Rudzi rutsva rweGram-negative, aerobic, inoshivirira munyu, inoshanda, yakaita shamhu, uye mabhakitiriya anodya ASxL5T yakaparadzaniswa nedziva rendove yemombe muNottinghamshire, England, uye yakashandisa Campylobacter semhuka yayo. Zvadaro, mamwe marudzi eCampylobacter uye nhengo dzemhuri yeEnterobacteriaceae dzakawanikwa semhuka. Mushure me subculture isina maseru anotambira, kusasimba kweaseptic kukura kwakawanikwa paBrain Heart Infusion Agar. Mamiriro ekukura akanyanya i 37 °C uye pH 7. Transmission electron microscopy yakaratidza zvimwe zvisingawanzo kujairika morphological maitiro ane chekuita nekuwanikwa kwemhuka. Kuongorora kwePhylogenetic uchishandisa 16S rRNA gene sequence yakaratidza kuti iyo yega yakabatana nenhengo yemhuri yeMarine Spirulina, asi haigoni kunyatsotsanangurwa senhengo yemhando ipi zvayo inozivikanwa. Yese-genome sequencing yeASxL5T yakasimbisa hukama nenhengo dzemarine spirochetes. Tsvagiridzo yedhatabhesi yakaratidza kuti akati wandei maASxL5Ts anogovera 16S rRNA gene kutevedzana nemabhakitiriya akati wandei asina kugadzirwa kubva mugungwa, pasi uye pasi pemvura. Isu tinopa zano kuti dhizaini ASxL5T inomiririra mhando nyowani mune itsva genus. Tinokurudzira zita rekuti Venatorbacter cucullus gen. Mbudzi, sp. MunaNovember, ASxL5T yakashandiswa semhando yemhando.
Predatory mabhakitiriya mabhakitiriya anoratidza kugona kuvhima nekuuraya mamwe mabhakitiriya mhenyu kuti awane biosynthetic zvinhu nesimba. Izvi zvakasiyana nekudzoserwa kwakawanda kwezvikafu kubva kune ma microorganisms akafa, uye zvakare zvakasiyana nekudyidzana kweparasitic, umo mabhakitiriya anoumba hukama hwepedyo neanovagamuchira pasina kuvauraya. Mabhakitiriya anoparadza akachinja mararamiro akasiyana kuti atore mukana wezvikafu zvakawanda munzvimbo dzavanowanikwa (senge nzvimbo dzemugungwa). Ivo iboka rakasiyana-siyana remitero, iro rinongobatanidzwa chete nehupenyu hwavo hwekuita sterilization1. Mienzaniso yehutachiona hwehutachiona yakawanikwa mune zvakasiyana-siyana zvephyla, kusanganisira: Proteobacteria, Bacteroides, uye Chlorella.3. Zvakadaro, mabhakitiriya anonyatso dzidza anokara iBdellovibrio neBdellovibrio-uye-sezvipenyu (BALOs4). Hutachiona hunotyisa inzvimbo inovimbisa yezvitsva zvebiologically active compounds uye antibacterial agents5.
Mabhakitiriya anouraya anotendwa kuti anowedzera kusiyanisa kwehutachiona uye ane mhedzisiro yakanaka pane ecosystem hutano, kugadzirwa uye kugadzikana6. Zvisinei nehunhu uhwu hwakanaka, pane zvidzidzo zvishoma pane mabhakitiriya matsva anodya nekuda kwekuomerwa kwekurima mabhakitiriya uye kukosha kwekunyatsocherechedza kupindirana kwesero kuti vanzwisise kutenderera kwehupenyu hwavo. Mashoko aya haasi nyore kuwana kubva kuongororo yekombuta.
Mune imwe nguva yekuwedzera antimicrobial resistance, nzira itsva dzekutarisa hutachiona hwehutachiona huri kuongororwa, zvakadai sekushandiswa kwebhakteriophages uye mabhakitiriya anoparadza7,8. Mabhakitiriya eASxL5T akaparadzaniswa muna 2019 achishandisa tekinoroji yekuzviparadzanisa nevamwe kubva pandove yemombe yakaunganidzwa kubva kuDairy Center yeYunivhesiti yeNottingham, Nottinghamshire. Chinangwa chekuferefeta ndechekusiyanisa zvipenyu zvine mukana seamiririri anodzora zvipenyu. Campylobacter hyointestinal is a zoonotic pathogen, iyo inowedzera kushamwaridzana nezvirwere zvevanhu zvemukati10. Iyo inowanikwa kwese mu serum uye inoshandiswa senge inotarirwa.
Bhakitiriya reASxL5T rakanga raparadzaniswa nejelly yemombe nokuti yakaonekwa kuti mapepa aakagadzira pamucheka weC. Uku ndiko kuwana kusingatarisirwi, nekuti chikamu cheiyo phage yekuzviparadzanisa nzira inosanganisira kusefa kuburikidza ne 0.2 µm sefa, iyo yakagadzirirwa kubvisa mabhakitiriya maseru. Ongororo yeMicroscopic yezvinhu zvakabviswa padombo yakaratidza kuti bhakitiriya diki giremu-negative curved rod haina kuunganidza polyhydroxybutyrate (PHB). Tsika yeaseptic yakazvimirira kubva kumasero emhuka inoonekwa pane yakapfuma yakasimba svikiro (senge brain heart infusion agar (BHI) uye blood agar (BA)), uye kukura kwayo hakuna kusimba. Inowanikwa mushure me subculture ine inorema inoculum inovandudza. Inokura zvakaenzana pasi pe microaerobic (7% v/v okisijeni) uye mamiriro okisijeni emuchadenga, asi kwete mumhepo inonzi anaerobic. Mushure memaawa makumi manomwe nemaviri, dhayamita yekoloni yaive diki kwazvo, yakasvika 2 mm, uye yaive beige, inopenya, yakatenderera, inopenya uye inopenya. Yakajairwa biochemical yekuongorora inokanganiswa nekuti ASxL5T haigone kugadzirwa yakavimbika mune yemvura midhiya, izvo zvinoratidza kuti inogona kuvimba nehupenyu hwakaoma kutenderera kwekuumbwa kwebiofilm. Zvisinei, kumiswa kweplate kwakaratidza kuti ASxL5T is aerobic, yakanaka kune oxidase uye catalase, uye inogona kushivirira 5% NaCl. ASxL5T inoshingirira ku10 µg streptomycin, asi inonzwa kune mamwe maantibioti ese akaongororwa. Iyo ASxL5T mabhakitiriya maseru akaongororwa neTEM (Mufananidzo 1). Kana yakura isina masero emhuka paBA, ASxL5T masero maduku Campylobacter, ane avhareji urefu hwe1.63 μm (± 0.4), hupamhi hwe0.37 μm (± 0.08), uye imwe chete yakareba (kusvika 5 μm) danda. Sexual flagella. Inenge 1.6% yemasero anoita seane hupamhi husingasviki 0.2 μm, iyo inobvumira kupfuura kuburikidza nesefa mudziyo. Kuwedzerwa kwezvimiro zvisina kujairika kwakaonekwa pamusoro pemamwe masero, akafanana neakanaka (Latin cucullus) (ona miseve mu1D, E, G). Izvi zvinoratidzika kunge zvakaumbwa nekunze kwekunze membrane, iyo inogona kunge iri nekuda kwekukurumidza kuderedzwa kwehukuru hwe periplasmic envelope, nepo kunze membrane inoramba yakasimba, ichiratidza "yakasununguka" kutaridzika. Kurima ASxL5T mukushayikwa kwezvokudya (muPBS) kwenguva yakareba pa 4 ° C yakaguma nemasero akawanda (asi kwete ose) anoratidza coccal morphology (Mufananidzo 1C). Apo ASxL5T inokura neCampylobacter jejuni semhuka ye48 maawa, chiyero chesero chepakati chakareba uye chakatetepa kudarika masero akakura asina muenzi (Table 1 uye Figure 1E). Kusiyana neizvi, apo ASxL5T inokura neE. coli semhuka ye48 maawa, chiyero chesero yakakura yakareba uye yakakura kupfuura iyo inokura isina mhuka (Table 1), uye urefu hwesero hunosiyana, kazhinji hunoratidza filamentous (Mufananidzo 1F). Kana yaiswa neCampylobacter jejuni kana E. coli semhuka ye48 maawa, ASxL5T masero haana kuratidza flagella zvachose. Tafura 1 inopfupisa kucherechedzwa kwekuchinja kwehukuru hwesero zvichienderana nekuvapo, kusavapo, uye mhuka yerudzi rweASxL5T.
TEM kuratidzwa kweASx5LT: (A) ASx5LT inoratidza tyava refu; (B) yakajairika ASx5LT bhatiri; (C) cocci ASx5LT masero mushure mekugara kwenguva refu pasina zvinovaka muviri; (D) boka reAsx5LT masero rinoratidza kusagadzikana (E) ASx5LT cell group yakavharwa neCampylobacter prey yakaratidza kuwedzera kwesero kureba kana ichienzaniswa nevaya vasina kukura kwemhuka (D) yakaratidzawo apical structure; (F) Yakakura Filamentous flagella, ASx5LT masero, mushure mekuiswa neE. coli mhuka; (G) Sero imwe chete yeASx5LT mushure mekuiswa neE. coli, ichiratidza chimiro chepamusoro chisina kujairika. Iyo bar inomiririra 1 μm.
Kusarudza iyo 16S rRNA gene sequence (nhamba yekupinda MT636545.1) inogonesa kutsvaga kwedhatabhesi kuti igadzire kutevedzana kwakafanana neiyo iri mukirasi yeGammaproteobacteria, uye iri padyo nemabhakitiriya emugungwa mumhuri yespirillum yegungwa (Mufananidzo 2), uye inhengo dzeThalassolituus genus genus. Hama yepedyo kuna Marine Bacillus. Iyo 16S rRNA gene sequence yakanyatso siyana kubva kune inodya mabhakitiriya emhuri yeBdelvibrionaceae (Deltaproteobacteria). Kuenzaniswa paviri kweB. bacteriovorus HD100T (type strain, DSM 50701) uye B. bacteriovorus DM11A yaive 48.4% ne47.7%, uye yeB. exovorus JSS yaive 46.7%. ASxL5T mabhakitiriya ane 3 makopi eiyo 16S rRNA gene, maviri ayo akafanana kune mumwe nemumwe, uye yechitatu mabhesi matatu akaparadzana. Mamwe maviri anoparadza mabhakitiriya akaparadzana (ASx5S uye ASx5O; 16S rRNA gene kuwanikwa nhamba ndeye MT636546.1 uye MT636547.1, zvichiteerana) ane akafanana morphological uye phenotypic maitiro kubva panzvimbo imwechete haana kufanana, asi iwo akasiyana neASxL5T uye uncultured Bact. Database sequences dzakabatanidzwa pamwe nemamwe genera mukati Oceanospirillaceae (Mufananidzo 2). Iyo yese genome sequence yeASxL5T yakatemerwa uye yakachengetwa muNCBI dhatabhesi, uye nhamba yekupinda ndeye CP046056. Iyo genome yeASxL5T ine denderedzwa chromosome ye2,831,152 bp ine G + C reshiyo ye56.1%. Iyo genome sequence ine 2653 CDS (yakazara), iyo 2567 inofanotaurwa kuti incode mapuroteni, ayo 1596 anogona kupihwa sekuisa mabasa (60.2%). Iyo genome ine 67 RNA-coding genes, kusanganisira 9 rRNAs (3 imwe neimwe ye5S, 16S, uye 23S) uye 57 tRNAs. Iyo genomic maitiro eASxL5T akafananidzwa neanowanikwa genomes emhando yehama yepedyo yakaonekwa kubva ku16S rRNA gene sequence (Table 2). Shandisa amino acid identity (AAI) kuenzanisa ese aripo Thalassolituus genomes kune ASxL5T. Iyo yepedyo inowanikwa (isina kukwana) genome sequence yakatemwa neAAI ndeye Thalassolituus sp. C2-1 (wedzera NZ_VNIL01000001). Iyi dhizaini yakaparadzaniswa kubva pakadzika-gungwa sediments yeMariana Trench, asi parizvino hapana ruzivo rwe phenotypic nezve iyi dhizaini yekuenzanisa. Kuenzaniswa neASxL5T's 2.82 Mb, genome yechipenyu yakakura pa4.36 Mb. Avhareji yegenome saizi yemarine spirochetes ingangoita 4.16 Mb (± 1.1; n = 92 yakazara mareferenzi genomes akaferefetwa kubva ku https://www.ncbi.nlm.nih.gov/assembly), saka genome yeASxL5T inoenderana ne odha Kana ichienzaniswa nedzimwe nhengo, idiki chaizvo. Shandisa GToTree 1.5.54 kugadzira genome-based inofungidzirwa yakanyanya mukana we phylogenetic muti (Mufananidzo 3A), uchishandisa yakarongwa uye yakabatana amino acid sequences ye172 single-copy genes yakanangana neGammaproteobacteria 11,12,13,14,15,16, 17 ,18. Ongororo yacho yakaratidza kuti yakabatana zvakanyanya neThalassolituus, Bacterial Plane, uye Marine Bacterium. Nekudaro, idzi data dzinoratidza kuti ASxL5T yakasiyana nehama dzayo muMarine Spirulina uye data yayo yekutevedzana kwegenome inowanikwa.
Muti we phylogenetic unoshandisa 16S rRNA gene sequence inosimbisa nzvimbo yeASxL5T, ASxO5, uye ASxS5 strains (ine guts) maererano neiyo isina kurima uye yegungwa mabhakitiriya anokonzera muMarine Spirulinaceae. Nhamba yekupinda yeGenbank inotevera zita rekutambudzika mumabarekani. Shandisa ClustalW kurongedza kutevedzana, uye shandisa yakanyanya mukana nzira uye Tamura-Nei modhi kuti infer phylogenetic hukama, uye kuita 1000 inotungamirwa replication muchirongwa cheMEGA X. Nhamba iri pabazi inoratidza kuti inotungamirwa kopi kukosha yakakura kupfuura 50%. Escherichia coli U/541T yakashandiswa seboka rekunze.
(A) Muti we phylogenetic wakavakirwa pajenome, unoratidza hukama pakati pegungwa reSpirospiraceae bacterium ASxL5T uye hama dzayo dzepedyo, E. coli U 5/41T seboka rekunze. (B) Kuenzaniswa neT. oleivorans MIL-1T, iyo inoshanda chikamu chekugovera majini inofanotaurwa zvichibva pane orthologous group (COG) cluster yeASx5LT protein. Mufananidzo uri kuruboshwe unoratidza huwandu hwemajini mune yega yega inoshanda COG chikamu mune yega genome. Girafu riri kurudyi rinoratidza huwandu hwema genomes ari muboka rega rega rinoshanda reCOG. (C) Kuenzaniswa neT. oleiverans MIL-1T, kuongororwa kweKEGG yakakwana (Kyoto Encyclopedia yeGenes uye Genomes) modular nzira ye ASxL5T.
Kushandisa iyo KEGG dhatabhesi kuongorora iyo chikamu majini aripo muASxL5T genome yakaratidza yakajairika metabolic nzira yeaerobic gamma Proteus. ASxL5T ine huwandu hwemakumi manomwe neshanu emajini akagoverwa kubhakitiriya mota mapuroteni, kusanganisira majini anobatanidzwa muchemotaxis, flagella assembly, uye type IV fimbriae system. Muchikamu chekupedzisira, 9 kubva mugumi majini ane basa rekufambisa kwekufamba kwemhando yezvimwe zvipenyu. Iyo genome yeASxL5T ine yakazara tetrahydropyrimidine biosynthetic nzira inotora chikamu mumhinduro yekudzivirira kune osmotic stress20, sezvinotarisirwa kune halophiles. Iyo genome inewo nzira dzakawanda dzakakwana dzemacofactors nemavhitamini, kusanganisira riboflavin synthesis nzira. Kunyangwe iyo alkane 1-monooxygenase (alkB2) gene iripo muASxL5T, nzira yekushandisa hydrocarbon haina kukwana. Mukutevedzana kwegenome kweASxL5T, homologues emajini akaonekwa seanonyanya kukonzera kuparara kwemahydrocarbon muT. oleiverans MIL-1T21, senge TOL_2658 (alkB) uye TOL_2772 (doro dehydrogenase) zviri pachena kuti asipo. Mufananidzo 3B inoratidza kuenzanisa kwekugoverwa kwemajini muchikamu cheCOG pakati peASxL5T neOlive Oiri MIL-1T. Pakazere, iyo diki ASxL5T genome ine zvishoma kubva kune yega yega COG chikamu kana ichienzaniswa neyakakura inoenderana genome. Kana huwandu hwemajini muchikamu chega chega chekushanda hunoratidzwa sechikamu chegenome, misiyano inocherechedzwa muchikamu chemajini mushanduro, ribosomal chimiro uye biogenesis zvikamu, uye kugadzirwa kwesimba nekushandura basa zvikamu, izvo zvinoumba yakakura ASxL5T. genome Chikamu chinofananidzwa neboka rimwechete riripo muT. oleiverans MIL-1T genome. Kusiyana neizvi, zvichienzaniswa neASxL5T genome, T. oleivorans MIL-1T ine chikamu chepamusoro chemajini mukudzokorora, kusanganisa uye kugadzirisa, uye zvikamu zvezvinyorwa. Sezvineiwo, musiyano mukuru pane zviri mukati mechikamu chega chega chinoshanda chemajini maviri ndiwo nhamba yemajini asingazivikanwe aripo muASxL5T (Mufananidzo 3B). Ongororo yekuvandudza yeKEGG modules yakaitwa, apo yega yega KEGG module inomiririra seti yemanyorerwo anotsanangurwa anoshanda mayuniti ekutsanangudza uye biological dudziro yegenome sequence data. Kuenzanisa kwekugoverwa kwemajini mune yakazara KOG module nzira yeASxL5T uye muorivhi MIL-1T inoratidzwa muMufananidzo 3C. Ongororo iyi inoratidza kuti kunyangwe ASxL5T ine sarufa yakakwana uye nitrogen metabolic nzira, T. oleiverans MIL-1T haina. Kusiyana neizvi, T. oleiverans MIL-1T ine yakazara cysteine ​​​​uye methionine metabolic nzira, asi haina kukwana muASxL5T. Naizvozvo, ASxL5T ine hunhu hwemodule ye sulfate assimilation (inotsanangurwa seti yemajini anogona kushandiswa se phenotypic markers, senge metabolic capacity kana pathogenicity; https://www.genome.jp/kegg/module.html) Mu T. Oleiverans MIL-1T. Kuenzanisa iyo gene yemukati yeASxL5T nerondedzero yemajini inoratidza mararamiro ehudzvinyiriri hazvigoneke. Kunyange zvazvo waaL gene encoding ligase yakabatana neO antigen polysaccharide kusvika pakati iripo muASxL5T genome (asi inowanikwa mumabhakitiriya akawanda eGram-negative), tryptophan 2,3-dioxygenase (TDO ) Genes inogona kusanganisira 60 amino. asidhi matunhu anowanzo kuwanikwa mumabhakitiriya anodya asipo. Iko hakuna mamwe emhando dzemajini ane hunhu muASxL5T genome, kusanganisira iwo encoding enzymes anobatanidzwa mune isoprenoid biosynthesis munzira mevalonate. Ziva kuti hapana geneti rinodzorwa regini gntR muboka rezvikara rakaongororwa, asi majini matatu akafanana negntR anogona kuonekwa muASxL5T.
Izvo phenotypic maitiro eASxL5T akapfupikiswa muTebhu 3 uye akaenzaniswa ne phenotypic maitiro eiyo genera 23, 24, 25, 26, uye 27 yakataurwa mumabhuku. Isolates kubva kuT. marinus, T. olevorans, B. sanyensis, uye Oceanobacter kriegii inoshanda, inoshivirira munyu, miviri yakaumbwa neoxidase-positive tsvimbo, asi inenge isina mamwe maitiro ephenotypic ane ASxL5T. Avhareji pH yegungwa i8.1 (https://ocean.si.edu/ocean-life/invertebrates/ocean-acidification#section_77), iyo inoratidzwa muT. marinus, T. olevorans, B. sanyensis uye O. kriegii. ASxL5T inokodzera iyo yakakura pH renji (4-9) yakajairwa yemhando dzisiri dzemugungwa. Phenotypic maitiro eThalassolituus sp. C2-1. Unknown. Iyo tembiricha yekukura yeASxL5T inowanzo fararira kupfuura yemhando dzemugungwa (4-42 ° C), kunyangwe mamwe asi asiri ese T. marinus akasarudzika anoshivirira kupisa. Kusakwanisa kukura ASxL5T mubroth media kwakadzivirira imwe phenotypic characterization. Shandisa API 20E kuti uedze zvinhu zvakarukwa kubva mundiro yeBA, ONPG, arginine dihydrolase, lysine decarboxylase, ornithine decarboxylase, citrate utilization, urease, tryptophan deaminase, gelatin hydrolysis Enzyme, mhinduro dzebvunzo dzese dzaive dzisina kunaka, asi hapana indole, acetoin uye H2S. dzakagadzirwa. Makabhohaidhiretsi asina kuviriswa anosanganisira: glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin uye arabinose. Kuenzaniswa neyakadhindwa yakabatana yereferenzi strains, iyo cellular fatty acid profile yeASxL5T strain inoratidzwa muTafura 4. Iyo huru cellular fatty acids ndeyeC16: 1ω6c uye / kana C16: 1ω7c, C16: 0 uye C18: 1ω9. Hydroxy fatty acids C12:0 3-OH uye C10:0 3-OH iripo zvakare. Chiyero cheC16: 0 muASxL5T chakakwirira kudarika kukosha kwakataurwa kwegenera rine hukama. Kusiyana neizvi, kana ichienzaniswa neyakataurwa T. marinus IMCC1826TT, chiyero cheC18: 1ω7c uye / kana C18: 1ω6c muASxL5T inoderedzwa. oleivorans MIL-1T uye O. kriegii DSM 6294T, asi haina kuonekwa muB. sanyensis KCTC 32220T. Kuenzanisa fatty acid profiles yeASxL5T uye ASxLS yakaburitsa misiyano isinganzwisisike muhuwandu hwemafuta ega ega pakati pemhando mbiri, inoenderana negenomic DNA kutevedzana kwemhando imwe chete. Hapana poly-3-hydroxybutyrate (PHB) zvimedu zvakaonekwa pachishandiswa bvunzo yeSudan nhema.
Iyo predation chiitiko cheASxL5T mabhakitiriya akaongororwa kuti aone huwandu hwemhuka. Utachiona uhwu hunogona kuumba maplaque pamarudzi eCampylobacter, anosanganisira: Campylobacter suis 11608T, Campylobacter jejuni PT14, Campylobacter jejuni 12662, Campylobacter jejuni NCTC 11168T; Escherichia coli NCTC 12667; C. helveticus NCTC 12472; C lari NCTC 11458 uye C. upsaliensis NCTC 11541T. Shandisa tsika dzakanyorwa muchikamu chekugadzirisa chiyero chenzira yekuedza huwandu hwakawanda hweGram-negative uye Gram-positive bacteria. Mhedzisiro yacho inoratidza kuti ASxL5T inogona zvakare kushandiswa muEscherichia coli NCTC 86 uye Citrobacter freundii NCTC 9750T. Plaques akaumbwa paKlebsiella oxytoca 11466. Kubatana kweTEM neE. coli NCTC 86 inoratidzwa muMufananidzo 4A-D, uye kuwirirana neCampylobacter jejuni PT14 uye Campylobacter suis S12 inoratidzwa muMufananidzo 4E-H pakati. Nzira yekurwisa inoita seinosiyana pakati pemhando dzemhuka dzakaedzwa, nemasero eE coli akabatanidzwa kune rimwe nerimwe ASxL5T sero uye akamiswa nechemberi pamwe nesero yakawedzerwa isati yadsorption. Kusiyana neizvi, ASxL5T inoita seinonamatira kuCampylobacter kuburikidza nenzvimbo imwe chete yekubatana, kazhinji inosangana nepamusoro pesero remhuka uye pedyo nepamusoro peCampylobacter cell (Mufananidzo 4H).
TEM inoratidza kudyidzana pakati peASx5LT nemhuka: (AD) uye E. coli mhuka; (EH) uye C. jejuni mhuka. (A) Yakajairika ASx5LT cell yakabatana kune imwechete E. coli (EC) sero; (B) A filamentous ASx5LT yakasungirirwa kune imwechete EC cell; (C) A filamentous ASx5LT cell yakabatanidzwa kune akawanda EC masero; (D) Attachment Diki ASx5LT masero pane imwe E. coli (EC) sero; (E) imwe chete ASx5LT cell yakabatana neCampylobacter jejuni (CJ) sero; (F) ASx5LT inorwisa C. hyointestinalis (CH) masero; (G) maviri One ASx5LT cell akarwisa CJ cell; (H) Maonero epedyo eiyo ASx5LT yekunamatira poindi, padhuze nepamusoro peiyo CJ cell (bar 0.2 μm). Iyo bhawa inomiririra 1 μm mukati (A-G).
Hutachiona hunotyisa hwakashanduka kuti hutore mukana wezvakawanda zvezvipfuyo. Zviripachena, ivo varipo zvakanyanya munzvimbo dzakawanda dzakasiyana. Nekuda kwehukuru hwakatetepa hwehuwandu hwevanhu, zvinokwanisika kutsaura ASxL5T mabhakitiriya kubva kune slurry uchishandisa nzira yekuparadzanisa phage. Iyo genomic relevance yeASxL5T kunhengo dze oceanospirillaceae mhuri yemabhakitiriya emugungwa inoshamisa, kunyangwe iyo mhenyu inoshivirira munyu uye inogona kukura pasvikiro rine 5% munyu. Kuongororwa kwemhando yemvura yeslurry kwakaratidza kuti sodium chloride yemukati yaive isingasviki 0.1%. Naizvozvo, madhaka ari kure nemhoteredzo yegungwa-zvese nzvimbo uye nemakemikari. Kuvapo kwezvitatu zvine hukama asi zvakasiyana kubva kunzvimbo imwechete kunopa humbowo hwekuti zvikara izvi zviri kubudirira munzvimbo iyi isiri yegungwa. Uye zvakare, kuongororwa kwemicrobiome (mafaira edata anowanikwa kubva ku https://www.ebi.ac.uk/ena/browser/view/PRJEB38990) yakaratidza kuti imwecheteyo 16S rRNA gene sequence inowanikwa mu50 yakanyanya kuwanda inoshanda mutero (OTU). ) Mune mashoma ekuenzanisa nguva dzedhaka. Mabhakitiriya akati wandei asina kugadzirwa akawanikwa mudura reGenbank, ane 16S rRNA gene sequences akafanana neASxL5T mabhakitiriya. Aya akatevedzana, pamwe chete neakatevedzana eASxL5T, ASxS5, uye ASxO5, anoita seanomiririra akasiyana akapatsanurwa kubva kuThalassolituus neOceanobacter (Mufananidzo 2). Mhando nhatu dzemabhakitiriya asina kugadzirwa (GQ921362, GQ921357 uye GQ921396) akaparadzaniswa kubva mumvura inoputika pakadzika makiromita 1.3 mumugodhi wegoridhe wekuSouth Africa muna 2009, uye mamwe maviri (DQ256320 uye DQ337006) aibva kuSouth Africa mumvura yepasi muna 2005). Iyo 16S rRNA gene sequence ine hukama zvakanyanya neASxL5T chikamu cheiyo 16S rRNA gene sequence yakawanikwa kubva mukupfumisa tsika yejecha sediments yakawanikwa kubva kumahombekombe ekuchamhembe kweFrance muna 2006 (nhamba yekuwana AM29240828). Imwe yakanyatsoenderana 16S rRNA gene kutevedzana kubva kuhutachiona husina tsika HQ183822.1 yakawanikwa kubva mutangi rekuunganidza rakanyudzwa kubva mudhorobha rinoraswa tsvina muChina. Zviripachena, mabhakitiriya eASxL5T haamiriri zvakanyanya mumadhatabheti etaxonomic, asi aya anoteedzana kubva kumabhakitiriya asina tsika anogona kumiririra zvipenyu zvakafanana neASxL5T, izvo zvinogoverwa pasirese, kazhinji munzvimbo dzakaoma. Kubva pane yese genome phylogenetic kuongororwa, hama yepedyo kune ASxL5T ndeye Thalassolituus sp. C2-1, T. marinus, T. oleivorans. Uye O. kriegii 23, 24, 25, 26, 27. Thalassolituus inhengo ye marine obligate hydrocarbon fragmentation bacteria (OHCB), iyo yakapararira munzvimbo dzegungwa nepasi, uye inowanzova iyo inotonga mushure mezviitiko zvekusvibiswa kwehydrocarbon30,31. Marine mabhakitiriya haasi nhengo dzeboka reOHCB, asi ari oga kubva kunharaunda yegungwa.
Phenotypic data inoratidza kuti ASxL5T imhando nyowani uye inhengo yeyaimbove isingazivikanwe genus mumarine spirospiraceae mhuri. Parizvino hapana chiyero chakajeka chekuisa mhando dzichangoparadzaniswa dzega kuita rudzi rutsva. Kuedza kwakaitwa kuona miganhu yegenera yepasirese, semuenzaniso, zvichibva pachikamu chegenome cheprotein inochengetedza (POCP), zvinokurudzirwa kuti kukosha kwekucheka-cheka kuve 50% kwakafanana nereferensi strain33. Vamwe vanokurudzira kushandisa AAI hunhu, idzo dzine zvakanakira POCP nekuti dzinogona kuwanikwa kubva kune isina kukwana genomes34. Munyori anotenda kuti kana kukosha kweAAI kuri pasi pe74% kana ichienzaniswa nemhando yemhando yemhando yemhando, dhizha rinomiririra rimwe genera. Iyo modhi genus mumarine spirillaceae is marine spirillum, uye mhando yemhando ndeye O. linum ATCC 11336T. Kukosha kweAAI pakati peASxL5T neO. linum ATCC 11336T ndeye 54.34%, uye kukosha kweAAI pakati peASxL5T neT. oleivorans MIL-1T (genus type strains) i67.61%, zvichiratidza kuti ASxL5T inomiririra rudzi rutsva rwakasiyana neThalassolituus. Uchishandisa 16S rRNA gene sequence seyakajairwa chiyero, yakakurudzirwa genus delimitation muganho ndeye 94.5% 35. ASxL5T inogona kuiswa muThalassolituus genus, ichiratidza 95.03% 16S rRNA sequence identity ine T. oleivorans MIL-1T uye 96.17%. marinus IMCC1826T. Zvisinei, ichaiswawo muBacteroides genus ine 94.64% 16S rRNA gene identity neB. sanyensis NV9, zvichiratidza kuti kushandiswa kwejini rimwe chete rakadai se16S rRNA gene rinogona kutungamirira mukusarudzwa kwemaitiro uye kugoverwa. Imwe nzira inokurudzirwa inoshandisa ANI uye Genome Alignment Score (AF) kuongorora kuunganidzwa kwemapoinzi edhata kubva kumarudzi ese uye asiri emhando yemhando iripo. Munyori anokurudzira kubatanidza muganho wegenus neinflection point yeinofungidzirwa genus chaiyo kune taxa iri kuongororwa. Nekudaro, kana pasina akakwana akazara genome sequences kubva kuThalassolituus isolates, hazvigoneke kuona kuti ASxL5T ndeyeThalassolituus genus neiyi nzira. Nekuda kwekuwanikwa kushoma kweakakwana genome sequences yekuongorora, iyo yose genome phylogenetic muti inofanira kududzirwa nekuchenjerera. Chechipiri, nzira dzese dzekuenzanisa genome hadzigone kuverengera misiyano yakakura muhukuru hweakaenzaniswa genomes. Vakayera kufanana kweakachengeterwa core single-copy genes pakati pegenera rine hukama, asi havana kufunga nezvehuwandu hwemajini asipo mune diki genome yeASxL5T. Zviripachena, ASxL5T nemapoka anosanganisira Thalassolituus, Oceanobacter, uye Bacterioplanes vane madzitateguru akafanana, asi kushanduka kwakatora nzira yakasiyana, zvichiita kuti kuderedzwa kwejenome, iyo inogona kunge iri yekuenderana nemararamiro emhuka. Izvi zvakasiyana ne T. oleivorans MIL-1T, iyo yakakura 28% uye yakashanduka pasi pekumanikidzwa kwakasiyana kwezvakatipoteredza kushandisa mahydrocarbons23,30. Kuenzanisa kunonakidza kunogona kuitwa neanosungirwa intracellular parasites uye symbionts, seRickettsia, Chlamydia, uye Buchnera. Saizi yavo yegenome inenge 1 Mb. Iko kugona kushandisa host cell metabolites inotungamira mukurasikirwa nemajini, saka Yakave yakakosha evolutionary genomic degradation. Shanduko dzekuchinja kubva kumakemikari emugungwa zvinovaka muviri kuenda kumararamiro emhuka anogona kukonzera kuderedzwa kwakafanana kwekukura kwegenome. Kuongororwa kweCOG neKEGG kunoratidza huwandu hwemajini anoshandiswa kune mamwe mabasa uye misiyano yepasi rose mugenomic nzira pakati peASxL5T neT. oleivorans MIL-1T, izvo zvisiri nekuda kwekuwanikwa kwakapararira kwe mobile genetic elements. Musiyano muchikamu cheG + C chejenome rose reASxL5T i56.1%, uye yeT. oleivorans MIL-1T ndeye 46.6%, iyo inoratidzawo kuti yakaparadzaniswa.
Kuongororwa kweiyo coding yemukati yeASxL5T genome inopa maitiro ekuona mune phenotypic maitiro. Kuvepo kwemajini encoding type IV fimbriae (Tfp) kunonyanya kufarirwa nekuti anokurudzira kufamba kwesero, kunonzi kusvetuka-svetuka kwevanhu kana kugwinha, pasina flagella pamusoro. Sekureva kwemishumo, Tfp ine mamwe mabasa, anosanganisira predation, pathogenesis, biofilm kuumbwa, yakasikwa DNA kutora, otomatiki masero aggregation uye budiriro38. Iyo ASxL5T genome ine 18 majini encoding diguanylate cyclase (enzyme inokonzeresa shanduko ye2 guanosine triphosphate kuita guanosine 2 phosphate uye cyclic diGMP) uye 6 majini encoding inoenderana diguanylate cyclase phosphate diguanylate. Jeni re esterase (rinokonzeresa kushatiswa kwe cyclic di-GMP kuenda kuguanosine monophosphate) rinonakidza nekuti cycl-di-GMP mutumwa akakosha wechipiri ane chekuita nekusimudzira biofilm uye kupatsanura, kufamba, kubatanidza sero uye virulence 39, 40 mukuita. Izvo zvinofanirwa kucherechedzwa kuti muBdellovibrio bacteriovorus, cyclic kaviri GMP yakaratidzwa kudzora shanduko pakati pehupenyu hwemahara nemararamiro ehutsinye41.
Tsvagiridzo zhinji pamusoro pemabhakitiriya anodya yakanangana neBdellovibrio, Bdellovibrio-senge zvipenyu, uye Myxococcus marudzi. Iyi nemimwe mienzaniso inozivikanwa yemabhakitiriya anodya inoumba boka rakasiyana. Pasinei nekusiyana uku, seti yemapuroteni emhuri anoratidza phenotypes e11 anozivikanwa mabhakitiriya anodya akaonekwa3,22. Zvisinei, majini chete encoding O antigen ligase (waaL) akaonekwa, ayo anonyanya kuwanda muGram-negative mabhakitiriya. Iyi nzira yekuongorora haibatsire pakusarudza ASxL5T sechikara, pamwe nekuti inoshandisa nzira yekurwisa. Kuvepo kweakawanda akasiyana-siyana anoparadza mabhakitiriya genomes kuchabatsira kugadzira ongororo yekugadziridza iyo inotora humbowo hwekushanda uye mutsauko wezvakatipoteredza pakati penhengo dzeboka. Mienzaniso yemabhakitiriya anodya asina kuisirwa muongororo iyi anosanganisira nhengo dzeCupriavidus necator42 uye Bradymonabacteria43, nekuti sezvo vaongorori vanoongorora nharaunda dzakasiyana siyana dzehutachiona, mutero wakawanda unotangwa unotangwa.
Chinhu chinonyanya kukosha cheASxL5T mabhakitiriya akabatwa neTEM mufananidzo ndeye yakasarudzika uye inoshanduka morphology, iyo inogona kukurudzira kudyidzana nemabhakitiriya emhuka. Rudzi rwekudyidzana rwakaonekwa rwakasiyana nemamwe mabhakitiriya anodya uye haasati ambowanikwa kana kutaurwa. Iyo yakarongwa ASxL5T yehupenyu hwehudzvinyiriri inoratidzwa muMufananidzo 5. Kune mienzaniso mishoma muzvinyorwa zvine akafanana apical zvimiro sezvatinoshuma pano, asi iyi mienzaniso inosanganisira Terasakiispira papahanaumokuakeensis, marine spirillum bhakitiriya ine dzimwe nguva apex kuwedzera 44, uye Alphaproteobacteria pusillasa, Terasakiispira papahanaumokuakeensis. , yaimbova yerudzi rweOceanospirillum, kuratidzira Izvo zvinonzi "polar film" 45. Mafomu eCocci anowanzoonekwa mutsika dzekare, kunyanya kune mabhakitiriya ane maitiro akakombama, akadai seVibrio, Campylobacter, uye Helicobacter 46, 47, 48, iyo inogona kumiririra nyika yakasvibiswa. Rimwe basa rinodiwa kujekesa kutenderera kwehupenyu hweASxL5T mabhakitiriya. Kuti uone kuti inobata sei uye inobata sei, uye kuti genome yayo inoisa biologically inoshanda komisheni inogona kushandiswa pakurapa kana biotechnological zvinangwa.
Tsanangudzo yeVenatorbacter gen. Mbudzi Venatorbacter (Ven.a.tor, ba'c.ter, L. inoumbwa nemavenator kubva kuL. n. venator,'hunter' uye Gr. n. bacter,'a rod'. Venatorbacter,'a hunting Rod' . Masero ane aerobic, anoshivirira munyu, akakombama Gram stain negative, maekisesaizi eCatalase uye oxidase anoita kuunganidza muhuwandu hwekushisa kwe4 kusvika ku42 °C Ingrown pH ye4-9 haina kujairika muhozhwa dzemugungwa, dzakawanda dzine acidic acids ndeyeC16: 1ω6c. C16:0 uye C18:1ω9 ; 3-OH inowanikwa se hydroxy fatty acids Haakuri muto we DNA G + C zvinyorwa zve 56.1 mol chinzvimbo cherudzi urwu chiri mumhuri.
Tsanangudzo yeVenatorbacter cuculus sp. Mbudzi Venatorbacter cucullus (cu'cull.us.; L. n. cucullus zvinoreva fairing).
Pamusoro pezvo, chinotsanangudza chejenasi ichi ndechekuti kana yakura paBA kana BHI, maseru akareba 1.63 µm uye 0.37 µm paupamhi. Makoroni ari paBHI agar madiki kwazvo, anosvika 2 mm mudhayamita mushure memaawa makumi manomwe nemaviri. Iwo ari beige, anopenya, akatenderera, anopenya uye anopenya. Nhengo dzerudzi urwu dzinogona kushandisa Escherichia coli uye Klebsiella. Campylobacter uye mamwe akawanda mabhakitiriya eGram-negative anoshanda semhuka.
Iyo yakajairika strain ASxL5T yakaparadzaniswa nemukaka wemombe muNottinghamshire, UK, uye yakaiswa muNational Type Culture Collection (UK): nhamba yekupinda NCTC 14397 uye Netherlands Bacterial Culture Collection (NCCB) nhamba yekupinda NCCB 100775. Iyo yakazara genome sequence yeASxL5T yakaiswa muGenbank maererano nekuwedzera kweCP046056.
ASxL5T mabhakitiriya akaparadzaniswa nemukaka wemombe uchishandisa phage isolation tekinoroji9,49. Slurry yakadiridzwa 1:9 (w/v) muSM buffer (50 mM Tris-HCl [pH 7.5], 0.1 M NaCl, 8 mM MgSO4.7H2O uye 0.01% gelatin; Sigma Aldrich, Gillingham, UK), Wozoisa pa 4°C kwemaawa makumi maviri nemana, ichitenderera zvishoma nezvishoma kuburitsa zvikara mubuffer. Kumiswa kwakaiswa centrifuged pa3000g kwemaminetsi matatu. Iyo supernatant yakaunganidzwa uye centrifuged pa13,000g kechipiri kwemaminitsi mashanu. Iyo supernatant yakabva yapfuudzwa ne 0.45 µm membrane sefa (Minisart; Sartorius, Gottingen, Germany) uye 0.2 µm membrane sefa (Minisart) kubvisa chero masero ebhakitiriya asara. ASxL5T inogona kupfuudza mafirita aya. Iyo yakapfava agar lawn yeCampylobacter enterosus S12 (NCBI yekuwedzera nhamba CP040464) kubva kune imwecheteyo slurry yakagadzirwa uchishandisa yakajairwa matekiniki. Iyo yakasefa slurry yakagovaniswa pane imwe neimwe yeaya magasi cell plate mu 10 µl madonhwe mumatatu uye akabvumidzwa kuoma. Iyo ndiro yakavharwa mu microaerophilic tank pa 37 ° C kwemaawa 48 pasi pemamiriro e microaerobic (5% O2, 5% H2, 10% CO2, uye 80% N2). Iyo yakawanikwa plaque inooneka yakabudiswa muSM buffer uye yakaendeswa kune itsva lawn yeC. hyointestinalis S12 kuti ienderere mberi kuparadzira zvipenyu zvine lysed. Pane imwe nguva yakatemwa kuti mabhakitiriya ndiwo anokonzera lytic plaque uye kwete phage, edza kukura zvisikwa zvakasununguka kubva kune muenzi uye kuwedzera kuzviratidza. The aerobic tsika yakaitwa pa 37 ° C ne 5% v / v defibrinated bhiza ropa (TCS Biosciences Lt, Buckingham, UK, kuwedzera). Zvinoenderana nemirairo yeNational Clinical Standards Committee, iyo disc diffusion nzira inoshandiswa kune antibacterial susceptibility test. BHI agar yakagadzirwa pa 37 ° C ichishandisa dhisiki rine zvinotevera antibiotics (Oxoid) ye aerobic tsika: amoxicillin uye clavulanic acid 30 µg; cefotaxime 30 µg; streptomycin 10 µg; ciprofloxacin 5 µg; Ceftazidime 30 µg Nalidixic asidhi 30 µg; Imipenem 10 µg; Azithromycin 15 µg; Chloramphenicol 30 µg; Cefoxitin 30 µg; Tetracycline 30 µg; Nitrofurantoin 300 µg; Aztreonam 30 µg; Ampicillin 10 µg; Cefpodoxime 10 µg; Trimethoprim-Sulfamethoxazole 25 µg. Kushivirira kwemunyu kwakatangwa neaerobic incubation paBHI agar plates pa37 °C. Yekuwedzera NaCl yakawedzerwa kumapureti eBHI agar kuti ape huwandu hwekutarisa hunosvika gumi muzana w/v. The pH range inotarwa ne aerobic culture paBHI agar plates pa 37°C, apo pH range yakagadziridzwa kusvika pakati pe 4 ne 9 ne sterile HCl kana sterile NaOH, uye chinangwa chepH kukosha kunosimbiswa tisati tadira ndiro. Kuongorora masero emafuta asidhi, ASxL5T yakagadzirwa paBHI agar kwemazuva matatu uye aerobic pa37 °C. Zvinoenderana neMIDI (Sherlock Microbial Identification System, vhezheni 6.10) yakajairwa protocol yeFERA Science Ltd, (York, UK), cell fatty acids yakatorwa, kugadzirirwa uye kuongororwa.
KuTEM, ASxL5T yakaitwa aerobic nekuparadzira zvakafanana paBA pa37 ° C kwemaawa makumi maviri nemana, ndokuzokohwewa kuita 1 ml ye3% (v/v) glutaraldehyde mu0.1 M cacodylate buffer pakupisa kwekamuri Gadzirisa kweawa imwe, wozoti centrifuge. pa10,000 g kwemaminitsi matatu. Wobva wamisa zvinyoro nyoro pellet mu600 μl 0.1 M cacodylate buffer. Chinja iyo yakamiswa ASxL5T kumiswa kuFormvar/carbon firimu pane mazana maviri mesh mhangura grid. Utachiona hwakasvibiswa ne0.5% (w / v) uranyl acetate kweminiti 1 uye yakaongororwa neTEM uchishandisa TEI Tecnai G2 12 Biotwin microscope. Sezvataurwa pamusoro, batanidza nhamba imwechete yemhuka uye mhuka inodya muNZCYM muto (BD Difco™, Fisher Scientific UK Ltd, Loughborough) uye incubate kwemaawa makumi mana nemasere pasi pe microaerobic mamiriro eCampylobacter kana Campylobacter pa 37 ° C , Kubatana kwechikara uye mhuka. yakaongororwawo neTEM. Aerobic mamiriro eEscherichia coli. Ongorora wakazvimirira mhuka uye mabhakitiriya anodya kuti aone chero shanduko musero morphology nekuda kwekupambwa. Nzira yeSudan nhema yakashandiswa kuoptical microscopy yePHB kuunganidza.
Kura ASxL5T tsika dzehusiku nekuzora kukura pamapureti eBHI kana BA ane swab isina utachiona. Unganidza masero eASxL5T woamisa muMRD (CM0733, Oxoid), uye woaisa pa4 ° C kwemazuva manomwe kuti auraye masero. The NCTC reference kana laboratory stock bacterial culture yakaiswa muBHI muto kana No. 2 nutrient broth (CM007, Oxoid), incubated overnight, centrifuged at 13,000g and resuspended in MRD until OD600 was 0.4. Tsika: Bacillus subtilis NCTC 3610T, Citrobacter freundii NCTC 9750T, Enterobacter aerogenes NCTC 10006T, Enterococcus faecalis NCTC 775T, Escherichia coli NCTC 86, Klebsiella 1uTC1co4 Oxysto6 10817. NCTC 10861, Staphylococcus aureus NCTC 8532T, Streptococcus pneumoniae NCTC 7465T, Yersinia enterocolitica NCTC 10460. The Campylobacter host was microaerobically incubated paBA plates pa 37 ° C uye suspendedMNZZ yakamiswa. Iwo akaedzwa Campylobacter mauto ndeaya: C. coli 12667 NCTC, C. jejuni 12662, C. jejuni PT14, C. jejuni NCTC 11168T, C. helveticus NCTC 12472, C. lari NCTC 114548, C. PT14, C... Unganidza masero muMRD, centrifuge pa 13,000g uye udzoke zvakare muMRD kusvikira OD600 iri 0.4. Wedzera aliquot ye0.5 ml kuturika kune 5 ml yakanyunguduka NZCYM yepamusoro agar (0.6% agar) uye uidururire pa1.2% NZCYM pasi ndiro. Mushure mekurapa nekuomeswa, iyo yakanyungudutswa ASxL5T yakagovaniswa semadonhwe makumi maviri µl pane yega lawn board mune katatu. Itsika tembiricha uye mhepo zvinoenderana nezvinodiwa bvunzo mabhakitiriya.
Shandisa GenElute ™ Bacterial Genomic DNA Kit (Sigma Aldridge) kugadzirira DNA kubva kune mabhakitiriya akaparadzaniswa. Maitiro akajairwa akashandiswa PCR kukwidziridzwa kwe16S rRNA gene uye chigadzirwa kutevedzana kutsunga uchishandisa dhayi kumisa chemistry (Eurofins Value Read Service, Germany). Shandisa chirongwa cheBLAST-N kuenzanisa aya anotevedzana nemamwe 16S rRNA gene sequences kuona uye kuunganidza zvine hukama mhando. Izvi zvakarongeka uchishandisa ClustalW muMEGA X chirongwa. Muti we phylogenetic wakavakwa patsva uchishandisa MEGA X uchishandisa iyo yakanyanya mukana nzira inoenderana neiyo Tamura-Nei modhi, ine 1000 inotungamirwa makopi54. Shandisa PureLink™ Genomic DNA Kit (Fisher Scientific, Loughborough, UK) kubvisa DNA yekutevedzana kwegenome. Iyo genome sequence yeASxL5T yakatemwa uchishandisa iyo Illumina MiSeq musanganiswa, iyo ine 250 bp inopera kaviri kuverenga inoumbwa neraibhurari yakagadzirwa uchishandisa Nextera labeling kit uye 2 kusvika 20 kb kureba kuverenga kubva paPacBio papuratifomu. Genomics DNA Sequencing Research Facility paSembia University. Iyo genome yakaunganidzwa uchishandisa CLC Genomics Workbench 12.0.3 (Qiagen, Aarhus, Denmark). ASxL5T tsika dzakaiswa muNational Type Culture Collection (UK) uye Netherlands Bacterial Culture Collection (NCCB). Iwo genomes ezvipenyu zvine hukama anoshandiswa pakuenzanisa ndeaya: Thalassolituus oleivorans MIL-1T (nhamba yekuwana HF680312, yakakwana); Bacterioplanes sanyensis KCTC 32220T (nhamba yekupinda BMYY01000001, isina kukwana); Oceanobacter kriegii DSM 6294T (nhamba yekuwana NZ_AUGV00000000, isina kukwana); Marinamonas nharaunda DSM 5604T (yakawedzera ASM436330v1, isina kukwana), Oceanospirullum linum ATCC 11336T (yakawedzera MTSD02000001, isina kukwana) uye Thalassolituus sp. C2-1 (wedzera NZ_VNIL01000001, isina kukwana). Shandisa JGI Genome Portal36 pa https://img.jgi.doe.gov//cgi-bin/mer/main.cgi?section=ANI&page= kuona iyo alignment mamakisi (AF) uye avhareji nucleic acid identity (ANI). Muvaviri. Nzira yeRodriguez-R & Konstantinidis55 yakashandiswa kugadzirisa amino acid identity (AAI). Shandisa GToTree 1.5.5411, 12, 13, 14, 15, 16, 17, 18 kugadzira fungidziro yemuti wephylogenetic. Iyo genome yekupinda inomiririra iripo referensi genome inosarudzwa kubva kune referenzi genera rinoonekwa seine hukama neASxL5T kubva ku16S rRNA phylogeny. Akazivisa muti uchishandisa muti unodyidzana wehupenyu online chishandiso (https://itol.embl.de/). Iyo inoshanda tsinhiro uye ongororo yeiyo ASxL5T genome inoitwa pachishandiswa BlastKOALA KEGG online chishandiso uchishandisa KEGG (Kyoto Encyclopedia yeGenes uye Genomes) module enrichment distribution. Kugoverwa kweCOG zvikamu (orthologous groups) kunotemerwa kushandisa eggNOG-mapper online tool.
Pérez, J., Moraleda-Muñoz, A., Marcos-Torres, FJ uye Muñoz-Dorado, J. Bacterial predation: 75 makore uye inoenderera mberi! . zvakatipoteredza. microorganism. 18, 766–779 (2016).
Linares-Otoya, L. etc. Kusiyana-siyana uye antibacterial simba remabhakitiriya anoparadza pamhenderekedzo yePeruvia. March zvinodhaka. 15. E308. https://doi.org/10.3390/md15100308 (2017).
Pasternak, Z. et al. Kuburikidza nemajini avo, unovanzwisisa: iwo genomic hunhu hwemabhakitiriya anodya. ISME J. 7, 756–769 (2013).
Sockett, RE Hupenyu hwekupamba hweiyo bacteriophage Bdellovibrio. install. Mufundisi ma microbes. 63, 523–539 (2009).
Korp, J., Vela Gurovic, MS & Nett, M. Mishonga inorwisa mabhakitiriya kubva kune mabhakitiriya anodya. Beilstein J. Histochemistry 12, 594-607 (2016).
Johnke, J., Fraune, S., Bosch, TCG, Hentschel, U. & Schulenburg, H. Bdellovibrio uye zvipenyu zvakafanana zvinofanotaura nezvekusiyana kwe microbiome mumapoka akasiyana-siyana. microorganism. Ecology. 79, 252–257 (2020).
Vila, J., Moreno-Morales, J. uye Ballesté-Delpierre, C. Ziva mamiriro ezvinhu ezvino emishonga inorwisa mabhakitiriya. kliniki. microorganism. Infect. https://doi.org/10.1016/j.cmi.2019.09.015 (2019).
Hobley, L. et al. Kudzvinyirirwa kwepage uye phage kunogona kubvisa E. coli mhuka pasina kana imwe nhapwa. J. Utachiona. 202, e00629-19. https://doi.org/10.1128/JB.00629-19 (2020).
El-Shibiny, A., Connerton, PL & Connerton, IF Kuverenga uye kusiyana-siyana kweCampylobacter uye bacteriophages yakaparadzaniswa panguva yekudyisa kwemahara-range uye organic huku. Nzvimbo yekushandisa. microorganism. 71, 1259–1266 (2005).
Wilkinson, DA etc. Gadzirisa iyo genomic taxonomy uye epidemiology yeCampylobacter swine. sainzi. Representative 8, 2393. https://doi.org/10.1038/s41598-018-20889-x (2018).
Lee, MD GToTree: Mushandisi-ane hushamwari mafambiro ehurongwa hwegenomics. Bioinformatics 35, 4162–4164 (2019).
Edgar, RC MUSCLE: Iyo yakawanda kutevedzana kurongeka nzira inoderedza nguva uye nzvimbo yakaoma. BMC biological ruzivo. 5, 113 (2004).
Capella-Gutiérrez, S., Silla-Martínez, JM & Gabaldón, T. TrimAl: Chishandiso chekugadzirisa otomatiki uye kuchekerera mukuongorora kukuru kwephylogenetic. Bioinformatics 25, 1972-1973 (2009).
Hyatt, D., LoCascio, PF, Hauser, LJ & Uberbacher, EC gene uye metagenomic sequence shanduro inotanga kufanotaura nzvimbo. Bioinformatics 28, 2223-2230 (2012).
Shen, W. & Xiong, J. TaxonKit: Cross-platform uye inoshanda NCBI classification toolkit. Bio Rxiv. (Yakasvika musi waChikumi 1, 2021); https://www.biorxiv.org/content/10.1101/513523v1 (2019).
Mutengo, MN, Dehal, PS & Arkin, AP FastTree 2-inofungidzira yakakura mukana wemuti une kurongeka kukuru. PLoS One 5, e9490 (2010).
Tange, O. GNU Parallel. (Yakasvika musi waChikumi 1, 2021); https://zenodo.org/record/1146014#.YOHaiJhKiUk (2018).
Kanehisa, M. & Goto, S. KEGG: Kyoto Encyclopedia yeGenes uye Genomes. Nucleic acid research. 28, 27-30 (2000).
Czech Republic, L. etc. Basa re extremolytes ectoine uye hydroxyectoine sezvidziviriro zvekushungurudzika uye zvinovaka muviri: genetics, system genomics, biochemistry, uye structural analysis. Gene (Basel). 9. E177. https://doi.org/10.3390/genes9040177 (2018).
Gregson, BH, Metodieva, G., Metodiev, MV, Golyshin, PN & McKew, BA Differential protein expression panguva yekukura kweiyo inosungirwa marine hydrocarbon-inosvibisa bhakitiriya Thalassolituus oleivorans MIL-1 panguva yekukura kwepakati uye refu cheni alkanes. pamberi. microorganism. 9, 3130 (2018).
Pasternak, Z., Ben Sasson, T., Cohen, Y., Segev, E., uye Jurkevitch, E. Imwe nzira itsva yekuenzanisa genomics yekutsanangura zviratidzo zve phenotypic-specific inotaridza nhaka chaiyo muchiratidzo chebhakitiriya rinodya. Public Sayenzi Raibhurari Imwe. 10. e0142933. https://doi.org/10.1371/journal.pone.0142933 (2015).
Yakimov, MM, etc. Thalassolituus oleivorans gene. Mbudzi, sp. nov., rudzi rutsva rwebhakitiriya remugungwa rinonyanya kushandisa mahydrocarbon. internationality. J. System. evolution. microorganism. 54, 141–148 (2004).
Wang, Y., Yu, M., Liu, Y., Yang, X. & Zhang, XH Bacterioplanoides pacificum gen. Mbudzi, sp. Muna November, yakaparadzana nemvura yomugungwa inotenderera muSouth Pacific. internationality. J. System. evolution. microorganism. 66, 5010–5015 (2016).