Propionic acid (PPA), ekirikurwanisa obukooko obu kandi ekirikugambwaho omu by‘okurya, kyorekwa ngu nikireetera okukura kw‘obutafaari bw‘omubiri okutari kwa buriijo omu mbeba okurikuhererukayo n‘okuremwa kukora gye omu nda, ekirikubaasa kureetwa okuremwa kukora gye kw‘omunda. Enkwatanisa ahagati y‘okukwatwa PPA omu by‘okurya n‘okukwatwaho kwa gut microbiota dysbiosis ekateekateekwaho, kwonka tekacondooziibwe butunu. Aha, tukacondooza aha mpindahinduka eziine akakwate na PPA omu nkora ya gut microbiota ezirikubaasa kureetera dysbiosis. Gut microbiomes z‘embeba ezaabaire ziriisiibwe eby‘okurya ebitaine bujanjabi (n=9) n‘eby‘okurya ebirimu PPA (n=13) bikakuratana barikukoresa okukuratana kw‘obwire buraingwa okwa metagenomic kukyebera entaaniso omu nkora ya microbial n‘emihanda y‘okuhindura eby‘okurya bya bacteria. PPA y’eby’okurya ekaba neekwatagana n’okwongyera aha bwingi bw’ebika by’ebinyonyi ebikuru, harimu ebika bya Bacteroides, Prevotella, na Ruminococcus bingi, ba memba baabyo babaire nibeejumba omu kuhanga PPA enyima. Microbiomes z‘embeba ezaabaire ziine PPA nazo zikaba ziine emihanda mingi erikukwatagana n‘okuhindura amashaju n‘okuhindura ebiriisa bya steroid. Ebirugire omu kucondooza kwaitu nibyoreka ngu PPA neebaasa kuhindura obutafaari bw‘omubiri oburikwetwa gut microbiota n‘emihanda yaayo ey‘okuhindura eby‘okurya. Empindahinduka ezi ezireebirwe nizooreka ngu ebirikurinda eby‘okurya ebiteirwe omu kicweka ky‘okurinda eby‘okurya nibibaasa kuhindura aha nkora y‘ebintu ebikye ebiri omu nda kandi, omu kugarukamu, n‘amagara g‘omuntu. Omuri ebyo, P, G nari S nibitooranwa kurugiirira aha rurengo rw‘okushoroora orurikushwijumwa. Okukyendeeza aha nkora y‘okushoroora ebishuba ebirungi, omutindo gw‘obungi bw‘ebintu ebikye munonga ogwa 1e-4 (1/10,000 okushoma) gukateebwaho. Okushwijuma kw‘embariirira kutakabaireho, obungi bw‘ebintu ebirikushushana ebyagambirweho Bracken (ekicweka_omubaro_gw‘okushoma) bukahindurwa barikukoresa okuhindura kw‘omubaro gw‘ebihandiiko (CLR) (Aitchison, 1982). Enkora ya CLR ekatooranwa ahabw‘okuhindura amakuru ahabw‘okuba neehinduka omu rulengo kandi neemara aha bihandiiko ebitari bikye (Gloor et al., 2017). Okuhindura kwa CLR nikukoresa logarithm y’obuhangwa. Ebihandiiko by‘okubara ebyagambirweho Bracken bikahindurwa eby‘obutoosha barikukoresa enkora y‘okubara ebiri omu nkora (RLE) (Anders na Huber, 2010). Ebishushani bikakorwa barikukoresa okuteerana kwa matplotlib v. 3.7.1, eby’omu nyanja v. 3.7.2 n’eby’okureeberaho by’okukuratana (Gloor na bagyenzi be, 2017). 0.12.2 n‘ebigambo ebitari bimwe na bimwe v. 0.5.0 (Omuhiigi, 2007; Waskom, 2021; Charlier n‘abandi, 2022). Omubaro gwa Bacillus/Bacteroidetes gukabarwa ahari buri sampo barikukoresa okubara kwa bacteria okwa buriijo. Emihendo erikugambwaho omu meeza neeyehinguririzibwa kuhika aha myanya 4 ya desimaali. Omubaro gw‘ebintu bitari bimwe na bimwe ogwa Simpson gukabarwa barikukoresa ekihandiiko kya alpha_diversity.py ekyaheebwa omu nkora ya KrakenTools v. Ripoota ya Bracken neeheebwa omu kihandiiko kandi Simpson index “Si” neeheebwa ahabw’ekicweka kya -an. Entaaniso z‘amaani omu bwingi zikashoboororwa nk‘entaaniso za CLR ≥ 1 nari ≤ -1. Entaaniso ya CLR eya ±1 neeyoreka okweyongyera kw‘emirundi 2.7 omu bwingi bw‘omuringo gw‘eky‘okureeberaho. Akabonero (+/-) nikooreka yaaba taxon erikukirayo omu sampo ya PPA n’omu sampo y’okukyebera, nk’oku biri. Obukuru bukamanyirwa aha kukoresa ekigyezo kya Mann-Whitney U (Virtanen na bagyenzi be, 2020). Eby‘okureeberaho by‘embariirira v. 0.14 (Benjamini na Hochberg, 1995; Seabold na Perktold, 2010) bikakoresibwa, kandi enkora ya Benjamini-Hochberg ekakoresibwa kushoboorora ahabw‘okukyebera kwingi. Omuhendo gwa p ogwahindurwa ≤ 0.05 gukakoresibwa nk‘omutindo gw‘okumanyisa obukuru bw‘embariirira.
Obuhangwa bw‘omuntu obukye nibukira kwetwa “ekicweka ky‘omubiri ekirikuhererukayo” kandi nibukora omurimo muhango omu magara g‘omuntu (Baquero na Nombela, 2012). Okukira munonga, obutafaari bw‘omubiri oburikwetwa gut microbiome nibumanyirwa aha kuhindura kwabwo omu nkora yoona n‘omurimo gwabwo omu mirimo mingi ekuru. Bacteria z‘okurya nizikanyira omu nda, nizitwara emyanya mingi y‘eby‘obuhangwa, nizikoresa ebiriisa, kandi nizihakanisa obukooko oburikubaasa kureeta endwara (Jandhyala et al., 2015). Ebicweka bya bacteria ebitari bimwe na bimwe eby‘omunda nibibaasa kureetaho ebiriisa ebikuru nka vitamini n‘okutunguura okugaaya eby‘okurya (Rowland et al., 2018). Ebirungo bya bacteria nabyo nibyorekwa ngu nibikwata aha kukura kw‘ebicweka by‘omubiri n‘okutunguura emihanda y‘okuhindura eby‘okurya n‘okurwanisa endwara (Heijtz et al., 2011; Yu et al., 2022). Ebirimu ebiri omunda y‘omuntu nibihinduka munonga kandi nibirugiirira aha nshonga z‘obuzaarwa n‘eby‘obuhangwa nk‘eby‘okurya, obuhangwa, emibazi, n‘embeera y‘amagara (Kumbhare et al., 2019).
Ebyokurya bya nyina omwana n‘ekintu kikuru omu kukura kw‘omwana ori omunda n‘omwana omuto kandi n‘entandikwa y‘ebintu ebirikuteekateekwaho ebirikubaasa kuhindura enkurakurana (Bazer et al., 2004; Innis, 2014). Ekimwe aha bikwatiraine n‘okukunda n‘ekirungo kya propionic acid (PPA), ekirungo ky‘ebishaju ekiine orujegyere rukye ekirikuruga omu kukyebera obukooko n‘ekintu ekirikugaita eby‘okurya (den Besten et al., 2013). PPA eine obushoboorozi bw’okurwanisa obukooko n’obukooko n’ahabw’ekyo neekoresibwa nk’ekintu ekirikurinda eby’okurya n’omu mirimo y’amakorero okuzibira obukooko n’okukura kw’obukooko (Wemmenhove et al., 2016). PPA eine ebirugiremu bitarikushushana omu bicweka by‘omubiri ebitarikushushana. Omu kifuba, PPA eine ebirikurugamu ebirikurwanisa okuzimba ahabw‘okuteganisa okureebeka kw‘ebicweka by‘omubiri ebirikwetwa cytokine omu bihaha ebihango (Kawasoe et al., 2022). Eki nikireetaho okutegyeka eki n‘omu butafaari obundi oburikurwanisa endwara, ekirikureetaho okukyendeeza aha kutegyeka okuzimba (Haase et al., 2021). Kwonka, ekirikushushana n’ekyo kireebirwe omu bwongo. Okucondooza okukozirwe enyima kworekire ngu okukoresa PPA nikureetaho emitwarize erikushushana n‘ey‘oburwaire bwa autism omu mbeba (El-Ansary et al., 2012). Ebindi bihandiiko byorekire ngu PPA neebaasa kureetera gliosis n‘okukora emihanda y‘okureetaho okuzimba omu bwongo (Abdelli et al., 2019). Ahabw‘okugira ngu PPA n‘asidi y‘amaani, neebaasa kujanjaara kurabira omu nkora y‘omu mara kuza omu shagama kandi obwo neehingura aha nshonga ezirikuzibira ezirimu enkora y‘okuzibira eshagama n‘obwongo hamwe n‘ekibunu ky‘okuzaara (Stinson et al., 2019), ekirikworeka obukuru bwa PPA nk‘ekintu ekirikutegyeka ebirikukorwa obukooko. N‘obu omurimo ogurikubaasa kukorwa PPA nk‘enshonga y‘akabi ahabw‘oburwaire bwa autism hati guri omu kucondooza, ebirikurugamu aha bantu abaine oburwaire bwa autism nibibaasa kurenga aha kureetaho okutaana kw‘obutafaari bw‘omubiri.
Obumanyiso bw‘omu nda nk‘okusheeshasheesha n‘okusheeshasheesha nibukira kubaho omu barwaire abaine oburemeezi bw‘okukura kw‘obutafaari bw‘omubiri (Cao et al., 2021). Okucondooza okukozirwe enyima kworekire ngu obutafaari bw‘omubiri oburikwetwa microbiome bw‘abarwaire abaine oburemeezi bw‘okuhurira (ASD) nibutandukaana n‘obw‘abantu abaine amagara marungi, ekirikumanyisa ngu hariho obutafaari bw‘omubiri oburikwetwa gut microbiota dysbiosis (Finegold et al., 2010). Omu muringo nigwo gumwe, emitwarize ya microbiome y‘abarwaire abaine endwara z‘okuzimba kw‘amara, okugira omubiri muhango, endwara ya Alzheimer, n‘ebindi nabyo nibitaana n‘eby‘abantu abaine amagara marungi (Turnbaugh na bagyenzi be, 2009; Vogt na bagyenzi be, 2017; Henke na bagyenzi be, 2019). Kwonka, kuhitsya hati, tihaine nkoragana y‘enshonga erikuteebwaho ahagati y‘ebicweka by‘omubiri ebikye eby‘omunda n‘endwara z‘ebihaha nari obumanyiso (Yap et al., 2021), n‘obu ebika bya bacteria bitari bimwe na bimwe biraabe nibiteekateekwaho kukora omurimo omu bimwe aha mbeera z‘endwara ezi. Nk‘eky‘okureeberaho, obukooko oburikwetwa Akkermansia, Bacteroides, Clostridium, Lactobacillus, Desulfovibrio n‘ebindi bika nibikira kuba bingi omu bihingwa ebikye eby‘abarwaire abaine oburwaire bwa autism (Tomova n‘abandi, 2015; Golubeva n‘abandi, 2017; Cristiano n‘abandi, 2018; Zurita20 na bagyenzi be. Ekikuru, ebika bya ba memba b’emwe aha miringo egi nibimanyirwa aha kugira ebiriisa ebiine akakwate n’okukora PPA (Reichardt na bagyenzi be, 2014; Yun na Lee, 2016; Zhang na bagyenzi be, 2019; Baur na Dürre, 2023). Ahakuba PPA neebaasa kurwanisa obukooko oburikureeta endwara, okwongyera aha bwingi bwayo nikibaasa kuba eky’omugasho aha kukura kw’obukooko oburikureeta PPA (Jacobson et al., 2018). Nahabwekyo, omwanya ogurimu PFA nigubaasa kureetera empindahinduka omu gut microbiota, obariiremu obukooko oburikureeta endwara omu nda, oburikubaasa kuba enshonga ezirikubaasa kureetera obumanyiso bw‘omu nda.
Ekibuuzo kikuru omu kucondooza kwa microbiome n‘okureeba yaaba entaaniso omu nkora ya microbial n‘enshonga nari akamanyiso k‘endwara eziri ahansi. Eidara ry‘okubanza omu kushoboorora akakwate kagumire akari ahagati y‘eby‘okurya, obukooko obukye omu nda, n‘endwara z‘ebihaha n‘okushwijuma ebirikuruga omu by‘okurya aha nkora y‘obukooko obukye. Okubaasa kuhikiiriza eki, tukakoresa okukuratana kwa metagenomic okushomirwe obwire buraingwa kugyeragyeranisa obutafaari bw’omunda bw’abaana b’embeba ezaabaire ziriisiibwe eby’okurya ebirimu PPA nari ebitarimu PPA. Abaana bakaba bariisibwa eby’okurya nk’ebya banyinabo. Twateekateeka ngu eby‘okurya ebirimu PPA nibiija kurugamu empindahinduka omu nkora y‘obukooko bw‘omunda n‘emihanda y‘okukora kw‘obukooko, namunonga ebirikukwata aha nkora ya PPA na/nari okukora PPA.
Okucondooza oku kukakoresa embeba eziine ebiriisa bya FVB/N-Tg(GFAP-GFP)14Mes/J (Jackson Laboratories) ezirikworeka munonga puroteyini y‘ekibira (GFP) ahansi y‘obushoboorozi bw‘omuhi w‘obuhwezi wa GFAP orikukuratira obuhabuzi bwa Yunivasite ya Central Florida 2000002). Bwanyima y’okuruga omu mabeere, embeba zikaba ziteebwa omu bishengye buri omwe n’embeba 1–5 eza buri kihama ahari buri kashengye. Embeba zikaba ziriisibwa ad libitum n‘eby‘okurya by‘okutegyeka ebishemeziibwe (eby‘okurya by‘omutindo ebihindwirwemu, 16 kcal% ebishaju) nari eby‘okurya ebirimu sodium propionate (eby‘okurya by‘omutindo ebihindwirwemu, 16 kcal% ebishaju, ebirimu 5,000 ppm sodium propionate). Omuhendo gwa sodium propionate ogwakozesiibwe gukaba nigwingana na 5,000 mg PFA/kg y‘oburemeezi bw‘eby‘okurya byona. Eki nikyo kirungo kya PPA ekirikukirayo obuhango ekiikiriziibwe kukoresibwa nk’ekintu ekirikurinda eby’okurya. Okwetebeekanisiza okucondooza oku, embeba z’abazaire zikaba ziriisibwa eby’okurya byombi kumara wiiki 4 zitakateeraine kandi zikagumizamu omu bwire bwona obu dam yaabaire aine enda. Embeba z‘abaana [embeba 22, embeba 9 ezaabaire nizitegyekwa (enshaija 6, enkazi 3) na PPA 13 (enshaija 4, enkazi 9)] zikaruga aha mabeere reero zaagumizamu n‘eby‘okurya nibyo bimwe nk‘ebiziba kumara ameezi 5. Embeba z’abaana zikaba ziheebwayo nk’ekitambo ziine ameezi 5 kandi ebiri omu nshaho zaabo z’omu mara bikatorotoorwaho kandi omu kubanza byabiikwa omu bikopo bya microcentrifuge ebya miriyoni 1.5 ahari -20°C reero byatwarwa omu firiiji ya -80°C kuhitsya obu DNA y’omugyenyi yaahwaho kandi microbial nucleic acids zaihwaho.
DNA y’omugyenyi ekaihwaho kurugiirira aha nkora ehindwirwemu (Charalampous na bagyenzi be, 2019). Omu bugufu, ebiri omu nshaho bikatwarwa omu 500 μl ya InhibitEX (Qiagen, Cat#/ID: 19593) kandi byabiikwa biri omu njura. Okukora ebihimba by’omubiri ebitarikurenga 1-2 buri kushohora. Ebirimu omu nshaho bikarugaho byahindurwa birikushushana omu muringo gw’ebyoma barikukoresa ekikopo ky’ekiveera omunda y’ekikopo kubaasa kukora ekirungo. Centrifuge eshampu aha 10,000 RCF kumara edakiika 5 nari kuhisya obu eshampu ziraabe ziine obujuma, reero oshushe ebiri aha mutwe kandi ogarukye oshukyemu obujuma omu 250 μl 1× PBS. Yongyera 250 μl 4.4% saponin omubazi (TCI, enamba y’ekintu S0019) aha sampo nk’ekintu ky’okushemeza okusheeshuura obutafaari bw’obutafaari bwa eukaryotic. Sampuli zikaba zitabura mpora mpora kuhitsya obu zaabaire zishemeire kandi ziteebwa omu bwire bw’obutagatsi bw’omu kishengye kumara edakiika 10. Ekindi, okutabura obutafaari bwa eukaryotic, amaizi 350 μl agatarimu nuclease gakateebwa omu sampo, gakateebwa omu nkora kumara 30 s, reero 12 μl 5 M NaCl gakateebwamu. Sampuli zikaba ziteebwa omu nkora ya centrifuge aha 6000 RCF kumara edakiika 5. Aspirate ebiri aha mutwe kandi ogarukye oshukye omu 100 μl 1X PBS. Okwihaho DNA y’omugyenyi, yongyeramu 100 μl y’ekirungo kya HL-SAN (12.8568 g NaCl, miriyoni 4 za MgCl2 1M, miriyoni 36 z’amaizi agatarimu nukiriya) n’ekirungo kya HL-SAN 10 μl (ArticZymes P/N 70910-202). Sampuli zikaba zitabura gye barikuziteera omu pipe kandi ziteebwa omu 37 °C kumara edakiika 30 aha 800 rpm aha EppendorfTM ThermoMixer C. Bwanyima y’okuziteeka, zikateebwa omu 6000 RCF kumara edakiika 3 kandi zaanaabwa emirundi ebiri na 800 μ0μl na P01. Eky’aha muheru, garukamu oshukye omu 100 μl 1X PBS.
DNA yoona ey’obukooko ekataanisibwa barikukoresa ekikwato ky’okushemeza DNA y’omugabe wa New England (New England Biolabs, Ipswich, MA, Cat# T3010L). Enkora y’okukora erikuheebwa n’ekikwato neehindurwamu kakye. Okufukaana n’okurinda amaizi agatarimu nuclease aha 60°C otakatandikire kukora ahabw’okusheeshasheesha okwahamuheru. Yongyera 10 μl ya Proteinase K na 3 μl ya RNase A ahari buri sampo. Reero otemu 100 μl ya Cell Lysis Buffer kandi otabwire mpora mpora. Sampuli bwanyima zikateebwa omu EppendorfTM ThermoMixer C aha 56°C na 1400 rpm kumara haakiri eshaaha 1 kuhika aha shaaha 3. Sampuli ezaabaire ziteirwe omu nkora y‘okukyebera zikaba ziteirwe omu nkora ya centrifuge aha 12,000 RCF kumara edakiika 3 kandi ebiri ahaiguru kuruga omu buri sampuli bikatwarwa omu nkora y‘okwetaanisaho eya 1.5 mL microcentrifuge tube erimu 400 μL y‘omubazi gw‘okukwatira hamwe. Ebipimo ebyo bikagaruka byateebwa omu nkora ya pulse vortex kumara sekondi 5–10 aha bwire bwa sekondi 1. Hindurira amaizi goona agari omu buri sampo (enkumi 600–700 μL) omu kacupa k‘okushwijuma akateirwe omu ntuumo y‘okushorooza erikugyendera omu maizi. Ebipimo bikateebwa omu nkora ya centrifuge aha 1,000 RCF kumara edakiika 3 kubaasa kuha orusa DNA y’okubanza kukwatira hamwe reero byateebwa omu centrifuge aha 12,000 RCF kumara edakiika 1 kwihaho amaizi agasigaireho. Enyomyo y‘eky‘okureeberaho ekatwarwa omu ntuumo nsya y‘okushorooza reero bwanyima yayozibwa emirundi ebiri. Aha kwozya kw‘okubanza, yongyera 500 μL y‘ekirungo ky‘okwozya ahari buri ntuumo. Hindura ekikopo emirundi 3–5 reero okore centrifuge aha 12,000 RCF kumara edakiika 1. Ote amaizi kuruga omu ntuumo y‘okushorooza kandi ote akacupa k‘okushwijuma ogarukye omu ntuumo niyo emwe ey‘okushorooza. Aha kwozya kwa kabiri, yongyera 500 μL y‘ekirungo ky‘okwozya omu kicupuri otarikuhindura. Sampuli zikaba ziteirwe omu 12,000 RCF kumara edakiika 1. Hindurira ekicweka ky‘okushwijuma omu 1.5 mL LoBind® tube kandi otemu 100 μL y‘amaizi agatarimu nuclease agabandize kwotsya. Ebishwijumirwe bikateebwa omu bwire bw’obutagatsi bw’omu kishengye kumara edakiika 1 reero byakyeberwa aha 12,000 RCF kumara edakiika 1. DNA eyashohoziibwe ekabiikwa aha -80°C.
Okukyebera kwa DNA kukakyeberwa barikukoresa ekipimo kya QubitTM 4.0 Fluorometer. DNA ekatebeekanisibwa barikukoresa ekikwato kya QubitTM 1X dsDNA (Cat. No. Q33231) kurugiirira aha biragiro by’omukozi. Okugabwa kw’oburaingwa bw’ebicweka bya DNA kukapimwa barikukoresa AglientTM 4150 nari 4200 TapeStation. DNA ekatebeekanisibwa barikukoresa ebikwato bya AgilentTM eby’okureeberaho bya DNA (Cat. No. 5067-5366) hamwe n’eby’okureeberaho bya DNA y’omubiri (Cat. No. 5067-5365). Okutebeekanisa eibiikiro ry’ebitabo kukakorwa barikukoresa ekikwato kya Oxford Nanopore TechnologiesTM (ONT) (SQK-RPB004) kurugiirira aha biragiro by’omukozi. DNA ekakuratana barikukoresa ekikwato kya ONT GridIONTM Mk1 ekirimu obutafaari bw’okusheesheka bwa Min106D (R 9.4.1). Okutebeekanisa kw‘okukuratana kukaba kuri: okuteera esimu y‘omusingi ey‘obuhikire bw‘ahaiguru, omuhendo gwa q oguri ahansi ya 9, okutebeekanisa kwa barcode, n‘okusharaho kwa barcode. Sampuli zikakuratana kumara eshaaha 72, bwanyima y‘ekyo base call data ekaheebwayo kubaasa kugaruka kukora n‘okushwijuma.
Okukora aha by’amagara g’abantu kukakorwa barikukoresa emiringo eyashobooroirwe enyima (Greenman et al., 2024). Fayiro za FASTQ ezatungirwe kuruga omu kukuratana zikabaganisibwamu endagiriro za buri sampo. Okushwijuma kwa bioinformatics kutakabaireho, ebihandiiko bikakorwa barikukoresa omuhanda ogu: eky’okubanza, fairo za FASTQ ez’eby’okureeberaho zikaba zigambwaho omu fairo emwe eya FASTQ. Bwanyima, okushoma okukye okukira ahari 1000 bp kukashwijumwa barikukoresa Filtlong v. 0.2.1, n‘ekicweka kyonka ekyahindukire kiri –min_length 1000 (Wick, 2024). 1.41.3 n’ebintu ebi: –fastq –plots dot –N50 -o .
Aha kushoroora omu nkora y’okushoroora, ebishomwa n’ebiteeraine bikashoroorwa barikukoresa Kraken2 v. 2.1.2 ( Wood et al., 2019 ). Okukora ripooti na fairo z‘okushohoza ahabw‘okushoma n‘okuteerana, nk‘oku biri. Koresa –use-names enkora y‘okushwijuma ebishomirwe n‘enteerane. Eby’okutooranamu –gzip-compressed na –paired nibishoboororwa ahabw’ebicweka by’okushoma. Obungi bw’ebicweka by’omubiri ebiri omu nkora y’ebicweka by’omubiri bukateberezibwa barikukoresa Bracken v. 2.8 ( Lu et al., 2017 ). Tukabanza twakora ekihandiiko kya kmer ekirimu ebicweka 1000 turikukoresa bracken-build n’ebintu ebi: -d .
Okushoboorora aha nkora y‘obuzaarwa n‘okutebereza aha bwingi bw‘ebintu bikakorwa barikukoresa omuringo oguhindwirwemu ogwashobooroirwe Maranga na bagyenzi be. (Maranga n’abandi, 2023). Eky‘okubanza, ebicweka by‘omubiri ebikye ebirikukira ahari 500 bp bikaihwaho omu nteerane zoona barikukoresa SeqKit v. 2.5.1 (Shen na bagyenzi be, 2016). Enteerane ezatoorainwe zikaba zigambwaho hamwe zikahinduka pan-metagenome. Ebihandiiko by‘okushoma ebikingire (ORFs) bikamanyirwa aha kukoresa Prodigal v. 1.0.1 (omuringo ogurikushushana n‘ogwa Prodigal v. 2.6.3) n‘ebicweka ebi: -d .
Enyuguta z‘omubiri zikabanza zateebwa omu guruupu kurugiirira aha bihandiiko bya Kyoto Encyclopedia of Genes and Genomes (KEGG) ortholog (KO) ebirikumanyisa eggNOG kugyeragyeranisa obungi bw‘emihanda y‘enganda. Enyuguta ezitaine knockouts nari genes eziine knockouts nyingi zikaba ziihwaho zitakashwijumirwe. Omubaro gw‘obungi bwa buri KO ahari buri sampo bwanyima gukabarwa kandi okucondooza kw‘embariirira kukakorwa. Enyuguta z‘okuhindura eby‘okurya bya PPA zikashoboororwa nk‘enkora yoona eyaabaire neeheebwa orunyiriri ko00640 omu nkora ya KEGG_Pathway, ekirikworeka omurimo omu kuhindura eby‘okurya bya propionate kurugiirira ahari KEGG. Enyuguta ezimanyirwe nk‘eziine akakwate n‘okukora PPA zihandiikirwe omu nkora y‘okwongyeraho 1 (Reichardt na bagyenzi be, 2014; Yang na bagyenzi be, 2017). Okukyebera kwa permutation kukakorwa kubaasa kumanya okuhindura kwa PPA n’okukora ebiriisa ebyabaire biri bingi munonga omu buri muringo gw’eky’okureeberaho. Okuhindura rukumi kukakorwa ahari buri nkora y’obuzaarwa eyashwijumirwe. Omuhendo gwa p ogwa 0.05 gukakoresibwa nk‘okusharaho kubaasa kumanya obukuru bw‘embariirira. Ebihandiiko ebirikukora bikaheebwa ebiriisa by’omuntu omwe omu kigombe kurugiirira aha bihandiiko by’ebiriisa ebirikujwekyera ebiri omu kigombe. Taxa eziine akakwate n‘okuhindura kwa PPA na/nari okukora PPA nizibaasa kumanyirwa aha kugyeragyeranisa IDs za contig omu fairo z‘okushohoza za Kraken2 na IDs za contig nizo zimwe ezibiikwa omu bwire bw‘okuhandiika ebirikukora barikukoresa eggNOG. Okukyebera obukuru kukakorwa barikukoresa ekigyezo kya Mann-Whitney U ekyashoboororwa enyima. Okutereeza ahabw‘okukyebera emirundi mingi kukakorwa barikukoresa enkora ya Benjamini-Hochberg. Omuhendo gwa p ogwa ≤ 0.05 gukakoresibwa nk‘okusharaho kubaasa kumanya obukuru bw‘embariirira.
Obutashushana bw’ebintu ebikye ebiri omu nda y’embeba bukakyeberwa barikukoresa enkora ya Simpson diversity index. Tihariho ntaaniso mpango eyaareebekire ahagati y‘eby‘okureeberaho by‘okukyebera n‘ebya PPA omu kureeba obutashushana bw‘ebika n‘ebika (p-omuhendo gw‘ebika: 0.18, p-omuhendo gw‘ebika: 0.16) (ekishushani 1). Bwanyima, ebicweka bya microbial bikagyeragyeranisibwa barikukoresa okucondooza kw‘ebicweka ebikuru (PCA). Ekishushani 2 nikyoreka okuteerana kw’eby’okureeberaho kurugiirira aha phyla yaabyo, ekirikworeka ngu hakaba hariho entaaniso omu nkora y’ebika by’ebintu ebikye ahagati y’eby’okureeberaho bya PPA n’eby’okukyebera. Okuteerana oku kukaba kutari kuhango aha rurengo rw‘ekika, ekirikumanyisa ngu PPA neekwata aha bacteria obumwe (Ekishushani ky‘okwongyeraho 1).
Ekishushani 1. Obutashushana bwa Alpha obw’ebika n’ebika by’ebihangirwe ebiri omunda y’embeba. Ebishushani by‘omu bokisi ebirikworeka obubonero bwa Simpson obw‘entaaniso y‘ebika (A) n‘ebika (B) omu PPA n‘eby‘okureeberaho by‘okutegyeka. Obukuru bukamanyirwa aha kukoresa ekigyezo kya Mann-Whitney U, kandi okutereeza emirundi mingi kukakorwa barikukoresa enkora ya Benjamini-Hochberg. ns, p-omuhendo gukaba gutari gw‘omugasho (p>0.05).
Ekishushani 2. Ebyarugamu omu kucondooza kw‘ebicweka ebikuru eby‘omubiri gw‘embeba aha rurengo rw‘ebika. Puloti y‘okushwijuma ebicweka ebikuru neeyoreka oku eby‘okureeberaho byagabwa omu bicweka byabyo bibiri ebikuru eby‘okubanza. Erangi nizooreka ekika ky‘eky‘okureeberaho: Embeba eziine PPA nizishushana n‘eza purpura kandi embeba ezirikutegyekwa nizishushana n‘eza kyenju. Ebicweka ebikuru 1 na 2 nibihandiikwa aha x-axis na y-axis, nk‘oku biri, kandi nibyorekwa nk‘omubaro gwabyo gw‘okuhinduka ogushobooroirwe.
Okukozesa RLE ehindwirwemu ebihandiiko by‘okubara, okukyendeera kw‘amaani omu nkora ya Bacteroidetes/Bacilli eya ahagati kukareebeka omu mbeba z‘okukyebera n‘eza PPA (okukyebera: 9.66, PPA: 3.02; p-omuhendo = 0.0011). Entaaniso egi ekaba neereetwa obungi bwa Bacteroidetes omu mbeba za PPA kugyeragyeranisa n’ezirikutegyekwa, n’obu entaaniso eraabe etari mpango (okukyebera nikumanyisa CLR: 5.51, PPA nikimanyisa CLR: 6.62; p-omuhendo = 0.054), obwo obungi bwa Bacteroidetes buri nk’obumwe (okukyebera nikumanyisa CLR7.6PA:7.7. p-omuhendo = 0.18).
Okushwijuma obungi bw’ebicweka by’omubiri eby’omunda (gut microbiome) kukashohoza ngu phylum 1 n’ebika 77 bikataanira kimwe ahagati ya PPA n’eby’okureeberaho by’okukyebera (Ekipande ky’okwongyeraho 2). Obungi bw’ebika 59 omu sampo za PPA bukaba buri ahaiguru munonga kukira obw’omu sampo z’okukyebera, obwo obungi bw’ebika 16 byonka omu sampo z’okukyebera bukaba buri ahaiguru kukira obw’omu sampo za PPA (ekishushani 3).
Ekishushani 3. Obungi bw‘ebicweka by‘omubiri ebitarikushushana omu gut microbiome ya PPA n‘embeba ezirikutegyeka. Ebishushani by’enshozi z’omuriro nibyoreka entaaniso omu bwingi bw’ebika (A) nari ebika (B) ahagati ya PPA n’eby’okureeberaho by’okukyebera. Obutonyezi oburikwiragura nibworeka ngu tihaine ntaaniso mpango omu bwingi bw’ebika. Obutonyezi oburimu erangi nibworeka entaaniso mpango omu bwingi (p-value ≤ 0.05). Ebika 20 eby‘ahaiguru ebiine entaaniso mpango omu bwingi ahagati y‘ebika by‘eby‘okureeberaho nibyorekwa omu rangi erikutukura n‘eya bururu erikutukura (eby‘okureeberaho by‘okutegyeka n‘ebya PPA), nk‘oku biri. Obutonyezi bwa kyenju na purple bukaba buri haakiri emirundi 2.7 omu kukyebera nari omu sampo za PPA kukira omu kukyebera. Obutonyezi oburikwiragura nibujwekyera taxa eziine obungi obutarikushushana munonga, n‘entaaniso za CLR eziri ahagati ya -1 na 1. Emihendo ya P ekabarwa barikukoresa ekigyezo kya Mann-Whitney U kandi ekaterezibwa ahabw‘okukyebera kwingi barikukoresa enkora ya Benjamini-Hochberg. Entaaniso za CLR eziri omu nkora ya Bold mean nizooreka entaaniso z‘amaani omu bwingi.
Bwanyima y‘okushwijuma ebiri omunda y‘obukooko obukye, tukakora okushoboorora okurikukoresibwa aha bukooko obukye. Bwanyima y’okushwijuma ebiriisa by’omutindo gwa hansi, ebiriisa by’omutaano 378,355 bikamanyirwa omu sampo zoona. Obungi obuhindukire obw‘ebicweka ebi bukakoresibwa omu kucondooza kw‘ebicweka ebikuru (PCA), kandi ebyarugamu bikooreka okuteerana kw‘ahaiguru kw‘ebika by‘eky‘okureeberaho kurugiirira aha nkora zaabo z‘emirimo (ekishushani 4).
Ekishushani 4. Ebirugire omu PCA barikukoresa enkora y‘okukora kw‘embeba gut microbiome. Puloti ya PCA neeyoreka oku sampuli zigabirwe omu bicweka byazo bibiri ebikuru eby‘okubanza. Erangi nizooreka ekika ky‘eky‘okureeberaho: Embeba eziine PPA nizishushana n‘eza purpura kandi embeba ezirikutegyekwa nizishushana n‘eza kyenju. Ebicweka ebikuru 1 na 2 nibihandiikwa aha x-axis na y-axis, nk‘oku biri, kandi nibyorekwa nk‘omubaro gwabyo gw‘okuhinduka ogushobooroirwe.
Tukakuratiraho kukyebera obungi bw‘okuteera kwa KEGG omu miringo y‘eky‘okureeberaho etari emwe n‘emwe. Omubaro gw‘ebihimba by‘omutaano 3648 bikamanyirwa, ahari ebyo 196 bikaba biri bingi munonga omu sampo z‘okukyebera kandi 106 bikaba biri bingi munonga omu sampo za PPA (ekishushani 5). Omubaro gw’ebicweka by’omubiri 145 bikashangwa omu sampo z’okukyebera kandi 61 omu sampo za PPA, n’obungi obutarikushushana munonga. Emihanda erikukwatagana n‘okuhindura kw‘amaizi g‘omubiri n‘eshagama y‘omubiri (aminosugar metabolism) ekaba erimu obugaiga bwingi omu sampo za PPA (Ekipande ky‘okwongyeraho 3). Emihanda erikukwatagana n‘okuhindura nitrogen n‘enkora z‘okujanjaaza sulfur zikaba ziri nyingi munonga omu sampo z‘okutegyeka (Ekipande ky‘okwongyeraho 3). Obungi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura ebiriisa by‘omubiri (ko:K21279) n‘okuhindura ebiriisa by‘omubiri (ko:K07291) bukaba buri ahaiguru munonga omu sampo za PPA (ekishushani 5). Sampuli z‘okukyebera zikaba ziine ebiriisa bingi munonga ebiine akakwate n‘okuhindura kwa benzoate (ko:K22270), okuhindura kwa nitrogen (ko:K00368), n‘okuhindura kwa shukaari/shukaari (ko:K00131) ( Ekishushani 5 ).
Ekishushani 5. Obungi bw‘entaaniso bwa KOs omu gut microbiome ya PPA n‘embeba ezirikutegyekwa. Puloti y‘orushozi rw‘omuriro neeyoreka entaaniso omu bwingi bw‘ebicweka by‘emirimo (KOs). Obutonyezi bw‘enzirugavu nibworeka KOs eziine obungi bwazo obutaine ntaaniso mpango ahagati y‘ebika by‘eky‘okureeberaho (p-value > 0.05). Obutonyezi oburimu erangi nibworeka entaaniso mpango omu bwingi (p-value ≤ 0.05). KO 20 eziine entaaniso mpango omu bwingi ahagati y‘ebika by‘eby‘okureeberaho nizoorekwa omu rangi erikutukura n‘eya bururu erikutukura, ezirikuhikaana n‘eky‘okureeberaho ky‘okutegyeka n‘ekya PPA, nk‘oku biri. Obutonyezi bwa kyenju na purple nibworeka KOs ezaabaire ziri haakiri emirundi 2.7 nyingi omu sampo z‘okukyebera n‘eza PPA, nk‘oku biri. Obutonyezi oburikwiragura nibworeka KOs eziine obungi obutarikushushana munonga, n‘entaaniso za CLR eziri ahagati ya -1 na 1. Emihendo ya P ekabarwa barikukoresa ekigyezo kya Mann-Whitney U kandi ekahindurwa ahabw‘okugyeragyeranisa kwingi barikukoresa enkora ya Benjamini-Hochberg. NaN neeyoreka ngu KO teri y‘omuhanda omu KEGG. Bold mean CLR emihendo y‘entaaniso neeyoreka entaaniso z‘amaani omu bwingi. Okumanya ebirikukiraho aha mihanda ei KO ezihandiikirwe zirimu, reeba ekipande ky‘okwongyeraho 3.
Omuri ebyo ebihandiikirwe, ebiriisa 1601 bikaba biine obungi obutarikushushana munonga ahagati y’ebika by’eky’okureeberaho (p ≤ 0.05), buri nkora y’obuzaarwa ekaba eri haakiri emirundi 2.7 y’obungi. Omuri ebyo, ebiriisa 4 bikaba biri bingi omu sampo z’okukyebera kandi ebiriisa 1597 bikaba biri bingi omu sampo za PPA. Ahabw‘okugira ngu PPA eine obushoboorozi bw‘okurwanisa obukooko, tukashwijuma obungi bw‘okuhindura PPA n‘okukora ebiriisa ahagati y‘ebika by‘eky‘okureeberaho. Omuri ebyo 1332 ebiine akakwate n‘okuhindura eby‘okurya bya PPA, ebiriisa 27 bikaba biri bingi munonga omu sampo z‘okukyebera kandi ebiriisa 12 bikaba biri bingi munonga omu sampo za PPA. Omuri ebyo 223 ebiine akakwate n’okukora PPA, ebiriisa 1 bikaba biri bingi munonga omu sampo za PPA. Ekishushani 6A nikigaruka kworeka obungi bw‘ahaiguru bw‘ebicweka by‘omubiri ebirikwejumba omu kuhindura kwa PPA, n‘obungi bw‘ahaiguru munonga omu sampo z‘okutegyeka n‘obuhango bw‘ebirikurugamu bwingi, obwo ekishushani 6B kirikworeka obungi bw‘ebicweka by‘omubiri eby‘omuntu ahabwe n‘obungi bw‘ahaiguru munonga obureebirwe omu sampo za PPA.
Ekishushani 6. Obungi bw‘entaaniso bw‘ebicweka by‘omubiri ebiine akakwate na PPA omu nkora y‘okukyebera ebiri omu nda y‘embeba. Ebishushani by‘enshozi z‘omuriro nibyoreka entaaniso omu bwingi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura PPA (A) n‘okukora PPA (B). Obutonyezi bw‘enzirugavu nibworeka ebiriisa ebiine obungi bwabyo obutaine ntaaniso mpango ahagati y‘ebika by‘eky‘okureeberaho (p-value > 0.05). Obutonyezi oburimu erangi nibworeka entaaniso mpango omu bwingi (p-value ≤ 0.05). Enyuguta 20 eziine entaaniso mpango omu bwingi nizoorekwa omu rangi erikutukura hamwe na bururu erikutukura (eby’okureeberaho by’okukyebera na PPA), nk’oku biri. Obungi bw‘obutonyezi bwa kyenju na purple bukaba buri haakiri emirundi 2.7 omu sampo z‘okukyebera na PPA kukira omu sampo z‘okukyebera. Obutonyezi oburikwiragura nibujwekyera ebiriisa ebiine obungi obutarikushushana munonga, n‘entaaniso za CLR eziri ahagati ya -1 na 1. Emihendo ya P ekabarwa barikukoresa ekigyezo kya Mann-Whitney U kandi ekaterezibwa ahabw‘okugyeragyeranisa kwingi barikukoresa enkora ya Benjamini-Hochberg. Enyuguta nizihikaana n’enyuguta ezirikujwekyera omu katabo k’enyuguta ezitarikugarukagaruka. Amaziina g’obuzaarwa garimu akamanyiso ka KEGG akarikworeka obuzaarwa bwa KO. Bold mean CLR entaaniso nizooreka obungi obutarikushushana munonga. Akabonero (-) nikooreka ngu tihaine kamanyiso k’obuzaarwa omu nkora ya KEGG.
Taxa eziine ebiriisa ebiine akakwate na PPA metabolism na/nari okukora bikamanyirwa aha kugyeragyeranisa obumanyiso bwa taxonomic bwa contigs na contig ID y’ebiriisa. Aha rurengo rw’ebika, ebika 130 bikashangwa biine ebiriisa ebiine akakwate n’okuhindura PPA kandi ebika 61 bikashangwa biine ebiriisa ebiine akakwate n’okukora PPA (Ekipande ky’okwongyeraho 4). Kwonka tihariho kika ekyayorekire entaaniso mpango omu bwingi (p > 0.05).
Aha rurengo rw’ebika, ebika bya bacteria 144 bikashangwa biine obutafaari oburikukwatagana n’okuhindura PPA kandi ebika bya bacteria 68 bikashangwa biine obutafaari oburikukwatagana n’okukora PPA (Ekipande ky’okwongyeraho 5). Omuri PPA metabolizers, bacteria munaana zikooreka okweyongyera kw‘amaani omu bwingi ahagati y‘ebika by‘eky‘okureeberaho, kandi zoona zikooreka empindahinduka z‘amaani omu kukora (Ekipande ky‘okwongyeraho 6). Ebirikuhindura PPA byona ebyamanyirwe ebiine entaaniso mpango omu bwingi bikaba biri bingi munonga omu sampo za PPA. Okushoroora aha rurengo rw’ebika kukashohoza abajwekyerwa b’ebika ebitarikutaana munonga ahagati y’ebika by’eky’okureeberaho, harimu ebika bya Bacteroides na Ruminococcus bingi, hamwe na Duncania dubois, Myxobacterium enterica, Monococcus pectinolyticus, na Alcaligenes polymorpha. Omuri za bacteria ezirikureeta PPA, bacteria ina zikworekire entaaniso mpango omu bwingi ahagati y’ebika bya sampo. Ebika ebiine entaaniso mpango omu bwingi bikaba birimu obukooko oburikwetwa Bacteroides novorossi, Duncania bois, Myxobacterium enteritidis, na Ruminococcus bovis.
Omu kucondooza oku, tukashwijuma ebirikuruga omu kunywa PPA aha gut microbiota y’embeba. PPA neebaasa kureetaho okugarukamu okutarikushushana omu bacteria ahabw‘okuba neekorwa ebika ebimwe, neekoresibwa nk‘entandikwa y‘eby‘okurya ebika ebindi, nari eine ebirikurwanisa obukooko. Nahabwekyo, okwongyeraho kwayo omu mbeera y‘omunda kurabira omu kugaita eby‘okurya nikibaasa kugira ebirugiremu bitarikushushana kurugiirira aha kugumisiriza, okukwatwaho, n‘obushoboorozi bw‘okugikozesa nk‘entandikwa y‘ebiriisa. Ebika bya bacteria ebirikuhurira gye nibibaasa kwihwaho kandi bigarurwemu ebirikubaasa kugumira PPA nari ebirikubaasa kugikozesa nk‘entandikwa y‘eby‘okurya, ekirikureetaho empindahinduka omu nkora ya gut microbiota. Ebirugire omu kucondooza kwaitu bikooreka entaaniso mpango omu nkora y‘obukooko obukye kwonka tihaine ekirikukwata aha nkora y‘obukooko obukye bwona. Ebirugiremu ebikuru bikareebeka aha rurengo rw’ebika, n’ebika ebirikurenga 70 ebyabaire bitarikushushana munonga omu bwingi ahagati ya PPA n’eby’okureeberaho by’okukyebera (Ekipande ky’okwongyeraho 2). Okwongyera kweshwijuma aha nkora y‘eby‘okureeberaho ebyabaire biri omu PPA kukashohoza obutashushana bwingi bw‘ebika by‘obukooko obukye kugyeragyeranisa n‘eby‘okureeberaho ebitarikworekwa, ekirikumanyisa ngu PPA neebaasa kwongyera aha mitwarize y‘okukura kwa bacteria n‘okukyendeeza aha bwingi bwa bacteria oburikubaasa kuguma omu myanya erimu PPA. Nahabwekyo, PPA neebaasa kutooranamu kureetaho empindahinduka omu mwanya gw‘okureetera okutabura okujanjaire kw‘obutafaari bw‘omubiri oburikwetwa gut microbiota.
Ebirungo by‘okurinda eby‘okurya nka PPA bikaba byorekwa enyimaho ngu nibihindura obungi bw‘ebicweka by‘omubiri ebiri omu nda hatariho kukwata aha nkora yoona (Nagpal et al., 2021). Aha, tukareeba entaaniso ezirikukirayo kutangaaza ahagati y’ebika bya Bacteroidetes omu kika kya Bacteroidetes (ekyabaire nikimanywa nka Bacteroidetes enyima), ebyabaire biine obugaiga bwingi omu mbeba ezaabaire ziine PPA. Okwongyera aha bwingi bw‘ebika bya Bacteroides nikukwatagana n‘okwongyera aha kusheeshasheesha kw‘ebizimba, ekirikubaasa kwongyera aha kabi k‘okukwatwa endwara n‘okureetaho okuzimba (Cornick na bagyenzi be, 2015; Desai na bagyenzi be, 2016; Penzol na bagyenzi be, 2019). Okucondooza kumwe kukashanga ngu empunu z‘abashaija ezizairwe zijanjabirwe na Bacteroides fragilis zikooreka emitwarize y‘abantu erikwijutsya oburemeezi bw‘okuhurira (ASD) (Carmel et al., 2023), kandi okucondooza okundi kworekire ngu ebika bya Bacteroides nibibaasa kuhindura emirimo y‘obutafaari bw‘omubiri kandi bireeteho okuzimba kw‘obutafaari bw‘omubiri. 2019). Ebika by’ebinyonyi ebiri omu kika kya Ruminococcus, Prevotella, na Parabacteroides nabyo bikeyongyera munonga omu mbeba ezaabaire ziine PPA (Coretti et al., 2018). Ebika bya Ruminococcus ebimwe nibikwatagana n‘endwara nk‘endwara ya Crohn kurabira omu kureetaho ebiriisa ebirikureetaho okuzimba (Henke et al., 2019), kandi ebika bya Prevotella nka Prevotella human nibikwatagana n‘endwara z‘okuhindura eby‘okurya nk‘okugira obuhuta bw‘omubiri n‘okuhurira insuliini.6. 2017). Eky’aha muheru, tukashanga ngu omubaro gwa Bacteroidetes (ogwabandize kumanyirwa nka Firmicutes) na Bacteroidetes gukaba guri ahansi munonga omu mbeba ezaabaire ziine PPA kukira omu mbeba ezirikutegyekwa ahabw’obungi bw’ebika bya Bacteroidetes bwona bwona. Omubaro ogu gukaba gworekwa enyima nk‘eky‘okureeberaho kikuru ky‘okuguma omu mara, kandi okutabura omu mubaro ogu kubaire nikukwatagana n‘embeera z‘endwara zitari zimwe na zimwe (Turpin na bagyenzi be, 2016; Takezawa na bagyenzi be, 2021; An na bagyenzi be, 2023), obariiremu endwara z‘okuzimba kw‘amara (Stov ,2.0). Omu kugaita, ebika bya phylum Bacteroidetes nibireebeka nk‘ebirikukirayo kukwatwaho PPA y‘eby‘okurya eri ahaiguru. Eki nikibaasa kuba nikireetwaho okugumisiriza munonga PPA nari obushoboorozi bw’okukozesa PPA nk’entandikwa y’amaani, ekirikworekwa kuba eky’amazima haakiri aha kika kimwe, Hoylesella enocea (Hitch et al., 2022). Ebindi, okukoresa PPA omu bakazi nikibaasa kwongyera aha kukura kw’omwana ori omunda obwo kirikuhindura omunda y’abaana b’embeba kubaasa kukwatwaho obutwa bwa Bacteroidetes; kwonka, enkora yaitu ey’okwega ekaba etarikwikiriza okushwijuma nk’okwo.
Okucondooza aha biri omu metagenomic kukashohoza entaaniso mpango omu bwingi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura PPA n‘okukora, n‘embeba ezaabaire ziine PPA zirikworeka obungi bw‘ebicweka by‘omubiri ebirikukora PPA, obwo embeba ezitaine PPA zirikworeka obungi bw‘ebicweka by‘omubiri ebirikukora PAA (Fig6). Ebirugire omu kucondooza oku nibyoreka ngu ebirikuruga omu PPA aha nkora y‘obukooko obukye nibibaasa kutabaho ahabw‘okubikozesa kwonka, kitari ekyo obungi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura PPA bukaba bushemerire kworeka obungi bw‘ahaiguru omu gut microbiome y‘embeba ezaabaire ziine PPA. Enshoboorora emwe n‘okugira ngu PPA neereetaho obungi bwa bacteria okukira munonga kurabira omu nkora yaayo ey‘okurwanisa obukooko okukira kurabira omu kukoresibwa kwayo bacteria nk‘ebiriisa. Okucondooza okukozirwe enyima kworekire ngu PPA neezibira okukura kwa Salmonella Typhimurium omu muringo ogurikurugiirira aha dozi (Jacobson na bagyenzi be, 2018). Okwemanyiza ebiriisa bya PPA bingi nikibaasa kutoorana obukooko obutarikubaasa kurwanisa obukooko bwabwo kandi nibubaasa kutabaasa kubuhindura nari kubukora. Nk’eky’okureeberaho, ebika bya Parabacteroides bingi bikooreka obungi bw’amaani omu sampo za PPA, kwonka tihaine nshonga z’obuzaarwa eziine akakwate n’okuhindura PPA nari okuzikora ezaashangirwe (Ebipande by’okwongyeraho 2, 4, na 5). Ekindi, okukora PPA nk’ekintu ekirikuruga omu kukyebera nikijanjaara munonga omu bacteria batari bamwe na bamwe (Gonzalez-Garcia et al., 2017). Obutashushana bwa bacteria oburi ahaiguru nibubaasa kuba enshonga y‘obungi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura kwa PPA omu sampo z‘okukyebera (Averina et al., 2020). Ekindi, ebicweka 27 byonka (2.14%) ahari 1332 eby‘obuzaarwa bikateberezibwa kuba biri eby‘obuzaarwa ebiine akakwate n‘okuhindura kwa PPA kwonka. Enyuguta nyingi eziine akakwate n‘okuhindura eby‘okurya bya PPA nazo nizejumba omu miringo endiijo y‘okuhindura eby‘okurya. Eki nikiyongyera kworeka ngu obungi bw‘ebicweka by‘omubiri ebirikwejumba omu kuhindura PPA bukaba buri ahaiguru omu sampo z‘okukyebera; obutafaari obu nibubaasa kukora omu miringo etarikureetaho okukoresa PPA nari okuhangwa nk‘ekirugiremu. Omu muringo ogu, oruzaaro rumwe rwonka orurikukwatagana n‘okuzaarwa kwa PPA rukooreka entaaniso mpango omu bwingi ahagati y‘ebika by‘eky‘okureeberaho. Okutaana n’ebicweka by’omubiri ebiine akakwate n’okuhindura PPA, ebicweka by’omubiri ebirikumanyisa okukora PPA bikatooranwa ahabw’okuba nibyejumba butoosha omu muhanda gwa bacteria ogw’okukora PPA. Omu mbeba ezaabaire ziine PPA, ebika byona bikashangwa biine okweyongyera kw’amaani omu bwingi n’obushoboorozi bw’okukora PPA. Eki nikishagika okuteebereza ngu PPAs niziija kutoorana abakozi ba PPA n‘ahabw‘ekyo niziteebereza ngu obushoboorozi bw‘okukora PPA nibwija kweyongyera. Kwonka, obungi bw’ebiriisa tiburikumanyisa ngu nibukwatagana n’okureebeka kw’ebiriisa; n‘ahabw‘ekyo, n‘obu obungi bw‘ebicweka by‘omubiri ebiine akakwate n‘okuhindura kwa PPA buraabe buri ahaiguru omu sampo z‘okutegyeka, omubaro gw‘okworeka nigubaasa kuba gutarikushushana (Shi et al., 2014). Okuhamya akakwate akari ahagati y‘okujanjaara kw‘ebicweka by‘omubiri ebirikureetaho PPA n‘okukora PPA, okucondooza aha kumanyisa kw‘ebicweka by‘omubiri ebirikwejumba omu kureetaho PPA nikwetengyesa.
Okushoboorora okurikukoresibwa PPA n‘okutegyeka metagenomes kukashohoza entaaniso ezimwe. Okushwijuma kwa PCA okwa ebiri omu nkora y‘obuzaarwa kukashohoza ebicweka by‘omubiri ebitarikushushana ahagati ya PPA n‘eby‘okureeberaho by‘okutegyeka (ekishushani 5). Okuteerana omunda y‘eby‘okureeberaho kukashohoza ngu ebiri omu nkora y‘okutegyeka ebiri omu nkora y‘obuzaarwa bikaba biri bingi, obwo eby‘okureeberaho bya PPA birikuteerana hamwe. Okuteerana kurugiirira aha biri omu nkora y’obuzaarwa kukaba nikugyeragyeranisibwa n’okuteerana kurugiirira aha nkora y’ebika. Nahabwekyo, entaaniso omu bwingi bw’emihanda nizikwatagana n’empindahinduka omu bwingi bw’ebika by’ebimera n’ebika by’ebimera ebimwe ebiri omunda yaabyo. Omu sampo za PPA, emihanda ebiri erimu obungi bw‘ahaiguru munonga ekaba neekwatagana n‘okuhindura shukaari y‘omubiri/nucleotide (ko:K21279) n‘emihanda mingi y‘okuhindura shukaari y‘omubiri (ko:K00647, ko:K03801; Ekitebyo ky‘okwongyeraho 3). Enyuguta eziine akakwate na ko:K21279 nizimanyirwa aha kukwatiraine n’ekika kya Bacteroides, kimwe aha kika ekiine omubaro gw’ebika bingi munonga omu sampo za PPA. Enzyme egi neebaasa kwetantara okugarukamu kw‘obutafaari bw‘omubiri obwo erikworeka ebiriisa by‘omubiri (Wang et al., 2008). Eki nikibaasa kuba nikyo kyareeteire okweyongyera kwa Bacteroidetes okureebirwe omu mbeba eziine PPA. Eki nikihikiiriza okweyongyera kw‘okukora kw‘ebishaju ebirikureebeka omu PPA microbiome. Bacteria nizikoresa omuhanda gwa FASIIko:K00647 (fabB) kureetaho asidi z‘ebishaju, ezirikubaasa kuhindura emihanda y‘okuhindura eby‘okurya omu bagyenyi (Yao na Rock, 2015; Johnson na bagyenzi be, 2020), kandi empindahinduka omu kuhindura kw‘ebishaju nizibaasa kukora kihango omu kukura kw‘obutafaari bw‘omubiri (Yu20 na bagyenzi be, 20). Omuhanda ogundi ogwabaire nigworeka okweyongyera kw‘obungi omu sampo za PPA gukaba guri okukora ebiriisa by‘omubiri (ko:K12343). Hariho obuhame oburikweyongyera ngu hariho akakwate akarikuhinduka ahagati y‘obushoboorozi bwa gut microbiota kuhindura aha rurengo rw‘ebiriisa n‘okuhindurwa aha rurengo rw‘eshagama, nk‘oku emihendo y‘eshagama eri ahaiguru erikubaasa kugira ebirugire omu magara g‘ahansi (Tetel et al., 2018).
Okwega oku tikurimu obugarukiro n’okuteekateekaho. Entaaniso nkuru eri ngu titukakora okukyebera kw’omubiri gw’enyamaishwa. Nahabwekyo, tikirikubaasika kuhendera butunu yaaba empindahinduka omu bihingwa ebikye nibikwatagana n’endwara yoona. Ekindi ekirikuteekateekwaho n’okugira ngu embeba omu kucondooza oku zikaba ziriisibwa eby’okurya nk’ebya banyinabo. Okucondooza okw‘omumaisho nikubaasa kumanyisa yaaba okuruga aha by‘okurya ebirimu PPA kuza aha by‘okurya ebitarimu PPA nikitunguura ebirikurugamu aha bihangirwe ebikye. Ekimwe aha bizibu by’okwega kwaitu, nk’ebindi bingi, n’obuhango bw’eky’okureeberaho obukye. N‘obu ebirugiremu ebihikire biraabe nibibaasa kutunga, obuhango bw‘eky‘okureeberaho kihango nibubaasa kuha amaani maingi g‘embariirira baaba nibashwijuma ebyarugamu. Kandi nitwegyendesereza aha kuhika aha nshonga z‘okuhendera aha nkoragana eri ahagati y‘empindahinduka omu nkora y‘okukyebera ebiri omu nda n‘endwara yoona (Yap et al., 2021). Enshonga ezirikuteganisa ezirimu emyaka, obuhangwa, n‘eby‘okurya nizibaasa kuhindura munonga aha nkora y‘obukooko obukye. Enshonga ezi nizibaasa kushoboorora obutaikirizana obureebirwe omu bihandiiko ebirikukwata aha nkoragana y‘obukooko bw‘omunda n‘endwara ezigumire (Johnson et al., 2019; Lagod na Naser, 2023). Nk‘eky‘okureeberaho, ab‘ekika kya Bacteroidetes boorekire ngu nibeyongyera nari nibukyendeera omu nyamaishwa n‘abantu abaine ASD (Angelis na bagyenzi be, 2013; Kushak na bagyenzi be, 2017). Omu muringo nigwo gumwe, okucondooza aha nkora y‘amara omu barwaire abaine endwara z‘okuzimba kw‘amara kushangire okweyongyera n‘okukyendeera omu nkora emwe (Walters na bagyenzi be, 2014; Forbes na bagyenzi be, 2018; Upadhyay na bagyenzi be, 2023). Okukyendeeza aha nkora y‘okushoroora omu buhangwa, tukagyezaho kureeba ngu abakazi n‘abashaija baajwekyerwa omu muringo gw‘okwingana kugira ngu entaaniso zibe nizikira kureetwa eby‘okurya. Ekizibu kimwe eky’okuhandiika ebirikukora n’okuihaho okukuratana kw’ebicweka by’omubiri ebitarikwetengyesa. Omuringo gwaitu gw’okuteerana ebiri omu nkora y’obuzaarwa nigwetengyesa 95% y’okumanya okukuratana kw’ebintu n’okushushana kw’oburaingwa 85%, hamwe n’okukingira 90% kw’okuteerana kw’ebicweka by’omubiri kubaasa kumaraho okuteerana kw’ebishuba. Kwonka omu mbeera ezimwe, tukareeba COGs eziine ebihandiiko ebirikushushana (nk‘eky‘okureeberaho, MUT) (ekishushani 6). Ebindi bihandiiko nibyetengyesa kureeba yaaba ebicweka ebi eby’omubiri (orthologs) nibitaana, biine akakwate n’ebika ebimwe, nari shi eki n’ekizibu ky’enkora y’okuteerana ebiriisa. Ekindi ekirikuremesa okushoboorora okurikukoresibwa n‘okubaasa kushoroora kubi; omubazi gwa bacteria mmdA n’enzigye emanyirwe erikwejumba omu kuhanga propionate, kwonka KEGG terikugikwatanisa n’omuhanda gw’okuhindura propionate. Okutaana n‘ekyo, ebicweka by‘omubiri ebya scpB na mmcD nibikwatagana. Omubaro muhango gw‘ebicweka by‘omubiri ebitaine knockouts ezitebeekanisiibwe nigubaasa kurugamu obutabaasa kumanya ebicweka by‘omubiri ebiine akakwate na PPA baaba nibashwijuma obungi bw‘ebicweka by‘omubiri. Okucondooza okw‘omumaisho nikwija kugasirwa omu kucondooza kwa metatranscriptome, okurikubaasa kuha okwetegyereza okurikukirayo aha mitwarize y‘emirimo y‘ebicweka by‘omubiri ebikye ebiri omu nda n‘okukwatiraine n‘okureebeka kw‘obuzaarwa n‘ebirikubaasa kuruga omu nkora y‘okugaruka ahansi. Aha kucondooza okurikukwata aha buremeezi bw‘omutaano obw‘okukura kw‘obutafaari bw‘omubiri nari endwara z‘okuzimba kw‘amara, okushwijuma kw‘omubiri n‘emitwarize y‘enyamaishwa nikwetengyesa kukwatisa empindahinduka omu nkora ya microbiome n‘oburemeezi obu. Okucondooza okundi okurikutaho obukooko obukye omu mbeba ezitarimu obukooko nakyo nikibaasa kuba eky’omugasho kumanya yaaba obukooko obu nibureetaho nari obumanyiso bw’endwara.
Omu bugufu, tukooreka ngu PPA y‘omu by‘okurya neekora nk‘enshonga y‘okuhindura ebiri omu nkora y‘ebintu ebikye ebiri omu nda. PPA n‘ekintu ekirikurinda eby‘okurya ekiikiriziibwe FDA ekirikushangwa munonga omu by‘okurya bitari bimwe na bimwe, kyamara obwire buraingwa, nikibaasa kureetera okutabura ebimera by‘omunda ebya butoosha. Twashanga empindahinduka omu bwingi bwa bacteria nyingi, ekirikumanyisa ngu PPA neebaasa kuhindura aha nkora ya gut microbiota. Empindahinduka omu microbiota nizibaasa kureetera empindahinduka omu mitwarize y‘emihanda emwe y‘okuhindura eby‘okurya, ekirikubaasa kureetera empindahinduka omu mubiri eziine akakwate n‘amagara g‘omukaikuru/omugurusi. Ebindi bihandiiko nibyetengyesa kureeba yaaba ebirikuruga omu by‘okurya bya PPA aha nkora y‘obukooko obukye nibibaasa kureetera dysbiosis nari endwara ezindi. Okucondooza oku nikutaho omusingi gw‘okucondooza okurikwija aha ku PPA erikukora aha nkora y‘omunda erikubaasa kukwata aha magara g‘omuntu.
Ebihandiiko ebyayorekwa omu kucondooza oku nibishangwa omu bitabo ebiri aha Intaneeti. Eiziina ry’ekibiina n’enamba y’okukitunga ni: https://www.ncbi.nlm.nih.gov/, PRJNA1092431.
Okucondooza oku aha nyamaishwa kukaikirizibwa akakiiko akarikureeberera n’okukozesa enyamaishwa omu yunivasite ya Central Florida (UCF-IACUC) (Enamba y’orusa rw’okukozesa enyamaishwa: PROTO202000002). Okwega oku nikukuratira ebiragiro by’omu kicweka, ebiragiro, n’ebirikwetengyesa omu kitongore.
NG: Okuteekateeka, Okutebeekanisa amakuru, Okushwijuma omu butongore, Okucondooza, Enkora, Puroguraamu, Okureeba, Okuhandiika (ekihandiiko ky‘okubanza), Okuhandiika (okushwijuma & okuhindura). LA: Okuteekateeka, okurinda amakuru, Enkora, Ebikwato, Okuhandiika (okushwijuma & okuhindura). SH: Okushwijuma omu butongore, Sofutiweya, Okuhandiika (okushwijuma & okuhindura). SA: Okucondooza, Okuhandiika (okushwijuma & okuhindura). Omuramuzi omukuru: Okucondooza, Okuhandiika (okushwijuma & okuhindura). SN: Okuteekateeka, okutegyeka purojekiti, eby‘okukoresa, okureeberera, okuhandiika (okushwijuma & okuhindura). TA: Okuteekateeka, okutegyeka purojekiti, okureeberera, okuhandiika (okushwijuma & okuhindura).
Abahandiiki bakarangirira ngu tibatungire buhwezi bw’esente bwona ahabw’okucondooza, okuhandiika, na/nari okushohoza ekihandiiko eki.
Abahandiiki nibarangirira ngu okucondooza oku kukakorwa hatariho nkoragana yoona ey’eby’obushuubuzi nari ey’esente erikubaasa kushoboororwa nk’erikubaasa kureetaho okutaikirizana. tikirikukoresibwa.
Ebiteekateeko byona ebiri omu kicweka eki n’eby’abahandiiki bonka kandi tibirikworeka ebiteekateeko by’ebitongore byabo, abahandiiki, abahandiiki, nari abarikubicondooza. Ebintu byona ebirikushwijumwa omu kicweka eki, nari ebi ababikozire barikugira, tibirikuhamibwa nari kuhamibwa omuhandiiki.
Ebihandiiko by’okwongyeraho eby’ekihandiiko eki noobaasa kubitunga aha Intaneeti: https://www.
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