of Manchester for the degree of Doctor of Philosophy in the Faculty of Life Sciences 2012 Alison Grace Wood Contents Abstract: ……………………………………………………………………………9 Declaration ................................................................................................................. 10 Chapter 1 .................................................................................................................... 15 General Introduction .................................................................................................. 15 1.0 Introduction ...................................................................................................... 16 1.1 Eukaryotic Protein Glycosylation .................................................................... 16 1.2 Protein Glycosylation in Archaea .................................................................... 19 1.3 O-linked Protein Glycosylation in Bacteria ..................................................... 20 1.4 Bacterial Cytoplasmic N-glycosylation Systems ............................................. 21 1.5 The C. jejuni N-linked Protein Glycosylation System ..................................... 22 1.6 The Bacterial OST and Amino Acid Sequon Specificity................................. 23 1.7 C. jejuni Glycosyltransferases and Assembly of the Heptasaccharide Glycan 26 1.8 Function of C. jejuni N-linked Glycoproteins.................................................. 28 1.9 Exploitating the C. jejuni N-linked Protein Glycosylation System ................. 29 1.10 Campylobacter N-linked Protein Glycosylation (pgl) Genes ........................ 32 1.11 The N-linked Protein Glycosylation System of W. succinogenes ................. 34 1.12 N-Linked Glycosylation Systems in Helicobacter Species ........................... 35 1.13 Summary ........................................................................................................ 38 1.14 Aims of This Study ........................................................................................ 39 Chapter 2 .................................................................................................................... 40 Materials and Methods ............................................................................................... 40 2.1 Bacterial Strains and Growth Conditions ......................................................... 41 2.2 Isolation of Plasmids and Genomic DNA ........................................................ 41 2.3 PCR Amplification ........................................................................................... 42 2.4 Reverse Transcriptase PCR .............................................................................. 42 2.5 Agarose Gel Electrophoresis ............................................................................ 42 2 2.6 DNA Manipulation .......................................................................................... 43 2.7 E. coli Transformation ..................................................................................... 43 2.7 DNA Sequencing. ............................................................................................ 44 2.8 Site Directed Mutagenesis................................................................................ 44 2.9 Helicobacter Transformation ........................................................................... 44 2.10 Purification of Glycoproteins from H. pullorum............................................ 45 2.11 SDS-PAGE and Western Blotting. ................................................................ 46 2.12 Mass Spectrometery for Characterisation of Proteins .................................... 47 2.13 Preparation of Bacterial Cell Membranes ...................................................... 48 2.14 Oligosaccharyltransferase Assay ................................................................... 48 2.15 Tricine SDS-PAGE ........................................................................................ 49 2.16 Purification of Biotinylated Peptide from the In vitro OST Assay ................ 49 Chapter 3 .................................................................................................................... 51 The Second Helicobacter pullorum Oligosaccharyltransferase (PglB2) is Neither Essential nor Required for N-linked Protein Glycosylation ...................................... 51 3.1 Background ...................................................................................................... 52 3.2 Results .............................................................................................................. 54 3.2.1 Construction of suicide plasmids for site specific integration of antibiotic resistance cassettes onto the H. pullorum chromosome. .................................... 54 3.2.2 Integration of antibiotic resistance cassettes onto the H. pullorum chromosome within the 23S rRNA gene. .......................................................... 56 3.2.3 Natural Transformation of H. pullorum NCTC 12824T with genomic DNA from H. pullorum 23::E and 23::Kn strains. ...................................................... 58 3.2.4 Expression of H. pullorum pglB1 and pglB2 genes .................................. 59 3.2.5 Insertional mutagenesis of H. pullorum pglB1 and pglB2 genes ............. 61 3.2.6 Role of PglB2 in the N-glycosylation of a small peptide in vitro ............. 64 3.2.7 The N-glycan structure of glycopeptide derived from the H. pullorum pglB2 mutant. ..................................................................................................... 66 3 3.3 Discussion ........................................................................................................ 69 Chapter 4 .................................................................................................................... 72 Identification of Putative H. pullorum Glycoproteins and Their Glycosylation in E. coli with the C. jejuni Heptasaccharide...................................................................... 72 4.1 Background ...................................................................................................... 73 4.2 Results .............................................................................................................. 75 4.2.1 Bioinformatic approach to predicting N-linked glycoproteins in Helicobacter species .......................................................................................... 75 4.2.2 Production of H. pullorum HgpA and Hp0114 putative glycoproteins in E. coli ...................................................................................................................... 78 4.2.3 Glycosylation of H. pullorum HgpA and Hp0114 in E. coli with a C. jejuni heptasaccharide glycan. ........................................................................... 80 4.2.4 Localisation of the site of glycosylation of Hp0114 ................................. 84 4.2.5 Amino acid site of glycosylation of H. pullorum HgpA ........................... 86 4.3 Discussion ........................................................................................................ 88 Chapter 5 .................................................................................................................... 91 Glycosylation of Hp0114 and HgpA in H. pullorum ................................................. 91 5.1 Background ...................................................................................................... 92 5.2 Results .............................................................................................................. 93 5.2.1 Production of hexa histidine tagged Hp0114 and HgpA proteins in H. pullorum ............................................................................................................. 93 5.2.3 Site of glycosylation of Hp0114 in H. pullorum ....................................... 98 5.2.4 Intact mass analysis of HgpA glycoprotein derived from H. pullorum .. 100 5.2.5 MALDI Mass Spectrometry of the HgpA glycoprotein derived from wild type H. pullorum. ............................................................................................. 103 5.3 Discussion ...................................................................................................... 110 Chapter 6 .................................................................................................................. 113 4 Characterization of the H. pullorum PglB1-Dependent N-Linked Protein Glycosylation System .............................................................................................. 113 6.1 Background .................................................................................................... 114 6.2 Results ............................................................................................................ 118 6.2.1 The effect of insertional mutagenesis of putative glycosyltransferase encoding genes on the electrophoretic mobility of H. pullorum HgpA protein .......................................................................................................................... 118 6.2.2 Intact mass analysis of HgpA derived from H. pullorum glycosyltransferase-encoding gene mutants. .................................................... 120 6.2.3 MALDI Mass Spectrometry of trypsin digested HgpA peptides from H. pullorum glycosyltransferase-encoding
Recommended publications
  • Kongre Kitabı
    Book of Abstracts International VETistanbul Group Congress 2014 28-30 April, 2014 Istanbul, Turkey Book of Abstracts www.vetistanbul2014.org International VETistanbul Group Congress 2014 28-30 April 2014 International VETistanbul Group Congress 2014 28-30 April, 2014 Istanbul, Turkey Organizing Committee Prof. Dr. Halil GÜNEŞ, Chair Prof. Dr. Bülent EKİZ Prof. Dr. Ali AYDIN Assoc. Prof. Dr. Serkan İKİZ Assoc. Prof. Dr. Hasret DEMİRCAN YARDİBİ Assoc. Prof. Dr. Gülsün PAZVANT Scientific Committee* Prof. Dr. Kemal AK, Turkey Prof. Dr. Anatoliy ALEXANDROVICH STEKOLNIKOV, Russia Prof. Dr. Bogdan AMINKOV, Bulgaria Prof. Dr. Geno ATASANOV ANGELOV, Bulgaria Prof. Dr. Hajrudin BESIROVIC, Bosnia and Herzegovina Prof. Dr. Nihad FEJZIC, Bosnia and Herzegovina Assoc. Prof. Dr. Plamen GEORGIEV, Bulgaria Prof. Dr. Zehra HAJRULAI MUSLIU, Macedonia Assoc. Prof. Dr. Afrim HAMIDI, Kosovo Prof. Dr. Telman ISKENDEROV, Azerbaijan Prof. Dr. Larisa KARPENKO, Russia Prof. Dr. Ismail KIRSAN, Turkey Prof. Dr. Mihni LYUTSKANOV, Bulgaria Assoc. Prof. Dr. Avni ROBAJ, Kosovo Prof. Dr. Velimir STOJKOVSKI, Macedonia Prof. Dr. Semsir VELIYEV, Azerbaijan *Alphabetically listed by the according to the family name Scientific Secreteria Prof. Dr. Bülent EKİZ, Turkey Dr. Karlo MURATOĞLU, Turkey International VETistanbul Group Congress 2014, 28-30 April, Istanbul, Turkey IV International VETistanbul Group Congress 2014 28-30 April 2014 Dear Respectable Colleagues and Guests, First of all, I greet you all with my heart. Also, I would like to thank you for taking place on our side due to the contribution given to the establishment of VETistanbul Group. Known as, VETistanbul Group was established, under the coordination of Istanbul University, with joint decision of Veterinary Faculty of the University of Sarajevo, Saint Petersburg State Academy of Veterinary Medicine, Stara Zagora Trakia University, Ss.
    [Show full text]
  • Supplemental Material S1.Pdf
    Phylogeny of Selenophosphate synthetases (SPS) Supplementary Material S1 ! SelD in prokaryotes! ! ! SelD gene finding in sequenced prokaryotes! We downloaded a total of 8263 prokaryotic genomes from NCBI (see Supplementary Material S7). We scanned them with the program selenoprofiles (Mariotti 2010, http:// big.crg.cat/services/selenoprofiles) using two SPS-family profiles, one prokaryotic (seld) and one mixed eukaryotic-prokaryotic (SPS). Selenoprofiles removes overlapping predictions from different profiles, keeping only the prediction from the profile that seems closer to the candidate sequence. As expected, the great majority of output predictions in prokaryotic genomes were from the seld profile. We will refer to the prokaryotic SPS/SelD !genes as SelD, following the most common nomenclature in literature.! To be able to inspect results by hand, and also to focus on good-quality genomes, we considered a reduced set of species. We took the prok_reference_genomes.txt list from ftp://ftp.ncbi.nlm.nih.gov/genomes/GENOME_REPORTS/, which NCBI claims to be a "small curated subset of really good and scientifically important prokaryotic genomes". We named this the prokaryotic reference set (223 species - see Supplementary Material S8). We manually curated most of the analysis in this set, while we kept automatized the !analysis on the full set.! We detected SelD proteins in 58 genomes (26.0%) in the prokaryotic reference set (figure 1 in main paper), which become 2805 (33.9%) when considering the prokaryotic full set (figure SM1.1). The difference in proportion between the two sets is due largely to the presence of genomes of very close strains in the full set, which we consider redundant.
    [Show full text]
  • Helicobacter Spp. — Food- Or Waterborne Pathogens?
    FRI FOOD SAFETY REVIEWS Helicobacter spp. — Food- or Waterborne Pathogens? M. Ellin Doyle Food Research Institute University of Wisconsin–Madison Madison WI 53706 Contents34B Introduction....................................................................................................................................1 Virulence Factors ...........................................................................................................................2 Associated Diseases .......................................................................................................................2 Gastrointestinal Disease .........................................................................................................2 Neurological Disease..............................................................................................................3 Other Diseases........................................................................................................................4 Epidemiology.................................................................................................................................4 Prevalence..............................................................................................................................4 Transmission ..........................................................................................................................4 Summary .......................................................................................................................................5
    [Show full text]
  • Helicobacter Pullorum As a Cause of Enterohepatic
    Table of contents _________________________________________________________________________________________ List of abbreviations 2 Scientific background 4 Enterohepatic Helicobacter species: a review of the literature 6 Scientific aims 48 Experimental studies Chapter 1 The agent Helicobacter pullorum: prevalence and in vitro susceptibility to different antimicrobial agents 50 1.1 Occurrence of Helicobacter pullorum in broiler chickens and comparison of isolates using amplified fragment length polymorphism profiling 52 1.2 Prevalence of Helicobacter pullorum among patients with gastrointestinal disease and clinically healthy persons 70 1.3 In vitro susceptibility of Helicobacter pullorum isolates to different antimicrobial agents 84 Chapter 2 Study of bacteria-host interactions 100 2.1 In vitro testing of virulence markers in Helicobacter pullorum 102 2.1.1 The cytolethal distending toxin among Helicobacter pullorum isolates from human and poultry origin 104 2.1.2 Cell death by mitotic catastrophe in mouse liver cells caused by Helicobacter pullorum 128 2.2 In vivo interaction of poultry and human Helicobacter pullorum isolates with broiler chickens 152 General discussion 174 Summary 194 Samenvatting 200 Author’s curriculum 206 Dankwoord 212 1 List of abbreviations _________________________________________________________________________________________ AFLP amplified fragment length polymorphism ATCC American Type Culture Collection ATM ataxia telangiectasia mutated ATP adenosine triphosphate ATR ATM and Rad3 related BHI brain heart
    [Show full text]
  • Comparison of the Culture and PCR Methods to Determine The
    Archive of SID Comparison of the Culture and PCR Methods to Determine the Prevalence and Antibiotic Resistance of Helicobacter pullorum Isolated from Chicken Thigh Samples in Semnan, Iran a b * c d Ashkan Jebellijavan | Seyed Hesamodin Emadi Chashmi | Hamid Staji | Hossein Akhlaghi a. Department of Food Hygiene and Quality Control, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran. b. Department of Clinical Science, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran. c. Department of Pathobiology, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran. d. DVM Student of Veterinary Medicine (D. V. M), Faculty of Veterinary Medicine, Semnan University, Semnan, Iran. *Corresponding author: Department of Clinical Science, Faculty of Veterinary Medicine, Semnan University, Semnan, Iran. Postal code: 3513119111. E-mail address: [email protected] ARTICLE INFO ABSTRACT Article type: Background: Helicobacter pullorum is among the most frequently reported pathogens Original article in poultry. The present study aimed to compare the culture and polymerase chain reaction (PCR) methods to assess the prevalence of H. pullorum isolated from chicken Article history: tight samples in Semnan, Iran. The antibiotic resistance pattern of the H. pullorum Received: 4 October 2020 isolates was also determined for the first time in Iran. Revised: 10 December 2020 Accepted: 29 December 2020 Methods: In total, 50 chicken thigh samples were collected from the local retail markets in Semnan city during January-September 2019. The samples were examined using the culture method and biochemical tests, and the final confirmation was based DOI: 10.29252/jhehp.6.4.3 on PCR with the 16S rRNA gene. In addition, antibiotic resistance test was performed using the disc-diffusion method.
    [Show full text]
  • Microbial and Mineralogical Characterizations of Soils Collected from the Deep Biosphere of the Former Homestake Gold Mine, South Dakota
    University of Nebraska - Lincoln DigitalCommons@University of Nebraska - Lincoln US Department of Energy Publications U.S. Department of Energy 2010 Microbial and Mineralogical Characterizations of Soils Collected from the Deep Biosphere of the Former Homestake Gold Mine, South Dakota Gurdeep Rastogi South Dakota School of Mines and Technology Shariff Osman Lawrence Berkeley National Laboratory Ravi K. Kukkadapu Pacific Northwest National Laboratory, [email protected] Mark Engelhard Pacific Northwest National Laboratory Parag A. Vaishampayan California Institute of Technology See next page for additional authors Follow this and additional works at: https://digitalcommons.unl.edu/usdoepub Part of the Bioresource and Agricultural Engineering Commons Rastogi, Gurdeep; Osman, Shariff; Kukkadapu, Ravi K.; Engelhard, Mark; Vaishampayan, Parag A.; Andersen, Gary L.; and Sani, Rajesh K., "Microbial and Mineralogical Characterizations of Soils Collected from the Deep Biosphere of the Former Homestake Gold Mine, South Dakota" (2010). US Department of Energy Publications. 170. https://digitalcommons.unl.edu/usdoepub/170 This Article is brought to you for free and open access by the U.S. Department of Energy at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in US Department of Energy Publications by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors Gurdeep Rastogi, Shariff Osman, Ravi K. Kukkadapu, Mark Engelhard, Parag A. Vaishampayan, Gary L. Andersen, and Rajesh K. Sani This article is available at DigitalCommons@University of Nebraska - Lincoln: https://digitalcommons.unl.edu/ usdoepub/170 Microb Ecol (2010) 60:539–550 DOI 10.1007/s00248-010-9657-y SOIL MICROBIOLOGY Microbial and Mineralogical Characterizations of Soils Collected from the Deep Biosphere of the Former Homestake Gold Mine, South Dakota Gurdeep Rastogi & Shariff Osman & Ravi Kukkadapu & Mark Engelhard & Parag A.
    [Show full text]
  • Emerging Pathogens in Meat and Poultry U.S
    A report from Sept 2016 Emerging Pathogens in Meat and Poultry U.S. must step up efforts to rapidly detect and control new foodborne hazards The Pew Charitable Trusts Susan K. Urahn, executive vice president Allan Coukell, senior director Safe food project Sandra Eskin, director Karin Hoelzer, officer External reviewers The report benefited from the insights and expertise of external peer reviewers Colin Parrish, Ph.D., the John M. Olin professor of virology and director of the Baker Institute for Animal Health and the Feline Health Center, both at Cornell University; and Ewen C.D. Todd, Ph.D., president of Ewen Todd Consulting, and former professor, Michigan State University. Neither the peer reviewers nor their organizations necessarily endorse the conclusions provided in this report. Acknowledgments The authors are grateful to J. Glenn Morris, Jr., M.D., M.P.H. and T.M., director of the Emerging Pathogens Institute and professor of infectious diseases, University of Florida; and Michael Batz, assistant director for food safety programs, Emerging Pathogens Institute, University of Florida. Both critically reviewed the topic and provided in-depth information regarding the risks posed by emerging pathogens in meat and poultry and the ways to address them. We appreciate the assistance of Betsy Towner Levine with fact-checking and Lisa Plotkin and Erika Compart with copy editing. Thank you to the following current and former Pew colleagues for their contributions to this report: Juliana Ruzante, Molly Mathews, Demetra Aposporos, Kimberly Burge, Matt Mulkey, and Elise Walter for their editorial input; and Kristin Centrella and Carol Conroy for their work preparing this report for publication.
    [Show full text]
  • WO 2012/055408 Al
    (12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date . 3 May 2012 (03.05.2012) WO 2012/055408 Al (51) International Patent Classification: DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, CI2Q 1/68 (2006.01) HN, HR, HU, ID, IL, IN, IS, JP, KE, KG, KM, KN, KP, KR, KZ, LA, LC, LK, LR, LS, LT, LU, LY, MA, MD, (21) International Application Number: ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, PCT/DK20 11/000120 NO, NZ, OM, PE, PG, PH, PL, PT, QA, RO, RS, RU, (22) International Filing Date: RW, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, 27 October 201 1 (27.10.201 1) TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (25) Filing Language: English (84) Designated States (unless otherwise indicated, for every (26) Publication Language: English kind of regional protection available): ARIPO (BW, GH, (30) Priority Data: GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, SZ, TZ, 61/407,122 27 October 2010 (27.10.2010) US UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, MD, PA 2010 70455 27 October 2010 (27.10.2010) DK RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, (71) Applicant (for all designated States except US): QUAN- LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, TIBACT A/S [DK/DK]; Kettegards Alle 30, DK-2650 SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, Hvidovre (DK).
    [Show full text]
  • Diversity of the Human Gastrointestinal Microbiota Novel Perspectives from High Throughput Analyses
    Diversity of the Human Gastrointestinal Microbiota Novel Perspectives from High Throughput Analyses Mirjana Rajilić-Stojanović Promotor Prof. Dr W. M. de Vos Hoogleraar Microbiologie Wageningen Universiteit Samenstelling Promotiecommissie Prof. Dr T. Abee Wageningen Universiteit Dr J. Doré INRA, France Prof. Dr G. R. Gibson University of Reading, UK Prof. Dr S. Salminen University of Turku, Finland Dr K. Venema TNO Quality for life, Zeist Dit onderzoek is uitgevoerd binnen de onderzoekschool VLAG. Diversity of the Human Gastrointestinal Microbiota Novel Perspectives from High Throughput Analyses Mirjana Rajilić-Stojanović Proefschrift Ter verkrijging van de graad van doctor op gezag van de rector magnificus van Wageningen Universiteit, Prof. dr. M. J. Kropff, in het openbaar te verdedigen op maandag 11 juni 2007 des namiddags te vier uur in de Aula Mirjana Rajilić-Stojanović, Diversity of the Human Gastrointestinal Microbiota – Novel Perspectives from High Throughput Analyses (2007) PhD Thesis, Wageningen University and Research Centre, Wageningen, The Netherlands – with Summary in Dutch – 214 p. ISBN: 978-90-8504-663-9 Abstract The human gastrointestinal tract is densely populated by hundreds of microbial (primarily bacterial, but also archaeal and fungal) species that are collectively named the microbiota. This microbiota performs various functions and contributes significantly to the digestion and the health of the host. It has previously been noted that the diversity of the gastrointestinal microbiota is disturbed in relation to several intestinal and not intestine related diseases. However, accurate and detailed defining of such disturbances is hampered by the fact that the diversity of this ecosystem is still not fully described, primarily because of its extreme complexity, and high inter-individual variability.
    [Show full text]
  • Isolation and Identification of Helicobacter Pullorum from Caecal Content of Broiler Chickens in Mashhad, Iran
    179 Iranian Journal of Veterinary Research, Shiraz University Scientific Report Isolation and identification of Helicobacter pullorum from caecal content of broiler chickens in Mashhad, Iran Jamshidi, A.1*; Bassami, M. R.2; Salami, H.3 and Mohammadi, S.3 1Department of Food Hygiene and Public Health, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran; 2Department of Clinical Sciences, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran, and Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran; 3Graduated from Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran *Correspondence: A. Jamshidi, Department of Food Hygiene and Public Health, Faculty of Veterinary Medicine, Ferdowsi University of Mashhad, Mashhad, Iran. E-mail: [email protected] (Received 4 Nov 2012; revised version 30 Jan 2014; accepted 19 Feb 2014) Summary The presence of Helicobacter pullorum in intestinal tract of broiler chickens may be a potential risk for human health. In this study, a total of 100 caecal samples of broilers carcasses from 20 flocks at a poultry abattoir in Mashhad suburb were tested for the presence of H. pullorum using modified conventional culture method by combination of culturing on Brucella sheep blood agar and a filtering technique. Suspected colonies were determined as H. pullorum using polymerase chain reaction (PCR) by amplifying a 447 bp fragment of the 16SrRNA gene of this bacteria. 41% of caecal content samples and samples from 12 broiler flocks (60%) were determined as positive for the presence of H. pullorum. This is the first report of H. pullorum in Iranian poultry flocks. The results showed high prevalence of this bacterium in broiler chickens in this area of Iran.
    [Show full text]
  • Microbial Source Tracking in Coastal Recreational Waters of Southern Maine
    University of New Hampshire University of New Hampshire Scholars' Repository Master's Theses and Capstones Student Scholarship Fall 2017 MICROBIAL SOURCE TRACKING IN COASTAL RECREATIONAL WATERS OF SOUTHERN MAINE: RELATIONSHIPS BETWEEN ENTEROCOCCI, ENVIRONMENTAL FACTORS, POTENTIAL PATHOGENS, AND FECAL SOURCES Derek Rothenheber University of New Hampshire, Durham Follow this and additional works at: https://scholars.unh.edu/thesis Recommended Citation Rothenheber, Derek, "MICROBIAL SOURCE TRACKING IN COASTAL RECREATIONAL WATERS OF SOUTHERN MAINE: RELATIONSHIPS BETWEEN ENTEROCOCCI, ENVIRONMENTAL FACTORS, POTENTIAL PATHOGENS, AND FECAL SOURCES" (2017). Master's Theses and Capstones. 1133. https://scholars.unh.edu/thesis/1133 This Thesis is brought to you for free and open access by the Student Scholarship at University of New Hampshire Scholars' Repository. It has been accepted for inclusion in Master's Theses and Capstones by an authorized administrator of University of New Hampshire Scholars' Repository. For more information, please contact [email protected]. MICROBIAL SOURCE TRACKING IN COASTAL RECREATIONAL WATERS OF SOUTHERN MAINE: RELATIONSHIPS BETWEEN ENTEROCOCCI, ENVIRONMENTAL FACTORS, POTENTIAL PATHOGENS, AND FECAL SOURCES BY DEREK ROTHENHEBER Microbiology BS, University of Maine, 2013 THESIS Submitted to the University of New Hampshire In Partial Fulfillment of the Requirements for the Degree of Master of Science in Microbiology September, 2017 This thesis has been examined and approved in partial fulfillment of the
    [Show full text]
  • The Interplay Between Campylobacter and Helicobacter Species And
    Kaakoush et al. Gut Pathogens 2014, 6:18 http://www.gutpathogens.com/content/6/1/18 RESEARCH Open Access The interplay between Campylobacter and Helicobacter species and other gastrointestinal microbiota of commercial broiler chickens Nadeem O Kaakoush1, Nidhi Sodhi1, Jeremy W Chenu2, Julian M Cox1,3,StephenMRiordan4,5 and Hazel M Mitchell1* Abstract Background: Poultry represent an important source of foodborne enteropathogens, in particular thermophilic Campylobacter species. Many of these organisms colonize the intestinal tract of broiler chickens as harmless commensals, and therefore, often remain undetected prior to slaughter. The exact reasons for the lack of clinical disease are unknown, but analysis of the gastrointestinal microbiota of broiler chickens may improve our understanding of the microbial interactions with the host. Methods: In this study, the fecal microbiota of 31 market-age (56-day old) broiler chickens, from two different farms, was analyzed using high throughput sequencing. The samples were then screened for two emerging human pathogens, Campylobacter concisus and Helicobacter pullorum, using species-specific PCR. Results: The gastrointestinal microbiota of chickens was classified into four potential enterotypes, similar to that of humans, where three enterotypes have been identified. The results indicated that variations between farms may have contributed to differences in the microbiota, though each of the four enterotypes were found in both farms suggesting that these groupings did not occur by chance. In addition to the identification of Campylobacter jejuni subspecies doylei and the emerging species, C. concisus, C. upsaliensis and H. pullorum, several differences in the prevalence of human pathogens within these enterotypes were observed. Further analysis revealed microbial taxa with the potential to increase the likelihood of colonization by a number of these pathogens, including C.
    [Show full text]