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Sithu Shein AM, Hongsing P, Khatib A, Phattharapornjaroen P, Miyanaga K, Cui L, Shibuya K, Amarasiri M, Monk PN, Kicic A, Chatsuwan T, Higgins PG, Abe S, Wannigama DL. Phage therapy could be key to conquering persistent bacterial lung infections in children. NPJ ANTIMICROBIALS AND RESISTANCE 2024; 2:31. [PMID: 39843534 PMCID: PMC11721074 DOI: 10.1038/s44259-024-00045-4] [Citation(s) in RCA: 1] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Subscribe] [Scholar Register] [Received: 05/15/2024] [Accepted: 08/29/2024] [Indexed: 01/24/2025]
Abstract
Persistent bacterial lung infections in children lead to significant morbidity and mortality due to antibiotic resistance. In this paper, we describe how phage therapy has shown remarkable efficacy in preclinical and clinical studies, demonstrating significant therapeutic benefits through various administration routes. Ongoing trials are evaluating its safety and effectiveness against different pathogens. Advancing phage therapy through systematic studies and international collaboration could provide a viable alternative to traditional antibiotics for persistent infections.
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Affiliation(s)
- Aye Mya Sithu Shein
- Department of Microbiology, Faculty of Medicine, Chulalongkorn University, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand
- Center of Excellence in Antimicrobial Resistance and Stewardship Research, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
| | - Parichart Hongsing
- Mae Fah Luang University Hospital, Chiang Rai, Thailand
- School of Integrative Medicine, Mae Fah Luang University, Chiang Rai, Thailand
| | - Aisha Khatib
- Department of Family & Community Medicine, University of Toronto, Toronto, ON, Canada
| | - Phatthranit Phattharapornjaroen
- Faculty of Health Science Technology, Chulabhorn Royal Academy, Bangkok, Thailand
- HRH Princess Chulabhorn Disaster and Emergency Medicine Center, Chulabhorn Royal Academy, Bangkok, Thailand
| | - Kazuhiko Miyanaga
- Division of Bacteriology, School of Medicine, Jichi Medical University, Tochigi, Japan
| | - Longzhu Cui
- Division of Bacteriology, School of Medicine, Jichi Medical University, Tochigi, Japan
| | - Kenji Shibuya
- Tokyo Foundation for Policy Research, Minato-ku, Tokyo, Japan
| | - Mohan Amarasiri
- Department of Civil and Environmental Engineering, Graduate School of Engineering, Tohoku University, Sendai, Miyagi, Japan
| | - Peter N Monk
- Department of Infection, Immunity & Cardiovascular Disease, University of Sheffield Medical School, Sheffield, UK
| | - Anthony Kicic
- Wal-yan Respiratory Research Centre, Telethon Kids Institute, University of Western Australia, Nedlands, 6009, WA, Australia.
- Centre for Cell Therapy and Regenerative Medicine, Medical School, The University of Western Australia, Nedlands, 6009, WA, Australia.
- Department of Respiratory and Sleep Medicine, Perth Children's Hospital, Nedlands, 6009, WA, Australia.
- School of Population Health, Curtin University, Bentley, 6102, WA, Australia.
| | - Tanittha Chatsuwan
- Department of Microbiology, Faculty of Medicine, Chulalongkorn University, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand.
- Center of Excellence in Antimicrobial Resistance and Stewardship Research, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
| | - Paul G Higgins
- Institute for Medical Microbiology, Immunology and Hygiene, Faculty of Medicine and University Hospital Cologne, University of Cologne, Cologne, Germany.
- German Centre for Infection Research, Partner site Bonn-Cologne, Cologne, Germany.
- Center for Molecular Medicine Cologne, Faculty of Medicine and University Hospital Cologne, University of Cologne, 50935, Cologne, Germany.
| | - Shuichi Abe
- Department of Infectious Diseases and Infection Control, Yamagata Prefectural Central Hospital, Yamagata, Japan
| | - Dhammika Leshan Wannigama
- Department of Microbiology, Faculty of Medicine, Chulalongkorn University, King Chulalongkorn Memorial Hospital, Thai Red Cross Society, Bangkok, Thailand.
- Center of Excellence in Antimicrobial Resistance and Stewardship Research, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand.
- Department of Infectious Diseases and Infection Control, Yamagata Prefectural Central Hospital, Yamagata, Japan.
- School of Medicine, Faculty of Health and Medical Sciences, The University of Western Australia, Nedland, WA, Australia.
- Biofilms and Antimicrobial Resistance Consortium of ODA receiving countries, The University of Sheffield, Sheffield, UK.
- Pathogen Hunter's Research Team, Department of Infectious Diseases and Infection Control, Yamagata Prefectural Central Hospital, Yamagata, Japan.
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Pradel N, Bartoli M, Koenen M, Bale N, Neumann-Schaal M, Spröer C, Bunk B, Rohde M, Pester M, Spring S. Description and genome analysis of a novel archaeon isolated from a syntrophic pyrite-forming enrichment culture and reclassification of Methanospirillum hungatei strains GP1 and SK as Methanospirillum purgamenti sp. nov. PLoS One 2024; 19:e0308405. [PMID: 39186748 PMCID: PMC11346949 DOI: 10.1371/journal.pone.0308405] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/15/2024] [Accepted: 07/23/2024] [Indexed: 08/28/2024] Open
Abstract
The archaeal isolate J.3.6.1-F.2.7.3T was obtained from an anaerobic enrichment culture, where it may play an important role in methane production during pyrite formation. The new isolate formed a species-level clade with Methanospirillum hungatei strains GP1 and SK, which is separate from the type strain JF-1T. Cultivation-independent surveys indicate the occurrence of this phylogenetic group in sediments and anaerobic digesters. The abundance of this clade appears to be negatively affected by high nitrogen loads, indicating a sensitivity to certain nitrogen compounds that is not known in M. hungatei JF-1T. The relatively large core genome of this Methanospirillum clade is indicative of niche specialization and efficient control of horizontal gene transfer. Genes for nitrogenase and F420-dependent secondary alcohol dehydrogenase contribute to the metabolic versatility of this lineage. Characteristics of the new isolate such as the ability to utilize 2-propanol as an electron donor or the requirement for acetate as a carbon source are found also in the strains GP1 and SK, but not in the type strain M. hungatei JF-1T. Based on the genomic differences to related species, a new species within the genus Methanospirillum is proposed with the name M. purgamenti sp. nov. The determined phenotypic characteristics support this proposal and indicate a metabolic adaptation to a separate ecological niche.
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Affiliation(s)
- Nathalie Pradel
- CNRS/INSU, IRD, MIO, UM 110, Aix-Marseille Université, Université du Sud Toulon-Var, Marseille, France
| | - Manon Bartoli
- CNRS/INSU, IRD, MIO, UM 110, Aix-Marseille Université, Université du Sud Toulon-Var, Marseille, France
| | - Michel Koenen
- Royal Netherlands Institute for Sea Research, Texel, Netherlands
| | - Nicole Bale
- Royal Netherlands Institute for Sea Research, Texel, Netherlands
| | - Meina Neumann-Schaal
- Research Group Metabolomics, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Cathrin Spröer
- Department Bioinformatics, Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Boyke Bunk
- Department Bioinformatics, Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Manfred Rohde
- Central Facility for Microscopy, Helmholtz Centre for Infection Research, HZI, Braunschweig, Germany
| | - Michael Pester
- Department Microorganisms, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
- Institute for Microbiology, Technical University of Braunschweig, Braunschweig, Germany
| | - Stefan Spring
- Department Microorganisms, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
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Guo M, Zhang Y, Wu L, Xiong Y, Xia L, Cheng Y, Ma J, Wang H, Sun J, Wang Z, Yan Y. Development and mouse model evaluation of a new phage cocktail intended as an alternative to antibiotics for treatment of Staphylococcus aureus-induced bovine mastitis. J Dairy Sci 2024; 107:5974-5987. [PMID: 38522833 DOI: 10.3168/jds.2024-24540] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2023] [Accepted: 02/19/2024] [Indexed: 03/26/2024]
Abstract
Bovine mastitis is a prevalent infectious disease in dairy herds worldwide, resulting in substantial economic losses. Staphylococcus aureus is a major cause of mastitis in animals, and its antibiotic resistance poses challenges for treatment. Recently, renewed interest has focused on the development of alternative methods to antibiotic therapy, including bacteriophages (phages), for controlling bacterial infections. In this study, 2 lytic phages, vB_SauM_JDYN (JDYN) and vB_SauM_JDF86 (JDF86), were isolated from the cattle sewage effluent samples collected from dairy farms in Shanghai. The 2 phages have a broad bactericidal spectrum against Staphylococcus of various origins. Genomic and morphological analyses revealed that the 2 phages belonged to the Myoviridae family. Moreover, JDYN and JDF86 remained stable under a wide temperature and pH range and were almost unaffected in chloroform. In this study, we prepared a phage cocktail (PHC-1) which consisted of a 1:1:1 ratio of JDYN, JDF86, and SLPW (a previously characterized phage). We found that PHC-1 showed the strongest bacteriolytic effect and the lowest frequency of emergence of bacteriophage insensitive mutants compared with monophages. Bovine mammary epithelial cells and lactating mice mastitis models were used to evaluate the effectiveness of PHC-1 in vitro and in vivo, respectively. The results demonstrated that PHC-1 treatment significantly reduced bacterial load, alleviated inflammatory response, and improved mastitis pathology. Altogether, these results suggest that PHC-1 has the potential to treat S. aureus-induced bovine mastitis and that phage cocktails can combat antibiotic-resistant S. aureus infections.
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Affiliation(s)
- Mengting Guo
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Yumin Zhang
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Lifei Wu
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Yangjing Xiong
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Lu Xia
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Yuqiang Cheng
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Jingjiao Ma
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Hengan Wang
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Jianhe Sun
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China
| | - Zhaofei Wang
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China.
| | - Yaxian Yan
- School of Agriculture and Biology, Shanghai Jiao Tong University, Shanghai Key Laboratory of Veterinary Biotechnology, Shanghai 201100, China.
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Jo IH, Ko SW. Acute cholangitis with Achromobacter xylosoxidans bacteremia after endoscopic retrograde cholangiopancreatography in hilar cholangiocarcinoma: A case report. World J Clin Cases 2024; 12:4377-4383. [PMID: 39015928 PMCID: PMC11235522 DOI: 10.12998/wjcc.v12.i20.4377] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 03/25/2024] [Revised: 05/14/2024] [Accepted: 05/27/2024] [Indexed: 06/30/2024] Open
Abstract
BACKGROUND Achromobacter xylosoxidans is a Gram-negative opportunistic aerobe, usually causing nosocomial infections in immunocompromised patients with manifestations including bacteremia, pneumonia, and catheter-related infections. However, A. xylosoxidans have not yet been reported to cause biliary system infections. CASE SUMMARY A 72-year-old woman presented to the outpatient department of our hospital with a chief complaint of jaundice. Computed tomography of her abdomen revealed the presence of a mass of approximately 2.4 cm in the hilar portion of the common hepatic duct, consistent with hilar cholangiocarcinoma. We performed endoscopic retrograde cholangiopancreatography (ERCP) to decompress the obstructed left and right intrahepatic ducts (IHDs) and placed 10 cm and 11 cm biliary stents in the left and right IHDs, respectively. However, the day after the procedure, the patient developed post-ERCP cholangitis as the length of the right IHD stent was insufficient for proper bile drainage. The blood culture of the patient tested positive for A. xylosoxidans. Management measures included the replacement of the right IHD stent (11 cm) with a longer one (12 cm) and administering culture-directed antibiotic therapy, solving the cholangitis-related complications. After the cholangitis had resolved, the patient underwent surgery for hilar cholangiocarcinoma and survived for 912 d without recurrence. CONCLUSION A. xylosoxidans-induced biliary system infections are extremely rare. Clinical awareness of physicians and endoscopists is required as this rare pathogen might cause infection after endoscopic procedures.
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Affiliation(s)
- Ik Hyun Jo
- Department of Internal Medicine, St. Vincent’s Hospital, College of Medicine, The Catholic University of Korea, Seoul 16471, South Korea
| | - Sung Woo Ko
- Department of Internal Medicine, Eunpyeong St. Mary’s Hospital, College of Medicine, The Catholic University of Korea, Seoul 03312, South Korea
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Vaezi A, Healy T, Ebrahimi G, Rezvankhah S, Hashemi Shahraki A, Mirsaeidi M. Phage therapy: breathing new tactics into lower respiratory tract infection treatments. Eur Respir Rev 2024; 33:240029. [PMID: 38925791 PMCID: PMC11216685 DOI: 10.1183/16000617.0029-2024] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/15/2024] [Accepted: 04/16/2024] [Indexed: 06/28/2024] Open
Abstract
Lower respiratory tract infections (LRTIs) present a significant global health burden, exacerbated by the rise in antimicrobial resistance (AMR). The persistence and evolution of multidrug-resistant bacteria intensifies the urgency for alternative treatments. This review explores bacteriophage (phage) therapy as an innovative solution to combat bacterial LRTIs. Phages, abundant in nature, demonstrate specificity towards bacteria, minimal eukaryotic toxicity, and the ability to penetrate and disrupt bacterial biofilms, offering a targeted approach to infection control. The article synthesises evidence from systematic literature reviews spanning 2000-2023, in vitro and in vivo studies, case reports and ongoing clinical trials. It highlights the synergistic potential of phage therapy with antibiotics, the immunophage synergy in animal models, and the pharmacodynamics and pharmacokinetics critical for clinical application. Despite promising results, the article acknowledges that phage therapy is at a nascent stage in clinical settings, the challenges of phage-resistant bacteria, and the lack of comprehensive cost-effectiveness studies. It stresses the need for further research to optimise phage therapy protocols and navigate the complexities of phage-host interactions, particularly in vulnerable populations such as the elderly and immunocompromised. We call for regulatory adjustments to facilitate the exploration of the long-term effects of phage therapy, aiming to incorporate this old-yet-new therapy into mainstream clinical practice to tackle the looming AMR crisis.
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Affiliation(s)
- Atefeh Vaezi
- Division of Pulmonary, Critical Care and Sleep Medicine, College of Medicine-Jacksonville, University of Florida, Jacksonville, FL, USA
| | - Thomas Healy
- Division of Pulmonary, Critical Care and Sleep Medicine, College of Medicine-Jacksonville, University of Florida, Jacksonville, FL, USA
| | | | | | - Abdolrazagh Hashemi Shahraki
- Division of Pulmonary, Critical Care and Sleep Medicine, College of Medicine-Jacksonville, University of Florida, Jacksonville, FL, USA
| | - Mehdi Mirsaeidi
- Division of Pulmonary, Critical Care and Sleep Medicine, College of Medicine-Jacksonville, University of Florida, Jacksonville, FL, USA
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Sarat N, Salim A, Pal S, Subhash S, Prasad M, Nair BG, Madhavan A. Mitigation of biogenic methanethiol using bacteriophages in synthetic wastewater augmented with Pseudomonas putida. Sci Rep 2023; 13:19480. [PMID: 37945592 PMCID: PMC10636157 DOI: 10.1038/s41598-023-46938-8] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/27/2023] [Accepted: 11/07/2023] [Indexed: 11/12/2023] Open
Abstract
Wastewater malodour is the proverbial 'elephant in the room' notwithstanding its severe implications on sanitation, health, and hygiene. The predominant malodorous compounds associated with wastewater treatment plants and toilets are volatile organic compounds, such as hydrogen sulphide, ammonia, methanethiol, and organic acids. Among them, methanethiol warrants more attention owing to its relatively low olfactory threshold and associated cytotoxicity. This requires an efficient odour-abatement method since conventional techniques are either cost-prohibitive or leave recalcitrant byproducts. Bacteriophage-based methodology holds promise, and the described work explores the potential. In this study, a non-lysogenous Pseudomonas putida strain is used as a model organism that produces methanethiol in the presence of methionine. Two double-stranded DNA phages of genome sizes > 10 Kb were isolated from sewage. ɸPh_PP01 and ɸPh_PP02 were stable at suboptimal pH, temperature, and at 10% chloroform. Moreover, they showed adsorption efficiencies of 53% and 89% in 12 min and burst sizes of 507 ± 187 and 105 ± 7 virions per cell, respectively. In augmented synthetic wastewater, ɸPh_PP01 and ɸPh_PP02 reduced methanethiol production by 52% and 47%, respectively, with the concomitant reduction in P. putida by 3 logs in 6 h. On extension of the study in P. putida spiked-sewage sample, maximum reduction in methanethiol production was achieved in 3 h, with 49% and 48% for ɸPh_PP01 and ɸPh_PP02, respectively. But at 6 h, efficiency reduced to 36% with both the phages. The study clearly demonstrates the potential of phages as biocontrol agents in the reduction of malodour in wastewater.
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Affiliation(s)
- Niti Sarat
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India
| | - Amrita Salim
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India
| | - Sanjay Pal
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India.
| | - Suja Subhash
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India
| | - Megha Prasad
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India
| | - Bipin G Nair
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India
| | - Ajith Madhavan
- School of Biotechnology, Amrita Vishwa Vidyapeetham, Clappana, Kerala, 690525, India.
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Abdelghafar A, El-Ganiny A, Shaker G, Askoura M. A novel lytic phage exhibiting a remarkable in vivo therapeutic potential and higher antibiofilm activity against Pseudomonas aeruginosa. Eur J Clin Microbiol Infect Dis 2023; 42:1207-1234. [PMID: 37608144 PMCID: PMC10511388 DOI: 10.1007/s10096-023-04649-y] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/25/2023] [Accepted: 08/07/2023] [Indexed: 08/24/2023]
Abstract
BACKGROUND Pseudomonas aeruginosa is a nosocomial bacterium responsible for variety of infections. Inappropriate use of antibiotics could lead to emergence of multidrug-resistant (MDR) P. aeruginosa strains. Herein, a virulent phage; vB_PaeM_PS3 was isolated and tested for its application as alternative to antibiotics for controlling P. aeruginosa infections. METHODS Phage morphology was observed using transmission electron microscopy (TEM). The phage host range and efficiency of plating (EOP) in addition to phage stability were analyzed. One-step growth curve was performed to detect phage growth kinetics. The impact of isolated phage on planktonic cells and biofilms was assessed. The phage genome was sequenced. Finally, the therapeutic potential of vB_PaeM_PS3 was determined in vivo. RESULTS Isolated phage has an icosahedral head and a contractile tail and was assigned to the family Myoviridae. The phage vB_PaeM_PS3 displayed a broad host range, strong bacteriolytic ability, and higher environmental stability. Isolated phage showed a short latent period and large burst size. Importantly, the phage vB_PaeM_PS3 effectively eradicated bacterial biofilms. The genome of vB_PaeM_PS3 consists of 93,922 bp of dsDNA with 49.39% G + C content. It contains 171 predicted open reading frames (ORFs) and 14 genes as tRNA. Interestingly, the phage vB_PaeM_PS3 significantly attenuated P. aeruginosa virulence in host where the survival of bacteria-infected mice was markedly enhanced following phage treatment. Moreover, the colonizing capability of P. aeruginosa was markedly impaired in phage-treated mice as compared to untreated infected mice. CONCLUSION Based on these findings, isolated phage vB_PaeM_PS3 could be potentially considered for treating of P. aeruginosa infections.
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Affiliation(s)
- Aliaa Abdelghafar
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Amira El-Ganiny
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Ghada Shaker
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Momen Askoura
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt.
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Cobián Güemes AG, Le T, Rojas MI, Jacobson NE, Villela H, McNair K, Hung SH, Han L, Boling L, Octavio JC, Dominguez L, Cantú VA, Archdeacon S, Vega AA, An MA, Hajama H, Burkeen G, Edwards RA, Conrad DJ, Rohwer F, Segall AM. Compounding Achromobacter Phages for Therapeutic Applications. Viruses 2023; 15:1665. [PMID: 37632008 PMCID: PMC10457797 DOI: 10.3390/v15081665] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/09/2023] [Revised: 07/27/2023] [Accepted: 07/27/2023] [Indexed: 08/27/2023] Open
Abstract
Achromobacter species colonization of Cystic Fibrosis respiratory airways is an increasing concern. Two adult patients with Cystic Fibrosis colonized by Achromobacter xylosoxidans CF418 or Achromobacter ruhlandii CF116 experienced fatal exacerbations. Achromobacter spp. are naturally resistant to several antibiotics. Therefore, phages could be valuable as therapeutics for the control of Achromobacter. In this study, thirteen lytic phages were isolated and characterized at the morphological and genomic levels for potential future use in phage therapy. They are presented here as the Achromobacter Kumeyaay phage collection. Six distinct Achromobacter phage genome clusters were identified based on a comprehensive phylogenetic analysis of the Kumeyaay collection as well as the publicly available Achromobacter phages. The infectivity of all phages in the Kumeyaay collection was tested in 23 Achromobacter clinical isolates; 78% of these isolates were lysed by at least one phage. A cryptic prophage was induced in Achromobacter xylosoxidans CF418 when infected with some of the lytic phages. This prophage genome was characterized and is presented as Achromobacter phage CF418-P1. Prophage induction during lytic phage preparation for therapy interventions require further exploration. Large-scale production of phages and removal of endotoxins using an octanol-based procedure resulted in a phage concentrate of 1 × 109 plaque-forming units per milliliter with an endotoxin concentration of 65 endotoxin units per milliliter, which is below the Food and Drugs Administration recommended maximum threshold for human administration. This study provides a comprehensive framework for the isolation, bioinformatic characterization, and safe production of phages to kill Achromobacter spp. in order to potentially manage Cystic Fibrosis (CF) pulmonary infections.
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Affiliation(s)
- Ana Georgina Cobián Güemes
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Tram Le
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Maria Isabel Rojas
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Nicole E. Jacobson
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Helena Villela
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
- Marine Microbiomes Lab, Red Sea Research Center, King Abdullah University of Science and Technology, Building 2, Level 3, Room 3216 WS03, Thuwal 23955-6900, Saudi Arabia
| | - Katelyn McNair
- Computational Sciences Research Center, San Diego State University, San Diego, CA 92182, USA
| | - Shr-Hau Hung
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Lili Han
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
- Research Centre for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing 100085, China
| | - Lance Boling
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Jessica Claire Octavio
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Lorena Dominguez
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Vito Adrian Cantú
- Computational Sciences Research Center, San Diego State University, San Diego, CA 92182, USA
| | - Sinéad Archdeacon
- College of Biological Sciences, University of California Davis, Davis, CA 95616, USA
| | - Alejandro A. Vega
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
- David Geffen School of Medicine, University of California Los Angeles, Los Angeles, CA 90025, USA
| | - Michelle A. An
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Hamza Hajama
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Gregory Burkeen
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Robert A. Edwards
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
- Computational Sciences Research Center, San Diego State University, San Diego, CA 92182, USA
- Flinders Accelerator for Microbiome Exploration, Flinders University, Sturt Road, Bedford Park 5042, Australia
| | - Douglas J. Conrad
- Department of Medicine, Division of Pulmonary, Critical Care and Sleep Medicine, University of California San Diego, San Diego, CA 9500, USA
| | - Forest Rohwer
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
| | - Anca M. Segall
- Department of Biology, Viral Information Institute, San Diego State University, San Diego, CA 92182, USA
- Computational Sciences Research Center, San Diego State University, San Diego, CA 92182, USA
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9
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Abdelghafar A, El-Ganiny A, Shaker G, Askoura M. Isolation of a bacteriophage targeting Pseudomonas aeruginosa and exhibits a promising in vivo efficacy. AMB Express 2023; 13:79. [PMID: 37495819 PMCID: PMC10371947 DOI: 10.1186/s13568-023-01582-3] [Citation(s) in RCA: 8] [Impact Index Per Article: 4.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2023] [Accepted: 07/07/2023] [Indexed: 07/28/2023] Open
Abstract
Pseudomonas aeruginosa is an important pathogen that causes serious infections. Bacterial biofilms are highly resistant and render bacterial treatment very difficult, therefore necessitates alternative antibacterial strategies. Phage therapy has been recently regarded as a potential therapeutic option for treatment of bacterial infections. In the current study, a novel podovirus vB_PaeP_PS28 has been isolated from sewage with higher lytic activity against P. aeruginosa. Isolated phage exhibits a short latent period, large burst size and higher stability over a wide range of temperatures and pH. The genome of vB_PaeP_PS28 consists of 72,283 bp circular double-stranded DNA, with G + C content of 54.75%. The phage genome contains 94 open reading frames (ORFs); 32 for known functional proteins and 62 for hypothetical proteins and no tRNA genes. The phage vB_PaeP_PS28 effectively inhibited the growth of P. aeruginosa planktonic cells and displayed a higher biofilm degrading capability. Moreover, therapeutic efficacy of isolated phage was evaluated in vivo using mice infection model. Interestingly, survival of mice infected with P. aeruginosa was significantly enhanced upon treatment with vB_PaeP_PS28. Furthermore, the bacterial load in liver and kidney isolated from mice infected with P. aeruginosa and treated with phage markedly decreased as compared with phage-untreated P. aeruginosa-infected mice. These findings support the efficacy of isolated phage vB_PaeP_PS28 in reducing P. aeruginosa colonization and pathogenesis in host. Importantly, the isolated phage vB_PaeP_PS28 could be applied alone or as combination therapy with other lytic phages as phage cocktail therapy or with antibiotics to limit infections caused by P. aeruginosa.
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Affiliation(s)
- Aliaa Abdelghafar
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Amira El-Ganiny
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Ghada Shaker
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt
| | - Momen Askoura
- Department of Microbiology and Immunology, Faculty of Pharmacy, Zagazig University, Zagazig, 44519, Egypt.
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10
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Laanto E, Oksanen HM. Three Phages from a Boreal Lake during Ice Cover Infecting Xylophilus, Caulobacter, and Polaromonas Species. Viruses 2023; 15:v15020307. [PMID: 36851521 PMCID: PMC9959647 DOI: 10.3390/v15020307] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/20/2022] [Revised: 01/16/2023] [Accepted: 01/18/2023] [Indexed: 01/24/2023] Open
Abstract
Although the important role of microbes in freshwater is well understood, studies on phage-host systems in such environments during ice cover are completely lacking. Here, we describe the isolation and characterization of three new bacteriophages infecting Xylophilus sp., Caudobacter sp., and Polaromonas sp. from freshwater samples taken under the ice cover of Lake Konnevesi, Finland. Lumi, Kuura, and Tiera bacteriophages have tailed icosahedral virions and double-stranded DNA. Lumi is a siphophage with a genome of 80,496 bp, and Kuura and Tiera are podophages, and their genomes are 43,205 and 45,327 bp in length, resembling viruses in the class Caudoviricetes. Their host ranges were very limited among the winter-isolated bacterial strains from Konnevesi, each infecting only their own hosts. They can infect efficiently at 4 °C, showing that they are adapted to living in lake water under ice cover. Analysis of the viral genome sequences showed that a significant number of the gene products of each virus are unique, indicating that there is unexplored viral diversity in freshwaters. To our knowledge, Lumi and Tiera are the first phages isolated on the Xylophilus sp. and Polaromonas sp. strains, allowing their exploitation in further studies of freshwater bacterial-phage interactions.
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Affiliation(s)
- Elina Laanto
- Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland
- Department of Biological and Environmental Science, Nanoscience Center, University of Jyväskylä, 40014 Jyväskylä, Finland
| | - Hanna M. Oksanen
- Molecular and Integrative Biosciences Research Programme, Faculty of Biological and Environmental Sciences, University of Helsinki, 00014 Helsinki, Finland
- Correspondence: ; Tel.: +358-2941-59104
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11
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Isolation and Characterization of a Novel Siphoviridae Phage, vB_AbaS_TCUP2199, Infecting Multidrug-Resistant Acinetobacter baumannii. Viruses 2022; 14:v14061240. [PMID: 35746711 PMCID: PMC9228384 DOI: 10.3390/v14061240] [Citation(s) in RCA: 16] [Impact Index Per Article: 5.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/12/2022] [Revised: 06/02/2022] [Accepted: 06/06/2022] [Indexed: 11/29/2022] Open
Abstract
Multidrug-resistant Acinetobacter baumannii (MDRAB) is a pathogen recognized as antimicrobial-resistant bacteria involved in healthcare-associated infections. Resistance to antibiotics has made alternative therapies necessary. Bacteriophage therapy is considered a potential solution to treat MDRAB. In this study, we isolated and characterized the phage vB_AbaS_TCUP2199 (TCUP2199), which can infect MDRAB. Morphological analysis revealed that TCUP2199 belongs to the Siphoviridae family. TCUP2199 has a wide host range, can adsorb rapidly (68.28% in 2 min), and has a burst size of 196 PFU/cell. At least 16 distinct structural proteins were visualized by SDS polyacrylamide gel electrophoresis. A stability test showed that TCUP2199 was stable at 37 °C and pH 7. Genome analysis of TCUP2199 showed that it consists of a double-stranded DNA genome of 79,572 bp with a G+C content of 40.39%, which contains 98 putative open reading frames, none of which is closely related to the bacteriophage genome sequence that was found in the public database. TCUP2199 shows similarity in genomic organization and putative packaging mechanism with Achromobacter phage JWF and Pseudoalteromonas phage KB12-38 based on protein BLAST and phylogenetic analysis. Because of those unique characteristics, we consider TCUP2199 to be a novel phage that is suitable for inclusion in a phage cocktail to treat A. baumannii infection.
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12
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Łubkowska B, Jeżewska-Frąckowiak J, Sobolewski I, Skowron PM. Bacteriophages of Thermophilic ' Bacillus Group' Bacteria-A Review. Microorganisms 2021; 9:1522. [PMID: 34361957 PMCID: PMC8303945 DOI: 10.3390/microorganisms9071522] [Citation(s) in RCA: 20] [Impact Index Per Article: 5.0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/03/2021] [Revised: 07/03/2021] [Accepted: 07/12/2021] [Indexed: 11/17/2022] Open
Abstract
Bacteriophages of thermophiles are of increasing interest owing to their important roles in many biogeochemical, ecological processes and in biotechnology applications, including emerging bionanotechnology. However, due to lack of in-depth investigation, they are underrepresented in the known prokaryotic virosphere. Therefore, there is a considerable potential for the discovery of novel bacteriophage-host systems in various environments: marine and terrestrial hot springs, compost piles, soil, industrial hot waters, among others. This review aims at providing a reference compendium of thermophages characterized thus far, which infect the species of thermophilic 'Bacillus group' bacteria, mostly from Geobacillus sp. We have listed 56 thermophages, out of which the majority belong to the Siphoviridae family, others belong to the Myoviridae and Podoviridae families and, apparently, a few belong to the Sphaerolipoviridae, Tectiviridae or Corticoviridae families. All of their genomes are composed of dsDNA, either linear, circular or circularly permuted. Fourteen genomes have been sequenced; their sizes vary greatly from 35,055 bp to an exceptionally large genome of 160,590 bp. We have also included our unpublished data on TP-84, which infects Geobacillus stearothermophilus (G. stearothermophilus). Since the TP-84 genome sequence shows essentially no similarity to any previously characterized bacteriophage, we have defined TP-84 as a new species in the newly proposed genus Tp84virus within the Siphoviridae family. The information summary presented here may be helpful in comparative deciphering of the molecular basis of the thermophages' biology, biotechnology and in analyzing the environmental aspects of the thermophages' effect on the thermophile community.
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Affiliation(s)
- Beata Łubkowska
- Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308 Gdansk, Poland; (J.J.-F.); (I.S.); (P.M.S.)
- The High School of Health in Gdansk, Pelplinska 7, 80-335 Gdansk, Poland
| | - Joanna Jeżewska-Frąckowiak
- Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308 Gdansk, Poland; (J.J.-F.); (I.S.); (P.M.S.)
| | - Ireneusz Sobolewski
- Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308 Gdansk, Poland; (J.J.-F.); (I.S.); (P.M.S.)
| | - Piotr M. Skowron
- Department of Molecular Biotechnology, Faculty of Chemistry, University of Gdansk, Wita Stwosza 63, 80-308 Gdansk, Poland; (J.J.-F.); (I.S.); (P.M.S.)
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13
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Imani S, Wijetunga A, Shumborski S, O’Leary E. Chronic osteomyelitis caused by Achromobacter xylosoxidans following orthopaedic trauma: A case report and review of the literature. IDCases 2021; 25:e01211. [PMID: 34277350 PMCID: PMC8267561 DOI: 10.1016/j.idcr.2021.e01211] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/19/2021] [Revised: 06/28/2021] [Accepted: 06/28/2021] [Indexed: 11/18/2022] Open
Abstract
Background Achromobacter xylosoxidans is an opportunistic environmental aerobe. In cases where A. xylosoxidans infects humans, it most commonly manifests as bacteraemia in the immunosuppressed. A. xylosoxidans causing chronic osteomyelitis is rare, particularly in the immunocompetent and young. Case We present the case of a 23-year-old man with chronic osteomyelitis of the right femur caused by co-infection of A. xylosoxidans and Staphylococcus aureus. Five years earlier, he had sustained a right femur fracture and was treated with intramedullary fixation at a peripheral hospital in a developing nation. Past medical history was otherwise unremarkable. Management comprised of surgical debridement and culture-directed antibiotic therapy, resulting in clinical cure. Conclusion In the context of local trauma and previous surgery, osteomyelitis caused by atypical pathogens must be considered. A multidisciplinary approach commensurate with duration and severity of infection and tailored to the causative organism is paramount.
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Affiliation(s)
- Sahand Imani
- Department of Orthopaedic Surgery, Hornsby Ku-Ring-Gai Hospital, Sydney, New South Wales, Australia
| | - Asanka Wijetunga
- School of Medicine, The University of Sydney, Sydney, New South Wales, Australia
- Corresponding author.
| | - Sarah Shumborski
- Department of Orthopaedic Surgery, Hornsby Ku-Ring-Gai Hospital, Sydney, New South Wales, Australia
| | - Edmund O’Leary
- Department of Orthopaedic Surgery, Hornsby Ku-Ring-Gai Hospital, Sydney, New South Wales, Australia
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Analysis of a Novel Bacteriophage vB_AchrS_AchV4 Highlights the Diversity of Achromobacter Viruses. Viruses 2021; 13:v13030374. [PMID: 33673419 PMCID: PMC7996906 DOI: 10.3390/v13030374] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/23/2021] [Revised: 02/18/2021] [Accepted: 02/24/2021] [Indexed: 12/12/2022] Open
Abstract
Achromobacter spp. are ubiquitous in nature and are increasingly being recognized as emerging nosocomial pathogens. Nevertheless, to date, only 30 complete genome sequences of Achromobacter phages are available in GenBank, and nearly all of those phages were isolated on Achromobacter xylosoxidans. Here, we report the isolation and characterization of bacteriophage vB_AchrS_AchV4. To the best of our knowledge, vB_AchrS_AchV4 is the first virus isolated from Achromobacter spanius. Both vB_AchrS_AchV4 and its host, Achromobacter spanius RL_4, were isolated in Lithuania. VB_AchrS_AchV4 is a siphovirus, since it has an isometric head (64 ± 3.2 nm in diameter) and a non-contractile flexible tail (232 ± 5.4). The genome of vB_AchrS_AchV4 is a linear dsDNA molecule of 59,489 bp with a G+C content of 62.8%. It contains no tRNA genes, yet it includes 82 protein-coding genes, of which 27 have no homologues in phages. Using bioinformatics approaches, 36 vB_AchrS_AchV4 genes were given a putative function. A further four were annotated based on the results of LC-MS/MS. Comparative analyses revealed that vB_AchrS_AchV4 is a singleton siphovirus with no close relatives among known tailed phages. In summary, this work not only describes a novel and unique phage, but also advances our knowledge of genetic diversity and evolution of Achromobacter bacteriophages.
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15
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Korf IHE, Kittler S, Bierbrodt A, Mengden R, Rohde C, Rohde M, Kroj A, Lehnherr T, Fruth A, Flieger A, Lehnherr H, Wittmann J. In Vitro Evaluation of a Phage Cocktail Controlling Infections with Escherichia coli. Viruses 2020; 12:v12121470. [PMID: 33352791 PMCID: PMC7768485 DOI: 10.3390/v12121470] [Citation(s) in RCA: 22] [Impact Index Per Article: 4.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/06/2020] [Revised: 12/11/2020] [Accepted: 12/16/2020] [Indexed: 12/13/2022] Open
Abstract
Worldwide, poultry industry suffers from infections caused by avian pathogenic Escherichia coli. Therapeutic failure due to resistant bacteria is of increasing concern and poses a threat to human and animal health. This causes a high demand to find alternatives to fight bacterial infections in animal farming. Bacteriophages are being especially considered for the control of multi-drug resistant bacteria due to their high specificity and lack of serious side effects. Therefore, the study aimed on characterizing phages and composing a phage cocktail suitable for the prevention of infections with E. coli. Six phages were isolated or selected from our collections and characterized individually and in combination with regard to host range, stability, reproduction, and efficacy in vitro. The cocktail consisting of six phages was able to inhibit formation of biofilms by some E. coli strains but not by all. Phage-resistant variants arose when bacterial cells were challenged with a single phage but not when challenged by a combination of four or six phages. Resistant variants arising showed changes in carbon metabolism and/or motility. Genomic comparison of wild type and phage-resistant mutant E28.G28R3 revealed a deletion of several genes putatively involved in phage adsorption and infection.
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Affiliation(s)
- Imke H. E. Korf
- Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures, Inhoffenstraße 7B, 38124 Braunschweig, Germany; (C.R.); (J.W.)
- Correspondence:
| | - Sophie Kittler
- Institute for Food Quality and Food Safety, University of Veterinary Medicine Hannover, Foundation, Bischofsholer Damm 15, 30173 Hannover, Germany;
| | | | - Ruth Mengden
- Food Inspection, Animal Welfare and Veterinary Service of the Land of Bremen, Border Control Post Bremerhaven, Senator-Borttscheller-Straße 8, 27568 Bremerhaven, Germany;
| | - Christine Rohde
- Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures, Inhoffenstraße 7B, 38124 Braunschweig, Germany; (C.R.); (J.W.)
| | - Manfred Rohde
- Central Facility for Microscopy, Helmholtz-Centre for Infection Research (HZI), Inhoffenstraße 7, 38124 Braunschweig, Germany;
| | - Andrea Kroj
- PTC Phage Technology Center GmbH, Siemensstraße 42, 59199 Bönen, Germany; (A.K.); (T.L.); (H.L.)
| | - Tatiana Lehnherr
- PTC Phage Technology Center GmbH, Siemensstraße 42, 59199 Bönen, Germany; (A.K.); (T.L.); (H.L.)
| | - Angelika Fruth
- Robert Koch Institute, Burgstraße 37, 38855 Wernigerode, Germany; (A.F.); (A.F.)
| | - Antje Flieger
- Robert Koch Institute, Burgstraße 37, 38855 Wernigerode, Germany; (A.F.); (A.F.)
| | - Hansjörg Lehnherr
- PTC Phage Technology Center GmbH, Siemensstraße 42, 59199 Bönen, Germany; (A.K.); (T.L.); (H.L.)
| | - Johannes Wittmann
- Leibniz Institute DSMZ—German Collection of Microorganisms and Cell Cultures, Inhoffenstraße 7B, 38124 Braunschweig, Germany; (C.R.); (J.W.)
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16
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Characterization of Novel Lytic Bacteriophages of Achromobacter marplantensis Isolated from a Pneumonia Patient. Viruses 2020; 12:v12101138. [PMID: 33049935 PMCID: PMC7600146 DOI: 10.3390/v12101138] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/20/2020] [Revised: 10/02/2020] [Accepted: 10/06/2020] [Indexed: 01/21/2023] Open
Abstract
Achromobacter spp. are becoming increasingly associated with lung infections in patients suffering from cystic fibrosis (CF). A. marplatensis, which is closely related to A. xylosoxidans, has been isolated from the lungs of CF patients and other human infections. This article describes the isolation, morphology and characterization of two lytic bacteriophages specific for an A. marplatensis strain isolated from a pneumonia patient. This host strain was the causal agent of hospital acquired pneumonia–the first clinical report of such an occurrence. Full genome sequencing revealed bacteriophage genomes ranging in size from 45901 to 46,328 bp. Transmission electron microscopy revealed that the two bacteriophages AMA1 and AMA2 belonged to the Siphoviridae family. Host range analysis showed that their host range did not extend to A. xylosoxidans. The possibility exists for future testing of such bacteriophages in the control of Achromobacter infections such as those seen in CF and other infections of the lungs. The incidence of antibiotic resistance in this genus highlights the importance of seeking adjuncts and alternatives in CF and other lung infections.
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17
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Essoh C, Vernadet JP, Vergnaud G, Coulibaly A, Kakou-N'Douba A, N'Guetta ASP, Ouassa T, Pourcel C. Characterization of sixteen Achromobacter xylosoxidans phages from Abidjan, Côte d'Ivoire, isolated on a single clinical strain. Arch Virol 2020; 165:725-730. [PMID: 31897726 DOI: 10.1007/s00705-019-04511-7] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/09/2019] [Accepted: 11/28/2019] [Indexed: 01/21/2023]
Abstract
Sixteen bacteriophages of Achromobacter xylosoxidans distributed into four genera have been isolated from sewage water in Abidjan, Côte d'Ivoire, using a single clinical strain, and their genomes have been sequenced. Three podoviruses belonged to the genus Phikmvvirus, and these represent the first A. xylosoxidans phages of this genus. Seven podoviruses, distributed into three groups, belonged to the genus Jwalphavirus. Among the siphoviruses, three revealed similarities to Pseudomonas phage 73 and members of the genus Septimatrevirus, and three were YuA-like phages. The virulence of these phages toward a panel of 10 genetically diverse strains was tested, with the phiKMV-like phages showing the broadest host range.
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Affiliation(s)
- Christiane Essoh
- Département de Biochimie-Génétique, UFR des Sciences Biologiques, Université Peleforo Gon- Coulibaly, Korhogo, Côte d'Ivoire
| | - Jean-Philippe Vernadet
- Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, 91198, Gif-sur-Yvette cedex, France
| | - Gilles Vergnaud
- Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, 91198, Gif-sur-Yvette cedex, France
| | - Adama Coulibaly
- Département de Biochimie-Génétique, UFR des Sciences Biologiques, Université Peleforo Gon- Coulibaly, Korhogo, Côte d'Ivoire
| | - Adèle Kakou-N'Douba
- Laboratoire de Bactériologie-Virologie, Département de Microbiologie, UFR des Sciences Médicales, Université Félix Houphouët-Boigny, Abidjan, Côte d'Ivoire
| | - Assavo S-P N'Guetta
- Laboratoire de Génétique, UFR Biosciences, Université Félix Houphouët-Boigny, Abidjan, Côte d'Ivoire
| | - Thimotée Ouassa
- Laboratoire de Microbiologie, UFR des Sciences Pharmaceutiques et Biologiques, Université Félix Houphouët-Boigny, Abidjan, Côte d'Ivoire
| | - Christine Pourcel
- Institute for Integrative Biology of the Cell (I2BC), Université Paris-Saclay, 91198, Gif-sur-Yvette cedex, France. .,Université Paris-Sud, I2BC, Bât 400, 91405, Orsay cedex, France.
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18
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Kohn T, Wiegand S, Boedeker C, Rast P, Heuer A, Jetten MSM, Schüler M, Becker S, Rohde C, Müller RW, Brümmer F, Rohde M, Engelhardt H, Jogler M, Jogler C. Planctopirus ephydatiae, a novel Planctomycete isolated from a freshwater sponge. Syst Appl Microbiol 2019; 43:126022. [PMID: 31785948 DOI: 10.1016/j.syapm.2019.126022] [Citation(s) in RCA: 47] [Impact Index Per Article: 7.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/29/2019] [Revised: 09/30/2019] [Accepted: 10/02/2019] [Indexed: 01/28/2023]
Abstract
The microbiome of freshwater sponges is rarely studied, and not a single novel bacterial species has been isolated and subsequently characterized from a freshwater sponge to date. A previous study showed that 14.4% of the microbiome from Ephydatia fluviatilis belong to the phylum Planctomycetes. Therefore, we sampled an Ephydatia sponge from a freshwater lake and employed enrichment techniques targeting bacteria from the phylum Planctomycetes. The obtained strain spb1T was subject to genomic and phenomic characterization and found to represent a novel planctomycetal species proposed as Planctopirus ephydatiae sp. nov. (DSM 106606 = CECT 9866). In the process of differentiating spb1T from its next relative Planctopirus limnophila DSM 3776T, we identified and characterized the first phage - Planctopirus phage vB_PlimS_J1 - infecting planctomycetes that was only mentioned anecdotally before. Interestingly, classical chemotaxonomic methods would have failed to distinguish Planctopirus ephydatiae strain spb1T from Planctopirus limnophila DSM 3776T. Our findings demonstrate and underpin the need for whole genome-based taxonomy to detect and differentiate planctomycetal species.
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Affiliation(s)
- T Kohn
- Department of Microbiology, Radboud University, Nijmegen, Netherlands
| | - S Wiegand
- Department of Microbiology, Radboud University, Nijmegen, Netherlands
| | - C Boedeker
- Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany
| | - P Rast
- Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany
| | - A Heuer
- Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany
| | - M S M Jetten
- Department of Microbiology, Radboud University, Nijmegen, Netherlands
| | - M Schüler
- Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
| | - S Becker
- University of Veterinary Medicine Hannover, Germany
| | - C Rohde
- Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany
| | - R-W Müller
- Institute of Biomaterials and Biomolecular Systems, University of Stuttgart, Germany
| | - F Brümmer
- Institute of Biomaterials and Biomolecular Systems, University of Stuttgart, Germany
| | - M Rohde
- Central Facility for Microscopy, Helmholtz-Centre for Infection Research (HZI), Braunschweig, Germany
| | - H Engelhardt
- Department of Molecular Structural Biology, Max Planck Institute of Biochemistry, Martinsried, Germany
| | - M Jogler
- Leibniz-Institut Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany
| | - C Jogler
- Department of Microbiology, Radboud University, Nijmegen, Netherlands; Department of Microbial Interactions, Friedrich Schiller Universität Jena, Germany.
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19
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de Melo ACC, da Mata Gomes A, Melo FL, Ardisson-Araújo DMP, de Vargas APC, Ely VL, Kitajima EW, Ribeiro BM, Wolff JLC. Characterization of a bacteriophage with broad host range against strains of Pseudomonas aeruginosa isolated from domestic animals. BMC Microbiol 2019; 19:134. [PMID: 31208333 PMCID: PMC6580649 DOI: 10.1186/s12866-019-1481-z] [Citation(s) in RCA: 28] [Impact Index Per Article: 4.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/23/2018] [Accepted: 05/07/2019] [Indexed: 12/16/2022] Open
Abstract
Background Pseudomonas aeruginosa is an opportunistic pathogen and one of the leading causes of nosocomial infections. Moreover, the species can cause severe infections in cystic fibrosis patients, in burnt victims and cause disease in domestic animals. The control of these infections is often difficult due to its vast repertoire of mechanisms for antibiotic resistance. Phage therapy investigation with P. aeruginosa bacteriophages has aimed mainly the control of human diseases. In the present work, we have isolated and characterized a new bacteriophage, named Pseudomonas phage BrSP1, and investigated its host range against 36 P. aeruginosa strains isolated from diseased animals and against P. aeruginosa ATCC strain 27853. Results We have isolated a Pseudomonas aeruginosa phage from sewage. We named this virus Pseudomonas phage BrSP1. Our electron microscopy analysis showed that phage BrSP1 had a long tail structure found in members of the order Caudovirales. “In vitro” biological assays demonstrated that phage BrSP1 was capable of maintaining the P. aeruginosa population at low levels for up to 12 h post-infection. However, bacterial growth resumed afterward and reached levels similar to non-treated samples at 24 h post-infection. Host range analysis showed that 51.4% of the bacterial strains investigated were susceptible to phage BrSP1 and efficiency of plating (EOP) investigation indicated that EOP values in the strains tested varied from 0.02 to 1.72. Analysis of the phage genome revealed that it was a double-stranded DNA virus with 66,189 bp, highly similar to the genomes of members of the genus Pbunavirus, a group of viruses also known as PB1-like viruses. Conclusion The results of our “in vitro” bioassays and of our host range analysis suggested that Pseudomonas phage BrSP1 could be included in a phage cocktail to treat veterinary infections. Our EOP investigation confirmed that EOP values differ considerably among different bacterial strains. Comparisons of complete genome sequences indicated that phage BrSP1 is a novel species of the genus Pbunavirus. The complete genome of phage BrSP1 provides additional data that may help the broader understanding of pbunaviruses genome evolution. Electronic supplementary material The online version of this article (10.1186/s12866-019-1481-z) contains supplementary material, which is available to authorized users.
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Affiliation(s)
- Anna Cristhina Carmine de Melo
- CCBS - Curso de Ciências Biológicas, Laboratório de Biologia Molecular e Virologia, Prédio 28, primeiro andar, Universidade Presbiteriana Mackenzie, Rua da Consolação, 896, Consolação, São Paulo, SP, CEP 01302-907, Brazil
| | - Amanda da Mata Gomes
- CCBS - Curso de Ciências Biológicas, Laboratório de Biologia Molecular e Virologia, Prédio 28, primeiro andar, Universidade Presbiteriana Mackenzie, Rua da Consolação, 896, Consolação, São Paulo, SP, CEP 01302-907, Brazil
| | - Fernando L Melo
- Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil
| | - Daniel M P Ardisson-Araújo
- Departamento de Bioquímica e Biologia Molecular, Universidade Federal de Santa Maria, Av. Roraima, 1000, Cidade Universitária, Santa Maria, RS, CEP 97105-900, Brazil
| | - Agueda Palmira Castagna de Vargas
- Departamento de Medicina Veterinária Preventiva (DMVP), Centro de Ciências Rurais (CCR)Avenida Roraima, Universidade Federal de Santa Maria, 1000. Prédio 44, Sala 5137, Santa Maria, RS, CEP 97105-900, Brazil
| | - Valessa Lunkes Ely
- Departamento de Medicina Veterinária Preventiva (DMVP), Centro de Ciências Rurais (CCR)Avenida Roraima, Universidade Federal de Santa Maria, 1000. Prédio 44, Sala 5137, Santa Maria, RS, CEP 97105-900, Brazil
| | - Elliot W Kitajima
- NAP/MEPA, Departamento de Fitopatologia e Nematologia, ESALQ, Universidade de São Paulo, Piracicaba, SP, Brazil
| | - Bergmann M Ribeiro
- Departamento de Biologia Celular, Instituto de Ciências Biológicas, Universidade de Brasília, Brasília, DF, Brazil
| | - José Luiz Caldas Wolff
- CCBS - Curso de Ciências Biológicas, Laboratório de Biologia Molecular e Virologia, Prédio 28, primeiro andar, Universidade Presbiteriana Mackenzie, Rua da Consolação, 896, Consolação, São Paulo, SP, CEP 01302-907, Brazil.
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20
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Akhwale JK, Rohde M, Rohde C, Bunk B, Spröer C, Klenk HP, Boga HI, Wittmann J. Comparative genomic analysis of eight novel haloalkaliphilic bacteriophages from Lake Elmenteita, Kenya. PLoS One 2019; 14:e0212102. [PMID: 30763364 PMCID: PMC6375668 DOI: 10.1371/journal.pone.0212102] [Citation(s) in RCA: 8] [Impact Index Per Article: 1.3] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/24/2018] [Accepted: 01/28/2019] [Indexed: 12/17/2022] Open
Abstract
We report complete genome sequences of eight bacteriophages isolated from Haloalkaline Lake Elmenteita found on the floor of Kenyan Rift Valley. The bacteriophages were sequenced, annotated and a comparative genomic analysis using various Bioinformatics tools carried out to determine relatedness of the bacteriophages to each other, and to those in public databases. Basic genome properties like genome size, percentage coding density, number of open reading frames, percentage GC content and gene organizations revealed the bacteriophages had no relationship to each other. Comparison to other nucleotide sequences in GenBank database showed no significant similarities hence novel. At the amino acid level, phages of our study revealed mosaicism to genes with conserved domains to already described phages. Phylogenetic analyses of large terminase gene responsible for DNA packaging and DNA polymerase gene for replication further showed diversity among the bacteriophages. Our results give insight into diversity of bacteriophages in Lake Elmenteita and provide information on their evolution. By providing primary sequence information, this study not only provides novel sequences for biotechnological exploitation, but also sets stage for future studies aimed at better understanding of virus diversity and genomes from haloalkaline lakes in the Rift Valley.
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Affiliation(s)
- Juliah Khayeli Akhwale
- Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, Braunschweig, Germany
- Department of Zoology, Jomo Kenyatta University of Agriculture and Technology, Nairobi, Kenya
| | - Manfred Rohde
- Helmholtz Centre for Infection Research, Central Facility for Microscopy, Inhoffenstrasse 7B, Braunschweig, Germany
| | - Christine Rohde
- Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, Braunschweig, Germany
| | - Boyke Bunk
- Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, Braunschweig, Germany
| | - Cathrin Spröer
- Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, Braunschweig, Germany
| | - Hans-Peter Klenk
- School of Natural and Environmental Sciences, Newcastle University, Newcastle upon Tyne, United Kingdom
| | | | - Johannes Wittmann
- Leibniz Institute DSMZ–German Collection of Microorganisms and Cell Cultures, Inhoffenstrasse 7B, Braunschweig, Germany
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21
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Phage therapy against Achromobacter xylosoxidans lung infection in a patient with cystic fibrosis: a case report. Res Microbiol 2018; 169:540-542. [DOI: 10.1016/j.resmic.2018.05.001] [Citation(s) in RCA: 64] [Impact Index Per Article: 9.1] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/31/2018] [Revised: 04/26/2018] [Accepted: 05/04/2018] [Indexed: 12/24/2022]
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22
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Aoyama S, Masaki A, Sakamoto Y, Takino H, Murase T, Ohshima K, Yoshino T, Kato S, Inagaki H. Achromobacter Infection Is Rare in Japanese Patients with Pulmonary B-cell Lymphoma. Intern Med 2018; 57:789-794. [PMID: 29151525 PMCID: PMC5891515 DOI: 10.2169/internalmedicine.9430-17] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Indexed: 12/17/2022] Open
Abstract
Objective Achromobacter xylosoxidans (A. xylosoxidans) has been recently reported to have an association with the development of pulmonary mucosa-associated lymphoid tissue (MALT) lymphoma in patients from European countries. However, the prevalence rates for A. xylosoxidans may vary significantly from country to country. To assess this association, the prevalence of A. xylosoxidans was analyzed in Japanese patients with pulmonary B-cell lymphoma. Methods DNA samples were obtained from formalin-fixed, paraffin-embedded sections of pulmonary MALT lymphomas (n=52), diffuse large B-cell lymphomas (DLBCLs, n=18), and benign pulmonary lesions (n=19). All samples were histopathologically reviewed by experienced hematopathologists, and the clonality of all MALT lymphoma cases was confirmed by a polymerase chain reaction (PCR)-based IGH rearrangement clonality assay. They were also tested for the API2-MALT1 fusion transcript. The presence of bacterial DNA was assessed with a nested PCR, and DNA sequencing was performed to confirm the PCR specificity. Results A. xylosoxidans DNA was detected in 1/52 cases of pulmonary MALT lymphoma, 2/18 cases of DLBCL, and 0/19 cases of benign pulmonary lesions. The prevalence of A. xylosoxidans in pulmonary lymphoma was not significantly higher than in benign lesions. Conclusion The present study shows that A. xylosoxidans infection may not be associated with pulmonary B-cell lymphoma in a Japanese case series. Large-scale international studies are needed to clarify the role of A. xylosoxidans in pulmonary lymphoma.
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Affiliation(s)
- Satsuki Aoyama
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
| | - Ayako Masaki
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
| | - Yuma Sakamoto
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
| | - Hisashi Takino
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
| | - Takayuki Murase
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
| | - Koichi Ohshima
- Department of Pathology, Kurume University School of Medicine, Japan
| | - Tadashi Yoshino
- Department of Pathology, Okayama University Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Japan
| | - Seiichi Kato
- Department of Pathology and Laboratory Medicine, Nagoya University Hospital, Japan
| | - Hiroshi Inagaki
- Department of Pathology and Molecular Diagnostics, Nagoya City University Graduate School of Medical Sciences, Japan
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23
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Dreiseikelmann B, Bunk B, Spröer C, Rohde M, Nimtz M, Wittmann J. Characterization and genome comparisons of three Achromobacter phages of the family Siphoviridae. Arch Virol 2017; 162:2191-2201. [PMID: 28357512 DOI: 10.1007/s00705-017-3347-8] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/22/2016] [Accepted: 03/17/2017] [Indexed: 11/30/2022]
Abstract
In this study, we present the characterization and genomic data of three Achromobacter phages belonging to the family Siphoviridae. Phages 83-24, JWX and JWF were isolated from sewage samples in Paris and Braunschweig, respectively, and infect Achromobacter xylosoxidans, an emerging nosocomial pathogen in cystic fibrosis patients. Analysis of morphology and growth parameters revealed that phages 83-24 and JWX have similar properties, both have nearly the same head and tail measurements, and both have a burst size between 85 and 100 pfu/cell. In regard to morphological properties, JWF had a much longer and more flexible tail compared to other phages. The linear double-stranded DNAs of all three phages are terminally redundant and not circularly permutated. The complete nucleotide sequences consist of 81,541 bp for JWF, 49,714 bp for JWX and 48,216 bp for 83-24. Analysis of the genome sequences showed again that phages JWX and 83-24 are quite similar. Comparison to the GenBank database via BLASTN revealed partial similarities to Roseobacter phage RDJL phi1 and Burkholderia phage BcepGomr. In contrast, BLASTN analysis of the genome sequence of phage JWF revealed only few similarities to non-annotated prophage regions in different strains of Burkholderia and Mesorhizobium.
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Affiliation(s)
- Brigitte Dreiseikelmann
- Department of Microbiology/Genetechnology, University of Bielefeld, 33615, Bielefeld, Germany
| | - Boyke Bunk
- Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, 38124, Brunswick, Germany
| | - Cathrin Spröer
- Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, 38124, Brunswick, Germany
| | - Manfred Rohde
- Central Facility for Microscopy, Helmholtz Centre for Infection Research, 38124, Brunswick, Germany
| | - Manfred Nimtz
- Protein Analytics Platform, Helmholtz Centre for Infection Research, 38124, Brunswick, Germany
| | - Johannes Wittmann
- Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures, 38124, Brunswick, Germany.
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24
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Kohn T, Heuer A, Jogler M, Vollmers J, Boedeker C, Bunk B, Rast P, Borchert D, Glöckner I, Freese HM, Klenk HP, Overmann J, Kaster AK, Rohde M, Wiegand S, Jogler C. Fuerstia marisgermanicae gen. nov., sp. nov., an Unusual Member of the Phylum Planctomycetes from the German Wadden Sea. Front Microbiol 2016; 7:2079. [PMID: 28066393 PMCID: PMC5177795 DOI: 10.3389/fmicb.2016.02079] [Citation(s) in RCA: 46] [Impact Index Per Article: 5.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/30/2016] [Accepted: 12/08/2016] [Indexed: 11/23/2022] Open
Abstract
Members of the phylum Planctomycetes are ubiquitous bacteria that dwell in aquatic and terrestrial habitats. While planctomycetal species are important players in the global carbon and nitrogen cycle, this phylum is still undersampled and only few genome sequences are available. Here we describe strain NH11T, a novel planctomycete obtained from a crustacean shell (Wadden Sea, Germany). The phylogenetically closest related cultivated species is Gimesia maris, sharing only 87% 16S rRNA sequence identity. Previous isolation attempts have mostly yielded members of the genus Rhodopirellula from water of the German North Sea. On the other hand, only one axenic culture of the genus Pirellula was obtained from a crustacean thus far. However, the 16S rRNA gene sequence of strain NH11T shares only 80% sequence identity with the closest relative of both genera, Rhodopirellula and Pirellula. Thus, strain NH11T is unique in terms of origin and phylogeny. While the pear to ovoid shaped cells of strain NH11T are typical planctomycetal, light-, and electron microscopic observations point toward an unusual variation of cell division through budding: during the division process daughter- and mother cells are connected by an unseen thin tubular-like structure. Furthermore, the periplasmic space of strain NH11T was unusually enlarged and differed from previously known planctomycetes. The complete genome of strain NH11T, with almost 9 Mb in size, is among the largest planctomycetal genomes sequenced thus far, but harbors only 6645 protein-coding genes. The acquisition of genomic components by horizontal gene transfer is indicated by the presence of numerous putative genomic islands. Strikingly, 45 “giant genes” were found within the genome of NH11T. Subsequent analysis of all available planctomycetal genomes revealed that Planctomycetes as such are especially rich in “giant genes”. Furthermore, Multilocus Sequence Analysis (MLSA) tree reconstruction support the phylogenetic distance of strain NH11T from other cultivated Planctomycetes of the same phylogenetic cluster. Thus, based on our findings, we propose to classify strain NH11T as Fuerstia marisgermanicae gen. nov., sp. nov., with the type strain NH11T, within the phylum Planctomycetes.
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Affiliation(s)
- Timo Kohn
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Anja Heuer
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Mareike Jogler
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - John Vollmers
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Christian Boedeker
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Boyke Bunk
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Patrick Rast
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Daniela Borchert
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Ines Glöckner
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Heike M Freese
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | | | - Jörg Overmann
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Anne-Kristin Kaster
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Manfred Rohde
- Helmholtz Centre for Infectious Disease Braunschweig, Germany
| | - Sandra Wiegand
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
| | - Christian Jogler
- Leibniz Institut Deutsche Sammlung Von Mikroorganismen und Zellkulturen Braunschweig, Germany
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Ramsay KA, Stockwell RE, Bell SC, Kidd TJ. Infection in cystic fibrosis: impact of the environment and climate. Expert Rev Respir Med 2016; 10:505-19. [PMID: 26949990 DOI: 10.1586/17476348.2016.1162715] [Citation(s) in RCA: 20] [Impact Index Per Article: 2.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/08/2022]
Abstract
In many countries numbers of adults with cystic fibrosis (CF) exceed that of children, with median survival predicted to surpass 50 years. Increasing longevity is, in part, due to intensive therapies including eradication of early infection and suppressive therapies and pulmonary exacerbations. Initial infections with common CF pathogens are thought to arise from the natural environment. We review the impact of climate and environment on infection in CF. Specifically, several studies indicate that higher ambient temperatures, proximity to the equator and the summer season may be linked to the increased prevalence of Pseudomonas aeruginosa in people with CF. The environment may also play an important role in the acquisition of Gram negative organisms other than P. aeruginosa. There is emerging data suggesting that climatic and environmental factors are likely to impact on the risk of infection with NTM and fungi in people which are found extensively throughout the natural environment.
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Affiliation(s)
- K A Ramsay
- a Lung Bacteria Group , QIMR Berghofer Medical Research Institute , Brisbane , Australia.,b Child Health Research Centre, The University of Queensland , Brisbane , Australia.,c School of Medicine , The University of Queensland , Brisbane , Australia
| | - R E Stockwell
- a Lung Bacteria Group , QIMR Berghofer Medical Research Institute , Brisbane , Australia
| | - S C Bell
- a Lung Bacteria Group , QIMR Berghofer Medical Research Institute , Brisbane , Australia.,c School of Medicine , The University of Queensland , Brisbane , Australia.,d Adult Cystic Fibrosis Centre , The Prince Charles Hospital , Brisbane , Australia
| | - T J Kidd
- b Child Health Research Centre, The University of Queensland , Brisbane , Australia.,e Centre for Infection and Immunity , Queen's University Belfast , Belfast , UK.,f School of Chemistry and Molecular Biosciences , The University of Queensland , Brisbane , Australia
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Li E, Zhao J, Ma Y, Wei X, Li H, Lin W, Wang X, Li C, Shen Z, Zhao R, Jiang A, Yang H, Yuan J, Zhao X. Characterization of a novel Achromobacter xylosoxidans specific siphoviruse: phiAxp-1. Sci Rep 2016; 6:21943. [PMID: 26908262 PMCID: PMC4764938 DOI: 10.1038/srep21943] [Citation(s) in RCA: 15] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/27/2015] [Accepted: 02/03/2016] [Indexed: 02/04/2023] Open
Abstract
Bacteriophages have recently been considered as an alternative biocontrol tool because of the widespread occurrence of antimicrobial-resistant Achromobacter xylosoxidans. Herein, we isolated a virulent bacteriophage (phiAxp-1) from a water sample of the Bohai sea of China that specifically infects A. xylosoxidans. Transmission electron microscopy revealed that phage phiAxp-1 belongs to the Siphoviridae. We sequenced the genome of phiAxp-1, which comprises 45,045 bp with 64 open reading frames. Most of the proteins encoded by phiAxp-1 have no similarity to sequences in the public databases. Twenty-one proteins with assigned functions share weak homology with those of other dsDNA bacteriophages infecting diverse hosts, such as Burkholderia phage KL1, Pseudomonas phage 73, Pseudomonas phage vB_Pae-Kakheti25, Pseudomonas phage vB_PaeS_SCH_Ab26, Acinetobacter phage IME_AB3 and Achromobacter phage JWX. The genome can be divided into different clusters for the head and tail structure, DNA replication and mazG. The sequence and genomic organization of bacteriophage phiAxp-1 are clearly distinct from other known Siphoviridae phages; therefore, we propose that it is a member of a novel genus of the Siphoviridae family. Furthermore, one-step growth curve and stability studies of the phage were performed, and the specific receptor of phiAxp-1 was identified as the lipopolysaccharide of A. xylosoxidans.
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Affiliation(s)
- Erna Li
- College of Food Science, South China Agricultural University, Guangzhou, China, 510642
| | - Jiangtao Zhao
- Emergency Department, the Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou, China, 450052
| | - Yanyan Ma
- College of Food Science, Henan Institute of Science and Technology, Xinxiang, China, 453003
| | - Xiao Wei
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
| | - Huan Li
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
| | - Weishi Lin
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
| | - Xuesong Wang
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
| | - Chao Li
- Key Laboratory of Risk Assessment and Control for Environment and Food Safety, Tianjin Institute of Health and Environmental Medicine, Tianjin, China, 300050
| | - Zhiqiang Shen
- Key Laboratory of Risk Assessment and Control for Environment and Food Safety, Tianjin Institute of Health and Environmental Medicine, Tianjin, China, 300050
| | - Ruixiang Zhao
- College of Food Science, Henan Institute of Science and Technology, Xinxiang, China, 453003
| | - Aimin Jiang
- College of Food Science, South China Agricultural University, Guangzhou, China, 510642
| | - Huiying Yang
- State Key Laboratory of Pathogen and Biosecurity, Beijing Institute of Microbiology and Epidemiology, Beijing, China, 100071
| | - Jing Yuan
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
| | - Xiangna Zhao
- Institute of Disease Control and Prevention, Academy of Military Medical Sciences, Beijing, China, 100071
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Essoh C, Latino L, Midoux C, Blouin Y, Loukou G, Nguetta SPA, Lathro S, Cablanmian A, Kouassi AK, Vergnaud G, Pourcel C. Investigation of a Large Collection of Pseudomonas aeruginosa Bacteriophages Collected from a Single Environmental Source in Abidjan, Côte d'Ivoire. PLoS One 2015; 10:e0130548. [PMID: 26115051 PMCID: PMC4482731 DOI: 10.1371/journal.pone.0130548] [Citation(s) in RCA: 56] [Impact Index Per Article: 5.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/26/2015] [Accepted: 05/22/2015] [Indexed: 12/12/2022] Open
Abstract
Twenty two distinct bacteriophages were isolated from sewage water from five locations in the city of Abidjan, Côte d'Ivoire over a two-year period, using a collection of Pseudomonas aeruginosa strains with diverse genotypes. The phages were characterized by their virulence spectrum on a panel of selected P. aeruginosa strains from cystic fibrosis patients and by whole genome sequencing. Twelve virions representing the observed diversity were visualised by electron microscopy. The combined observations showed that 17 phages, distributed into seven genera, were virulent, and that five phages were related to temperate phages belonging to three genera. Some showed similarity with known phages only at the protein level. The vast majority of the genetic variations among virulent phages from the same genus resulted from seemingly non-random horizontal transfer events, inside a population of P. aeruginosa phages with limited diversity. This suggests the existence of a single environmental reservoir or ecotype in which continuous selection is taking place. In contrast, mostly point mutations were observed among phages potentially capable of lysogenisation. This is the first study of P. aeruginosa phage diversity in an African city and it shows that a large variety of phage species can be recovered in a limited geographical site at least when different bacterial strains are used. The relative temporal and spatial stability of the Abidjan phage population might reflect equilibrium in the microbial community from which they are released.
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Affiliation(s)
- Christiane Essoh
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
| | - Libera Latino
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
| | - Cédric Midoux
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
| | - Yann Blouin
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
| | - Guillaume Loukou
- Laboratoire National de Santé Publique, Abidjan, Côte d’Ivoire
- Laboratoire de Bactériologie-Virologie, département de Sciences pharmaceutiques et Biologiques, Univ Félix Houphouët-Boigny, Abidjan, Côte d’Ivoire
| | - Simon-Pierre A. Nguetta
- Laboratoire de Génétique, Département des Biosciences, Univ Félix Houphouet-Boigny, Abidjan, Côte d’Ivoire
| | - Serge Lathro
- Laboratoire National de Santé Publique, Abidjan, Côte d’Ivoire
| | | | | | - Gilles Vergnaud
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
- ENSTA ParisTech, Université Paris-Saclay, Palaiseau, France
| | - Christine Pourcel
- Institute for Integrative Biology of the Cell, CEA, CNRS, Univ Paris-Sud, Université Paris-Saclay, Orsay, France
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Jeske O, Schüler M, Schumann P, Schneider A, Boedeker C, Jogler M, Bollschweiler D, Rohde M, Mayer C, Engelhardt H, Spring S, Jogler C. Planctomycetes do possess a peptidoglycan cell wall. Nat Commun 2015; 6:7116. [PMID: 25964217 PMCID: PMC4432640 DOI: 10.1038/ncomms8116] [Citation(s) in RCA: 126] [Impact Index Per Article: 12.6] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/29/2014] [Accepted: 04/07/2015] [Indexed: 11/28/2022] Open
Abstract
Most bacteria contain a peptidoglycan (PG) cell wall, which is critical for
maintenance of shape and important for cell division. In contrast, Planctomycetes
have been proposed to produce a proteinaceous cell wall devoid of PG. The apparent
absence of PG has been used as an argument for the putative planctomycetal ancestry
of all bacterial lineages. Here we show, employing multiple bioinformatic methods,
that planctomycetal genomes encode proteins required for PG synthesis. Furthermore,
we biochemically demonstrate the presence of the sugar and the peptide components of
PG in Planctomycetes. In addition, light and electron microscopic experiments reveal
planctomycetal PG sacculi that are susceptible to lysozyme treatment. Finally,
cryo-electron tomography demonstrates that Planctomycetes possess a typical PG cell
wall and that their cellular architecture is thus more similar to that of other
Gram-negative bacteria. Our findings shed new light on the cellular architecture and
cell division of the maverick Planctomycetes. Planctomycetes appear to differ from all other bacteria in their
cellular organization and their apparent lack of a peptidoglycan (PG) cell wall. Here
Jeske et al. show that Planctomycetes do possess a typical PG cell wall and that
their cellular architecture resembles that of Gram-negative bacteria.
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Affiliation(s)
- Olga Jeske
- Independent Junior Research Group Microbial Cell Biology and Genetics, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
| | - Margarete Schüler
- Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18, Martinsried 82152, Germany
| | - Peter Schumann
- Department of Microbiology, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
| | - Alexander Schneider
- Department of Microbiology and Biotechnology, University of Tübingen, Auf der Morgenstelle 28, Tübingen 72076, Germany
| | - Christian Boedeker
- Independent Junior Research Group Microbial Cell Biology and Genetics, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
| | - Mareike Jogler
- Independent Junior Research Group Microbial Cell Biology and Genetics, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
| | - Daniel Bollschweiler
- Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18, Martinsried 82152, Germany
| | - Manfred Rohde
- Research Group Molecular Mechanisms of Streptococci, Helmholtz Center for Infection Research GmbH, Inhoffenstraße 7, Braunschweig 38124, Germany
| | - Christoph Mayer
- Department of Microbiology and Biotechnology, University of Tübingen, Auf der Morgenstelle 28, Tübingen 72076, Germany
| | - Harald Engelhardt
- Department of Molecular Structural Biology, Max-Planck-Institute for Biochemistry, Am Klopferspitz 18, Martinsried 82152, Germany
| | - Stefan Spring
- Department of Microbiology, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
| | - Christian Jogler
- Independent Junior Research Group Microbial Cell Biology and Genetics, Leibniz Institute-DSMZ, Inhoffenstraße 7b, Braunschweig 38124, Germany
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Khan Mirzaei M, Nilsson AS. Isolation of phages for phage therapy: a comparison of spot tests and efficiency of plating analyses for determination of host range and efficacy. PLoS One 2015; 10:e0118557. [PMID: 25761060 PMCID: PMC4356574 DOI: 10.1371/journal.pone.0118557] [Citation(s) in RCA: 266] [Impact Index Per Article: 26.6] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/22/2014] [Accepted: 01/20/2015] [Indexed: 11/18/2022] Open
Abstract
Phage therapy, treating bacterial infections with bacteriophages, could be a future alternative to antibiotic treatment of bacterial infections. There are, however, several problems to be solved, mainly associated to the biology of phages, the interaction between phages and their bacterial hosts, but also to the vast variation of pathogenic bacteria which implies that large numbers of different phages are going to be needed. All of these phages must under present regulation of medical products undergo extensive clinical testing before they can be applied. It will consequently be of great economic importance that effective and versatile phages are selected and collected into phage libraries, i.e., the selection must be carried out in a way that it results in highly virulent phages with broad host ranges. We have isolated phages using the Escherichia coli reference (ECOR) collection and compared two methods, spot testing and efficiency of plating (EOP), which are frequently used to identify phages suitable for phage therapy. The analyses of the differences between the two methods show that spot tests often overestimate both the overall virulence and the host range and that the results are not correlated to the results of EOP assays. The conclusion is that single dilution spot tests cannot be used for identification and selection of phages to a phage library and should be replaced by EOP assays. The difference between the two methods can be caused by many factors. We have analysed if the differences and lack of correlation could be caused by lysis from without, bacteriocins in the phage lysate, or by the presence of prophages harbouring genes coding for phage resistance systems in the genomes of the bacteria in the ECOR collection.
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Affiliation(s)
- Mohammadali Khan Mirzaei
- Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden
| | - Anders S. Nilsson
- Department of Molecular Biosciences, The Wenner-Gren Institute, Stockholm University, Stockholm, Sweden
- * E-mail:
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Wittmann J, Dreiseikelmann B, Rohde M, Meier-Kolthoff JP, Bunk B, Rohde C. First genome sequences of Achromobacter phages reveal new members of the N4 family. Virol J 2014; 11:14. [PMID: 24468270 PMCID: PMC3915230 DOI: 10.1186/1743-422x-11-14] [Citation(s) in RCA: 43] [Impact Index Per Article: 3.9] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/08/2013] [Accepted: 01/21/2014] [Indexed: 12/30/2022] Open
Abstract
BACKGROUND Multi-resistant Achromobacter xylosoxidans has been recognized as an emerging pathogen causing nosocomially acquired infections during the last years. Phages as natural opponents could be an alternative to fight such infections. Bacteriophages against this opportunistic pathogen were isolated in a recent study. This study shows a molecular analysis of two podoviruses and reveals first insights into the genomic structure of Achromobacter phages so far. METHODS Growth curve experiments and adsorption kinetics were performed for both phages. Adsorption and propagation in cells were visualized by electron microscopy. Both phage genomes were sequenced with the PacBio RS II system based on single molecule, real-time (SMRT) technology and annotated with several bioinformatic tools. To further elucidate the evolutionary relationships between the phage genomes, a phylogenomic analysis was conducted using the genome Blast Distance Phylogeny approach (GBDP). RESULTS In this study, we present the first detailed analysis of genome sequences of two Achromobacter phages so far. Phages JWAlpha and JWDelta were isolated from two different waste water treatment plants in Germany. Both phages belong to the Podoviridae and contain linear, double-stranded DNA with a length of 72329 bp and 73659 bp, respectively. 92 and 89 putative open reading frames were identified for JWAlpha and JWDelta, respectively, by bioinformatic analysis with several tools. The genomes have nearly the same organization and could be divided into different clusters for transcription, replication, host interaction, head and tail structure and lysis. Detailed annotation via protein comparisons with BLASTP revealed strong similarities to N4-like phages. CONCLUSIONS Analysis of the genomes of Achromobacter phages JWAlpha and JWDelta and comparisons of different gene clusters with other phages revealed that they might be strongly related to other N4-like phages, especially of the Escherichia group. Although all these phages show a highly conserved genomic structure and partially strong similarities at the amino acid level, some differences could be identified. Those differences, e.g. the existence of specific genes for replication or host interaction in some N4-like phages, seem to be interesting targets for further examination of function and specific mechanisms, which might enlighten the mechanism of phage establishment in the host cell after infection.
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Affiliation(s)
- Johannes Wittmann
- Department of Microorganisms, Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Brigitte Dreiseikelmann
- Department of Microbiology/Genetechnology, Faculty of Biology, University of Bielefeld, Bielefeld, Germany
| | - Manfred Rohde
- Helmholtz Centre for Infection Research, Department of Medical Microbiology, Central Facility for Microscopy, Braunschweig, Germany
| | - Jan P Meier-Kolthoff
- Department of Microorganisms, Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Boyke Bunk
- Bioinformatics, Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
| | - Christine Rohde
- Department of Microorganisms, Leibniz Institute DSMZ – German Collection of Microorganisms and Cell Cultures, Braunschweig, Germany
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