Ubwoko bushya bwa Gram-negative, aerobic, bwihanganira umunyu, bukora, bumeze nkinkoni, hamwe na bagiteri ASxL5T yitaruye icyuzi cy’inka cy’inka i Nottinghamshire, mu Bwongereza, kandi ikoresha Campylobacter nk'umuhigo wacyo. Nyuma, andi moko ya Campylobacter hamwe nabagize umuryango wa Enterobacteriaceae bavumbuwe nkumuhigo. Nyuma ya subculture idafite selile yakira, gukura kwa aseptic kwagaragaye kuri Brain Heart Infusion Agar. Imiterere myiza yo gukura ni 37 ° C na pH ni 7. Ikwirakwizwa rya electron microscopi ryerekanye ibintu bimwe na bimwe bidasanzwe byimiterere ijyanye no kubona umuhigo. Isesengura rya phylogeneque ukoresheje urutonde rwa 16S rRNA rwerekanaga ko kwigunga bifitanye isano numuryango wumuryango wa Marine Spirulina, ariko ntibishobora gushyirwa mubikorwa nkumunyamuryango wubwoko bwose buzwi. Urutonde rwa genome yose ya ASxL5T yemeje umubano nabanyamuryango ba marine spirochetes. Ubushakashatsi bwakozwe bwerekanye ko ASxL5Ts nyinshi zisangiye 16S rRNA ikurikirana hamwe na bagiteri nyinshi zidafite umuco ziva mu nyanja, hejuru yubutaka n’amazi yo mu butaka. Turasaba ko amoko ASxL5T agereranya ubwoko bushya mubwoko bushya. Turasaba izina Venatorbacter cucullus gen. Ugushyingo, sp. Ugushyingo, ASxL5T yakoreshejwe nkubwoko bwubwoko.
Indwara ya bagiteri ni bagiteri zigaragaza ubushobozi bwo guhiga no kwica izindi bagiteri nzima kugirango zibone ibikoresho bya biosintetike n'imbaraga. Ibi bitandukanye no gukira kwintungamubiri muri mikorobe zapfuye, kandi biratandukanye kandi n’imikoranire ya parasitike, aho bagiteri zigirana umubano wa hafi na nyirazo utabishe. Indwara ziterwa na bagiteri zahinduye ubuzima butandukanye kugirango zungukire ku biribwa byinshi mu bibanza biboneka (nk'ahantu ho mu nyanja). Ni itsinda ryamatagisi atandukanye, ahujwe gusa nubuzima bwabo bwihariye bwo kuboneza urubyaro1. Ingero za bagiteri zangiza zabonetse muri phyla zitandukanye, harimo: Proteobacteria, Bacteroide, na Chlorella.3. Nyamara, bacteri zize cyane zize cyane ni Bdellovibrio na Bdellovibrio-n’ibinyabuzima bisa (BALOs4). Indwara ya bagiteri ni isoko itanga ibyiringiro bishya bikoresha ibinyabuzima hamwe na antibacterial agent5.
Indwara ya bagiteri yizera ko izamura mikorobe kandi ikagira ingaruka nziza kubuzima bwibidukikije, umusaruro n’umutekano6. Nubwo iyi mico myiza, hariho ubushakashatsi buke kuri bagiteri nshya zangiza bitewe ningorabahizi zo guhinga bagiteri no gukenera kwitegereza neza imikoranire ya selile kugirango twumve ubuzima bwabo bugoye. Aya makuru ntabwo yoroshye kubona mubisesengura rya mudasobwa.
Mu gihe cyo kongera imiti igabanya ubukana bwa mikorobe, harigwa ingamba nshya zo kurwanya indwara ziterwa na bagiteri, nko gukoresha bacteriofage na bagiteri zangiza 7,8. Bagiteri ya ASxL5T yitaruye mu mwaka wa 2019 ikoresheje ikoranabuhanga ryo kwigunga ry’amase yakuwe mu kigo cy’amata cya kaminuza ya Nottingham, Nottinghamshire. Intego yiperereza ni ugutandukanya ibinyabuzima bifite ubushobozi bwo kugenzura ibinyabuzima. Campylobacter hyointestinalis ni zoonotic pathogen, igenda ifitanye isano n'indwara zo munda z'abantu10. Iraboneka hose muri serumu kandi ikoreshwa nkintego yakiriye.
Bagiteri ya ASxL5T yitandukanije na jelly y'inka kubera ko byagaragaye ko icyapa cyakoze ku cyatsi cya C. hyointestinalis gisa n'icyakozwe na bagiteri. Ubu ni ubushakashatsi butunguranye, kuko igice cyibikorwa byo kwigunga kirimo gushungura binyuze muri 0.2 µm muyunguruzi, igenewe gukuraho selile. Isuzuma rya Microscopique ryibikoresho byakuwe kuri plaque ryerekanye ko bagiteri ntoya ya garama-mbi igoramye ya bagiteri itagizwe na polyhydroxybutyrate (PHB). Umuco wa aseptic udashingiye ku ngirabuzimafatizo uhiga uboneka muburyo bukomeye (nk'ubwonko bwo mu bwonko bwo mu bwonko (BHI) na agar y'amaraso (BA)), kandi imikurire yayo ni ntege. Iraboneka nyuma ya subculture hamwe na inoculum iremereye. Irakura neza kimwe na microaerobic (7% v / v ogisijeni) hamwe na ogisijeni yo mu kirere, ariko ntabwo iri mu kirere cya anaerobic. Nyuma yamasaha 72, diameter ya koloni yari nto cyane, igera kuri mm 2, kandi yari beige, yoroheje, izengurutse, convex kandi irabagirana. Igeragezwa ryibinyabuzima risanzwe rirabangamiwe kubera ko ASxL5T idashobora guhingwa mu buryo bwizewe mu bitangazamakuru byamazi, ibyo bikaba byerekana ko ishobora gushingira ku mibereho igoye yo kubaho kwa biofilm. Nyamara, guhagarika isahani byerekanaga ko ASxL5T ari aerobic, nziza kuri okiside na catalase, kandi ishobora kwihanganira 5% NaCl. ASxL5T irwanya 10 µg streptomycine, ariko irumva izindi antibiyotike zose zapimwe. Ingirabuzimafatizo za ASxL5T zasuzumwe na TEM (Ishusho 1). Iyo ikuze idafite selile zihiga kuri BA, selile ASxL5T ni ntoya ya Campylobacter, ifite uburebure bwa metero 1,63 (± 0.4), ubugari bwa 0.37 mm (± 0.08), hamwe na pole ndende (kugeza kuri 5 mm). Imibonano mpuzabitsina. Hafi ya 1,6% ya selile bigaragara ko ifite ubugari buri munsi ya 0.2 mm, bizemerera kunyura mubikoresho byo kuyungurura. Kwagura imiterere idasanzwe byagaragaye hejuru ya selile zimwe, bisa no kurenganura (latine cucullus) (reba imyambi muri 1D, E, G). Ibi bisa nkaho bigizwe nibice byo hanze birenze, bishobora guterwa no kugabanuka byihuse mubunini bw ibahasha ya periplasme, mugihe membrane yo hanze ikomeza kuba ntamakemwa, yerekana isura "idakabije". Guhinga ASxL5T mugihe habuze intungamubiri (muri PBS) igihe kinini kuri 4 ° C byaviriyemo selile nyinshi (ariko sibyose) zerekana morphologie ya coccal (Ishusho 1C). Iyo ASxL5T ikuze hamwe na Campylobacter jejuni nk'umuhigo w'amasaha 48, impuzandengo y'utugari ni ndende cyane kandi ndende kuruta selile ikura idafite nyiricyubahiro (Imbonerahamwe 1 na Ishusho 1E). Ibinyuranye, iyo ASxL5T ikuze hamwe na E. coli nk'umuhigo w'amasaha 48, impuzandengo y'utugari ni ndende kandi yagutse kuruta iyo ikura idafite umuhigo (Imbonerahamwe 1), kandi uburebure bw'akagari burahinduka, ubusanzwe bwerekana ububi (Ishusho 1F). Iyo ushyizwemo na Campylobacter jejuni cyangwa E. coli nk'umuhigo amasaha 48, selile ASxL5T yerekanaga nta flagella namba. Imbonerahamwe 1 yerekana incamake yibyerekeranye nimpinduka mubunini bwakagari hashingiwe kuboneka, kubura, nubwoko bwa ASxL5T.
TEM yerekana ASx5LT: (A) ASx5LT yerekana ikiboko kirekire; (B) bateri isanzwe ya ASx5LT; (C) cocci ASx5LT selile nyuma yubushakashatsi burebure butagira intungamubiri; . (F) Flagment nini ya flagella, selile ASx5LT, nyuma yubushakashatsi hamwe na E. coli umuhigo; (G) Akagari kamwe ASx5LT nyuma yubushakashatsi hamwe na E. coli, yerekana imiterere yo hejuru idasanzwe. Akabari kagereranya 1 mm.
Kumenya urutonde rwa 16S rRNA (kwinjira nomero MT636545.1) ituma ubushakashatsi bwububiko bwerekana urutonde nkurwo mu cyiciro cya Gammaproteobacteria, kandi rukaba hafi ya bagiteri zo mu nyanja zo mu muryango wa marine spirillum (Ishusho 2), kandi ni abanyamuryango ba Thalassolituus. Umuvandimwe wa hafi na Marine Bacillus. Urutonde rwa 16S rRNA rutandukanye rwose na bagiteri zangiza zo mu muryango wa Bdelvibrionaceae (Deltaproteobacteria). Kugereranya byombi B. bacteriovorus HD100T (ubwoko bwubwoko, DSM 50701) na B. bacteriovorus DM11A yari 48.4% na 47.7%, naho kuri B. exovorus JSS yari 46.7%. Bagiteri ya ASxL5T ifite kopi 3 za gen 16S rRNA, ebyiri muri zo zisa nizindi, naho iya gatatu ni 3 shingiro. Izindi bagiteri ebyiri zangiza (ASx5S na ASx5O; 16S rRNA nimero yinjira ni MT636546.1 na MT636547.1) hamwe nibiranga morfologiya na fenotipiki bisa ahantu hamwe ntabwo ari bimwe, ariko bitandukanye na ASxL5T na Bakteri zidafite umuco. Ububikoshingiro bukusanyirijwe hamwe hamwe nizindi genera muri Oceanospirillaceae (Ishusho 2). Urutonde rwa genome yose ya ASxL5T yaramenyekanye kandi ibikwa muri data base ya NCBI, kandi numero yinjira ni CP046056. Genome ya ASxL5T igizwe na chromosome izenguruka ya 2,831.152 bp hamwe na G + C ya 56.1%. Urutonde rwa genome rurimo CDS 2653 (zose), muri zo 2567 ziteganijwe kuzuza poroteyine, muri zo 1596 zishobora guhabwa imirimo yo gushyira (60.2%). Genome irimo genes 67 za RNA-code, harimo 9 rRNAs (3 imwe kuri 5S, 16S, na 23S) na tRNA 57. Ibiranga genomicike ya ASxL5T byagereranijwe na genome ziboneka zubwoko bwubwoko bugereranijwe bwagaragaye kuva 16S rRNA ikurikirana (Imbonerahamwe 2). Koresha indangamuntu ya amino (AAI) kugirango ugereranye genome zose za Thalassolituus na ASxL5T. Urutonde rwa hafi (rutuzuye) genome ikurikirana yagenwe na AAI ni Thalassolituus sp. C2-1 (ongeraho NZ_VNIL01000001). Iyi miterere yatandukanijwe n’imisozi miremire yo mu nyanja ya Mariana, ariko kuri ubu nta makuru ya fenotipiki yerekeye iyi miterere yo kugereranya. Ugereranije na ASxL5T ya 2.82 Mb, genome y'ibinyabuzima nini kuri 4.36 Mb. Impuzandengo ya genome ya marine spirochette igera kuri 4.16 Mb (± 1.1; n = 92 genome yuzuye yerekanwe kuri https://www.ncbi.nlm.nih.gov/assemble), bityo genome ya ASxL5T ijyanye na gutondekanya Ugereranije nabandi banyamuryango, ni nto rwose. Koresha GToTree 1.5.54 kugirango ubyare genome igereranijwe ishobora kuba igiti cyitwa phylogeneque (Ishusho 3A), ukoresheje aside amine ikomatanya kandi ihuza amoko 172 ya kopi imwe yihariye ya Gammaproteobacteria 11,12,13,14,15,16, 17, 18. Isesengura ryerekanye ko rifitanye isano rya hafi na Thalassolituus, Indege ya Bagiteri, na Bacterium ya Marine. Nyamara, aya makuru yerekana ko ASxL5T itandukanye na benewabo muri Marine Spirulina kandi amakuru yayo akurikirana arahari.
Igiti cya phylogeneque ukoresheje gene ya 16S rRNA yerekana umwanya wa ASxL5T, ASxO5, na ASxS5 (hamwe ninda) ugereranije na bagiteri zidahinzwe na marine zo muri Marine Spirulinaceae. Umubare winjira muri Genbank ukurikira izina rinini mumutwe. Koresha ClustalW kugirango uhuze urutonde, kandi ukoreshe uburyo bushoboka bwose hamwe na moderi ya Tamura-Nei kugirango umenye isano ya phylogeneque, kandi ukore inshuro 1000 ziyobowe muri gahunda ya MEGA X. Umubare ku ishami werekana ko kopi yayoboye agaciro karenze 50%. Escherichia coli U / 541T yakoreshejwe nkitsinda.
. . Igishushanyo kiri ibumoso cyerekana umubare wa gen muri buri cyiciro cya COG gikora muri buri genome. Igishushanyo kiri iburyo cyerekana ijanisha rya genome zikubiye muri buri tsinda rikora COG. .
Ukoresheje ububiko bwa KEGG kugirango usuzume ibice bigize gen zigaragara muri genome ya ASxL5T byagaragaje inzira isanzwe ya metabolike ya aerobic gamma Proteus. ASxL5T ikubiyemo genes zose hamwe 75 zahawe poroteyine za moteri ya bagiteri, harimo gen zigira uruhare muri chemotaxis, guteranya flagella, hamwe na sisitemu ya IV fimbriae. Mu cyiciro giheruka, 9 kuri 10 zifite inshingano zo gukurura urujya n'uruza rw'ibindi binyabuzima. Genome ya ASxL5T ikubiyemo inzira yuzuye ya tetrahydropyrimidine biosynthetic yinzira igira uruhare mukurinda umutekano wa osmotic20, nkuko byari byitezwe kuri halofile. Genome ikubiyemo kandi inzira nyinshi zuzuye za cofactors na vitamine, harimo na synthesis ya riboflavin. Nubwo gene ya alkane 1-monooxygenase (alkB2) iboneka muri ASxL5T, inzira yo gukoresha hydrocarubone ntabwo yuzuye. Muburyo bwa genome ya ASxL5T, homologues ya genes yagaragaye nkinshingano zo kwangirika kwa hydrocarbone muri T. oleiverans MIL-1T21, nka TOL_2658 (alkB) na TOL_2772 (alcool dehydrogenase) biragaragara ko idahari. Igicapo 3B cerekana kugereranya ikwirakwizwa rya gene murwego rwa COG hagati ya ASxL5T namavuta ya Olive MIL-1T. Muri rusange, genome ntoya ya ASxL5T irimo genes nkeya ugereranije na buri cyiciro cya COG ugereranije na genome nini ifitanye isano. Iyo umubare wa gen muri buri cyiciro cyibikorwa ugaragajwe nkijanisha rya genome, itandukaniro rigaragara mwijanisha rya genes mubisobanuro, imiterere ya ribosomal hamwe na biogenezi, hamwe ningufu zibyara ingufu nibikorwa byo guhindura, bigize ASxL5T nini genome Ijanisha rigereranwa nitsinda rimwe riri muri T. oleiverans MIL-1T genome. Ibinyuranye, ugereranije na genoside ya ASxL5T, T. oleivorans MIL-1T ifite ijanisha ryinshi rya gen mu kwigana, kwiyubaka no gusana, no mu byiciro byandikirwa. Igishimishije, itandukaniro rinini mubiri muri buri cyiciro cyibikorwa bya genome zombi ni umubare wa genes zitazwi ziboneka muri ASxL5T (Ishusho 3B). Isesengura ryitunganyirize ryamasomo ya KEGG ryakozwe, aho buri module ya KEGG igereranya urutonde rwibikorwa byasobanuwe nintoki kugirango bisobanurwe kandi bisobanure ibinyabuzima byamakuru bikurikirana. Kugereranya gukwirakwiza gene muburyo bwuzuye bwa KOG module yinzira ya ASxL5T na olive MIL-1T irerekanwa mumashusho 3C. Iri sesengura ryerekana ko nubwo ASxL5T ifite inzira ya sulfure yuzuye na azote ya metabolike, T. oleiverans MIL-1T ntabwo. Ibinyuranye, T. oleiverans MIL-1T ifite sisitemu yuzuye na methionine metabolike inzira, ariko ntabwo yuzuye muri ASxL5T. Kubwibyo, ASxL5T ifite module iranga assimilasiyo ya sulfate (isobanurwa nkurutonde rwa genes zishobora gukoreshwa nkibimenyetso bya fenotipiki, nkubushobozi bwa metabolike cyangwa indwara ziterwa na virusi; https://www.genome.jp/kegg/module.html) Muri T oleiverans MIL-1T. Kugereranya ibirimo gene ya ASxL5T nurutonde rwa genes zerekana ubuzima bwinyamaswa ntibishoboka. Nubwo gene yaaa irimo ligase ifitanye isano na O antigen polysaccharide ya O yibanze kuri genome ya ASxL5T (ariko ikunze kugaragara muri bagiteri nyinshi za Gram-negative), tryptophan 2,3-dioxygenase (TDO) irashobora kuba irimo amine 60 uturere twa aside dukunze kuboneka muri bagiteri zangiza zidahari. Nta yandi moko aranga inyamaswa ziri muri genome ya ASxL5T, harimo na enzymes enoding zifite uruhare muri isoprenoid biosynthesis mu nzira ya mevalonate. Menya ko nta genzura ya gene gntR iri mumatsinda yinyamanswa yasuzumwe, ariko genes eshatu zimeze nka gntR zishobora kumenyekana muri ASxL5T.
Ibiranga fenotipiki biranga ASxL5T byavuzwe mu mbonerahamwe ya 3 kandi ugereranije na fenotipiki iranga genera bifitanye isano 23, 24, 25, 26, na 27 byavuzwe mu bitabo. Kwigunga kuva T. marinus, T. olevorans, B. sanyensis, na Oceanobacter kriegii birakora, byihanganira umunyu, umubiri wa okiside-nziza, ariko nta bindi bintu biranga fenotipiki bifite ASxL5T. Impuzandengo ya pH yinyanja ni 8.1 (https://ocean.si.edu/inyanja-life/invertebrates/inyanja-acidification#section_77), ibyo bikaba bigaragara muri T. marinus, T. olevorans, B. sanyensis na O. kriegii. ASxL5T ibereye kurwego runini rwa pH (4-9) rusanzwe rwibinyabuzima bitari inyanja. Ibiranga fenotypic biranga Thalassolituus sp. C2-1. Ntazwi. Ubushyuhe bwo gukura bwa ASxL5T muri rusange ni bugari kuruta ubw'amazi yo mu nyanja (4-42 ° C), nubwo bamwe ariko atari T. marinus yonyine yihanganira ubushyuhe. Kudashobora gukura ASxL5T mubitangazamakuru byumuswa byabujije ibindi biranga fenotipiki. Koresha API 20E kugirango ugerageze ibikoresho byakuwe ku isahani ya BA, ONPG, dihydrolase ya arginine, lysine decarboxylase, ornithine decarboxylase, gukoresha citrate, urease, tryptophan deaminase, gelatin hydrolysis Enzyme, ibisubizo byikizamini byose byari bibi, ariko nta indole, acetoin na H2S. byakozwe. Carbohydrates idasembuye irimo: glucose, mannose, inositol, sorbitol, rhamnose, sucrose, melibiose, amygdalin na arabinose. Ugereranije n’ibisobanuro byashyizwe ahagaragara, imiterere ya aside irike ya selile ya ASxL5T igaragara mu mbonerahamwe ya 4. Acide nyamukuru ya selile selile ni C16: 1ω6c na / cyangwa C16: 1ω7c, C16: 0 na C18: 1ω9. Hydroxy fatty acide C12: 0 3-OH na C10: 0 3-OH nayo irahari. Ikigereranyo cya C16: 0 muri ASxL5T kiri hejuru kurenza agaciro kavuzwe na genera ifitanye isano. Ibinyuranye, ugereranije na T. marinus IMCC1826TT ivugwa, igipimo cya C18: 1ω7c na / cyangwa C18: 1ω6c muri ASxL5T cyaragabanutse. oleivorans MIL-1T na O. kriegii DSM 6294T, ariko ntibigaragara muri B. sanyensis KCTC 32220T. Ugereranije imyirondoro ya acide ya ASxL5T na ASxLS yerekanye itandukaniro rito mubunini bwa acide yibinure ya buri bwoko hagati yubwoko bubiri, ibyo bikaba bihuye na genomic ADN ikurikirana yubwoko bumwe. Nta bice bya poly-3-hydroxybutyrate (PHB) byagaragaye hakoreshejwe ikizamini cyirabura cya Sudani.
Igikorwa cyo guhiga bagiteri ya ASxL5T cyizwe kugirango hamenyekane aho umuhigo uhwanye. Iyi bagiteri irashobora gukora plaque kumoko ya Campylobacter, harimo: 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 na C. upsaliensis NCTC 11541T. Koresha imico iri kurutonde rwabashinzwe kugena uburyo bwo kugerageza intera nini ya Gram-mbi na Gram-nziza ya bagiteri. Ibisubizo byerekana ko ASxL5T ishobora no gukoreshwa muri Escherichia coli NCTC 86 na Citrobacter freundii NCTC 9750T. Icyapa cyakozwe kuri Klebsiella oxytoca 11466. Imikoranire ya TEM na E. coli NCTC 86 irerekanwa ku gishushanyo cya 4A-D, kandi imikoranire na Campylobacter jejuni PT14 na Campylobacter suis S12 irerekanwa ku gishushanyo cya 4E-H hagati. Uburyo bwo gutera busa nkaho butandukanye hagati yubwoko bwinyamanswa bwageragejwe, hamwe na selile imwe cyangwa nyinshi E. coli zifatanije na buri selire ASxL5T hanyuma igashyirwa kuruhande hafi ya selile yagutse mbere ya adsorption. Ibinyuranye, ASxL5T isa nkaho ihurira na Campylobacter binyuze mumwanya umwe wo guhuza, mubisanzwe uhura na apex ya selile yinyamanswa ndetse no hafi yisonga rya selire ya Campylobacter (Ishusho 4H).
TEM yerekana imikoranire hagati ya ASx5LT n'umuhigo: (AD) na E. coli umuhigo; (EH) na C. jejuni umuhigo. (A) Akagari gasanzwe ASx5LT gahujwe na selile imwe E. coli (EC); (B) ASx5LT yuzuye ifatanye na selile imwe ya EC; (C) Akagari ka ASx5LT kagizwe na selile nyinshi za EC; (D) Umugereka Utugingo duto ASx5LT selile imwe ya E. coli (EC); (E) selile imwe ya ASx5LT ihujwe na selire ya Campylobacter jejuni (CJ); (F) ASx5LT yibasiye selile C. hyointestinalis (CH); (G) selile ebyiri ASx5LT yibasiye selile CJ; . Umurongo ugereranya 1 mm muri (A - G).
Indwara ziterwa na bagiteri zahindutse kugirango zungukire ku masoko menshi y’inyamanswa. Biragaragara, zirahari henshi mubidukikije bitandukanye. Bitewe n'ubunini buke bw'abaturage, birashoboka gutandukanya bacteri za ASxL5T ziva mubitotsi hakoreshejwe uburyo bwo gutandukanya fage. Ibyerekeranye na genomic ya ASxL5T kubanyamuryango ba Oceanospirillaceae ya bagiteri zo mu nyanja biratangaje, nubwo ibinyabuzima byihanganira umunyu kandi bishobora gukura muburyo burimo umunyu 5%. Isesengura ry’amazi y’amazi ryerekanye ko sodium ya chloride iri munsi ya 0.1%. Kubwibyo, ibyondo biri kure y’ibidukikije byo mu nyanja - haba mu turere ndetse no mu rwego rwa shimi. Kuba hari ibintu bitatu bifitanye isano ariko bitandukanye bitandukanya isoko imwe bitanga ibimenyetso byerekana ko izo nyamaswa zitera imbere muri ibi bidukikije bitari inyanja. Hiyongereyeho, isesengura rya microbiome (dosiye zamakuru ziboneka kuri https://www.ebi.ac.uk/ena/browser/view/PRJEB38990) zerekanye ko 16S rRNA ikurikirana imwe iri muri tagisi 50 yambere ikora cyane (OTU) ) Mugihe gito cyo gutoranya intera yicyondo. Bagiteri nyinshi zidafite umuco zabonetse mububiko bwa Genbank, zifite 16S rRNA zikurikirana nka bagiteri ASxL5T. Uru rutonde, hamwe nurutonde rwa ASxL5T, ASxS5, na ASxO5, bisa nkaho bihagarariye ibice bitandukanye bitandukanijwe na Thalassolituus na Oceanobacter (Ishusho 2). Ubwoko butatu bwa bagiteri zidafite umuco (GQ921362, GQ921357 na GQ921396) bwatandukanijwe n’amazi yatoboye ku bujyakuzimu bwa kilometero 1.3 mu kirombe cya zahabu yo muri Afurika yepfo mu 2009, naho andi abiri (DQ256320 na DQ337006) yavaga mu mazi y’ubutaka (no muri Afurika yepfo) muri 2005). Urutonde rwa 16S rRNA rufitanye isano rya bugufi na ASxL5T ni igice cya 16S rRNA ikurikirana yakuwe mu muco wo gutunganyiriza ubutayu bw’umucanga wabonetse ku nkombe z’amajyaruguru y’Ubufaransa mu 2006 (nimero ya AM29240828). Iyindi mikorere ya 16S rRNA ikurikirana ya bagiteri idafite umuco HQ183822.1 yakuwe mu kigega cyo gukusanya cyasohotse mu myanda ya komini mu Bushinwa. Ikigaragara ni uko bagiteri ya ASxL5T idahagarariwe cyane mububiko bwimisoro, ariko izi nteruro ziva muri bagiteri zidafite umuco zishobora kugereranya ibinyabuzima bisa na ASxL5T, bikwirakwizwa kwisi yose, mubisanzwe mubidukikije bigoye. Uhereye kuri genome yose ya phylogeneque, hafi ya ASxL5T ni Thalassolituus sp. C2-1, T. marinus, T. oleivorans. Naho O. kriegii 23, 24, 25, 26, 27. Thalassolituus ni umwe mu bagize bacteri zo mu bwoko bwa hydrocarubone zigabanywa (OHCB), zikwirakwizwa cyane mu bidukikije byo mu nyanja no ku isi, kandi ubusanzwe ziba ziganje nyuma y’ibyuka bihumanya hydrocarubone 30,31. Bagiteri zo mu nyanja ntabwo ziri mu itsinda rya OHCB, ariko zitandukanijwe n’ibidukikije byo mu nyanja.
Amakuru ya fenotypic yerekana ko ASxL5T ari ubwoko bushya kandi akaba umwe mubagize ubwoko butaramenyekana mumuryango wa marine spirospiraceae. Kugeza ubu nta gipimo gisobanutse cyo gushyira ubwoko bushya bwitaruye mu bwoko bushya. Bagerageje kumenya imbibi za genera ku isi hose, urugero, hashingiwe ku ijanisha rya genome ya poroteyine igabanya ubukana (POCP), birasabwa ko agaciro kaciwe kangana na 50% bihuye n’ibisobanuro 33. Abandi batanga inama yo gukoresha indangagaciro za AAI, zifite ibyiza kurenza POCP kuko zishobora kuboneka muri genome zuzuye34. Umwanditsi yizera ko niba agaciro ka AAI kari munsi ya 74% ugereranije nubwoko bwikitegererezo bwubwoko bwikitegererezo, ubwoko bwerekana ubwoko butandukanye. Ubwoko bw'icyitegererezo muri marine spirillaceae ni marine spirillum, naho ubwoko bw'icyitegererezo ni O. linum ATCC 11336T. Agaciro AAI hagati ya ASxL5T na O. linum ATCC 11336T ni 54.34%, naho agaciro ka AAI hagati ya ASxL5T na T. oleivorans MIL-1T (ubwoko bwubwoko) ni 67.61%, byerekana ko ASxL5T ihagarariye ubwoko bushya butandukanye na Thalassolituus. Ukoresheje genS ya 16S rRNA nkurwego rwo gutondekanya ibyiciro, igitekerezo cyo gutandukanya ubwoko ni 94.5% 35. ASxL5T irashobora gushyirwa mubwoko bwa Thalassolituus, ikerekana 95.03% 16S rRNA ikurikirana hamwe na T. oleivorans MIL-1T na 96.17%. marinus IMCC1826T. Icyakora, izashyirwa kandi mubwoko bwa Bacteroide ifite 94,64% 16S rRNA ya gene hamwe na B. sanyensis NV9, byerekana ko gukoresha gene imwe nka 16S rRNA bishobora kuganisha mubyiciro no kugenwa uko bishakiye. Ubundi buryo bwatekerejweho bukoresha amanota ya ANI na Genome Alignment (AF) kugirango isuzume ihuriro ryamakuru yatanzwe muburyo bwose nubwoko butari ubwoko bwa genera iriho. Umwanditsi arasaba guhuza imbibi zubwoko n’ahantu ho guhinduranya ubwoko bugereranijwe bwihariye kuri tagisi isesengurwa. Ariko, niba nta genome zuzuye zihagije ziva muri Thalassolituus zitandukanya, ntibishoboka kumenya niba ASxL5T ari iy'ubwoko bwa Thalassolituus hakoreshejwe ubu buryo. Bitewe no kuboneka kwa genome yuzuye kugirango isesengurwe, igiti cyose cyitwa phylogeneque igiti kigomba gusobanurwa neza. Icya kabiri, uburyo bwo kugereranya genome yose ntishobora kubara itandukaniro rinini mubunini bwa genome ugereranije. Bapimye isano ya genes yabitswe imwe-imwe ikoporora hagati ya genera ifitanye isano, ariko ntibitaye ku mubare munini wa gen zitagaragara muri genome ntoya ya ASxL5T. Ikigaragara ni uko ASxL5T hamwe nitsinda ririmo Thalassolituus, Oceanobacter, na Bacterioplanes bafite abakurambere basangiye, ariko ubwihindurize bwafashe indi nzira, bituma igabanuka rya genome, rishobora kuba rihuza nubuzima bwangiza. Ibi bitandukanye na T. oleivorans MIL-1T, nini 28% kandi yagiye ihinduka bitewe n’ibitutu bitandukanye by’ibidukikije kugirango ikoreshe hydrocarbone 23,30. Ikigereranyo gishimishije kirashobora kugereranywa na parasite idasanzwe hamwe na sibionts, nka Rickettsia, Chlamydia, na Buchnera. Ingano ya genome igera kuri 1 Mb. Ubushobozi bwo gukoresha selile metabolite yakira biganisha ku gutakaza gene, bityo hakabaho kwangirika kwa genomic kwangirika. Imihindagurikire y’ubwihindurize kuva mu binyabuzima byintungamubiri zo mu nyanja zijya mu mibereho y’inyamaswa zishobora gutuma igabanuka risa n’ubunini bwa genome. Isesengura rya COG na KEGG ryerekana umubare wa gen zikoreshwa mu mirimo yihariye n’itandukaniro ry’isi yose mu nzira ya genomic hagati ya ASxL5T na T. oleivorans MIL-1T, ibyo bikaba bidatewe nuko haboneka ibintu byinshi bigendanwa. Itandukaniro mu kigereranyo cya G + C cya genome yose ya ASxL5T ni 56.1%, naho iya T. oleivorans MIL-1T ni 46,6%, ibyo bikaba byerekana kandi ko itandukanijwe.
Isuzuma rya code ya genome ya ASxL5T itanga ubushishozi mubikorwa biranga fenotipiki. Kubaho gen zigizwe n'ubwoko bwa IV fimbriae (Tfp) birashimishije cyane kuko biteza imbere ingirabuzimafatizo, bita glide social cyangwa guhungabana, nta flagella hejuru. Nk’uko amakuru abitangaza, Tfp ifite indi mirimo, harimo guhiga, gutera indwara, gukora biofilm, gufata ADN karemano, guteranya ingirabuzimafatizo no gutera imbere38. Genome ya ASxL5T ikubiyemo genes 18 zikubiyemo diguanylate cyclase (enzyme itera ihinduka rya guanosine triphosphate 2 muri guanosine 2 fosifate na cyclic diGMP) hamwe na genes 6 zikubiyemo diguanylate cyclase fosifate diguanylate. Gene ya esterase (itera kwangirika kwa cyclic di-GMP kuri guanosine monophosphate) irashimishije kuko cycl-di-GMP nintumwa ya kabiri ikomeye igira uruhare mu iterambere rya biofilm no gutandukana, kugenda, kwizirika hamwe na virusi 39, 40 muriki gikorwa. Twabibutsa kandi ko muri Bdellovibrio bacteriovorus, cyikubye kabiri GMP yerekanwe kugenzura inzibacyuho hagati yubuzima bwubuzima nubuzima bw’inyamaswa41.
Ubushakashatsi bwinshi kuri bagiteri zangiza bwibanze kuri Bdellovibrio, ibinyabuzima bisa na Bdellovibrio, nubwoko bwa Myxococcus. Izi nizindi ngero zizwi za bagiteri zangiza zigize itsinda ritandukanye. Nuburyo butandukanye, hamenyekanye urutonde rwimiryango ya poroteyine iranga phenotypes ya bagiteri 11 zizwi zangiza. Nyamara, hamenyekanye gusa gen zigizwe na O antigen ligase (waaL), zikunze kugaragara cyane muri bagiteri-mbi. Ubu buryo bwo gusesengura ntabwo bufasha mu kwerekana ASxL5T nk'inyamanswa, birashoboka kuko ikoresha ingamba zo gutera udushya. Kuboneka kwa genoside zinyuranye zinyamanswa zizafasha guteza imbere isesengura ryiza ryita kubimenyetso byerekana itandukaniro ryimikorere nibidukikije hagati yabagize itsinda. Ingero za bagiteri zangiza zitashyizwe muri iri sesengura zirimo abanyamuryango ba Cupriavidus necator42 na Bradymonabacteria43, kubera ko nkuko abashakashatsi bakora iperereza ku mikorobe itandukanye, hashyirwaho tagisi nyinshi.
Ikintu kigaragara cyane muri bagiteri ya ASxL5T yafashwe nishusho ya TEM ni imiterere yihariye kandi yoroheje, ishobora guteza imbere imikoranire na bagiteri zihiga. Ubwoko bw'imikoranire bwagaragaye butandukanye nizindi bagiteri zangiza kandi ntizigeze zivumburwa cyangwa ngo zitangwe raporo. Icyifuzo cyubuzima bwa ASxL5T cyerekanwe mubishusho 5. Hano hari ingero nke mubitabo bifite imiterere isa nkiyi nkuko tubitangaza hano, ariko izi ngero zirimo Terasakiispira papahanaumokuakeensis, bacterium marine spirillum ifite rimwe na rimwe kwaguka 44, na Alphaproteobacteria, Terasakiella pusilla , ahahoze ari ubwoko bwa Oceanospirillum, bwerekana icyo bita "polar film" 45. Imiterere ya Cocci ikunze kugaragara mu mico ishaje, cyane cyane kuri bagiteri zifite imiterere igoramye, nka Vibrio, Campylobacter, na Helicobacter 46, 47, 48, zishobora kugereranya leta yangiritse. Ibindi bikorwa birakenewe kugirango tumenye neza ubuzima bwa bagiteri ya ASxL5T. Kugirango umenye uko ifata ninyambo, kandi niba genome yayo ikubiyemo ibinyabuzima bikora biologiya bishobora gukoreshwa mubuvuzi cyangwa ibinyabuzima.
Ibisobanuro bya Venatorbacter gen. Ugushyingo Venatorbacter (Ven.a.tor, ba'c.ter, L. igizwe n'abashitsi ba L. n. Venator, 'umuhigi' na Gr. . Ingirabuzimafatizo ni aerobic, itihangana-umunyu intera ya 4-9 ntisanzwe. Mubisimba byo mu nyanja, ibyinshi ntibihanganira aside irike ni C16: 1ω6c na / cyangwa C16: 1ω7c, C16: 0 na C18: 1ω9; : 0 3-OH iboneka nka aside irike ya hydroxy. Ntibakura mubitangazamakuru byumunyu Kandi imyitwarire yo guhiga abagize umuryango wa Enterobacteriaceae.
Ibisobanuro bya Venatorbacter cucullus sp. Ugushyingo Venatorbacter cucullus (cu'cull.us.; L. n. Cucullus bisobanura kurenganura).
Mubyongeyeho, ibiranga ubu bwoko ni uko iyo bihinzwe kuri BA cyangwa BHI, selile zifite uburebure bwa 1,63 µm na 0.37 µm z'ubugari. Ubukoroni kuri BHI agar ni buto cyane, bugera kuri mm 2 z'umurambararo nyuma yamasaha 72. Ni beige, bisobanutse, bizengurutse, convex kandi birabagirana. Abagize ubu bwoko barashobora gukoresha Escherichia coli na Klebsiella. Campylobacter hamwe na bagiteri nyinshi za Gram-mbi zitanga umuhigo.
Ubwoko busanzwe bwa ASxL5T bwatandukanijwe n’amata y’inka i Nottinghamshire, mu Bwongereza, kandi bushyirwa mu cyegeranyo cy’igihugu cy’umuco (UK): nomero yinjira NCTC 14397 hamwe n’Ubuholandi bw’umuco wa bagiteri (NCCB) nomero NCCB 100775. yashyizwe muri Genbank ukurikije inyongera ya CP046056.
Bagiteri ya ASxL5T yitandukanije n'amata y'inka ikoresheje tekinoroji yo kwigunga ya fage9,49. Amashanyarazi yavomwe 1: 9 (w / v) muri buffer ya SM (50 mM Tris-HCl [pH 7.5], 0.1 M NaCl, 8 mM MgSO4.7H2O na 0.01% gelatine; Sigma Aldrich, Gillingham, UK), Hanyuma incubate kuri 4 ° C kumasaha 24, kuzunguruka buhoro kugirango ucyure inyamanswa muri buffer. Ihagarikwa ryashyizwe kuri 3000g muminota 3. Supernatant yakusanyirijwe hamwe hanyuma igera kuri 13,000g kunshuro ya kabiri muminota 5. Supernatant yahise inyuzwa kuri 0.45 µm membrane filter (Minisart; Sartorius, Gottingen, Ubudage) hamwe na 0.2 µm membrane filter (Minisart) kugirango ikureho selile zose zisigaye. ASxL5T irashobora kunyuza muyunguruzi. Icyatsi cyoroshye cya agar ya Campylobacter enterosus S12 (NCBI yinjira numero CP040464) uhereye kumurongo umwe wateguwe hakoreshejwe tekinoroji isanzwe. Akayunguruzo kayunguruzo kagabanijwe kuri buri cyapa cyakiriye muri selile 10 µl ibitonyanga bitatu kandi byemewe gukama. Isahani yashyizwe mu kigega cya microaerophilique kuri 37 ° C mu masaha 48 mu bihe bya mikorobe (5% O2, 5% H2, 10% CO2, na 80% N2). Icyapa kiboneka cyabonetse cyakuwe muri buffer ya SM hanyuma cyimurirwa muri nyakatsi nshya ya C. hyointestinalis S12 kugirango irusheho gukwirakwiza ibinyabuzima byanduye. Bimaze kwemezwa ko bagiteri arizo nyirabayazana ya plaque ya litike ntabwo ari fage, gerageza gukura ibinyabuzima utigengwa na nyirubwite hanyuma urusheho kubiranga. Umuco wo mu kirere wakorewe kuri 37 ° C hamwe na 5% v / v amaraso y’ifarashi (TCS Biosciences Lt, Buckingham, UK, inyongera). Dukurikije umurongo ngenderwaho wa komite y’igihugu ishinzwe amavuriro, uburyo bwo gukwirakwiza disiki bukoreshwa mu gupima antibacterial suseptibilité. BHI agar yarezwe kuri 37 ° C ikoresheje disiki irimo antibiyotike ikurikira (Oxoid) kumuco windege: amoxicillin na aside clavulanic 30 µg; cefotaxime 30 µg; streptomycine 10 µg; ciprofloxacin 5 µg; Ceftazidime 30 µg Acide Nalidixic 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. Kwihanganira umunyu byashyizweho nubushakashatsi bwindege kuri plaque ya BHI kuri 37 ° C. Inyongera NaCl yongewe kumasahani ya BHI agar kugirango itange umurongo wa 10% w / v. Urwego rwa pH rugenwa numuco windege kuri plaque ya BHI kuri 37 ° C, aho pH yahinduwe kugeza hagati ya 4 na 9 hamwe na sterile HCl cyangwa sterile NaOH, kandi intego pH igenzurwa mbere yo gusuka isahani. Kubisesengura rya selile ya selile, ASxL5T yarezwe kuri BHI agar iminsi 3 na aerobic kuri 37 ° C. Dukurikije MIDI (Sherlock Microbial Identification Sisitemu, verisiyo 6.10) protocole isanzwe ya FERA Science Ltd, (York, UK), acide fatty selile yakuwe, itegurwa kandi irasesengurwa.
Kuri TEM, ASxL5T yari ifite umuco wo mu kirere ikwirakwiza kimwe kuri BA kuri 37 ° C mu masaha 24, hanyuma igasarurwa muri ml 1 ya 3% (v / v) glutaraldehyde muri 0.1 M ya cacodylate yubushyuhe bwo mucyumba Gukosora isaha 1, hanyuma centrifuge kuri 10,000 g muminota 3. Noneho uhagarike witonze pellet muri 600 μl 0.1 M ya cacodylate buffer. Hindura ihagarikwa rya ASxL5T kuri firime ya Formvar / karubone kuri gride ya mesh 200 mesh. Bagiteri yandujwe na 0.5% (w / v) uranyl acetate kumunota 1 hanyuma isuzumwa na TEM ikoresheje microscope ya TEI Tecnai G2 12 Biotwin. Nkuko byavuzwe haruguru, komatanya umubare w’inyamanswa n’inyamanswa mu muyoboro wa NZCYM (BD Difco ™, Fisher Scientific UK Ltd, Loughborough) hanyuma ukore amasaha 48 mu bihe bya microaerobic ya Campylobacter cyangwa Campylobacter kuri 37 ° C, Imikoranire y’inyamanswa n’inyambo. yasuzumwe kandi na TEM. Imiterere yindege ya Escherichia coli. Wigenga usuzume bagiteri zihiga ninyamaswa zangiza kugirango umenye impinduka zose zijyanye na morphologie bitewe nubwoko. Uburyo bwirabura bwa Sudani bwakoreshejwe kuri microscopi optique yo kwirundanya kwa PHB.
Kura ASxL5T ijoro ryose imico isiga imikurire kuri plaque ya BHI cyangwa BA hamwe na sterile swab. Kusanya selile ASxL5T hanyuma uyihagarike muri MRD (CM0733, Oxoid), hanyuma ubishyire kuri 4 ° C muminsi 7 kugirango ushonje inzara. Umuco wa bagiteri wa NCTC cyangwa laboratoire yatewe mu muyoboro wa BHI cyangwa No 2 wintungamubiri zintungamubiri (CM007, Oxoid), winjijwe ijoro ryose, ushyirwa kuri 13,000g hanyuma uhagarikwa muri MRD kugeza OD600 yari 0.4. Umuco: Bacillus subtilis NCTC 3610T, Citrobacter freundii NCTC 9750T, Enterobacter aerogenes NCTC 10006T, Enterococcus faecalis NCTC 775T, Escherichia coli NCTC 86, Klebsiella oxytoca 11466, Leuconostoc NCTC 10817, Listeron stuartsii NCTC 10318, Pseudomonas fluorescens SMDL, Rhodococcus submarine hamburger NCTC 1621T, Salmonella yo mu nda ya Mondeville NCTC 5747, mucilage NCTC 10861, Staphylococcus aureus NCTC 8532T, Streptococcus pneumoniaol NCTC 7465. Umucumbitsi wa Campylobacter yashizwemo microaerobic kuri plaque ya BA kuri 37 ° C hanyuma ihagarikwa mumitsi ya NZCYM. Ikizamini cya Campylobacter cyapimwe ni: C. coli 12667 NCTC, C. jejuni 12662, C. jejuni PT14, C. jejuni NCTC 11168T, C. helveticus NCTC 12472, C. lari NCTC 11458, C. lari NCTC 11458, C. jejuni PT14 , C ... Kusanya selile muri MRD, centrifuge kuri 13,000g hanyuma uhagarare muri MRD kugeza OD600 ari 0.4. Ongeramo aliquot ya 0,5 ml ihagarikwa kuri ml 5 yashonze NZCYM agar hejuru (0,6% agar) hanyuma uyisuke kuri plaque yo hasi ya 1.2% NZCYM. Nyuma yo gukiza no gukama, ASxL5T yakuwe muburyo bukwirakwizwa nka 20 µl ibitonyanga kuri buri kibaho cya nyakatsi muri bitatu. Ubushyuhe bwumuco nikirere biterwa nibisabwa na bagiteri yipimishije.
Koresha GenElute Kit Bakteri ya Genomic ADN Kit (Sigma Aldridge) kugirango utegure ADN ivuye muri bagiteri. Uburyo busanzwe bwakoreshejwe mugukwirakwiza PCR ya 16S rRNA no kugena ibicuruzwa bikurikirana ukoresheje chimie yo kurangiza irangi (Eurofins Agaciro Soma Serivisi, Ubudage). Koresha gahunda ya BLAST-N kugirango ugereranye uru rutonde nizindi 16S rRNA zikurikirana kugirango umenye kandi ukusanyirize hamwe amoko afitanye isano. Ibi bihujwe ukoresheje ClustalW muri gahunda ya MEGA X. Igiti cya phylogeneque cyongeye kubakwa hifashishijwe MEGA X hakoreshejwe uburyo bushoboka bushoboka bushingiye ku cyitegererezo cya Tamura-Nei, hamwe na kopi 1000 ziyobowe54. Koresha PureLink Kit Kitike ya ADN (Fisher Scientific, Loughborough, UK) kugirango ukure ADN kugirango ukurikirane genome yose. Urutonde rwa genome ya ASxL5T rwagenwe hifashishijwe Illumina MiSeq ikomatanya, igizwe na 250 bp isomwa kabiri-isomwe igizwe nibitabo byateguwe hifashishijwe ibikoresho bya label ya Nextera hamwe na 2 kugeza kuri 20 kb bisomwa kuva kuri platifomu ya PacBio. Ikirangantego cya ADN Ikurikirana Ubushakashatsi muri kaminuza ya Sembia. Genome yakusanyirijwe hamwe ikoresheje CLC Genomics Workbench 12.0.3 (Qiagen, Aarhus, Danimarike). Imico ya ASxL5T ishyirwa mu cyegeranyo cy’umuco w’igihugu (UK) no mu cyegeranyo cy’umuco wa bacteri w’Ubuholandi (NCCB). Imiterere y'ibinyabuzima bifitanye isano ikoreshwa mu kugereranya ni: Thalassolituus oleivorans MIL-1T (nomero yinjira HF680312, yuzuye); Bacterioplanes sanyensis KCTC 32220T (nimero yo kwinjira BMYY01000001, ituzuye); Oceanobacter kriegii DSM 6294T (nomero yo kwinjira NZ_AUGV00000000, ituzuye); Umuryango wa Marinamonas DSM 5604T (wongeyeho ASM436330v1, utuzuye), Oceanospirullum linum ATCC 11336T (wongeyeho MTSD02000001, utuzuye) na Thalassolituus sp. C2-1 (ongeraho NZ_VNIL01000001, ituzuye). Koresha JGI Genome Port36 kuri https://img.jgi.doe.gov//cgi-bin/mer/main.cgi?section=ANI&page= kugirango umenye amanota yo guhuza (AF) hamwe nimpuzandengo ya acide nucleic (ANI). Muri babiri. Uburyo bwa Rodriguez-R & Konstantinidis55 bwakoreshejwe kugirango hamenyekane aside amine (AAI). Koresha GToTree 1.5.5411,12,13,14,15,16,17,18 kugirango ubyare igiti kinini cya phylogeneque. Iyinjiza genome yerekana genome iboneka yatoranijwe muri genera yerekanwe ko ifitanye isano na ASxL5T kuva muri 16S rRNA phylogene. Annotated igiti ukoresheje igiti cyimikorere yubuzima kumurongo (https://itol.embl.de/). Gutanga amakuru no gusesengura genome ya ASxL5T bikorwa hakoreshejwe igikoresho cya interineti cya BlastKOALA KEGG ukoresheje KEGG (Kyoto Encyclopedia ya Gen na Genomes) ikwirakwizwa ryuzuye. Ikwirakwizwa ryibyiciro bya COG (amatsinda ya orthologique) bigenwa ukoresheje igiNOG-mapper igikoresho cyo kumurongo.
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