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  • 1
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    Oligotyping reveals community level habitat selection within the genus Vibrio (2014)
    Frontiers Media
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    Publication Date: 2014-11-13
    Electronic ISSN: 1664-302X
    Topics: Biology
    Published by Frontiers Media
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  • 2
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    A method for selectively enriching microbial DNA from contaminating vertebrate host DNA (2013)
    Public Library of Science
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    Publication Date: 2022-05-25
    Description: © The Author(s), 2013. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS ONE 8 (2013): e76096, doi:10.1371/journal.pone.0076096.
    Description: DNA samples derived from vertebrate skin, bodily cavities and body fluids contain both host and microbial DNA; the latter often present as a minor component. Consequently, DNA sequencing of a microbiome sample frequently yields reads originating from the microbe(s) of interest, but with a vast excess of host genome-derived reads. In this study, we used a methyl-CpG binding domain (MBD) to separate methylated host DNA from microbial DNA based on differences in CpG methylation density. MBD fused to the Fc region of a human antibody (MBD-Fc) binds strongly to protein A paramagnetic beads, forming an effective one-step enrichment complex that was used to remove human or fish host DNA from bacterial and protistan DNA for subsequent sequencing and analysis. We report enrichment of DNA samples from human saliva, human blood, a mock malaria-infected blood sample and a black molly fish. When reads were mapped to reference genomes, sequence reads aligning to host genomes decreased 50-fold, while bacterial and Plasmodium DNA sequences reads increased 8–11.5-fold. The Shannon-Wiener diversity index was calculated for 149 bacterial species in saliva before and after enrichment. Unenriched saliva had an index of 4.72, while the enriched sample had an index of 4.80. The similarity of these indices demonstrates that bacterial species diversity and relative phylotype abundance remain conserved in enriched samples. Enrichment using the MBD-Fc method holds promise for targeted microbiome sequence analysis across a broad range of sample types.
    Description: LAZ and VS were funded by a Brown University Office of the Vice President of Research SEED grant (LAZ) entitled “Tracking disease spread through the wildlife trade: New techniques to identify infectious microbes in aquarium fishes.” SOO and MQ were funded by Wellcome Trust Sanger Institute grant numbers 098051 and 079355/Z/06/Z.
    Repository Name: Woods Hole Open Access Server
    Type: Article
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  • 3
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    Microbial diversity and potential pathogens in onamental fish aquarium water (2012)
    Public Library of Science
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    Publication Date: 2022-05-26
    Description: © The Author(s), 2012. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in PLoS ONE 7 (2012): e39971, doi:10.1371/journal.pone.0039971.
    Description: Ornamental fishes are among the most popular and fastest growing categories of pets in the United States (U.S.). The global scope and scale of the ornamental fish trade and growing popularity of pet fish in the U.S. are strong indicators of the myriad economic and social benefits the pet industry provides. Relatively little is known about the microbial communities associated with these ornamental fishes or the aquarium water in which they are transported and housed. Using conventional molecular approaches and next generation high-throughput amplicon sequencing of 16S ribosomal RNA gene hypervariable regions, we characterized the bacterial community of aquarium water containing common goldfish (Carassius auratus) and Chinese algae eaters (Gyrinocheilus aymonieri) purchased from seven pet/aquarium shops in Rhode Island and identified the presence of potential pathogens. Our survey identified a total of 30 phyla, the most common being Proteobacteria (52%), Bacteroidetes (18%) and Planctomycetes (6%), with the top four phyla representing 〉80% of all sequences. Sequences from our water samples were most closely related to eleven bacterial species that have the potential to cause disease in fishes, humans and other species: Coxiella burnetii, Flavobacterium columnare, Legionella birminghamensis, L. pneumophila, Vibrio cholerae, V. mimicus. V. vulnificus, Aeromonas schubertii, A. veronii, A. hydrophila and Plesiomonas shigelloides. Our results, combined with evidence from the literature, suggest aquarium tank water harboring ornamental fish are an understudied source for novel microbial communities and pathogens that pose potential risks to the pet industry, fishes in trade, humans and other species.
    Repository Name: Woods Hole Open Access Server
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  • 4
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    The influence of fishmeal-free diets on microbial communities in Atlantic salmon Salmo salar recirculation aquaculture systems (2016)
    American Society for Microbiology
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    Publication Date: 2022-05-26
    Description: © The Author(s), 2016. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Applied and Environmental Microbiology 82 (2016): 4470-4481, doi:10.1128/AEM.00902-16.
    Description: Reliance on fishmeal as a primary protein source is among the chief economic and environmental concerns in aquaculture today. Fishmeal-based feeds often require harvest from wild fish stocks, placing pressure on natural ecosystems and causing price instability. Alternative diet formulations without the use of fishmeal provide a potential solution to this challenge. Although the impact of alternative diets on fish performance, intestinal inflammation, palatability, and gut microbiota has been a topic of recent interest, less is known about how alternative feeds impact the aquaculture environment as a whole. The recent focus on recirculating aquaculture systems (RAS) and the closed containment approach to raising food fish highlights the need to maintain stable environmental and microbiological conditions within a farm environment. Microbial stability in RAS biofilters is particularly important, given its role in nutrient processing and water quality in these closed systems. If and how the impacts of alternative feeds on microbial communities in fish translate into changes to the biofilters is not known. We tested the influence of a fishmeal-free diet on the microbial communities in RAS water, biofilters and salmon microbiomes using high-throughput 16S rRNA gene V6 hypervariable region amplicon sequencing. We grew Atlantic salmon (Salmo salar) to market-size in six replicate RAS tanks, three with traditional fishmeal diets and three with alternative protein, fishmeal-free diets. We sampled intestines and gills from market-ready adult fish, water and biofilter media in each corresponding RAS unit. Our results provide the first data on how fish diet influences the RAS environment, and corroborates previous findings that diet has a clear influence on microbiome structure of the salmon intestine, particularly within the order Lactobacillales (lactic acid bacteria). We conclude that strong stability of taxa likely involved in water quality processing regardless of diet (e.g. Nitrospira) may further alleviate concerns regarding the use of alternative feeds in RAS operations.
    Description: This work was supported by USDA ARS agreement 59-1930-5-510 with the Freshwater Institute, Shepherdstown, WV. V.S. was supported during this work by an NSF EPSCoR grant (EPS-1004057) and an NSF Doctoral Dissertation Improvement Grant (DDIG) to L.A.-Z. and V.S. (DEB-1402051).
    Repository Name: Woods Hole Open Access Server
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  • 5
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    Subtle microbiome manipulation using probiotics reduces antibiotic-associated mortality in fish (2018)
    American Society for Microbiology
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    Publication Date: 2022-05-26
    Description: © The Author(s), 2017. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in mSystems 2 (2017): e00133-17, doi:10.1128/mSystems.00133-17.
    Description: Prophylactic antibiotics in the aquaculture and ornamental fish industry are intended to prevent the negative impacts of disease outbreaks. Research in mice and humans suggests that antibiotics may disturb microbiome communities and decrease microbiome-mediated disease resistance, also known as “colonization resistance.” If antibiotics impact fish as they do mice and humans, prophylactic administrations on aquaculture farms may increase downstream disease susceptibility in target hosts, despite short-term pathogen control benefits. We tested the effects of antibiotics on mortality after a pathogen challenge in the Poecilia sphenops black molly and subsequently tested if probiotic inoculations could reverse any antibiotic-induced losses of disease resistance. We found that antibiotic treatment significantly increased fish mortality. We further found that our two candidate probiotic bacterial species, Phaeobacter inhibens S4Sm and Bacillus pumilus RI06-95Sm, were able to colonize black molly microbiomes and reverse the negative impacts of antibiotics. Despite the positive impact on survival, probiotic treatment did not influence overall microbiome community structure or diversity. Our results suggest that subtle manipulations of microbiome composition can have dramatic impacts on host phenotype. The results of this study have implications for how antibiotic-treated microbiomes can be restored and suggest that small-scale additions may be as effective as wholesale transplants.
    Description: L.A.-Z., V.S., and K.S. acknowledge support from National Science Foundation Doctoral Dissertation Improvement Grant no. 1402051 for V.S.
    Keywords: Antibiotics ; Aquaculture ; Bacillus ; Colonization resistance ; Microbial ecology ; Microbiome ; Phaeobacter ; Probiotics ; Vibrio
    Repository Name: Woods Hole Open Access Server
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  • 6
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    Oligotyping reveals community level habitat selection within the genus Vibrio (2015)
    Frontiers Media
    Details
    Publication Date: 2022-05-26
    Description: © The Author(s), 2014. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Frontiers in Microbiology 5 (2014): 563, doi:10.3389/fmicb.2014.00563.
    Description: The genus Vibrio is a metabolically diverse group of facultative anaerobic bacteria, common in aquatic environments and marine hosts. The genus contains several species of importance to human health and aquaculture, including the causative agents of human cholera and fish vibriosis. Vibrios display a wide variety of known life histories, from opportunistic pathogens to long-standing symbionts with individual host species. Studying Vibrio ecology has been challenging as individual species often display a wide range of habitat preferences, and groups of vibrios can act as socially cohesive groups. Although strong associations with salinity, temperature and other environmental variables have been established, the degree of habitat or host specificity at both the individual and community levels is unknown. Here we use oligotyping analyses in combination with a large collection of existing Vibrio 16S ribosomal RNA (rRNA) gene sequence data to reveal patterns of Vibrio ecology across a wide range of environmental, host, and abiotic substrate associated habitats. Our data show that individual taxa often display a wide range of habitat preferences yet tend to be highly abundant in either substrate-associated or free-living environments. Our analyses show that Vibrio communities share considerable overlap between two distinct hosts (i.e., sponge and fish), yet are distinct from the abiotic plastic substrates. Lastly, evidence for habitat specificity at the community level exists in some habitats, despite considerable stochasticity in others. In addition to providing insights into Vibrio ecology across a broad range of habitats, our study shows the utility of oligotyping as a facile, high-throughput and unbiased method for large-scale analyses of publically available sequence data repositories and suggests its wide application could greatly extend the range of possibilities to explore microbial ecology.
    Description: This work was supported by an NSF Collaborative grant to Erik Zettler (OCE-1155379), Tracy J. Mincer (OCE-1155671) and Linda A. Amaral-Zettler (OCE-1155571), NSF TUES grant to Erik Zettler and Linda A. Amaral-Zettler (DUE-1043468). Additional support came from the Woods Hole Center for Oceans and Human Health from the National Institutes of Health and National Science Foundation (NIH/NIEHS 1 P50 ES012742-01 and NSF/OCE 0430724-J: Linda A. Amaral-Zettler and Leslie Murphy) and an NSF/OCE-1128039 award (Linda A. Amaral-Zettler and Leslie Murphy). Victor Schmidt was supported during this work by an NSF IGERT fellowship (DGE 0966060, Dr. David Rand, PI).
    Keywords: Oligotyping ; Vibrio ecology ; Host-microbe interactions ; Illumina sequencing ; 16S rRNA analysis ; Plastisphere ; Aquaculture pathogens ; Meta-analysis
    Repository Name: Woods Hole Open Access Server
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