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Accepted Article 1 Norway Breivika, N-9037, Norway, of University Arctic The Institute ofHydrobiology,Chinese Institute China Wuhan, Sciences, of Academy Chinese Institute, Research River This article This protectedis by Allcopyright. rights reserved. cite this article as an 'Accepted Article', doi: 10.1111/jam.12626 whichmay lead to differences betweenthis beennot throughthe copyediting,typesetting, pagination and proofreading process This article hasbeen accepted for publicationand undergone fullpeer review but has 3 2 LiXuemei pond? spathala theDo intestinal microbiotas differbetween paddlefish ( Article : type Original Article Accepted Date : 15-Aug-2014 Revised Date : 11-Aug-2014 Received: Date 20-May-2014 Key Laboratory of Freshwater Biodiversity KeyCons Biodiversity Laboratory ofFreshwater Norwegian College ofFisheryFacultNorwegian College Science, Key Laboratory of Aquatic Biodiversity and Conservation of Chinese Academy of ofChineseSciences, of Academy LaboratoryKey and Conservation ofAquatic Biodiversity 1 ) and bighead ( carp bighead and ) , Yongjiu Zhu , Yongjiu 1 , Qingyun Yan Qingyun , Academy of Sciences,Wuhan, China Aristichthys nobilis Aristichthys 2 , Einar , Einar Ringø y of Biosciences, Fisher y ofBiosciences, version Version the and of Record. Please ervation, Ministry ofAgriculture Chinaof 3 , DeguoYang ) reared in the same same the in ) reared 1 * ies and Economics, UiT ies and Economics, Polyodon Polyodon Accepted Article prey items in the same pond (20 m lengths of 43.39±2.78 cm and 19.33± 3.68 cm, respectively were reared with natural Methods and Results assumed to be involved in the intestinal bacterial compositions. hostpotential factors, such as the genetic background, gut histology and physiology are paddlefish andbighead carpreared inthe samepond when fedsimilarnatureThe food. Conclusions: (12.6%). Bacteroidetes(37.0%), Fusobacteria (35.1% detected in bigheadthe carp, whiledominan (7.3%), Planctomycetes (6.0%)andActinobacteria (3.2%) Verrucomicrobia were species. Proteobacteria (57.3%), Firm paddlefish. Moreover, different core intestin carp ( This article This protectedis by Allcopyright. rights reserved. Aims: Abstract China 430223, Hubei, Wuhan, Zone, Development Hi-Tech Donghu Road, Wudayuan 1st No.8, Sciences, Fishery of Academy Chinese Institute, Research Fisheries Yangtze River Address: [email protected]; E-mail: Fax: +86-27-81780108; Tel: +86-27-81780108; * author: Corresponding specieswith similar ( correspondence analysis (DCA). Shannon diversity ( species according tothe alpha- and beta-diversity measurements anddetrended inthe were observed deviations of microbialcommunities the two Interestingly, fish of thetwo fish specieswere assessed by pyrosequencingof 16SrRNA genes. P =0.035) revealed significant=0.035) lower diversityof the intestinalmicrobiota of Aristichthys nobilis

a studywas a conducted to compare the intestinal microbial compositions oftwo our results revealed our results revealed intestinal betweenthat differed microbiotathe : age-0 paddlefish and bighead carp with mean average body

feeding strategy;paddlefish ( ) reared in the same pond. 2 ). ). After 30-days of rearing the intestinal microbiota icutes Fusobacteria(11.9%), (8.9%), t phyla in paddlefish intestines were almicrobiota was noticed in the twofish ),Firmicutes (14.8%)and Proteobacteria

P Polyodon spathala =0.015) and Pielou.evenness=0.015)

) and bighead Accepted Article metabolism, development,metabolism, immunity nutritional statusand (Rawls indicates thatintestinal fish microbiota hasa tight and coordinated connection with fish Actinobacteria, and Cyanobacteria (Romer 2010), and is dominated mainly by four bacter Key words: Key microbial composition. The intestinal fish microbiota canreach 10 2006; Ward environment, soil/sediment or live feed (Ringø et al andBirkbeck 1999). The gutmi 2008; Nayak 2010). Fish intestine are colonized by bacteria at the yolk sac stage (Ringø host and have beenstudied innumerous studies fish (Sugita bacterial playThe intestinalcommunities rolesin the important health nutrition of and Introduction ( of the importanceof the of paddlefish andbighead thestudycarp, present presents basic knowledgefor evaluation This article This protectedis by Allcopyright. rights reserved. the andImpact Study: of Significance the distinct growth environments and feeding strategies of different fishspecies, studies (Ringø compositionsuch diet, as; environments, developmental stage fish gut structure and al ncmaio fteritsia irboaaelcig on comparison of their intestinal microbiota are lacking. Polyodon spathala Polyodon . 2012). However, manyexogenous and endogenous factors can theaffect microbial . 1996;Ringø 1999) andBirkbeck and becan derived from the surroundingaquatic et al . 1996;. Ringø Olsenand 1999; Ringø Filter-feeding fish, bighead bighead fish, Filter-feeding carp ( et al 09. . 2009). ), intestinal pyrosequencing.microbiota,

host factors (genetic background and gut anatomy) on intestinal crobiotas are established during initialfeeding (Ringø considering the similar 7 o and Navarrete 2006). Moreover,evidence -10 ial phyla: Proteobacteria, Fusobacterium, Aristichthys nobilis Aristichthys 11 et al. bacteria g et al 1996; Romero andNavarrete . 2012a; 2012b). Therefore, given -1 in the in(Nayak digesta et al et

feeding strategy of of strategy feeding . 1998; Kuz`mina ), paddlefish et al et . 2004; Ray et Accepted Article treatment ponds.treatment Thestockingdensity of eachpond was similar and included 14 This article This protectedis by Allcopyright. rights reserved. species.Two ponds (20 m enoughnaturalprey items(cladocera, copepod,rotifer protozoa) and the for twofish fertilizer(Huang Jintai, China)used was pr averagebody lengthsof 43.39±2.78 cm and 19.33±3.68 respectively.cm, compoundA base inJingzhou provincein China, using age-0 paddlefish andbighead with carp mean management Fish Materials and Methods environments as revealed in an earlier study (Li the wholeduring They trial. basically are competitivetheirin dietinsame the growth reared in samethe pond fedonlyonand plankton natural organisms in water column paddlefish and bighead using carp pyrosequencing16SrRNA of genes. The fishwere (Weisellength) 1973). in contrast to paddlefish having stomach and a short intestine (RIL = one third the body intestine (relative intestinal length (RIL) = five times the body length) (Li comparativestudy.Thisis of importance to evaluate as bighead carphas along microbial of compositions paddlefish andbighead not havecarp beeninvestigated in a two fish species were similar. However, (2002) and Li from the water column, a hypothesis later confirmed by Kozfkay and Scarnecchia Hoxmeiercarp: 85mm), DeVriesand (1997)suggestedthat they filter similar prey Due1999). Wu to theirsimilar spacing gillraker bighead 60-90mmand (paddlefish: important filter-feeding fish species in poly-culture ponds in China (Zhang Paddlefish ( A 30-day feeding experiment was conducted at the Guanyin Dangfish farming presentthe In study, wethe evaluatedintestinal microbial of juvenile composition et al. Polyodon spathala Polyodon (2013a) showing that the food compositions in the digesta of the 2 each) rich in cladocera and copepod were selected as ) and bighead carp ( carp bighead ) and to the knowledgeauthors’ theintestinal ior toin theexperiment toprovide order et al. et al. Aristichthys nobilis Aristichthys 2013a). ) are two two ) are et al. et al 2013a) . 1989; Accepted Article This article This protectedis by Allcopyright. rights reserved. Sample collection days. 15 after of trial, the middle paddlefish and 14bighead The carp. compound wasfertilizer usedonce more in the variations in the gut microbiota (Ringø procedurenormal usedwhen investigating gut the microbiota to avoid interindividual sterile tube according to MacDonald intestinal contents thesefrom sectionsthree were collected pooledand together into a was divided intothree segments: 1) stomach/foregut, 2)midgut 3) and hindgut, and (scissors tweezers). and instruments The excisedintestine eachfrom individual fish before withswabbed 75%dissection of ethanol the usingintestine whole sterile pond), and anesthetized with an overdose ofMS 222.exterior Fishsurfaces were randomly sampledthe from two(3 paddlefish ponds and3 bighead carp each from 4°C. Pellets were washed with 70% ethanol and air dried for 30 min before precipitated with cold ethanol and pelleted by centrifugation at 13.000g for 20 min at extraction phenol/chloroform as previouslydescribed (Li K,proteinase 0.05 mg RNaseA) overnight55°C, at followed bystandard solution (30 mM EDTA, 10 mM TrisHCl, 0.5% sodium dodecyl sulfate (SDS), 0.1 mg DNA extraction extraction. tothethe where laboratory transportation samples were stored at−20 °CDNA until DNA. All intestinal - and water samples we throughglassfiltered fiber filters (Whatman type GF/C, 1.2 At theend ofthe 30-day period,culture DNA preparation wasperformed byincubatingpreparation DNA intestinal lysishomogenates in 1ml

At thesame as time gut sampling, 1 Lwater from pondeach was collected and

et al et al . (1986).Analysesof pooled samplesa are redry on temporarily stored during ice 6 paddlefish and 6bighead were carp 06. . 2006). et al μ m pore size) to extract . 2012). DNA was Accepted Article calculated according tothe procedur al. The taxonomic dissimilarities/similaritiesThe taxonomic between paireach of communities were for assignment using Ribosomal Da analysis Statistical thiscontrol wereutilized in study. werereads conducted on the Galaxy pipeline, and thesequences passedthat quality accordingto the manufacturer’s instructions.Quality filtering andprocessing of MiSeq Thepurification. DNA library was sequencedusingMiSeq platform Illumina the PicoGreen dsDNA Assay kit (invitogen),equally combined and followedby gel suremake thereno was Th contamination. visualizedwere on 1% agarose gels, andnegativewere controls always to performed sequenceprimer included adap appropriate fortemplate the second PCRamplification with the same primerset but the reverse This article This protectedis by Allcopyright. rights reserved. withpurified Agencout 2011). An initial 10 cycles of PCR amplification were performed, the products were amplifications were performedinwith triplicate 515F/806Rprimers the (Bates considered to yield more accurate taxonomic information (Liu 16S rRNAgene was used as it shows few biasesdifferentbacterial for taxa, and is samples were assessedMiSeq usingIllumina sequencingplatform. The V4regionof Bacterial 16S rRNA Gene Pyrosequencing use. further TEbuffer solution.DNA solutionwasstored re-suspensionat in 50µl −20°C until 2003). Inorderto the compare bacterial

Representativesequences of eachoperational taxonomic units (OTUs)was used A total of 14 samples including 6 paddlefish and 6 bighead carp and 2 water ® Ampure ®

XP (Beckman Coulter,Inc.), andthenused as es previously described(Caporaso e positiveamplicons werequantified using ters and different barcodes.PCR products communities, alpha-diversity indices were tabase Project (RDP) classifier (Cole et al . 2007). PCR et al . 2011). et al . et Accepted Article microbial assemblages alsodiffered paddlefish between bighead and carp(Fig.3). microbialharbored assemblages distinct fr compositions revealedthat samples from fish species (Fig. 2). Detrended correspondence analysis (DCA) of all microbial thewithinfrom same samples fish similar Bray-Curtis indices revealed that the al 1). (Fig. intestine the microbiota measurements, indiversity water was similar to that in bighead carp This article This protectedis by Allcopyright. rights reserved. and Pielou.evenness; Simpson.evenness were apparent among paddlefish, inShannonespecially diversity carp,bighead reductions diversity, inShannon Simpson’s values,Pielou.evenness and habitat and the differentiation respectively habitats,among 2011). (PeterCompared to measurements were first applied, to describespecies compositionin one specific paddlefish, 6 bighead carp 2and pond water samples).Alpha- andbeta-diversity sequencing platform after quality filtering and normalization of 14(6 samples sequencing theV4ofregions the 16SrRNA genes usingIllumina MiSeq the carp andbighead paddlefish intestines in communities microbial Allochthonous RESULTS 3.0.1) package. (version 18.0 and R SPSS the with software performed were analyses between bacterial paddlefish intestinal communities Statisticalbighead and carp. One way ANOVA and two-tailed Student’s t-test were used to assess the differences of correspondence analysis (DCA) was also performed to visually depict the differences. determined using the Jaccarddistance and Bray-Curtis indices, and detrended In addition to the beta diversity, to thetheIn addition betaJaccardaccording results to the and

A totalof 364,724rRNA 16S sequence gene were reads analyzed following

P =0.015 and P= 0.035, respectively. According to alpha-diversity lochthonous intestinal microbiota wasmore paddlefish andbighead carp intestines ompond water. Meanwhile, the intestinal species than between than species samples from the two

Accepted Article higher in bighead carp intestine (5.78%) compared withsample (5.78%) water (0.57%). compared intestine carp inbighead higher compared with paddlefish. The proportion of phylum Actinobacteria was considerably significantwere higher in abundance amongwater andbighead carpintestinesamples interesting tonoticethat Actinobacteria, Verrucomicrobiaand unclassified bacteria This article This protectedis by Allcopyright. rights reserved. andothers) the comprising “ Chlamydiae, Chloroflexi, Spirochaetes, Synergistetes, Tenericutes, Verrucomicrobia 4A). (Fig. water (1.46%) carpand bighead intestine (1.23%) than in paddlefish (0.04%)intestine Similarly, samples. paddlefish accounting 7.3%for and 2.0%, respectively, whilethis phylum was absent in all samples water and intestine carp bighead from detected were Planctomycetes with proportions of 37.0%, 35.1%, 14.8% and 12.6%, respectively. Phylum paddlefish intestinewere Bacteroidetes, Fusobacteria, Firmicutes andProteobacteria, 11.9%, 8.9%and 9.9%,respectively.In contrast, the dominant phyla detected in FusobacteriaFirmicutes, along with other le abundance, respectively. In bighead intestinalcarp segments, Proteobacteria, accounting for 25.1%42.9%, and 24.0% of the total Bacteroidetes and Proteobacteria waters to be similar, with the dominant phyla Cyanobacteria/ Chloroplast, bedominantto (Fig.4A). Indetail, results the showedmicrobiota the oftwo pond intheidentified different fish andpondwaterof samples which, 6phyla were identified representativesequences were classified using RDPclassifier. A total of17 phylawere carp intestines andbighead phyla inpaddlefish Dominant

Read number of the rare phyla (Actinobacteria, Acidobacteria, Armatimonadetes, To provide an overview of the sequence reads associated with fish species, the others the unclassifiedbacteria had ina higher representation ” category in Fig. 4A displayed in Fig. 4B. It is It is 4B. Fig. in 4A displayed Fig. in ” category ssabundant phyla weredominant: 57.3%, Accepted Article intestine (Weisel 1973). The carnivorous stat resembles othercarnivorous fish species, having apouch-likestomach and a short were similar such preys are available, although food c the otherhand,it is reportedthat young the factthat more bacterial phyla were detectedin bighead carp intestine (Fig. 4). On comparedrespectively) tobigheaddifference(Fig. carp1). may This be attributable to intestinal microbiota significantly lower in paddlefish( al indices of OTUs at a 97% sequence similarity (species level) as suggested by Rausch alpha-diversity, while beta-diversityweremeasured basedon Jaccard andBray-Cutis Shannon, inverse Simpson’s values, Pielou.evenness andSimpson.evenness for severalmeasurementsdifferentTherefore, were used in the present study, including the diversity measures commonly used in community ecology (Hughes known that microbial communities are highlydiverseand areoften poorly amenable to alpha-diversity and beta-diversity tobehave (Gering evaluated andCrist2002). Itis community structureanalysis, two important parametersof communitystructure: microbiota oftwofilter fishfeeding species:paddlefish bighead and carp. For DISCUSSION Bacteroidetes was significantly higher in paddlefish. significantly higherabundances inbigheadcarp. Genus This article This protectedis by Allcopyright. rights reserved. Aeromonas 1).(Table carpAmong intestine bighead the phylum Proteobacteria:genera and Bacteroidetes were significant different in paddlefish intestine compared to in genus Dominant paddlefish and bighead carp intestines . (2011). Ourresults revealed that the Shannon diversity Pielou.evenness and of In the present study, we thedetermine structure and composition of intestinal At genus taxonomysevenbacterial level, genera belongingto theProteobacteria , Aurantimonas, (Michaletz et

al.

Nitrobacter 1982).Besides, the anatomy ofpaddlefish intestine , paddlefish preferredlargewhen onsumption of paddlefish bigheadand carp Rhodobacter e may partly explain the dominance of a and P

Paludibacter =0.015 and 0.035, and=0.015 Roseomonas within the had et al . 2001).

et et Accepted Article environment (Lienvironment for for the microbiota (Weisel 1973). Studies of rainbow ( of paddlefish is more complex than bighead carp, and thus may provide different niches a role in shaping the fish intestinal microbiota in the The study. present digestive tract carp bluntsnoutand bream ( foundin intestines (Austin2006; Navarrete (14.8%), Firmicutes and (1 Proteobacteria paddlefish identifiedin intestine were Bact Verrucomicrobia (3.2%) asrevealedin bighead carp,while the dominantphyla (11.9%), Fusobacteria (8.9%),Planctomycetes (7.3%), Actinobacteria (6.0%) and previousin a investigation (Holben gastrointestinal shaping, long vs. short intestine and with or without stomach, as stated derived from water and/or diet and a new microbiota is established depending on the Bacteroidetes (Fig. 4). These results revealed that the fish intestinal microbiota are phyla comprisedmainly ofCyanobacteria/Chloroplast,Proteobacteria and toCompared gut microbiota thewater bacterial community structurewas different and compositionis influenced bythe host(Navarrete transgenic ( carp ( distinctthat microbiota dominant intestinal we thein same pondwith similar plankton available, withconsistentstudy ourprevious andbighead carptheir have specific intestinalmicrobiota even when they werereared differentfrom fish species (Fig. 2 and 3). The results indicated that juvenile paddlefish microbialintestinal communities fromthesame fishspecies much was higher than that This article This protectedis by Allcopyright. rights reserved. species notothenioid Antarctic (Ley fish increase from carnivores omnivoresto to herbivores both inhuman,mice andtwo of low number taxain gut, paddlefish becausebacterialdiversity been has to observed molitrix According to the beta-diversity measures and DCA analysis, the similarity of A core intestinal microbiota comprising et al. 2012). Hostsuch 2012). maybeplay factors, as gutstructure claimed to Cyprinus carpio Megalobrama amblycephala ), ( et al etal . 2002). 2.6%) 4).Proteobacteria, (Fig. a commonly eroidetes eroidetes (37.0%), Fusobacteria (35.1%), L.) revealedalso thatgut the microbiota . 2008; Ward of(57.3%), Proteobacteria Firmicutes re revealed in four fish species (silver (silver species in fish four revealed re Ctenopharyngodon idella et al . 2012;Li )) reared))the same in et al et al Oncorhynchus mykiss . 2009). . 2012) was most et al . 2013b). ), bighead ), bighead ) and ) and Accepted Article studyis unknown and meritsinvestigation. further Whether these features have any relevance for the fish species evaluated in the present reported to bein involved polysaccharide degradation (van Kessel the phylum Planctomycetes dominant inbighead carphas intestine previously been (Ueki have a and metabolismfermentative isable (RaftiloseP95) to degradeoligofructose Bacteroidetes was numerous in paddlefish intestine. well represented genus pesticide- degrading activity (van Kessel carpio carp intestine have similar beneficial effect as revealed of isolates from carp ( studies. studies. intestinal bacterial compositions, and these issuesshould be the subject offuture such as; the genetic background, gut histology and physiology areassumed to theaffect are present in bothcarp bigheadand paddlefish intestine.potentialThe host factors it shouldbeHowever, emphasized thatdifferent bacterialphyla withsimilar functions the presenceof similar naturalprey items possessed different intestinal microbiota. This article This protectedis by Allcopyright. rights reserved. genera the Whether enteritis as reported elsewhere (Chopra and Houston 1999)merits investigation.further bacterial of rate incident high have carp bighead why explain can finding our Whether pathogens well-known ofcorals and finfish (Denner findingsthat the two fish speciespossess inbigheadprevalent carp, while inpaddlefish. common Thesesupport the findings Our results revealedOur results that paddlefish and bigheadreared carp in the same pondin

The genera The genera et al L.) intestine where they are suggested to have nitrate-decomposing and . 2006; Van der Meulen Aeromonas Nitrobacter

Paludibacter and , Rhodobacter Aurantimonas et al , within classphylum, Bacteroidia and 06.Frhroe ti fitrs ont ht itis.Furthermore, ofinterest 2006). tonote that etal hi pcfcitsia irboa theirspecific intestinal microbiota. . 2011) deserve further attention. Another and detected in bighead carp included included carp bighead in detected Roseomonas Paludibacter et al . 2003; Ringø detected in bighead has been to reported et al . 2011). et al

. 2014). Cyprinus

Accepted Article This article This protectedis by Allcopyright. rights reserved. Austin, bacterialB. (2006)The References Special Fund Agrofor Scientific Research in the Public of Interest China (201003055). (2012BAD25B05), The Postdoctoral Innovation FundinHubei Province and the (2013M542099) work was This financially supportedby Acknowledgments The author declares no conflictof interest. Conflict of Interest Cole,J.R., Chai, B., Marsh, T.L.,Farris, R.J., Wang, Q., Kulam, S.A.Chandra, S., Caporaso, J.G.,Lauber, C.L., Walters, W.A., Berg-Lyons,D.,Lozupone, C.A, Bates,S.T., Berg-Lyons, D., Caporaso, J.G., Walters, W.A., Knight, R. and Fierer, N. 931-945.

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sp. strain NM strain of sp. bacterium12, intestinal an anaerobic, Gram-negative, propionate- 165 , 39-44. , S.K., (2012b)Use ofchitin and krill in quaculture: possibilities andpossibilities challenges. quaculture: 6-8. , 269-280. K., Adriany, T. andDe Vuyst, L. (2006) . (2006) Cyprinus carpio Paludibacter propionicigenes Bacteroides Oncorhynchus L.). L.). Appl Environ J Mar Sci: Res Res Sci: J Mar AMB Express AMB and Aquacult Rev

Accepted Article Wu, Y.B. (1999) Practice Y.B.Wu, and ofdevelopmentanalysis of This article This protectedis by Allcopyright. rights reserved. Paddlefish at intestine. levelinfish thebacteriagenus mostabundant taxonomy of deviation and standard per treatment) (%ofsequences abundances Table relative Average 1. Figures and Tables Weisel, G.F. (1973) Anatomy and histology of N.,Ward, Steven, B., Penn, K., Methé, B.andDetrich, W. (2009)Characterization of Zhang, Y.Z.,Zhang,Tan, Y.J. andO-Yang, H. (1989)Pond fish culture inChina.pp. 318-353. Genus Genus ( the intestinal microbiota twoof Antarcticnotothenioid fishspecies. Beijing: Science Publisher. Beijing: China. 13 Unclassified Unclassified 10.3±3.3 27.2±13.0 0.023

Polyodon spathula Polyodon , 679-685. Chinese Fish Econ Resh Econ Fish Chinese Aurantimonas Aurantimonas Paludibacter Paludibacter Rhodobacter Rhodobacter Roseomonas Roseomonas Nitrobacter Nitrobacter Aeromonas Aeromonas ).

J Morphol

3 140 , 18-19. , 243–255. (mean% ±SD) 0.05±0.08 2.33±1.91 0.032 0.032 2.33±1.91 0.05±0.08 0.016 0.02±0.038 30.5±21.1 0.05±0.09 2.14+1.55 0.021 0.021 2.14+1.55 0.05±0.09 0.02±0.01 0.65±0.54 0.035 0.035 0.65±0.54 0.02±0.01 -- 1.22±0.56 0.003 0.003 1.22±0.56 -- -- 0.29±0.23 0.036 0.036 0.29±0.23 -- the digestive system ofthepaddlefish (mean% ±SD) Bighead carp carp Bighead polyodon spathula polyodon Extremophiles Student’s t-test t-test Student’s P value culture in culture

Accepted Article This article This protectedis by Allcopyright. rights reserved. (OTUs). Error bars indicate SD, * (0.005< significant indicates association axonomic units operational onspecies-level Simpson.evenness based and(d) Pielou.evenness Figure me Meanalphadiversity 1. ( a ( ) c

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asures: (a) (b) Shannon, inverse. ( ( b d ) )

Simpson, (c) Simpson, P <0.05).

Accepted Article This article This protectedis by Allcopyright. rights reserved. bigheadcarp. species and represents BC paddlefish species based species-levelon axonomic operational (OTUs).units Errorbars indicate PrepresentsSD, distances Bray-Curtis (b) and measures: distances Jaccard Figure(a) Meanbeta 2. diversity (a) (b)

Accepted Article This article This protectedis by Allcopyright. rights reserved. samples. (DCA analysis correspondence Figure Detrended 3.

) of fish intestinal samplesand water pond ) offish intestinal

Accepted Article numbers indicate numbers last the respectively; bighead carp, and paddlefish species P and BC represents letters second This article This protectedis by Allcopyright. rights reserved. A. in and unclassifi rare phyla the among number Read ofFigure (B) read (A)Distribution insampled4. phyla number among the individuals. major

For each sample For eachsample code, (B) (A)

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the first letter and number p1andp2representsfirst the the ponds; two ed bacteria ed bacteria comprising the “Others” category