Addition of insoluble fiber to isolation media allows for increased metabolite diversity of lab-cultivable microbes derived from zebrafish gut samples

There is a gap in measured microbial diversity when comparing genomic sequencing techniques versus cultivation from environmental samples in a laboratory setting. Standardized methods in artificial environments may not recapitulate the environmental conditions that native microbes require for optimal growth. For example, the intestinal tract houses microbes at various pH values as well as minimal oxygen and light environments. These microbes are also exposed to an atypical source of carbon: dietary fiber compacted in fecal matter. To investigate how the addition of insoluble fiber to isolation media could affect the cultivation of microbes from zebrafish intestines, an isolate library was built and analyzed using the bioinformatics pipeline IDBac. The addition of fiber led to an increase in bacterial growth and encouraged the growth of species from several phyla. Furthermore, fiber addition altered the metabolism of the cultivated gut-derived microbes and induced the production of unique metabolites that were not produced when microbes were otherwise grown on standard isolation media. Addition of this inexpensive carbon source to media supported the cultivation of a diverse community whose specialized metabolite production may more closely replicate their metabolite production in vivo.

[1]  Laura M. Sanchez,et al.  Using the Open-Source MALDI TOF-MS IDBac Pipeline for Analysis of Microbial Protein and Specialized Metabolite Data. , 2019, Journal of visualized experiments : JoVE.

[2]  A. Ross,et al.  Culturing marine bacteria from the genus Pseudoalteromonas on a cotton scaffold alters secondary metabolite production , 2018, MicrobiologyOpen.

[3]  Yongchao Zhang,et al.  Possible association of Firmicutes in the gut microbiota of patients with major depressive disorder , 2018, Neuropsychiatric disease and treatment.

[4]  Mohammad Alanjary,et al.  Assessing the Efficiency of Cultivation Techniques To Recover Natural Product Biosynthetic Gene Populations from Sediment. , 2018, ACS chemical biology.

[5]  A. Gasbarrini,et al.  Actinobacteria: A relevant minority for the maintenance of gut homeostasis. , 2018, Digestive and liver disease : official journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver.

[6]  Roger G. Linington,et al.  Marine Mammal Microbiota Yields Novel Antibiotic with Potent Activity Against Clostridium difficile. , 2018, ACS infectious diseases.

[7]  Laura M. Sanchez,et al.  Coupling MALDI-TOF mass spectrometry protein and specialized metabolite analyses to rapidly discriminate bacterial function , 2017, Proceedings of the National Academy of Sciences.

[8]  Darren J. Martin,et al.  Environmental Screening of Aeromonas hydrophila, Mycobacterium spp., and Pseudocapillaria tomentosa in Zebrafish Systems , 2017, Journal of visualized experiments : JoVE.

[9]  R. Mahajan,et al.  Cost-effective screening and isolation of xylano-cellulolytic positive microbes from termite gut and termitarium , 2017, 3 Biotech.

[10]  Zhixin Wu,et al.  Effect of Bacillus subtilis on Aeromonas hydrophila-induced intestinal mucosal barrier function damage and inflammation in grass carp (Ctenopharyngodon idella) , 2017, Scientific Reports.

[11]  William H. Gerwick,et al.  Retrospective analysis of natural products provides insights for future discovery trends , 2017, Proceedings of the National Academy of Sciences.

[12]  B. Bohannan,et al.  The composition of the zebrafish intestinal microbial community varies across development , 2015, The ISME Journal.

[13]  Richard H. Baltz,et al.  Natural product discovery: past, present, and future , 2016, Journal of Industrial Microbiology & Biotechnology.

[14]  James B. Munro,et al.  The Microbiota of Freshwater Fish and Freshwater Niches Contain Omega-3 Fatty Acid-Producing Shewanella Species , 2015, Applied and Environmental Microbiology.

[15]  R. Talukdar,et al.  Role of the normal gut microbiota. , 2015, World journal of gastroenterology.

[16]  S. Stephenson,et al.  Evaluation of Physarum polycephalum plasmodial growth and lipid production using rice bran as a carbon source , 2015, BMC Biotechnology.

[17]  Didier Raoult,et al.  Current and Past Strategies for Bacterial Culture in Clinical Microbiology , 2015, Clinical Microbiology Reviews.

[18]  D. Powell,et al.  Polyketide Glycosides from Bionectria ochroleuca Inhibit Candida albicans Biofilm Formation , 2014, Journal of natural products.

[19]  Rachel J. Dutton,et al.  Cheese Rind Communities Provide Tractable Systems for In Situ and In Vitro Studies of Microbial Diversity , 2014, Cell.

[20]  Pedro Farias,et al.  Persistence of microbial communities including Pseudomonas aeruginosa in a hospital environment: a potential health hazard , 2014, BMC Microbiology.

[21]  S. Yadav,et al.  Simultaneous production of alkaline lipase and protease by antibiotic and heavy metal tolerant Pseudomonas aeruginosa , 2013, Journal of basic microbiology.

[22]  J. Hamedi,et al.  Screening of Antibacterial Producing Actinomycetes from Sediments of the Caspian Sea , 2013, International journal of molecular and cellular medicine.

[23]  J. Davies,et al.  Specialized microbial metabolites: functions and origins , 2013, The Journal of Antibiotics.

[24]  Roger G Linington,et al.  Development of antibiotic activity profile screening for the classification and discovery of natural product antibiotics. , 2012, Chemistry & biology.

[25]  G. Siuzdak,et al.  XCMS Online: a web-based platform to process untargeted metabolomic data. , 2012, Analytical chemistry.

[26]  Roger G. Linington,et al.  Examining the Fish Microbiome: Vertebrate-Derived Bacteria as an Environmental Niche for the Discovery of Unique Marine Natural Products , 2012, PloS one.

[27]  Ralf Tautenhahn,et al.  Meta-analysis of untargeted metabolomic data from multiple profiling experiments , 2012, Nature Protocols.

[28]  Roger G. Linington,et al.  An image-based 384-well high-throughput screening method for the discovery of biofilm inhibitors in Vibrio cholerae. , 2011, Molecular bioSystems.

[29]  E. Mittge,et al.  Evidence for a core gut microbiota in the zebrafish , 2011, The ISME Journal.

[30]  U. Kalawat,et al.  Emerging Infections: Shewanella – A Series of Five Cases , 2010, Journal of Laboratory Physicians.

[31]  S. Sauer,et al.  Phylogenetic classification and identification of bacteria by mass spectrometry , 2009, Nature Protocols.

[32]  K. Hong,et al.  Actinomycetes for Marine Drug Discovery Isolated from Mangrove Soils and Plants in China , 2009, Marine drugs.

[33]  L. Zon,et al.  Transparent adult zebrafish as a tool for in vivo transplantation analysis. , 2008, Cell stem cell.

[34]  H. Flint,et al.  Cultivable bacterial diversity from the human colon , 2007, Letters in applied microbiology.

[35]  S. Shamoun,et al.  The effects of culture media, solid substrates, and relative humidity on growth, sporulation and conidial discharge of Valdensinia heterodoxa. , 2006, Mycological research.

[36]  A. Henriques,et al.  The Intestinal Life Cycle of Bacillus subtilis and Close Relatives , 2006, Journal of bacteriology.

[37]  Yifan Hu,et al.  Efficient, High-Quality Force-Directed Graph Drawing , 2006 .

[38]  T. Mincer,et al.  Culturable marine actinomycete diversity from tropical Pacific Ocean sediments. , 2005, Environmental microbiology.

[39]  William Fenical,et al.  Widespread and Persistent Populations of a Major New Marine Actinomycete Taxon in Ocean Sediments , 2002, Applied and Environmental Microbiology.

[40]  J. Mcchesney,et al.  Cotton fiber as a substitute for agar support in tissue culture , 1995 .

[41]  C. Hirsch,et al.  Method for Isolation and Purification of Cyanobacteria , 1991, Applied and environmental microbiology.

[42]  D. Lane 16S/23S rRNA sequencing , 1991 .

[43]  J. T. Staley,et al.  Measurement of in situ activities of nonphotosynthetic microorganisms in aquatic and terrestrial habitats. , 1985, Annual review of microbiology.

[44]  R. E. Hungate,et al.  Effect of alfalfa fiber substrate on culture counts of rumen bacteria , 1976, Applied and environmental microbiology.