Aside Propionic (PPA), umuti urwanya icyorezo n'inyongeramusaruro isanzwe mu mirire, byagaragaye ko itera imikurire idasanzwe mu mbeba hamwe n'imikorere mibi y'igifu, ishobora guterwa na dysbiosis y'igifu. Hagaragajwe isano iri hagati yo guhura na PPA mu mirire na dysbiosis y'igifu, ariko ntabwo yasuzumwe mu buryo butaziguye. Aha, twasuzumye impinduka zijyanye na PPA mu miterere ya microbiota mu gifu zishobora gutera dysbiosis. Microbiome zo mu gifu cy'imbeba zagaburirwaga indyo itavuwe (n = 9) n'indyo ikungahaye kuri PPA (n = 13) byakurikiranywe hakoreshejwe uburyo bwo gusuzuma ihinduka ry'imiterere y'udukoko mu gihe kirekire kugira ngo harebwe itandukaniro riri hagati y'imiterere ya mikorobe n'inzira za bagiteri. PPA mu mirire yajyanye no kwiyongera k'ubwinshi bw'udukoko twinshi, harimo ubwoko bwinshi bwa Bacteroides, Prevotella, na Ruminococcus, amoko yabwo akaba yaranagize uruhare mu ikorwa rya PPA. Microbiome z'imbeba zahuye na PPA nazo zari zifite inzira nyinshi zijyanye na metabolism ya lipide na hormone ya steroide. Ibisubizo byacu bigaragaza ko PPA ishobora guhindura microbiota mu gifu n'inzira zayo za metabolisme. Izi mpinduka zagaragaye zigaragaza ko imiti irinda ibiryo yashyizwe mu byiciro by’umutekano wo kuribwa ishobora kugira ingaruka ku miterere ya mikorobe zo mu mara, ndetse no ku buzima bw’abantu. Muri byo, P, G cyangwa S bitoranywa bitewe n'urwego rw'ishyirwa mu byiciro riri gusesengurwa. Kugira ngo hagabanywe ingaruka zo gushyira mu byiciro ibinyoma, hashyizweho ingano ntoya y'ubwinshi bw'ibipimo ya 1e-4 (1/10.000 byasomwe). Mbere y'isesengura ry'ibarurishamibare, ubwinshi bw'ibipimo byavuzwe na Bracken (ibice_bipimo_byose_byasomwe) byahinduwe hakoreshejwe impinduka ya log-ratio (CLR) (Aitchison, 1982). Uburyo bwa CLR bwatoranyijwe mu guhindura amakuru kuko buhinduka mu buryo busanzwe kandi buhagije ku makuru atari make (Gloor et al., 2017). Ihinduka rya CLR rikoresha logarithm karemano. Amakuru y'ibarura yatangajwe na Bracken yahinduwe hakoreshejwe imvugo ya log (RLE) (Anders na Huber, 2010). Imibare yakozwe hakoreshejwe uruvange rwa matplotlib v. 3.7.1, seaborn v. 3.7.2 na logarithms zikurikiranye (Gloor et al., 2017). 0.12.2 na stantanotations v. 0.5.0 (Hunter, 2007; Waskom, 2021; Charlier et al., 2022). Igipimo cya Bacillus/Bacteroidetes cyabazwe kuri buri ngero hakoreshejwe imibare ya bagiteri isanzwe. Agaciro kavuzwe mu mbonerahamwe kazengurutswe ahantu hagera kuri 4 decimal. Igipimo cya Simpson diversity kibazwe hakoreshejwe alpha_diversity.py script yatanzwe muri paki ya KrakenTools v. 1.2 (Lu et al., 2022). Raporo ya Bracken iratangwa muri script naho igipimo cya Simpson "Si" gitangwa kuri parametre ya -an. Itandukaniro rikomeye mu bwinshi ryasobanuwe nk'itandukaniro ry'ikigereranyo cya CLR ≥ 1 cyangwa ≤ -1. Itandukaniro ry'ikigereranyo cya CLR cya ±1 rigaragaza ukwiyongera kwikubye 2.7 mu bwinshi bw'ubwoko bw'icyitegererezo. Ikimenyetso (+/-) kigaragaza niba taxon ari nyinshi mu gipimo cya PPA n'icyitegererezo cyo kugenzura, uko bikurikirana. Akamaro kagaragajwe hakoreshejwe ikizamini cya Mann-Whitney U (Virtanen et al., 2020). Statsmodels v. 0.14 (Benjamini na Hochberg, 1995; Seabold na Perktold, 2010) niho hakoreshejwe, kandi uburyo bwa Benjamini-Hochberg bwakoreshejwe mu gukosora ibizamini byinshi. Agaciro ka p kahinduwe ≤ 0.05 ni ko kakoreshejwe nk'umupaka wo kumenya akamaro k'ibarurishamibare.
Mikorobe y'umuntu ikunze kwitwa "urugingo rwa nyuma rw'umubiri" kandi igira uruhare runini mu buzima bw'umuntu (Baquero na Nombela, 2012). By'umwihariko, mikorobe y'igifu izwiho ingaruka zayo mu buryo bwose n'uruhare rwayo mu mirimo myinshi y'ingenzi. Bagiteri zisanzwe ziba nyinshi mu mara, zigafata ahantu henshi hatandukanye mu bidukikije, zikoresha intungamubiri, kandi zigahangana n'udukoko dushobora gutera indwara (Jandhyala et al., 2015). Ibice bitandukanye bya bagiteri bya mikorobe y'igifu bishobora gukora intungamubiri z'ingenzi nka vitamine no guteza imbere igogora (Rowland et al., 2018). Ibinyabutabire bya bagiteri nabyo byagaragaye ko bigira ingaruka ku mikurire y'ingingo no kongera uburyo bwo gukora imikorere y'umubiri n'ubwirinzi bw'umubiri (Heijtz et al., 2011; Yu et al., 2022). Imiterere ya mikorobe y'igifu cy'umuntu iratandukanye cyane kandi iterwa n'ibintu bigize imiterere y'umubiri n'ibidukikije nk'imirire, igitsina, imiti, n'ubuzima (Kumbhare et al., 2019).
Indyo y'umubyeyi ni ingenzi mu mikurire y'umwana uri mu nda n'umwana ukiri muto, ikaba ari isoko y'ibintu bishobora kugira ingaruka ku mikurire (Bazer et al., 2004; Innis, 2014). Kimwe muri ibyo bintu bishimishije ni aside propionic (PPA), aside irike y'uruhererekane rugufi ikomoka ku gushyushya bagiteri n'inyongera ku biryo (den Besten et al., 2013). PPA ifite ubushobozi bwo kurwanya bagiteri n'imiti yica udukoko, bityo ikoreshwa nk'uburyo bwo kubungabunga ibiryo no mu nganda kugira ngo ikumire imyuka n'imikurire ya bagiteri (Wemmenhove et al., 2016). PPA ifite ingaruka zitandukanye mu ngingo zitandukanye. Mu mwijima, PPA igira ingaruka ku bushyuhe mu kugira ingaruka ku mikorere ya cytokine muri macrophages (Kawasoe et al., 2022). Iyi ngaruka yo kugenzura yagaragaye no mu zindi ngirabuzimafatizo z'ubudahangarwa bw'umubiri, bigatuma ubushyuhe bugabanuka (Haase et al., 2021). Ariko, ingaruka zitandukanye zagaragaye mu bwonko. Ubushakashatsi bwakozwe mbere bwagaragaje ko PPA itera imyitwarire nk'iya autism mu mbeba (El-Ansary et al., 2012). Ubundi bushakashatsi bwagaragaje ko PPA ishobora gutera gliosis no gukora inzira zitera ububyimbirwe mu bwonko (Abdelli et al., 2019). Kubera ko PPA ari aside idakomeye, ishobora gukwirakwira mu mara ikagera mu maraso bityo ikarenga inzitizi zirimo urubibi rw'amaraso n'ubwonko ndetse n'uruvange rw'inyuma (Stinson et al., 2019), bigaragaza akamaro ka PPA nk'ikintu kigenga kivangwa na bagiteri. Nubwo uruhare rwa PPA nk'ikintu gishobora gutera autism rurimo gukorwaho iperereza, ingaruka zayo ku bantu bafite autism zishobora kurenga guteza imiterere y'imitsi.
Ibimenyetso byo mu gifu nko gucibwamo no kutagira impatwe bikunze kugaragara ku barwayi bafite indwara z’imitsi (Cao et al., 2021). Ubushakashatsi bwakozwe mbere bwagaragaje ko mikorobe z’abarwayi bafite indwara ya autism spectrum disorders (ASD) zitandukanye n’iz’abantu bazima, bigaragaza ko hari indwara ya mikorobe z’igifu (Finegold et al., 2010). Mu buryo nk’ubwo, imiterere ya mikorobe z’abarwayi bafite indwara z’amara ziterwa n’ububyibuho bukabije, indwara ya Alzheimer, n’ibindi bitandukanye n’iz’abantu bazima (Turnbaugh et al., 2009; Vogt et al., 2017; Henke et al., 2019). Ariko, kugeza ubu, nta sano iragaragara hagati ya mikorobe z’igifu n’indwara cyangwa ibimenyetso by’imitsi (Yap et al., 2021), nubwo hari ubwoko bwinshi bwa bagiteri bufatwa nk’ubugira uruhare muri zimwe muri izi ndwara. Urugero, Akkermansia, Bacteroides, Clostridium, Lactobacillus, Desulfovibrio n'izindi genera ni nyinshi mu mikorobe z'abarwayi bafite autism (Tomova et al., 2015; Golubeva et al., 2017; Cristiano et al., 2018; Zurita et al., 2020). Ikigaragara ni uko amoko amwe y'iyi genera azwiho kugira gene zijyanye no gukora PPA (Reichardt et al., 2014; Yun na Lee, 2016; Zhang et al., 2019; Baur na Dürre, 2023). Bitewe n'ubushobozi bwa PPA bwo kurwanya mikorobe, kongera ubwinshi bwayo bishobora kuba ingirakamaro mu gukura kwa bagiteri zikora PPA (Jacobson et al., 2018). Bityo, ibidukikije bikungahaye kuri PFA bishobora gutuma habaho impinduka mu mikorobe z'amara, harimo n'udukoko two mu gifu, dushobora kuba impamvu zishobora gutuma habaho ibimenyetso by'igifu.
Ikibazo cy'ingenzi mu bushakashatsi ku binyabuzima ni ukumenya niba itandukaniro mu miterere ya mikorobe ari impamvu cyangwa ikimenyetso cy'indwara zishingiyeho. Intambwe ya mbere yo gusobanura isano iri hagati y'imirire, mikorobe yo mu nda, n'indwara zo mu bwonko ni ukureba ingaruka z'imirire ku miterere ya mikorobe. Muri urwo rwego, twakoresheje uburyo bwa metagenomic sequencing bwasomwe igihe kirekire kugira ngo tugereranye mikorobe zo mu nda z'abana b'imbeba zagaburirwaga indyo ikungahaye kuri PPA cyangwa idafite PPA. Abana bagaburirwaga indyo imwe na ba nyina. Twatekereje ko indyo ikungahaye kuri PPA yatuma habaho impinduka mu miterere ya mikorobe zo mu nda n'imikorere yazo, cyane cyane izijyanye na metabolism ya PPA cyangwa umusaruro wa PPA.
Ubu bushakashatsi bwakoresheje imbeba za FVB/N-Tg (GFAP-GFP)14Mes/J transgenic (Jackson Laboratories) zikoresha poroteyine y’icyatsi kibisi (GFP) igenzurwa n’umutezi wa GFAP wihariye wa glia hakurikijwe amabwiriza ya Komite Ishinzwe Kwita ku Matungo ya Kaminuza ya Central Florida (UCF-IACUC) (Nomero y’Uruhushya rwo Gukoresha Amatungo: PROTO202000002). Nyuma yo konka, imbeba zashyizwe ukwazo mu tuzu hamwe n’imbeba 1-5 za buri gitsina kuri buri tuzu. Imbeba zagaburirwaga indyo yuzuye (indyo isanzwe yahinduwe, 16 kcal% ibinure) cyangwa indyo yuzuyemo sodium propionate (indyo isanzwe yahinduwe, 16 kcal% ibinure, irimo 5,000 ppm sodium propionate). Ingano ya sodium propionate yakoreshejwe yari ingana na 5,000 mg PFA/kg uburemere bw’ibiribwa byose. Iki ni cyo gipimo kinini cya PPA cyemewe gukoreshwa nk'uburyo bwo kubungabunga ibiryo. Mu kwitegura ubu bushakashatsi, imbeba z’ababyeyi zagaburiwe indyo zombi mu byumweru 4 mbere yo gutwita kandi zikomeza mu gihe cyose inda y’imbeba yari itwite. Imbeba z’imbeba [imbeba 22, imbeba 9 zigenzura (abagabo 6, abagore 3) na PPA 13 (abagabo 4, abagore 9)] zarakuwe ku ibere hanyuma zikomeza kurya indyo imwe n’imbeba mu gihe cy’amezi 5. Imbeba z’imbeba zaratanzweho igitambo ku mezi 5 maze umwanda wazo wo mu mara ukusanywa hanyuma ubanza kubikwa mu miyoboro ya microcentrifuge ya 1.5 ml kuri -20°C hanyuma zijyanwa muri firigo ya -80°C kugeza igihe ADN y’imbeba yakuweho kandi aside nucleic mikorobe zikavanwamo.
ADN y’umurwayi yakuweho hakurikijwe protocole yahinduwe (Charalampous et al., 2019). Muri make, ibyari mu mwenda byimuwe kuri 500 µl InhibitEX (Qiagen, Cat#/ID: 19593) bibikwa muri firigo. Hakorwa uduce tw’umwenda ntarengwa 1-2 kuri buri kimwe cyo gukuramo. Ibyari mu mwenda byahinduwe mu buryo bwa mekanike hakoreshejwe agace ka pulasitiki kari mu muyoboro kugira ngo hakorwe ururenda. Shyira ingero kuri 10.000 RCF mu gihe cy’iminota 5 cyangwa kugeza igihe ingero zimaze gushonga, hanyuma usukemo supernatant hanyuma wongere ushyiremo agace muri 250 µl 1 × PBS. Ongeraho 250 µl 4.4% saponin (TCI, nimero y’umusaruro S0019) ku gipimo nk’isabune yo koroshya uturemangingo twa eukaryotic. Ingero zavanzwe buhoro buhoro kugeza zimeze neza kandi zishyirwa ku bushyuhe bw’icyumba mu gihe cy’iminota 10. Hanyuma, kugira ngo habeho guhungabanya uturemangingo twa eukaryotike, hongewemo amazi 350 μl adafite nuclease mu gipimo, ashyirwa mu gihe cy'amasegonda 30, hanyuma hongerwamo 12 μl ya NaCl ya 5 M. Ingero zashyizwe muri centrifuge kuri 6000 RCF mu gihe cy'iminota 5. Shyira umwuka wa supernatant hanyuma wongere ushyiremo agace gato muri 100 μl ya 1X PBS. Kugira ngo ukureho ADN y'umuntu, ongeramo 100 μl ya HL-SAN buffer (12.8568 g NaCl, 4 ml ya 1M MgCl2, 36 ml y'amazi adafite nuclease) na 10 μl ya HL-SAN enzyme (ArticZymes P/N 70910-202). Ingero zavanzwe neza hakoreshejwe imiyoboro y'amazi, hanyuma zishyirwa ku bushyuhe bwa 37 °C mu minota 30 kuri 800 rpm kuri Eppendorf™ ThermoMixer C. Nyuma yo gukaraba, hashyirwa kuri 6000 RCF mu minota 3 hanyuma hakaraba kabiri hakoreshejwe 800 µl na 1000 µl 1X PBS. Amaherezo, ongera ushyiremo agace ka 100 µl 1X PBS.
ADN yose ya bagiteri yakuwemo hakoreshejwe New England Biolabs Monarch Genomic DNA Purification Kit (New England Biolabs, Ipswich, MA, Cat# T3010L). Uburyo busanzwe bwo gukora bwatanzwe n'iyi kit bwahinduwe gato. Shyira amazi adafite nuclease kuri 60°C mbere yo kubagwa kugira ngo ubashe kuyakuraho burundu. Ongeramo 10 µl Proteinase K na 3 µl RNase A kuri buri kitegererezo. Hanyuma ongeramo 100 µl Cell Lysis Buffer hanyuma uvange buhoro buhoro. Ingero zashyizwe muri Eppendorf™ ThermoMixer C kuri 56°C na 1400 rpm byibuze isaha 1 kugeza ku masaha 3. Ingero zashyizwe muri centrifuge kuri 12.000 RCF mu gihe cy'iminota 3, hanyuma supernatant yavuye muri buri kitegererezo yimurirwa mu muyoboro wa microcentrifuge wa 1.5 mL urimo 400 µL y'umuti uhambiriye. Hanyuma imiyoboro yashyizwemo umwuka mu gihe cy'amasegonda 5-10 mu gihe cy'isegonda 1. Hindura ibikomoka ku mazi byose bya buri gipimo (hafi 600-700 µL) ku gasanduku k'urufunguzo kashyizwe mu muyoboro w'ikusanyirizo. Imiyoboro yashyizwe kuri centrifuge kuri 1.000 RCF mu gihe cy'iminota 3 kugira ngo ADN ifatanye bwa mbere hanyuma ishyirwa kuri centrifuge kuri 12.000 RCF mu gihe cy'umunota 1 kugira ngo amazi asigaye akurweho. Inkingi y'icyitegererezo yimuriwe mu muyoboro mushya w'ikusanyirizo hanyuma ikarabwa kabiri. Ku nshuro ya mbere, ongeramo 500 µL y'agasanduku k'ikusanyirizo kuri buri muyoboro. Hindura umuyoboro inshuro 3-5 hanyuma ushyire kuri centrifuge kuri 12.000 RCF mu gihe cy'umunota 1. Kuramo amazi mu muyoboro w'ikusanyirizo hanyuma ushyiremo agasanduku k'ikusanyirizo mu muyoboro umwe w'ikusanyirizo. Ku nshuro ya kabiri, ongeramo 500 µL y'agasanduku k'ikusanyirizo ku muyoboro w'ikusanyirizo nta guhinduranya. Ingero zashyizwe kuri centrifuge kuri 12.000 RCF mu gihe cy'umunota 1. Shyira akayunguruzo mu muyoboro wa LoBind® wa 1.5 mL hanyuma wongeremo 100 µL y'amazi yashyushye mbere adafite nuclease. Akayunguruzo kashyizwe mu bushyuhe bw'icyumba mu gihe cy'umunota 1 hanyuma gashyirwa kuri centrifuge kuri 12.000 RCF mu gihe cy'umunota 1. ADN ya Eluted yabikwaga kuri -80°C.
Igipimo cya ADN cyapimwe hakoreshejwe Qubit™ 4.0 Fluorometer. ADN yateguwe hakoreshejwe Qubit™ 1X dsDNA High Sensitivity Kit (Cat. No. Q33231) hakurikijwe amabwiriza y'uwakoze. Ikwirakwizwa ry'uburebure bw'ibice bya ADN ryapimwe hakoreshejwe Aglient™ 4150 cyangwa 4200 TapeStation. ADN yateguwe hakoreshejwe Agilent™ Genomic DNA Reagents (Cat. No. 5067-5366) na Genomic DNA ScreenTape (Cat. No. 5067-5365). Gutegura isomero byakozwe hakoreshejwe Oxford Nanopore Technologies™ (ONT) Rapid PCR Barcoding Kit (SQK-RPB004) hakurikijwe amabwiriza y'uwakoze. ADN yakurikiranywe hakoreshejwe ONT GridION™ Mk1 sequencer hamwe na Min106D flow cell (R 9.4.1). Imiterere y'ikurikirana yari: guhamagara kw'ibanze kw'ubuziranenge bwo hejuru, agaciro ka q ka 9, gushyiraho barcode, no kugabanya barcode. Ingero zakurikiranywe mu masaha 72, hanyuma amakuru y’ibanze yoherejwe kugira ngo akomeze gutegurwa no gusesengura.
Gutunganya Bioinformatics byakozwe hakoreshejwe uburyo bwavuzwe mbere (Greenman et al., 2024). Dosiye za FASTQ zabonetse mu gutondeka zagabanijwemo ububiko bwa buri ngero. Mbere yo gusesengura bioinformatics, amakuru yatunganyijwe hakoreshejwe uburyo bukurikira: ubwa mbere, dosiye za FASTQ z'ibipimo zahujwe muri dosiye imwe ya FASTQ. Hanyuma, abasomyi bari munsi ya 1000 bp bayungururwa hakoreshejwe Filtlong v. 0.2.1, aho parameter imwe rukumbi yahindutse ikaba –min_length 1000 (Wick, 2024). Mbere yo gusesengura bundi bushya, ireme ry'isomwa ryagenzurwaga hakoreshejwe NanoPlot v. 1.41.3 hamwe n'ibipimo bikurikira: –fastq –plots dot –N50 -o
Ku bijyanye no gushyira mu byiciro taxonomike, abasomyi n'ibice byakusanyijwe byashyizwe mu byiciro hakoreshejwe Kraken2 v. 2.1.2 (Wood et al., 2019). Kora raporo n'amadosiye yo gusohora abasomyi n'ibice, uko bikurikirana. Koresha uburyo bwa –use-names kugira ngo usesengure abasomyi n'ibice. Amahitamo ya –gzip-compressed na –paired yagenwe ku bice byasomwe. Ubwinshi bw'ibice bya taxa muri metagenome bwagereranyijwe hakoreshejwe Bracken v. 2.8 (Lu et al., 2017). Twabanje gukora database ya kmer irimo ishingiro 1000 dukoresheje bracken-build hamwe n'ibipimo bikurikira: -d
Isesengura ry’uturemangingo n’igereranya ry’ubwinshi bw’uturemangingo byakozwe hakoreshejwe verisiyo yahinduwe ya protocole yasobanuwe na Maranga et al. (Maranga et al., 2023). Ubwa mbere, ibice biri munsi ya 500 bp byakuwe mu matsinda yose hakoreshejwe SeqKit v. 2.5.1 (Shen et al., 2016). Amatsinda yatoranijwe yahujwe muri pan-metagenome. Amafuremu yo gusoma afunguye (ORFs) yamenyekanye hakoreshejwe Prodigal v. 1.0.1 (verisiyo isa na Prodigal v. 2.6.3) hamwe n’ibipimo bikurikira: -d
Ingirabuzimafatizo zabanje gushyirwa mu matsinda hakurikijwe indangamuntu za Kyoto Encyclopedia of Genes and Genomes (KEGG) ortholog (KO) zatanzwe na eggNOG kugira ngo hagereranywe ubwinshi bw'inzira y'ingirabuzimafatizo. Ingirabuzimafatizo zitarimo knockouts cyangwa ingirabuzimafatizo zifite knockouts nyinshi zakuweho mbere yo gusesengura. Ubunini bw'ikigereranyo cya buri KO kuri buri gipimo bwarabanje kubarwa maze hakorwa isesengura ry'imibare. Ingirabuzimafatizo za metabolism ya PPA zasobanuwe nk'ingirabuzimafatizo iyo ari yo yose yatanzwe umurongo wa ko00640 mu nkingi ya KEGG_Pathway, bigaragaza uruhare mu metabolism ya propionate nk'uko KEGG ibivuga. Ingirabuzimafatizo zagaragajwe ko zifitanye isano n'umusaruro wa PPA zanditswe mu mbonerahamwe y'inyongera ya 1 (Reichardt et al., 2014; Yang et al., 2017). Ibizamini bya permutation byakozwe kugira ngo hamenyekane metabolism ya PPA na ingirabuzimafatizo z'umusaruro zari nyinshi cyane muri buri bwoko bw'icyitegererezo. Permutations igihumbi zakozwe kuri buri ngirabuzimafatizo yasesenguwe. Agaciro ka p ka 0.05 kakoreshejwe nk'ishingiro kugira ngo hamenyekane akamaro k'ingirabuzimafatizo. Inyandiko z'imikorere zatanzwe ku ngirabuzimafatizo imwe mu itsinda hashingiwe ku nyandiko z'ingirabuzimafatizo zihagarariye iri mu itsinda. Taxa ifitanye isano na metabolism ya PPA cyangwa umusaruro wa PPA ishobora kumenyekana hakoreshejwe guhuza contig ID muri dosiye za Kraken2 zisohoka hamwe na contig ID zimwe zabitswe mu gihe cyo gusobanura imikorere hakoreshejwe eggNOG. Isuzuma ry'akamaro ryakozwe hakoreshejwe ikizamini cya Mann-Whitney U cyavuzwe mbere. Gukosora ibizamini byinshi byakozwe hakoreshejwe uburyo bwa Benjamini-Hochberg. Agaciro ka p ka ≤ 0.05 kakoreshejwe nk'ishingiro kugira ngo hamenyekane akamaro k'imibare.
Ubudasa bw'uturemangingo tw'inda tw'imbeba bwapimwe hakoreshejwe igipimo cya Simpson diversity. Nta tandukaniro rinini ryagaragaye hagati y'ibipimo bya PPA mu kugenzura no mu gutandukanya ubwoko n'ubwoko (agaciro ka p kuri genus: 0.18, agaciro ka p kuri genus: 0.16) (Ishusho ya 1). Imiterere ya mikorobe yagereranyijwe hakoreshejwe isesengura ry'ibice by'ingenzi (PCA). Ishusho ya 2 igaragaza ishyirwa mu byiciro ry'ibipimo hakurikijwe phyla yabyo, bigaragaza ko hari itandukaniro mu miterere y'ubwoko bwa mikorobe hagati ya PPA n'ibipimo bya régime. Uku gukusanya ntibyagaragaye cyane ku rwego rw'ubwoko, bigaragaza ko PPA igira ingaruka kuri bagiteri zimwe na zimwe (Ishusho y'inyongera ya 1).
Ishusho ya 1. Ubudasa bw'ubwoko bwa Alpha bw'ubwoko n'imiterere y'ubwoko bwa mikorobe y'imbeba. Ibishushanyo by'udusanduku bigaragaza ibimenyetso by'ubudasa bwa Simpson bw'ubwoko (A) n'ubwoko (B) muri PPA n'ingero zo kugenzura. Akamaro kagaragajwe hakoreshejwe ikizamini cya Mann-Whitney U, kandi hakosowe inshuro nyinshi hakoreshejwe uburyo bwa Benjamini-Hochberg. ns, agaciro ka p ntabwo kari gafite agaciro (p>0.05).
Ishusho ya 2. Ibisubizo by'isesengura ry'ibice by'ingenzi by'imiterere ya mikorobe zo mu nda y'imbeba ku rwego rw'ubwoko bw'ibinyabuzima. Isesengura ry'ibice by'ingenzi ryerekana uko ingero zikwirakwira mu bice bibiri by'ingenzi byazo. Amabara agaragaza ubwoko bw'icyitegererezo: Imbeba zagaragaye kuri PPA ni umuhengeri naho imbeba zo kugenzura ni umuhondo. Ibice by'ingenzi bya 1 na 2 byashushanyijwe ku murongo wa x na y, uko bikurikirana, kandi bigaragazwa nk'ikigereranyo cyazo cy'impinduka.
Hakoreshejwe amakuru ya RLE yahinduwe, hagaragaye igabanuka rikomeye ry’ikigereranyo cya Bacteroidetes/Bacilli mu mbeba zo mu bwoko bwa control na PPA (igenzura: 9.66, PPA: 3.02; agaciro ka p = 0.0011). Iri tandukaniro ryatewe n’ubwinshi bwa Bacteroidetes mu mbeba zo mu bwoko bwa PPA ugereranije n’izo mu bwoko bwa control, nubwo itandukaniro ritari rinini (ikigereranyo cya CLR: 5.51, ikigereranyo cya PPA CLR: 6.62; agaciro ka p = 0.054), mu gihe ubwinshi bwa Bacteroidetes bwari busa (ikigereranyo cya CLR: 7.76, ikigereranyo cya PPA CLR: 7.60; agaciro ka p = 0.18).
Isesengura ry’ubwinshi bw’ibice bigize taxonomic bya mikorobe yo mu nda ryagaragaje ko ubwoko bumwe n’ubwoko 77 bitandukanye cyane hagati ya PPA n’ibipimo byo kugenzura (Imbonerahamwe y’inyongera ya 2). Ubwinshi bw’ubwoko 59 mu bipimo bya PPA bwari bwinshi cyane ugereranyije n’ubwo mu bipimo byo kugenzura, mu gihe ubwinshi bw’ubwoko 16 gusa mu bipimo byo kugenzura bwari bwinshi ugereranyije n’ubwo mu bipimo bya PPA (Ishusho ya 3).
Ishusho ya 3. Ubwinshi butandukanye bw'udukoko mu mara ya mikorobe ya PPA n'imbeba zigenzura. Imiterere y'ibirunga igaragaza itandukaniro mu bwinshi bw'ubwoko (A) cyangwa ubwoko (B) hagati ya PPA n'ingero zigenzura. Utudomo tw'ikijuju nta tandukaniro rigaragara mu bwinshi bw'udukoko. Utudomo tw'amabara tugaragaza itandukaniro rigaragara mu bwinshi (agaciro ka p ≤ 0.05). Utudomo 20 twa mbere dufite itandukaniro rinini mu bwinshi hagati y'ubwoko bw'ingero tugaragazwa mu ibara ry'umutuku n'ubururu bwerurutse (ingero zigenzura n'izigenzura na PPA), uko bikurikirana. Utudomo tw'umuhondo n'umuhengeri twari twinshi byibuze inshuro 2.7 mu ingero zigenzura cyangwa izigenzura cyangwa PPA kurusha mu igenzura. Utudomo tw'umukara tugaragaza udukoko dufite ubwinshi butandukanye cyane, hamwe n'itandukaniro ry'ikigereranyo cya CLR hagati ya -1 na 1. Agaciro ka P kabazwe hakoreshejwe ikizamini cya Mann-Whitney U kandi gakosorwa kugira ngo hakosorwe ibizamini byinshi hakoreshejwe uburyo bwa Benjamini-Hochberg. Itandukaniro ry'ikigereranyo cya CLR rigaragara neza rigaragaza itandukaniro rigaragara mu bwinshi.
Nyuma yo gusesengura imiterere ya mikorobe zo mu mara, twakoze isesengura ry’imikorere ya mikorobe. Nyuma yo gusesengura jeni zitari nziza, habonetse jeni zihariye 378.355 mu bipimo byose. Ubwinshi bw’izi jeni bwahinduwe bwakoreshejwe mu gusesengura ibice by’ingenzi (PCA), kandi ibyavuye mu bushakashatsi byagaragaje urwego rwo hejuru rwo gukusanya ubwoko bw’ingero hashingiwe ku mikorere yazo (Ishusho ya 4).
Ishusho ya 4. PCA ikora ikoresheje imiterere y'imikorere ya mikorobe y'imbeba. Igishushanyo cya PCA kigaragaza ikwirakwizwa ry'ibipimo mu bice bibiri bya mbere by'ingenzi. Amabara agaragaza ubwoko bw'icyitegererezo: Imbeba za PPA-exposed ni umuhengeri naho imbeba zo kugenzura ni umuhondo. Ibice by'ingenzi bya 1 na 2 bishushanyije ku murongo wa x na y, uko bikurikirana, kandi bigaragazwa nk'ikigereranyo cyazo cy'impinduka.
Hanyuma twasuzumye ubwinshi bwa KEGG knockouts mu bwoko butandukanye bw'ibipimo. Habonetsemo knockouts zidasanzwe 3648, muri zo 196 zari nyinshi cyane mu bipimo byo kugenzura naho 106 zari nyinshi cyane mu bipimo bya PPA (Ishusho ya 5). Habonetsemo genes 145 mu bipimo byo kugenzura na genes 61 mu bipimo bya PPA, hamwe n'ubwinshi butandukanye cyane. Inzira zijyanye na metabolism ya lipide na aminosugar zakungahaye cyane mu bipimo bya PPA (Imbonerahamwe y'inyongera ya 3). Inzira zijyanye na metabolism ya azote na sisitemu yo kujyana sulfure zakungahaye cyane mu bipimo byo kugenzura (Imbonerahamwe y'inyongera ya 3). Ubwinshi bwa genes zijyanye na metabolism ya aminosugar/nucleotide (ko:K21279) na metabolism ya phosphate ya inositol (ko:K07291) bwari bwinshi cyane mu bipimo bya PPA (Ishusho ya 5). Ingero zo kugenzura zari zifite imiterere myinshi cyane ijyanye na metabolism ya benzoate (ko: K22270), metabolism ya azote (ko: K00368), na glycolysis / gluconeogenesis (ko: K00131) (Ishusho ya 5).
Ishusho ya 5. Ubwinshi butandukanye bwa KO mu mara ya mikorobe ya PPA n'imbeba zigenzura. Igishushanyo cy'ikirunga kigaragaza itandukaniro mu bwinshi bw'amatsinda y'imikorere (KOs). Utudomo tw'imvi tugaragaza KOs ubwinshi bwazo butari butandukanye cyane hagati y'ubwoko bw'ingero (agaciro ka p > 0.05). Utudomo tw'amabara tugaragaza itandukaniro rikomeye mu bwinshi (agaciro ka p ≤ 0.05). KOs 20 zifite itandukaniro rinini mu bwinshi hagati y'ubwoko bw'ingero zigaragazwa mu ibara ry'umutuku n'ubururu bwerurutse, bihuye n'ingero za PPA, uko bikurikirana. Utudomo tw'umuhondo n'umuhengeri tugaragaza KOs zari nyinshi byibuze inshuro 2.7 mu kugenzura n'ingero za PPA, uko bikurikirana. Utudomo tw'umukara tugaragaza KOs zifite ubwinshi butandukanye cyane, hamwe n'itandukaniro ry'impuzandengo ya CLR hagati ya -1 na 1. Agaciro ka P kabazwe hakoreshejwe ikizamini cya Mann-Whitney U kandi gahindurwa kugira ngo hagereranywe byinshi hakoreshejwe uburyo bwa Benjamini-Hochberg. NaN igaragaza ko KO atari mu nzira muri KEGG. Itandukaniro ry'impuzandengo ya CLR igaragara neza rigaragaza itandukaniro rikomeye mu bwinshi. Kugira ngo ubone amakuru arambuye ku nzira KO zanditse zigomba kujyamo, reba Imbonerahamwe y'Inyongera ya 3.
Muri za gene zavuzweho, gene 1601 zari zifite ubwinshi butandukanye cyane hagati y’ubwoko bw’ingero (p ≤ 0.05), aho buri gene yari ifite byibuze inshuro 2.7 z’ubwinshi. Muri izi gene, gene 4 zari nyinshi mu bipimo byo kugenzura naho gene 1597 zari nyinshi mu bipimo bya PPA. Kubera ko PPA ifite ubushobozi bwo kurwanya mikorobe, twasuzumye ubwinshi bwa PPA metabolisme n’ingero z’umusaruro hagati y’ubwoko bw’ingero. Muri gene 1332 zijyanye na metabolisme ya PPA, gene 27 zari nyinshi cyane mu bipimo byo kugenzura naho gene 12 zari nyinshi mu bipimo bya PPA. Muri gene 223 zijyanye na PPA production, gene 1 yari nyinshi cyane mu bipimo bya PPA. Ishusho ya 6A ikomeza kwerekana ubwinshi bwa genes zigize uruhare mu metabolisme ya PPA, hamwe n’ubwinshi bwinshi mu bipimo byo kugenzura n’ingano nini y’ingaruka, mu gihe Ishusho ya 6B igaragaza genes imwe ifite ubwinshi bwinshi cyane bwagaragaye mu bipimo bya PPA.
Ishusho ya 6. Ubwinshi butandukanye bw'uturemangingo dufitanye isano na PPA mu mikorobe y'imbeba. Imiterere y'ibirunga igaragaza itandukaniro mu bwinshi bw'uturemangingo dufitanye isano na metabolism ya PPA (A) n'umusaruro wa PPA (B). Utudomo tw'imvi tugaragaza uturemangingo dufite ubwinshi butatandukanye cyane hagati y'ubwoko bw'ingero (agaciro ka p > 0.05). Utudomo tw'amabara tugaragaza itandukaniro rikomeye mu bwinshi (agaciro ka p ≤ 0.05). Uturemangingo 20 dufite itandukaniro rinini mu bwinshi tugaragazwa mu ibara ry'umutuku n'ubururu bwerurutse (ingero za PPA zo kugenzura n'izigaragaza PPA), uko bikurikirana. Ubwinshi bw'utudomo tw'umuhondo n'umuhengeri bwari bwikubye nibura inshuro 2.7 mu ingero za PPA zo kugenzura n'izigaragaza PPA ugereranyije n'ingero za PPA zo kugenzura. Utudomo tw'umukara tugaragaza uturemangingo dufite ubwinshi butandukanye cyane, hamwe n'itandukaniro ry'ikigereranyo cya CLR hagati ya -1 na 1. Agaciro ka P kabazwe hakoreshejwe ikizamini cya Mann-Whitney U kandi gakosorwa kugira ngo hagereranywe byinshi hakoreshejwe uburyo bwa Benjamini-Hochberg. Uturemangingo duhuye n'uturemangingo duhagarariye mu kataloge y'uturemangingo tudasubirwaho. Amazina y'uturemangingo agizwe n'ikimenyetso cya KEGG kigaragaza gene ya KO. Itandukaniro ry'ikigereranyo cya CLR rigaragara mu buryo bw'inyuguti rigaragaza ubwinshi butandukanye cyane. Akarongo (-) kerekana ko nta kimenyetso cy'ingirabuzimafatizo kiri muri KEGG database.
Taxa ifite jeni zijyanye na metabolism ya PPA cyangwa umusaruro wayo yamenyekanye binyuze mu guhuza indangamuntu y’uturemangingo n’indangamuntu y’uturemangingo. Ku rwego rw’ubwoko, jeni 130 zagaragaye ko zifite jeni zijyanye na metabolism ya PPA naho jeni 61 zagaragaye ko zifite jeni zijyanye na PPA (Imbonerahamwe y’inyongera ya 4). Ariko, nta jeni na rimwe ryagaragaje itandukaniro rinini mu bwinshi (p > 0.05).
Ku rwego rw'ubwoko, ubwoko 144 bwa bagiteri byagaragaye ko bufite imiterere ijyanye na metabolism ya PPA naho ubwoko 68 bwa bagiteri byagaragaye ko bufite imiterere ijyanye na PPA (Imbonerahamwe y'inyongera ya 5). Mu bagiteri ba PPA, bagiteri umunani zagaragaje ukwiyongera gukomeye mu bwinshi hagati y'ubwoko bw'ibipimo, kandi zose zagaragaje impinduka zikomeye mu mikorere yazo (Imbonerahamwe y'inyongera ya 6). Bagiteri zose za PPA zagaragaye zifite itandukaniro rikomeye mu bwinshi zari nyinshi mu bipimo bya PPA. Ibyiciro by'ubwoko byagaragaje imiterere idatandukanye cyane hagati y'ubwoko bw'ibipimo, harimo ubwoko bwinshi bwa Bacteroides na Ruminococcus, ndetse na Duncania dubois, Myxobacterium enterica, Monococcus pectinolyticus, na Alcaligenes polymorpha. Muri bagiteri zikora PPA, bagiteri enye zagaragaje itandukaniro rikomeye mu bwinshi hagati y'ubwoko bw'ibipimo. Ubwoko bufite itandukaniro rikomeye mu bwinshi bwarimo Bacteroides novorossi, Duncania dubois, Myxobacterium enteritidis, na Ruminococcus bovis.
Muri ubu bushakashatsi, twasuzumye ingaruka zo kwanduzwa na PPA ku mikorobe zo mu nda y'imbeba. PPA ishobora gutanga ibisubizo bitandukanye muri bagiteri kuko ikorwa n'ubwoko bumwe na bumwe, igakoreshwa nk'isoko y'ibiribwa n'ubundi bwoko bw'ibinyabuzima, cyangwa ikaba ifite ingaruka zo kurwanya mikorobe. Kubwibyo, kongeramo kwayo mu mara binyuze mu nyongeramusaruro bishobora kugira ingaruka zitandukanye bitewe n'uburyo bwo kwihanganira, kwifata, n'ubushobozi bwo kuyikoresha nk'isoko y'intungamubiri. Ubwoko bwa bagiteri bushobora gukurwaho bugasimburwa n'ubworoshye kurwanya PPA cyangwa bushobora kuyikoresha nk'isoko y'ibiribwa, bigatera impinduka mu miterere ya mikorobe zo mu nda. Ibisubizo byacu byagaragaje itandukaniro rikomeye mu miterere ya mikorobe ariko nta ngaruka ku budasa bw'udukoko muri rusange. Ingaruka nini zagaragaye ku rwego rw'ubwoko, aho amoko arenga 70 atandukanye cyane hagati ya PPA n'ingero zo kugenzura (Imbonerahamwe y'inyongera ya 2). Isuzuma ryinshi ry'imiterere ya mikorobe zagaragaye muri PPA ryagaragaje ubudasa bwinshi bw'ubwoko bwa mikorobe ugereranije n'ingero zitaragaragaye, bigaragaza ko PPA ishobora kongera imiterere y'imikurire ya bagiteri no kugabanya umubare wa bagiteri zishobora kubaho mu duce twinshi twa PPA. Bityo, PPA ishobora guteza impinduka mu buryo bwihariye aho guteza ihungabana rikomeye ry'ubudasa bw'udukoko two mu nda.
Ibiryo bibungabunga ubuzima nka PPA byagaragaye mbere ko bihindura ubwinshi bw'ibice bya mikorobe mu mara bitagize ingaruka ku budasa rusange (Nagpal et al., 2021). Aha, twabonye itandukaniro rikomeye hagati y'ubwoko bwa Bacteroidetes muri phylum Bacteroidetes (yari izwi nka Bacteroidetes), yari ikunzwe cyane mu mbeba zagaragaye muri PPA. Ubwinshi bw'ubwoko bwa Bacteroides bujyana no kwangirika kw'ibicurane, bishobora kongera ibyago byo kwandura no gutera ububyimbirwe (Cornick et al., 2015; Desai et al., 2016; Penzol et al., 2019). Ubushakashatsi bumwe bwagaragaje ko imbeba z'abagabo zivutse zavuwe na Bacteroides fragilis zagaragaje imyitwarire yo mu muryango isa n'indwara ya autism spectrum disorder (ASD) (Carmel et al., 2023), kandi ubundi bushakashatsi bwagaragaje ko ubwoko bwa Bacteroides bushobora guhindura imikorere y'ubudahangarwa bw'umubiri no gutera indwara y'umutima iterwa n'ubwiyongere bw'umubiri (Gil-Cruz et al., 2019). Ubwoko bw’ubwoko bwa Ruminococcus, Prevotella, na Parabacteroides nabwo bwiyongereye cyane mu mbeba zagaragayeho PPA (Coretti et al., 2018). Amoko amwe ya Ruminococcus afitanye isano n’indwara nka Crohn's disease binyuze mu gukora cytokines zitera ubushyuhe (Henke et al., 2019), mu gihe ubwoko bwa Prevotella nka Prevotella humani bufitanye isano n’indwara za metabolisme nka hypertension na insuline sensitivity (Pedersen et al., 2016; Li et al., 2017). Amaherezo, twasanze igipimo cya Bacteroidetes (cyahoze kizwi nka Firmicutes) kuri Bacteroidetes cyari gito cyane mu mbeba zagaragayeho PPA ugereranyije n’imbeba zigenzurwa bitewe n’ubwinshi bw’ubwoko bwa Bacteroidetes. Iyi ratio yagaragaye mbere ko ari ikimenyetso cy'ingenzi cy'uko amara ahora mu mubiri, kandi ibibazo muri iyi ratio byagiye bifitanye isano n'indwara zitandukanye (Turpin et al., 2016; Takezawa et al., 2021; An et al., 2023), harimo n'indwara zo mu mara ziterwa n'ububyimbirwe (Stojanov et al., 2020). Muri rusange, ubwoko bwa Bacteroidetes phylum busa nkaho bugira ingaruka zikomeye ku mirire ya PPA. Ibi bishobora guterwa no kwihanganira cyane PPA cyangwa ubushobozi bwo gukoresha PPA nk'isoko y'ingufu, byagaragaye ko ari ukuri ku bwoko bumwe nibura, Hoylesella enocea (Hitch et al., 2022). Ubundi buryo, PPA ya nyina ishobora kongera imikurire y'umwana w'uruhinja ituma amara y'imbeba ashobora kwangirika cyane no kwibasirwa na Bacteroidetes; icyakora, ubushakashatsi bwacu ntibwatumye isuzuma nk'iryo rikorwa.
Isuzuma ry’ibikubiye mu miterere ryagaragaje itandukaniro rikomeye mu bwinshi bw’uturemangingo dufitanye isano n’imikorere ya PPA n’umusaruro, aho imbeba zagaragaye ko zifite PPA zigaragaza ubwinshi bw’uturemangingo dutuma PPA ikorwa, mu gihe imbeba zitari iza PPA zigaragaza ubwinshi bw’uturemangingo dutuma PAA ikorwa (Ishusho ya 6). Ibi byavuye mu bushakashatsi bigaragaza ko ingaruka za PPA ku miterere ya mikorobe zishobora kuba zitaterwa gusa no gukoreshwa kwayo, bitabaye ibyo ubwinshi bw’uturemangingo dufitanye isano na PPA bwagombaga kugaragaza ubwinshi bwinshi mu mikorobe zo mu mara z’imbeba zagaragaye ko zifite PPA. Kimwe mu bisobanuro ni uko PPA ihuza ubwinshi bwa bagiteri ahanini binyuze mu ngaruka zayo zo kurwanya mikorobe aho gukoresha bagiteri nk’intungamubiri. Ubushakashatsi bwakozwe mbere bwagaragaje ko PPA ibuza gukura kwa Salmonella Typhimurium mu buryo bushingiye ku gipimo (Jacobson et al., 2018). Kugaragara ku bwinshi bwa PPA bishobora guhitamo bagiteri zirwanya imiterere yayo yo kurwanya mikorobe kandi zishobora kuba zidashobora kuyihindura cyangwa kuyikora. Urugero, ubwoko bwinshi bwa Parabacteroides bwagaragaje ubwinshi bukabije mu bipimo bya PPA, ariko nta gene zijyanye na metabolism ya PPA cyangwa umusaruro zabonetse (Imbonerahamwe y'inyongera 2, 4, na 5). Byongeye kandi, umusaruro wa PPA nk'umusaruro ukomoka ku guhinga ukwirakwira cyane muri bagiteri zitandukanye (Gonzalez-Garcia et al., 2017). Ubwinshi bw'uturemangingo twa bagiteri bushobora kuba impamvu y'ubwinshi bw'uturemangingo tujyanye na metabolism ya PPA mu bipimo byo kugenzura (Averina et al., 2020). Byongeye kandi, 27 (2.14%) muri gene 1332 gusa ni zo zavuzwe ko ari gene zifitanye isano na metabolism ya PPA gusa. Gene nyinshi zifitanye isano na metabolism ya PPA nazo zifitanye isano n'izindi nzira za metabolism. Ibi byerekana kandi ko ubwinshi bw'uturemangingo twinjira muri metabolism ya PPA bwari bwinshi mu bipimo byo kugenzura; izi gene zishobora gukora mu nzira zidatuma PPA ikoreshwa cyangwa ngo ikore nk'umusaruro. Muri iki gihe, gene imwe gusa ifitanye isano no gukora PPA yagaragaje itandukaniro rikomeye mu bwinshi hagati y'ubwoko bw'ingero. Bitandukanye na gene zifitanye isano na metabolism ya PPA, gene z'ibimenyetso byo gukora PPA zatoranyijwe kuko zigira uruhare rutaziguye mu nzira ya bagiteri yo gukora PPA. Mu mbeba zagaragayemo PPA, amoko yose yagaragaye ko yiyongereye cyane mu bwinshi n'ubushobozi bwo gukora PPA. Ibi bishyigikira igitekerezo cy'uko PPA zahitamo abakora PPA bityo zigateganya ko ubushobozi bwo gukora PPA buziyongera. Ariko, ubwinshi bw'uturemangingo ntabwo bufitanye isano n'imikorere y'uturemangingo; bityo, nubwo ubwinshi bw'uturemangingo dufitanye isano na metabolism ya PPA buri hejuru mu bipimo byo kugenzura, igipimo cyo gukora gishobora kuba gitandukanye (Shi et al., 2014). Kugira ngo hemezwe isano iri hagati y'ubwinshi bw'uturemangingo dukora PPA n'ikorwa rya PPA, hakenewe inyigo ku mikorere y'uturemangingo dufitanye isano no gukora PPA.
Isesengura ry’imikorere ya PPA na metagenome zo kugenzura ryagaragaje itandukaniro rimwe na rimwe. Isesengura rya PCA ry’ibiri mu ngirabuzimafatizo ryagaragaje amatsinda atandukanye hagati ya PPA n’ingero zo kugenzura (Ishusho ya 5). Guhuza ingero byagaragaje ko ingano y’uturemangingo two kugenzura yari itandukanye cyane, mu gihe ingero za PPA zakusanyijwe hamwe. Guhuza ingero hakurikijwe ingano y’uturemangingo byagereranywa no guhuza ingero z’ubwoko. Bityo, itandukaniro mu bwinshi bw’inzira rihura n’impinduka mu bwinshi bw’ubwoko bwihariye n’ubwoko buri muri bwo. Mu ngero za PPA, inzira ebyiri zifite ubwinshi bwinshi cyane zari zifitanye isano na metabolism y’isukari ya aminosugar/nucleotide (ko:K21279) n’inzira nyinshi za metabolism y’ibinure (ko:K00647, ko:K03801; Imbonerahamwe y’inyongera ya 3). Uturemangingo dufitanye isano na ko:K21279 tuzwiho kuba dufitanye isano na genus Bacteroides, imwe mu ngirabuzimafatizo ifite umubare munini w’ubwoko mu ngero za PPA. Iyi enzyme ishobora kwirinda ubudahangarwa bw’umubiri binyuze mu kugaragaza polysaccharides za capsular (Wang et al., 2008). Ibi bishobora kuba impamvu y’ubwiyongere bwa Bacteroidetes zagaragaye mu mbeba zagaragaye muri PPA. Ibi byuzuza ubwiyongere bw’imisemburo ya aside irike igaragara muri mikorobe ya PPA. Bagiteri zikoresha inzira ya FASIIko: K00647 (fabB) kugira ngo zikore aside irike, ishobora kugira ingaruka ku nzira za metabolic z’umuntu (Yao na Rock, 2015; Johnson et al., 2020), kandi impinduka mu mikorere ya lipide zishobora kugira uruhare mu iterambere ry’ubwonko (Yu et al., 2020). Indi nzira igaragaza ubwiyongere bw’ubwinshi mu bipimo bya PPA ni hormone ya steroide biosynthesis (ko: K12343). Hari ibimenyetso byinshi bigaragaza ko hari isano iri hagati y’ubushobozi bwa mikorobe yo mu mara bwo kugira ingaruka ku rugero rw’imisemburo no kugira ingaruka ku misemburo, ku buryo urugero rwa steroide rwinshi rushobora kugira ingaruka ku buzima (Tetel et al., 2018).
Ubu bushakashatsi ntabwo budafite imbogamizi n'ibitekerezo. Itandukaniro rikomeye ni uko tutakoze isuzuma ry’imiterere y’inyamaswa. Kubwibyo, ntabwo bishoboka kwemeza neza niba impinduka muri mikorobe zifitanye isano n’indwara iyo ari yo yose. Ikindi kintu cyatekerezwaho ni uko imbeba zo muri ubu bushakashatsi zagaburiwe indyo imwe na ba nyina. Ubushakashatsi bw’ejo hazaza bushobora kwemeza niba guhindura indyo ikungahaye kuri PPA ukajya ku ndyo idafite PPA byongera ingaruka zayo kuri mikorobe. Imwe mu mbogamizi z’ubushakashatsi bwacu, kimwe n’izindi nyinshi, ni ingano nto y’icyitegererezo. Nubwo imyanzuro yumvikana ishobora gufatwa, ingano nini y’icyitegererezo yatanga imbaraga nyinshi mu mibare mu gusesengura ibyavuye mu bushakashatsi. Turitondera kandi gufata imyanzuro ku isano iri hagati y’impinduka muri mikorobe zo mu mara n’indwara iyo ari yo yose (Yap et al., 2021). Ibintu bivangavanze birimo imyaka, igitsina, n’imirire bishobora kugira ingaruka zikomeye ku miterere ya mikorobe. Ibi bintu bishobora gusobanura ukudahuza kwagaragaye mu nyandiko ku bijyanye n’isano ya mikorobe zo mu mara n’indwara zikomeye (Johnson et al., 2019; Lagod na Naser, 2023). Urugero, byagaragaye ko amoko ya Bacteroidetes yiyongereye cyangwa igabanuka mu nyamaswa no ku bantu bafite ASD (Angelis et al., 2013; Kushak et al., 2017). Mu buryo nk'ubwo, ubushakashatsi ku miterere y'amara ku barwayi bafite indwara zo kubyimba mu mara bwagaragaje ko kwiyongera no kugabanuka mu rugero rumwe (Walters et al., 2014; Forbes et al., 2018; Upadhyay et al., 2023). Kugira ngo tugabanye ingaruka z'ivangura rishingiye ku gitsina, twagerageje kwemeza ko ibitsina byombi bihagarariwe kimwe kugira ngo itandukaniro rishobore guterwa n'imirire. Imwe mu mbogamizi zo gusobanura imikorere ni ugukuraho uruhererekane rw'uturemangingo tw'uturemangingo. Uburyo bwacu bwo guhuza uturemangingo busaba 95% by'imiterere y'uruhererekane n'uburebure busa na 85%, ndetse no guhuza 90% kugira ngo dukureho uruhererekane rw'amakosa. Ariko, mu bihe bimwe na bimwe, twabonye COG zifite ibisobanuro bimwe (urugero, MUT) (Ishusho ya 6). Hakenewe ubundi bushakashatsi kugira ngo hamenyekane niba izi orthologs zitandukanye, zifitanye isano n'ubwoko runaka, cyangwa niba iyi ari imbogamizi ku buryo bwo gukusanya gene. Indi mbogamizi ku mikorere ni ukwivanga mu byiciro; gene ya bagiteri mmdA ni enzyme izwiho kugira uruhare mu gukora propionate, ariko KEGG ntabwo iyihuza n'inzira ya propionate metabolism. Ibinyuranye n'ibyo, scpB na mmcD orthologs zirafitanye isano. Umubare munini wa genes zidafite knockouts zagenwe ushobora gutuma umuntu atabasha kumenya genes zijyanye na PPA mu gihe cyo gusuzuma ubwinshi bwa genes. Ubushakashatsi bw'ejo hazaza buzakungukira mu isesengura rya metatranscriptome, rishobora gutanga ubumenyi bwimbitse ku mikorere ya gene microbiota mu mara no guhuza imiterere ya gene n'ingaruka zishobora kubaho. Ku bushakashatsi bujyanye n'indwara zihariye z'imikurire y'ubwonko cyangwa indwara zo mu mara zitera ububabare, hakenewe isuzuma ry'umubiri n'imyitwarire y'inyamaswa kugira ngo hahuzwe impinduka mu miterere ya gene na izi ndwara. Ubushakashatsi bw'inyongera bwo gutera microbiome mu mbeba zidafite inzoka nabwo bwaba ingirakamaro mu kumenya niba microbiome ari yo itera cyangwa iranga indwara.
Muri make, twerekanye ko PPA y’imirire igira uruhare mu guhindura imiterere ya mikorobe zo mu nda. PPA ni imiti irinda indwara yemewe na FDA iboneka cyane mu biribwa bitandukanye, iyo ikoreshejwe igihe kirekire, ishobora gutuma ibinyabuzima bisanzwe byo mu nda bihungabana. Twabonye impinduka mu bwinshi bwa bagiteri nyinshi, bigaragaza ko PPA ishobora kugira ingaruka ku miterere ya mikorobe zo mu nda. Impinduka muri mikorobe zishobora gutuma habaho impinduka mu buryo bumwe na bumwe bw’imikorere y’umubiri, bishobora gutera impinduka mu miterere y’umubiri zijyanye n’ubuzima bw’umuntu. Hakenewe ubundi bushakashatsi kugira ngo hamenyekane niba ingaruka za PPA y’imirire ku miterere ya mikorobe zishobora gutera dysbiosis cyangwa izindi ndwara. Ubu bushakashatsi bushyiraho urufatiro rw’ubushakashatsi bw’ejo hazaza ku buryo ingaruka za PPA ku miterere y’amara zishobora kugira ingaruka ku buzima bw’abantu.
Amakuru yagaragajwe muri ubu bushakashatsi araboneka mu bubiko bwo kuri interineti. Izina ry'ububiko n'inomero yabwo ni: https://www.ncbi.nlm.nih.gov/, PRJNA1092431.
Ubu bushakashatsi bw’inyamaswa bwemejwe na Komite ishinzwe kwita no gukoresha inyamaswa muri Kaminuza ya Central Florida (UCF-IACUC) (Nomero y’uruhushya rwo gukoresha inyamaswa: PROTO202000002). Ubu bushakashatsi bwubahiriza amategeko, amabwiriza n’ibisabwa n’inzego.
NG: Gusesengura ibitekerezo, Gusesengura amakuru, Gusesengura ku mugaragaro, Iperereza, Uburyo bwo gukora, Porogaramu, Gushushanya, Kwandika (umushinga w'umwimerere), Kwandika (isuzuma n'ikosora). LA: Gusesengura ibitekerezo, Gusesengura amakuru, Uburyo bwo gukora, Amasoko, Kwandika (isuzuma n'ikosora). SH: Isesengura ryemewe, Porogaramu, Kwandika (isuzuma n'ikosora). SA: Iperereza, Kwandika (isuzuma n'ikosora). Umucamanza Mukuru: Iperereza, Kwandika (isuzuma n'ikosora). SN: Gusesengura ibitekerezo, Imicungire y'imishinga, Amasoko, Gukurikirana, Kwandika (isuzuma n'ikosora). TA: Gusesengura ibitekerezo, Imicungire y'imishinga, Gukurikirana, Kwandika (isuzuma n'ikosora).
Abanditsi batangaje ko nta nkunga y'amafaranga babonye ku bushakashatsi, ubwanditsi, cyangwa igisohoka cy'iyi nkuru.
Abanditsi batangaza ko ubushakashatsi bwakozwe mu gihe nta mibanire y’ubucuruzi cyangwa imari yari ihari ishobora gufatwa nk’igongana ry’inyungu.
Ibitekerezo byose bivugwa muri iyi nkuru ni iby’abanditsi gusa kandi ntibigaragaza ibitekerezo by’ibigo byabo, abasohora ibitabo, abanditsi, cyangwa abasesengura. Ibicuruzwa byose byasuzumwe muri iyi nkuru, cyangwa ibyavuzwe n’ababikora, ntabwo byemejwe cyangwa ngo byemezwe n’uwabisohoye.
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Igihe cyo kohereza: 18 Mata 2025