Open Yonghua Luo.Pdf

Total Page:16

File Type:pdf, Size:1020Kb

Open Yonghua Luo.Pdf The Pennsylvania State University The Graduate School Department of Crop and Soil Sciences CHARACTERIZATION OF H2-PRODUCING BACTERIAL COMMUNITIES FROM HEAT-TREATED SOIL AND ISOLATION OF DOMINANT CLOSTRIDIUM SPP. A Thesis in Soil Science by Yonghua Luo © 2007 Yonghua Luo Submitted in Partial Fulfillment of the Requirements for the Degree of Doctor of Philosophy August 2007 The thesis of Yonghua Luo was reviewed and approved* by the following: Mary Ann Bruns Associate Professor of Soil Science Thesis Advisor Chair of Committee John M.Regan Assistant Professor of Civil and Environmental Engineering David R.Huff Associate Professor of Turfgrass Breeding and Genetics Hery Lin Assistant Professor of Hydropedology/Soil Hydrology Curtis J.Dell Soil Scientist and Adjunct Professor of Soil Science David M.Sylvia Professor of Soil Microbiology Head of the Department of Crop and Soil Sciences *Signatures are on file in the Graduate School iii ABSTRACT Hydrogen is an attractive alternative to conventional fossil fuels. Currently most hydrogen is produced from nonrenewable natural gas, oil and coal. However, from the perspectives of economics and environmental quality, hydrogen production from renewable resources is gaining more and more attention. Biohydrogen production from the fermentation of wastewater containing carbohydrates is one approach for exploiting renewable resources. Much literature has been published on the effects of external factors such as substrate concentration, temperature, pH, etc. on hydrogen production by pure cultures or defined mixed cultures. However, only a few studies have been conducted on the effects of these external factors on H2-producing microbial communities from uncharacterized, mixed inocula. The objectives of this research were threefold: to determine the effect of glucose concentration on H2-producing bacterial community composition in the fermentation of synthetic wastewater; to determine the effect of L- cysteine and successive transfer on H2 producing bacterial community composition in batch cultures; and to isolate and characterize a new strain of H2-producing Clostridium spp. from mixed soil inocula using L-cysteine as reducing agent. In the first study, a continuous flow bioreactor was continuously fed with synthetic wastewater for periods of 7-14 days. Heat-treated soil was introduced into the reactor as a mixed inoculum and L- cysteine was added to a final concentration of 0.5g/L as reducing agent. PCR-based ribosomal RNA intergenic spacer analysis (RISA) was used to characterize the bacterial community composition in the bioreactor. It was revealed that RISA profiles of bacterial communities grown in media with 5.0, 7.5 or 10g/L glucose yielded partial 16S rRNA iv sequences most related to Clostridium spp. In contrast, RISA profiles of cultures grown in 2.5 g/L glucose yielded more diverse DNA sequences. The lowest glucose concentration gave rise to sequences representing two bacterial families, Clostridiaceae and Acidaminococcaeae, in the low G+C Gram-positive Firmicutes division, and one family, Enterobacteraceae in the gamma Proteobacteria division. Using an oligonucleotide probe “LYHI”, complementary to rRNA of an isolate obtained from the reactor containing 10 g/L glucose, it was shown that about 90% percent of the bacterial cells in microscope fields from bioreactor samples at 10 g/L glucose hybridized with the LYH1 probe. In samples from the bioreactor fed with 2.5 g/L glucose, only 26% of bacterial cells hybridized to LYH1 probe. These results further confirmed the effect of glucose on the composition of bacterial communities as judged by RISA. In the second study, batch experiments were carried out with the same nutrient levels used in the continuous flow bioreactor. Regardless of the glucose concentration and number of serial transfers, RISA profiles of bacterial communities grown without L-cysteine yielded partial 16S rRNA sequences closely related to Enterobacteriaceae. In contrast, RISA profiles of batch cultures with L-cysteine yielded results similar to those observed with the continuous flow bioreactor, which indicated that 5g/L appeared to be the lowest favorable glucose concentration for the successful competition of Clostridium spp. over other bacterial species. L-cysteine could have affected the composition of bacterial communities in batch experiments either due to its function as a quick oxygen scavenger or its role as a spore germinant. In the third study, a new Clostridium sp. strain LYH2 was isolated from a silty clay loam soil using L-cysteine as a reducing agent in the growth medium. Closest cultured relatives of strain LYH2 were Clostridium acidisoli, v Clostridium akagii and Clostridium pasteurianum. This dissertation showed that heat- treated soil provided a useful source of Clostridium spp. for producing biohydrogen from renewable wastes, and that glucose concentrations greater than 5g/L and L-cysteine enhanced their growth and H2 production. vi TABLE OF CONTENTS ABSTRACT…………………………………………………………………………....iii TABLE OF CONTENTS……………………………………………………………......vi LIST OF FIGURES…………………………………... ...............................................vviiii LIST OF TABLES........................................................................................................xiii ACKNOWLEDGEMENTS..........................................................................................xiv Chapter 1 Introduction…………………………………………………………………...1 REFERENCES…………………………………………………………………………..9 Chapter 2 The effect of glucose concentration on composition of H2- producing bacterial communities in a continuous flow bioreactor………………….………...…....14 ABSTRACT…………………………………………………………………………… 15 INTRODUCTION…………………………………………………………………….. ...17 MATERIALS AND METHODS……………………………………………………….19 RESULTS……………………………………………………………………………..... .26 DISCUSSION………………………………………………………………………......30 REFERENCES………………………………………………………………………......37 Chapter 3 The effect of L-cysteine on bacterial community composition and H2 production in batch cultures of heat-treated soil inocula……………………………….57 ABSTRACT………………………………………………………………………........58 INTRODUCTION…………………………………………………………….……. …...60 MATERIALS AND METHODS……………………………………………………......61 RESULTS…………………………………………………………………………….....66 DISCUSSION…………………………………………………………………………..69 vii REFERENCE………………………………………………………………………….74 Chapter 4 Strain LYH2, a N2-fixing, ferric-iron reducing Clostridium sp. isolated from a silty clay loam soil………………………………………………………….………….85 ABSTRACT……………………………………………………………………………...86 INTRODUCTION……………………………………………………………………….87 MATERIALS AND METHODS……………………………………………………….88 RESULTS……………………………………………………………………………...93 DISCUSSION……………………………………………………………………….....96 REFERENCE…………………………………………………………………………..99 Chapter 5 GENERAL CONCLUSIONS……………………………………………..110 Conclusion Appendix A…………………………………………………………………………..114 Appendix B…………………………………………………………………………... 116 viii LIST OF FIGURES Figure 1-1: H2 production pathways…………………………………………………..3 Figure 1-2: Pathway to hydrogen production by fermentation………………………...4 Figure 1-3 Structural formula of L-cysteine. The thiol group can effectively bind oxygen in liquid medium.………………………...................................................7 Figure 2-1: RISA profiles showing the bacterial community fingerprints of bioreactor samples with different glucose concentrations and hydraulic retention times. The glucose concentrations and hydraulic retention times are indicated at the top. The sizes of DNA marker bands are indicated by the arrows. The different DNA bands in bioreactor samples are indicated with arrows in their respective lanes ..………………………………………………………………………………....44 Figure 2-2: Petri plate after 6 days of incubation on anaerobic medium containing L-cysteine and 10g/L glucose, showing colonies of H2 producing bacteria...…...47 Figure 2-3: RISA fingerprint of genomic DNA of isolate from 10g/L glucose culture. The sizes of DNA bands are indicated by the arrows………………………….. 48 Figure 2-4: Neighbor-joining dendrogram depicting phylogenetic relationships of strain LYH1 with other Clostridium spp. based on nearly complete 16S rRNA sequences, using Bacillus subtilis as outgroup. GenBank accession numbers are in parentheses. The numbers at the nodes represent bootstrap values of 1,000 replicates. The scale bar represents the calculated number of changes per nucleotide position………………………………………………………………49 Figure 2-5: Comparison of biogas production (A) and H2 percentage in biogas (B) in ix samples grown with 0.5g/L L-cysteine and without L-cysteine. Column heights represent means and error bars indicate standard deviations. …………………....50 Figure 2-6: Accumulated gas production during growth of strain LYH1in batch cultures with and without L-cysteine (squares) and without L-cysteine (diamonds) …………………………………………………………………………………...51 Figure 2-7: Alignments of the LYH1 probe sequence, its target site, and sequences of corresponding sites in reference organisms. 17 related clostridia, Selenomonas strain SB90, Enterobacter cloacae strain CP1 and Bacillus subtilis were selected as reference organisms. The target sequence is displayed in the upper row; mismatches between target and sequences of corresponding sites in reference organisms are shadowed……………………………………………...52 Figure 2-8: Confocal Scanning Laser Microscope images of sample culture at 10g/L glucose concentration with 10-h HRT: (A) The hybridization of sample cells with universal EUB probe labeled Cy-3 and LYH1 probe labeled by Cy-5. (B) Morphology of sample cells examined by differential interference
Recommended publications
  • Structure and Dynamics of the Microbial Communities Underlying the Carboxylate Platform for Biofuel Production
    Structure and dynamics of the microbial communities underlying the carboxylate platform for biofuel production E.B. Hollister 1, A.K. Forrest 2, H.H. Wilkinson 3, D.J. Ebbole 3, S.A. Malfatti 4, S.G. Tringe 4, M.T. Holtzapple 2, T.J. Gentry 1 1 Department of Soil and Crop Sciences, Texas A&M University, College Station, TX, USA 77843-2474 2 Department of Chemical Engineering, Texas A&M University, College Station, TX, USA 77843-3122 3 Department of Plant Pathology and Microbiology, Texas A&M University, College Station, TX USA 77843-2132 4 DOE Joint Genome Institute, Walnut Creek, CA, USA 94598 July 31, 2010 ACKNOWLEDGMENT The work conducted by the U.S. Department of Energy Joint Genome Institute is supported by the Office of Science of the U.S. Department of Energy under Contract No. DE-AC02-05CH11231 DISCLAIMER This document was prepared as an account of work sponsored by the United States Government. While this document is believed to contain correct information, neither the United States Government nor any agency thereof, nor The Regents of the University of California, nor any of their employees, makes any warranty, express or implied, or assumes any legal responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by its trade name, trademark, manufacturer, or otherwise, does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof, or The Regents of the University of California.
    [Show full text]
  • Genomic and Evolutionary Insights
    CORE Metadata, citation and similar papers at core.ac.uk Provided by Apollo GBE Preterm Infant-Associated Clostridium tertium, Clostridium cadaveris,andClostridium paraputrificum Strains: Genomic and Evolutionary Insights Raymond Kiu1,2, Shabhonam Caim1, Cristina Alcon-Giner1, Gusztav Belteki3,PaulClarke4, Derek Pickard5, Gordon Dougan5,andLindsayJ.Hall1,* 1The Gut Health and Food Safety Programme, Quadram Institute Bioscience, Norwich Research Park, Norwich, United Kingdom 2Norwich Medical School, Norwich Research Park, University of East Anglia, Norwich, United Kingdom 3Neonatal Intensive Care Unit, The Rosie Hospital, Cambridge University Hospitals NHS Foundation Trust, United Kingdom 4Neonatal Intensive Care Unit, Norfolk and Norwich University Hospitals NHS Foundation Trust, Norwich, United Kingdom 5Wellcome Trust Sanger Institute, Wellcome Genome Campus, Hinxton, United Kingdom *Corresponding author: E-mail: [email protected]. Accepted: September 28, 2017 Data deposition: This project has been deposited at European Nucleotide Archive (EMBL-EBI) under the accession PRJEB22142. Bacterial strain deposition: Newly sequenced strains are deposited at National Collection of Type Cultures (NCTC; a culture depository of Public Health England). Abstract Clostridium species (particularly Clostridium difficile, Clostridium botulinum, Clostridium tetani and Clostridium perfringens)are associated with a range of human and animal diseases. Several other species including Clostridium tertium, Clostridium cadaveris, and Clostridium paraputrificum have also been linked with sporadic human infections, however there is very limited, or in some cases, no genomic information publicly available. Thus, we isolated one C. tertium strain, one C. cadaveris strain and three C. paraputrificum strains from preterm infants residing within neonatal intensive care units and performed Whole Genome Sequencing (WGS) using Illumina HiSeq. In this report, we announce the open availability of the draft genomes: C.
    [Show full text]
  • Supplementary Materials
    SUPPLEMENTARY MATERIALS Table S1. Chemical characteristics of the two digestate forms (SD and WD). Values quoted are expressed as % of air-dry digestate (means followed by standard error in brackets). References for the employed methods used for determination of each chemical characteristic is also reported. SD WD Reference Org C % 44.4 (0.33) 1.1 (0.01) [81] Tot N % 1.4 (0.01) 0.4 (0.01) [82] C/N 31.4 (0.17) 3.1 (0.04) NH4-N % n.d 0.2 (0.00) [83] K % 1.7 (0.00) n.d. [84] P % 0.9 (0.01) n.d. [84] S % 0.23 (0.02) n.d. [85] SD = solid digestate; WD = whole digestate. Table S2. Soil physical and chemical characteristics at the beginning of trial (t0) (means from 9 observations followed by standard errors in brackets). Clay (%) 41.9 (1.22) Silt (%) 47.8 (2.13) Moisture (%) 24.46 (1.24) Bulk density (g cm-3) 1.39 (0.04) pH 8.3 (0) CaCO3 (%) 11.4 (0.7) TOC (g kg-1) 12.8 (0.3) TN (g kg-1) 1.4 (0) C/N 9.4 (0.3) CEC (cmol(+) kg-1) 21.0 (0.7) Exchangeable Bases (mg kg-1) K 278.7 (8.0) Na 22.0 (3.0) Mg 201.7 (25.6) Ca 3718.6 (176.3) Available Microelements (mg kg-1) Cu 28.0 (5.0) Zn 1.7 (0.2) Fe 15.4 (0.5) Mn 16.3 (0.5) TOC = total organic C; TN = total N; CEC = cation exchange capacity Table S3.
    [Show full text]
  • Botulism: Guidance for Health Care Providers Key Medical and Public Health Interventions After Identification of a Suspected Case
    Virginia Department of Health Botulism: Guidance for Health Care Providers Key Medical and Public Health Interventions after Identification of a Suspected Case Table of Contents 1. Epidemiology ........................................................................................................................................ 1 2. Clinical Manifestations .......................................................................................................................... 2 3. Laboratory Testing and Diagnosis ......................................................................................................... 3 4. Treatment ............................................................................................................................................. 6 5. Post-exposure Prophylaxis .................................................................................................................... 6 6. Vaccination ........................................................................................................................................... 6 7. Infection Control ................................................................................................................................... 6 8. Decontamination .................................................................................................................................. 7 9. Postmortem Practices ........................................................................................................................... 7 10. Public Health
    [Show full text]
  • WO 2018/064165 A2 (.Pdf)
    (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 WO 2018/064165 A2 05 April 2018 (05.04.2018) W !P O PCT (51) International Patent Classification: Published: A61K 35/74 (20 15.0 1) C12N 1/21 (2006 .01) — without international search report and to be republished (21) International Application Number: upon receipt of that report (Rule 48.2(g)) PCT/US2017/053717 — with sequence listing part of description (Rule 5.2(a)) (22) International Filing Date: 27 September 2017 (27.09.2017) (25) Filing Language: English (26) Publication Langi English (30) Priority Data: 62/400,372 27 September 2016 (27.09.2016) US 62/508,885 19 May 2017 (19.05.2017) US 62/557,566 12 September 2017 (12.09.2017) US (71) Applicant: BOARD OF REGENTS, THE UNIVERSI¬ TY OF TEXAS SYSTEM [US/US]; 210 West 7th St., Austin, TX 78701 (US). (72) Inventors: WARGO, Jennifer; 1814 Bissonnet St., Hous ton, TX 77005 (US). GOPALAKRISHNAN, Vanch- eswaran; 7900 Cambridge, Apt. 10-lb, Houston, TX 77054 (US). (74) Agent: BYRD, Marshall, P.; Parker Highlander PLLC, 1120 S. Capital Of Texas Highway, Bldg. One, Suite 200, Austin, TX 78746 (US). (81) Designated States (unless otherwise indicated, for every kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW.
    [Show full text]
  • Clostridium Amazonitimonense, Clostridium Me
    ORIGINAL ARTICLE Taxonogenomic description of four new Clostridium species isolated from human gut: ‘Clostridium amazonitimonense’, ‘Clostridium merdae’, ‘Clostridium massilidielmoense’ and ‘Clostridium nigeriense’ M. T. Alou1, S. Ndongo1, L. Frégère1, N. Labas1, C. Andrieu1, M. Richez1, C. Couderc1, J.-P. Baudoin1, J. Abrahão2, S. Brah3, A. Diallo1,4, C. Sokhna1,4, N. Cassir1, B. La Scola1, F. Cadoret1 and D. Raoult1,5 1) Aix-Marseille Université, Unité de Recherche sur les Maladies Infectieuses et Tropicales Emergentes, UM63, CNRS 7278, IRD 198, INSERM 1095, Marseille, France, 2) Laboratório de Vírus, Departamento de Microbiologia, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil, 3) Hopital National de Niamey, BP 247, Niamey, Niger, 4) Campus Commun UCAD-IRD of Hann, Route des pères Maristes, Hann Maristes, BP 1386, CP 18524, Dakar, Senegal and 5) Special Infectious Agents Unit, King Fahd Medical Research Center, King Abdulaziz University, Jeddah, Saudi Arabia Abstract Culturomics investigates microbial diversity of the human microbiome by combining diversified culture conditions, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and 16S rRNA gene identification. The present study allowed identification of four putative new Clostridium sensu stricto species: ‘Clostridium amazonitimonense’ strain LF2T, ‘Clostridium massilidielmoense’ strain MT26T, ‘Clostridium nigeriense’ strain Marseille-P2414T and ‘Clostridium merdae’ strain Marseille-P2953T, which we describe using the concept of taxonogenomics. We describe the main characteristics of each bacterium and present their complete genome sequence and annotation. © 2017 Published by Elsevier Ltd. Keywords: ‘Clostridium amazonitimonense’, ‘Clostridium massilidielmoense’, ‘Clostridium merdae’, ‘Clostridium nigeriense’, culturomics, emerging bacteria, human microbiota, taxonogenomics Original Submission: 18 August 2017; Revised Submission: 9 November 2017; Accepted: 16 November 2017 Article published online: 22 November 2017 intestine [1,4–6].
    [Show full text]
  • Clostridium Tertium Isolated from Gas Gangrene Wound; Misidentified As Lactobacillus Spp Initially Due to Aerotolerant Feature
    ARTICLE IN PRESS Anaerobe 13 (2007) 161–165 www.elsevier.com/locate/anaerobe Short communication — Anaerobiosis: Molecular Biology, Genetics and Other Aspects Clostridium tertium isolated from gas gangrene wound; misidentified as Lactobacillus spp initially due to aerotolerant feature Shigeki Fujitania,Ã, Chengxu X. Liua, Sydney M. Finegolda, Yuli L. Songa, Glenn E. Mathisenb aInfectious Diseases Section, VA Greater Los Angeles Healthcare System, 11301 Wilshire Blvd., Los Angeles, CA 90073, USA bInfectious Disease Service, UCLA-Olive View Medical Center, 14445 Olive View Dr., Sylmar, CA 91342, USA Received 19 January 2007; received in revised form 24 February 2007; accepted 5 March 2007 Available online 12 March 2007 Abstract Clostridium tertium has been increasingly reported as a human pathogen. This organism is an aerotolerant Gram-positive rod that is often mistaken for other organisms, such as Lactobacillus or Bacillus species. We describe a case of a patient with a history of intravenous drug use presenting to UCLA-Olive View Medical Center with gas gangrene of both upper extremities. The organism was initially misidentified as a Lactobacillus species on aerobic culture plates. However, terminal spore formation was detected in this isolate on a sub- cultured anaerobic culture plate and this isolate was confirmed as C. tertium biochemically and genetically by 16S rDNA sequencing. Additional DNA cloning libraries made from the formalin-fixed specimen revealed Peptoniphilus species and an uncultured Clostridium clone, but not C. tertium. C. tertium might be a causative organism of gas-producing myonecrosis but such an association has never been described. Clinicians should be aware of the phenomenon of aerotolerance of some anaerobes and need to clarify the identification of organisms if the clinical picture does not fit the isolated organism.
    [Show full text]
  • 974-Form.Pdf
    California Association for Medical Laboratory Technology Distance Learning Program ANAEROBIC BACTERIOLOGY FOR THE CLINICAL LABORATORY by James I. Mangels, MA, CLS, MT(ASCP) Consultant Microbiology Consulting Services Santa Rosa, CA Course Number: DL-974 3.0 CE/Contact Hour Level of Difficulty: Intermediate © California Association for Medical Laboratory Technology. Permission to reprint any part of these materials, other than for credit from CAMLT, must be obtained in writing from the CAMLT Executive Office. CAMLT is approved by the California Department of Health Services as a CA CLS Accrediting Agency (#0021) and this course is is approved by ASCLS for the P.A.C.E. ® Program (#519) 1895 Mowry Ave, Suite 112 Fremont, CA 94538-1766 Phone: 510-792-4441 FAX: 510-792-3045 Notification of Distance Learning Deadline All continuing education units required to renew your license must be earned no later than the expiration date printed on your license. If some of your units are made up of Distance Learning courses, please allow yourself enough time to retake the test in the event you do not pass on the first attempt. CAMLT urges you to earn your CE units early!. CAMLT Distance Learning Course # DL-974 1 © California Association for Medical Laboratory Technology Outline A. Introduction B. What are anaerobic bacteria? Concepts of anaerobic bacteriology C. Why do we need to identify anaerobes? D. Normal indigenous anaerobic flora; the incidence of anaerobes at various body sites E. Anaerobic infections; most common anaerobic infections F. Specimen collection and transport; acceptance and rejection criteria G. Processing of clinical specimens 1. Microscopic examination 2.
    [Show full text]
  • A Report of 22 Unrecorded Bacterial Species in Korea, Isolated from Namhangang
    Journal114 of Species Research 7(2):114-122, 2018JOURNAL OF SPECIES RESEARCH Vol. 7, No. 2 A report of 22 unrecorded bacterial species in Korea, isolated from Namhangang Chaeyun Baek1 and Hana Yi1,2,* 1Department of Public Health Sciences, Graduate School, Korea University, Seoul 02841, Republic of Korea 2School of Biosystem and Biomedical Science, Korea University, Seoul 02841, Republic of Korea *Correspondent: [email protected] As part of a larger study of indigenous prokaryotic species diversity in South Korea, various samples from Namhangang were subjected to analyses. Fresh water, underwater sediment, and moss-inhabiting aerobic and anaerobic bacteria were isolated. 22 of the isolates were identified as unrecorded bacterial species in Korea that had ≥98.7% 16S rRNA gene sequence similarity with published species. The aerobic strains isolated were Kurthia gibsonii and Massilia plicata. Also identified were four facultative anaerobic strains: Bacillus hisashii, Enterococcus rotai, Paenibacillus vini, and Pediococcus pentosaceus. 16 strictly anaerobic strains were identified as Bacteroides xylanolyticus, Carnobacterium maltaromaticum, Clostridium argentinense, Clostridium beijerinckii, Clostridium butyricum, Clostridium cavendishii, Clostridium diolis, Clostridium frigidicarnis, Clostridium perfringens, Clostridium saccharoperbutylacetonicum, Clostridium sphenoides, Clostridium subterminale, Cutibacterium acnes, Paraclostridium bifermentans, Prevotella paludivivens, and Romboutsia lituseburensis. Based on the examination of morphological, cultural, physiological, and biochemical properties of the isolates, descriptive information of these previously unrecorded species is provided here. Keywords: anaerobes, Namhangang, unrecorded species Ⓒ 2018 National Institute of Biological Resources DOI:10.12651/JSR.2018.7.2.114 INTRODUCTION and Jungnyeongcheon, are freshwater rivers with 10.19 km2, 69.11 km2, 150.5 km2, and 130.19 km2 areas, respec- While molecular methods have supplanted traditional tively.
    [Show full text]
  • Comparison of Sampling Techniques and Different Media for The
    Systematic and Applied Microbiology 42 (2019) 481–487 Contents lists available at ScienceDirect Systematic and Applied Microbiology jou rnal homepage: http://www.elsevier.com/locate/syapm Comparison of sampling techniques and different media for the enrichment and isolation of cellulolytic organisms from biogas fermentersଝ a a b a,∗ Regina Rettenmaier , Carina Duerr , Klaus Neuhaus , Wolfgang Liebl , a,c,∗ Vladimir V. Zverlov a Department of Microbiology, Technical University of Munich, Emil-Ramann-Str. 4, 85354 Freising, Germany b Core Facility Microbiome/NGS, ZIEL — Institute for Food & Health, Technical University of Munich, Weihenstephaner Berg 3, 85354 Freising, Germany c Institute of Molecular Genetics, RAS, Kurchatov Sq. 2, 123128 Moscow, Russia a r t i c l e i n f o a b s t r a c t Article history: Biogas plants achieve its highest yield on plant biomass only with the most efficient hydrolysis of cellulose. Received 27 March 2019 This is driven by highly specialized hydrolytic microorganisms, which we have analyzed by investigating Received in revised form 15 May 2019 enrichment strategies for the isolation of cellulolytic bacteria out of a lab-scale biogas fermenter. We Accepted 17 May 2019 compared three different cultivation media as well as two different inoculation materials: Enrichment on filter paper in nylon bags (in sacco) or raw digestate. Next generation sequencing of the V3/V4 region Keywords: of the bacterial 16S rRNA of metagenomic DNA from six different enrichment cultures, each in biolog- Biogas fermenter ical triplicates, revealed an average richness of 48 different OTU’s with an average evenness of 0.3 in Cellulose degradation each sample.
    [Show full text]
  • How Do Pesticides Influence Gut Microbiota? a Review
    International Journal of Environmental Research and Public Health Review Toxicology and Microbiota: How Do Pesticides Influence Gut Microbiota? A Review Federica Giambò 1,†, Michele Teodoro 1,† , Chiara Costa 2,* and Concettina Fenga 1 1 Department of Biomedical and Dental Sciences and Morphofunctional Imaging, Occupational Medicine Section, University of Messina, 98125 Messina, Italy; [email protected] (F.G.); [email protected] (M.T.); [email protected] (C.F.) 2 Clinical and Experimental Medicine Department, University of Messina, 98125 Messina, Italy * Correspondence: [email protected]; Tel.: +39-090-2212052 † Equally contributed. Abstract: In recent years, new targets have been included between the health outcomes induced by pesticide exposure. The gastrointestinal tract is a key physical and biological barrier and it represents a primary site of exposure to toxic agents. Recently, the intestinal microbiota has emerged as a notable factor regulating pesticides’ toxicity. However, the specific mechanisms related to this interaction are not well known. In this review, we discuss the influence of pesticide exposure on the gut microbiota, discussing the factors influencing gut microbial diversity, and we summarize the updated literature. In conclusion, more studies are needed to clarify the host–microbial relationship concerning pesticide exposure and to define new prevention interventions, such as the identification of biomarkers of mucosal barrier function. Keywords: gut microbiota; microbial community; pesticides; occupational exposure; dysbiosis Citation: Giambò, F.; Teodoro, M.; Costa, C.; Fenga, C. Toxicology and Microbiota: How Do Pesticides Influence Gut Microbiota? A Review. 1. Introduction Int. J. Environ. Res. Public Health 2021, 18, 5510. https://doi.org/10.3390/ In recent years, the demand for food has risen significantly in relation to the world ijerph18115510 population’s increase.
    [Show full text]
  • Molecular Characterization of Bacterial Populations of Different Soils
    Brazilian Journal of Microbiology (2006) 37:439-447 ISSN 1517-8382 MOLECULAR CHARACTERIZATION OF BACTERIAL POPULATIONS OF DIFFERENT SOILS Rodrigo Matheus Pereira; Érico Leandro da Silveira; Denilson César Scaquitto; Eliamar Aparecida Nascimbém Pedrinho; Silvana Pompéia Val-Moraes; Ester Wickert; Lúcia Maria Carareto-Alves; Eliana Gertrudes de Macedo Lemos* Departamento de Tecnologia, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista, Jaboticabal (SP), Brazil Submitted: January 01, 2006; Returned to authors for corrections: April 17, 2006; Approved: October 13, 2006 ABSTRACT Until recently, few studies were carried out in Brazil about diversity of bacterial soil communities. Aiming to characterize the bacterial population in the soil through 16S rRNA analysis, two types of soil have been analyzed: one of them characterized by intensive use where tomato, beans and corn were cultivated (CS); the other analyzed soil was under forest (FS), unchanged by man; both located in Guaíra, São Paulo State, Brazil. Using specific primers, 16S rRNA genes from metagenomic DNA in both soils were amplified by PCR, amplicons were cloned and 139 clones from two libraries were partially sequenced. The use of 16S rRNA analysis allowed identification of several bacterial populations in the soil belonging to the following phyla: Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria Verrucomicrobia in addition to the others that were not classified, beyond Archaea domain. Differences between FS and CS libraries were observed in size phyla. A larger number of phyla and, consequently, a greater bacterial diversity were found in the under-forest soil. These data were confirmed by the analyses of genetic diversity that have been carried out. The characterization of bacterial communities of soil has made its contribution by providing facts for further studies on the dynamics of bacterial populations in different soil conditions in Brazil.
    [Show full text]