An nuna cewa Propionic acid (PPA), wani sinadari mai kashe ƙwayoyin cuta da kuma ƙarin abinci mai gina jiki, yana haifar da rashin ci gaban jijiyoyi mara kyau a cikin beraye tare da rashin aikin narkewar abinci, wanda ƙila yana iya faruwa ne sakamakon dysbiosis na hanji. An ba da shawarar alaƙa tsakanin fallasa PPA a cikin abinci da dysbiosis na microbiota na hanji, amma ba a bincika kai tsaye ba. A nan, mun binciki canje-canjen da PPA ke da alaƙa da su a cikin tsarin microbiota na hanji wanda zai iya haifar da dysbiosis. An tsara ƙwayoyin microbiomes na hanji na beraye da aka ciyar da abinci mara magani (n=9) da abinci mai wadatar PPA (n=13) ta amfani da jerin metagenomic na dogon lokaci don tantance bambance-bambance a cikin tsarin microbiodes da hanyoyin metabolism na ƙwayoyin cuta. PPA na abinci yana da alaƙa da ƙaruwar yawan muhimman abubuwa, gami da nau'ikan Bacteroides, Prevotella, da Ruminococcus da yawa, waɗanda a baya aka sanya membobinsu a cikin samar da PPA. Ƙananan ƙwayoyin beraye na PPA da aka fallasa suma suna da ƙarin hanyoyin da suka shafi metabolism na lipid da biosynthesis na hormone steroid. Sakamakonmu ya nuna cewa PPA na iya canza ƙwayoyin microbiota na hanji da hanyoyin metabolism da ke da alaƙa da shi. Waɗannan canje-canjen da aka lura sun nuna cewa abubuwan kiyayewa da aka ware a matsayin masu aminci don amfani na iya yin tasiri ga tsarin ƙwayoyin cuta na hanji da kuma lafiyar ɗan adam. Daga cikinsu, ana zaɓar P, G ko S dangane da matakin rarrabuwa da ake nazari a kai. Don rage tasirin rarrabuwar ƙarya mai kyau, an ɗauki mafi ƙarancin ma'aunin yawan dangi na 1e-4 (karanta 1/10,000). Kafin nazarin ƙididdiga, an canza yawan dangi da Bracken ya ruwaito (karanta_ƙasa_ƙasa_ƙasa) ta amfani da canjin rabon log-ratio na tsakiya (CLR) (Aitchison, 1982). An zaɓi hanyar CLR don canza bayanai saboda ba ta canzawa kuma ta isa ga bayanan da ba su da yawa (Gloor et al., 2017). Sauyin CLR yana amfani da logarithm na halitta. Bayanan ƙidaya da Bracken ya ruwaito an daidaita su ta amfani da bayanin log na dangi (RLE) (Anders da Huber, 2010). An samar da alkaluma ta amfani da haɗin matplotlib v. 3.7.1, seaborn v. 3.7.2 da logarithms masu jere (Gloor et al., 2017). 0.12.2 da kuma stantanotations v. 0.5.0 (Hunter, 2007; Waskom, 2021; Charlier et al., 2022). An ƙididdige rabon Bacillus/Bacteoroidetes ga kowane samfurin ta amfani da ƙididdigar ƙwayoyin cuta da aka daidaita. Ƙimar da aka ruwaito a cikin tebura an zagaye ta zuwa wurare 4 na decimal. An ƙididdige ma'aunin bambancin Simpson ta amfani da rubutun alpha_diversity.py da aka bayar a cikin fakitin KrakenTools v. 1.2 (Lu et al., 2022). An bayar da rahoton Bracken a cikin rubutun kuma an bayar da ma'aunin Simpson "Si" don ma'aunin -an. An bayyana manyan bambance-bambance a cikin yawa a matsayin matsakaicin bambance-bambancen CLR ≥ 1 ko ≤ -1. Matsakaicin bambancin CLR na ±1 yana nuna ƙaruwa sau 2.7 a cikin yawan nau'in samfurin. Alamar (+/-) tana nuna ko taxon ya fi yawa a cikin samfurin PPA da samfurin sarrafawa, bi da bi. An tantance mahimmanci ta amfani da gwajin Mann-Whitney U (Virtanen et al., 2020). An yi amfani da Statsmodels v. 0.14 (Benjamini da Hochberg, 1995; Seabold da Perktold, 2010), kuma an yi amfani da hanyar Benjamini-Hochberg don gyara don gwaji da yawa. An yi amfani da ƙimar p da aka daidaita ≤ 0.05 a matsayin maƙasudin tantance mahimmancin ƙididdiga.
Ana kiran ƙwayoyin cuta na ɗan adam da "gabobi na ƙarshe na jiki" kuma suna taka muhimmiyar rawa a lafiyar ɗan adam (Baquero da Nombela, 2012). Musamman ma, ƙwayoyin cuta na hanji an san su da tasirinsu a faɗin tsarinsu da kuma rawar da suke takawa a cikin ayyuka masu mahimmanci da yawa. Kwayoyin cuta na gama gari suna da yawa a cikin hanji, suna mamaye wurare daban-daban na muhalli, suna amfani da abubuwan gina jiki, kuma suna fafatawa da ƙwayoyin cuta masu yuwuwa (Jandhyala et al., 2015). Abubuwa daban-daban na ƙwayoyin cuta na hanji suna iya samar da muhimman abubuwan gina jiki kamar bitamin da haɓaka narkewar abinci (Rowland et al., 2018). An kuma nuna cewa ƙwayoyin cuta suna yin tasiri ga ci gaban nama da haɓaka hanyoyin rayuwa da garkuwar jiki (Heijtz et al., 2011; Yu et al., 2022). Tsarin ƙwayoyin cuta na hanjin ɗan adam ya bambanta sosai kuma ya dogara da abubuwan gado da muhalli kamar abinci, jinsi, magunguna, da matsayin lafiya (Kumbhare et al., 2019).
Abincin uwa muhimmin bangare ne na ci gaban tayi da jarirai kuma tushen sinadaran da za su iya yin tasiri ga ci gaba (Bazer et al., 2004; Innis, 2014). Ɗaya daga cikin irin wannan sinadarin da ke da ban sha'awa shine propionic acid (PPA), wani abu mai kama da kitse mai ɗan gajeren sarka da aka samo daga fermentation na ƙwayoyin cuta da kuma wani ƙarin abinci (den Besten et al., 2013). PPA yana da kaddarorin kashe ƙwayoyin cuta da na fungal kuma saboda haka ana amfani da shi azaman abin kiyaye abinci da kuma a aikace-aikacen masana'antu don hana ci gaban mold da ƙwayoyin cuta (Wemmenhove et al., 2016). PPA yana da tasiri daban-daban a cikin kyallen takarda daban-daban. A cikin hanta, PPA yana da tasirin hana kumburi ta hanyar shafar bayyanar cytokine a cikin macrophages (Kawasoe et al., 2022). An kuma lura da wannan tasirin tsari a cikin wasu ƙwayoyin rigakafi, wanda ke haifar da raguwar kumburi (Haase et al., 2021). Duk da haka, an lura da akasin haka a cikin kwakwalwa. Nazarin da aka yi a baya ya nuna cewa fallasa PPA yana haifar da halayyar autism a cikin beraye (El-Ansary et al., 2012). Wasu bincike sun nuna cewa PPA na iya haifar da gliosis da kunna hanyoyin kumburi a cikin kwakwalwa (Abdelli et al., 2019). Saboda PPA acid ne mai rauni, yana iya yaɗuwa ta cikin epithelium na hanji zuwa cikin jini don haka ya ketare shinge masu tsauri, gami da shingen jini-kwakwalwa da kuma mahaifa (Stinson et al., 2019), yana nuna mahimmancin PPA a matsayin metabolite mai tsari wanda ƙwayoyin cuta ke samarwa. Kodayake ana binciken yuwuwar rawar da PPA ke takawa a matsayin abin da ke haifar da autism a halin yanzu, tasirinsa ga mutanen da ke da autism na iya wuce gona da iri wajen haifar da bambancin jijiyoyi.
Alamomin ciki kamar gudawa da maƙarƙashiya sun zama ruwan dare a cikin marasa lafiya da ke fama da cututtukan ci gaban jijiyoyi (Cao et al., 2021). Nazarin da aka yi a baya ya nuna cewa ƙwayoyin cuta na marasa lafiya da ke fama da cututtukan autism (ASD) sun bambanta da na mutanen da ke da lafiya, wanda ke nuna kasancewar ƙwayoyin cuta na hanji (Finegold et al., 2010). Hakazalika, halayen ƙwayoyin cuta na marasa lafiya da ke fama da cututtukan hanji masu kumburi, kiba, cutar Alzheimer, da sauransu suma sun bambanta da na mutanen da ke da lafiya (Turnbaugh et al., 2009; Vogt et al., 2017; Henke et al., 2019). Duk da haka, har zuwa yau, ba a kafa wata alaƙa tsakanin ƙwayoyin cuta na hanji da cututtukan jijiyoyi ko alamu ba (Yap et al., 2021), kodayake ana tsammanin nau'ikan ƙwayoyin cuta da yawa suna taka rawa a wasu daga cikin waɗannan yanayin cututtuka. Misali, Akkermansia, Bacteroides, Clostridium, Lactobacillus, Desulfovibrio da sauran nau'ikan halittu sun fi yawa a cikin ƙwayoyin cuta na marasa lafiya da ke fama da autism (Tomova et al., 2015; Golubeva et al., 2017; Cristiano et al., 2018; Zurita et al., 2020). Abin lura shi ne, nau'ikan wasu daga cikin waɗannan nau'ikan halittu an san su da suna da kwayoyin halitta da ke da alaƙa da samar da PPA (Reichardt et al., 2014; Yun da Lee, 2016; Zhang et al., 2019; Baur da Dürre, 2023). Ganin cewa PPA tana da ƙwayoyin cuta masu kashe ƙwayoyin cuta, ƙara yawanta na iya zama da amfani ga ci gaban ƙwayoyin cuta masu samar da PPA (Jacobson et al., 2018). Don haka, yanayin da ke da wadataccen PFA na iya haifar da canje-canje a cikin ƙwayoyin cuta na hanji, gami da ƙwayoyin cuta na ciki, waɗanda ƙila su zama abubuwan da za su iya haifar da alamun cututtukan ciki.
Babban tambaya a binciken ƙwayoyin cuta ita ce ko bambance-bambancen da ke cikin ƙwayoyin cuta suna da alaƙa ko kuma alamun cututtuka da ke haifar da su. Mataki na farko don bayyana alaƙar da ke tsakanin abinci, ƙwayoyin cuta na hanji, da cututtukan jijiyoyi shine a tantance tasirin abinci akan ƙwayoyin cuta. Don wannan dalili, mun yi amfani da jerin metagenomic da aka karanta na dogon lokaci don kwatanta ƙwayoyin cuta na cikin 'ya'yan beraye da aka ciyar da abinci mai wadataccen PPA ko abinci mai ƙarancin PPA. An ciyar da 'ya'yan iri iri kamar yadda uwayensu suka ci. Mun yi hasashen cewa abinci mai wadataccen PPA zai haifar da canje-canje a cikin tsarin ƙwayoyin cuta na hanji da hanyoyin aiki na ƙwayoyin cuta, musamman waɗanda ke da alaƙa da metabolism na PPA da/ko samar da PPA.
Wannan binciken ya yi amfani da berayen da aka canza launinsu zuwa FVB/N-Tg(GFAP-GFP)14Mes/J (Jackson Laboratories) waɗanda ke ɗauke da furotin mai haske mai haske (GFP) a ƙarƙashin ikon mai haɓaka GFAP na musamman ta hanyar bin ƙa'idodin Kwamitin Kula da Dabbobi da Amfani da su na Jami'ar Central Florida (UCF-IACUC) (Lambar Izinin Amfani da Dabbobi: PROTO202000002). Bayan an yaye beraye, an ajiye su daban-daban a cikin keji tare da berayen 1-5 na kowane jinsi a kowane keji. An ciyar da berayen ad libitum tare da ko dai abincin da aka tsarkake (abincin da aka gyara na buɗaɗɗen lakabin, 16 kcal% mai) ko abincin da aka ƙara na sodium propionate (abincin da aka gyara na buɗaɗɗen lakabin, 16 kcal% mai, wanda ke ɗauke da 5,000 ppm sodium propionate). Adadin sodium propionate da aka yi amfani da shi yayi daidai da jimlar nauyin abinci na 5,000 mg na PFA/kg. Wannan shine mafi girman yawan PPA da aka amince da shi don amfani azaman abin kiyaye abinci. Domin shirya wannan binciken, an ciyar da beraye biyu na tsawon makonni 4 kafin su haɗu kuma an ci gaba da yin ciki a cikin dam ɗin. An yaye beraye [beraye 22, masu kula da 9 (maza 6, mata 3) da kuma 13 PPA (maza 4, mata 9)] sannan aka ci gaba da cin abinci iri ɗaya da dam ɗin na tsawon watanni 5. An yanka beraye 'ya'yan da suka haifa suna da watanni 5 kuma an tattara abubuwan da ke cikin najasar hanjinsu kuma an fara adana su a cikin bututun microcentrifuge mai girman 1.5 ml a -20°C sannan aka mayar da su cikin injin daskarewa mai matsakaicin -80°C har sai an cire DNA ɗin mai masaukin baki kuma an cire ƙwayoyin cuta masu ƙwayoyin cuta.
An cire DNA na mai masaukin baki bisa ga wata yarjejeniya da aka gyara (Charalampous et al., 2019). A takaice, an canja abin da ke cikin najasa zuwa 500 µl InhibitEX (Qiagen, Cat#/ID: 19593) kuma an adana shi a daskare. A sarrafa aƙalla ƙwayoyin najasa 1-2 a kowane cirewa. Daga nan aka haɗa abubuwan da ke cikin najasa ta hanyar injiniya ta amfani da ƙwayar filastik a cikin bututu don samar da slurry. A sanya samfuran a cikin injin centrifuge a 10,000 RCF na tsawon minti 5 ko har sai samfuran sun yi pellet, sannan a zuba ruwan da ke cikin su sannan a sake sanya pellet ɗin a cikin 250 µl 1× PBS. A ƙara 250 µl 4.4% saponin bayani (TCI, lambar samfurin S0019) zuwa samfurin a matsayin sabulun wanki don sassauta membranes na ƙwayoyin eukaryotic. An haɗa samfuran a hankali har sai sun yi laushi kuma an rufe su a zafin ɗaki na minti 10. Bayan haka, don wargaza ƙwayoyin eukaryotic, an ƙara ruwa mara nuclease 350 μl a cikin samfurin, an saka shi na tsawon daƙiƙa 30, sannan aka ƙara NaCl 12 μl 5 M. Sannan aka sanya samfuran a centrifuge a 6000 RCF na tsawon mintuna 5. A shafa ruwan da ke cikin ruwan sannan a sake dasa pellet ɗin a cikin 100 μl 1X PBS. Don cire DNA mai masaukin baki, a ƙara 100 μl HL-SAN buffer (12.8568 g NaCl, 4 ml 1M MgCl2, 36 ml ruwa mara nuclease) da 10 μl HL-SAN enzyme (ArticZymes P/N 70910-202). An gauraya samfuran sosai ta hanyar bututun bututu sannan aka saka su a zafin jiki na 37 °C na tsawon minti 30 a 800 rpm a kan Eppendorf™ ThermoMixer C. Bayan an saka su a cikin injin Eppendorf™ ThermoMixer C, sai a saka su a centrifuge a 6000 RCF na tsawon minti 3 sannan a wanke su sau biyu da 800 µl da 1000 µl PBS. A ƙarshe, a sake mayar da pellet ɗin a cikin 100 µl PBS.
An ware jimillar DNA na ƙwayoyin cuta ta amfani da New England Biolabs Monarch Genomic DNA Purification Kit (New England Biolabs, Ipswich, MA, Cat# T3010L). An ɗan gyara tsarin aiki na yau da kullun da aka bayar tare da kayan aikin. A saka ruwa mara nuclease kuma a kula da shi a zafin 60°C kafin a yi aiki don cirewa na ƙarshe. A ƙara 10 µl Proteinase K da 3 µl RNase A ga kowane samfurin. Sannan a ƙara 100 µl Cell Lysis Buffer a gauraya a hankali. Daga nan aka saka samfuran a cikin Eppendorf™ ThermoMixer C a zafin 56°C da 1400 rpm na akalla awa 1 zuwa 1400 rpm na akalla awa 1 har zuwa awanni 3. An saka samfuran da aka saka a centrifuge a 12,000 RCF na tsawon mintuna 3 kuma an canja wurin supernatant daga kowane samfurin zuwa wani bututun microcentrifuge na 1.5 mL wanda ke ɗauke da 400 µL na maganin ɗaurewa. Daga nan aka yi ta bugun bututun na tsawon daƙiƙa 5-10 a tazara ta daƙiƙa 1. A mayar da dukkan ruwan da ke cikin kowanne samfurin (kimanin 600–700 µL) zuwa wani akwati mai tacewa da aka sanya a cikin bututun tattarawa. An sanya bututun a centrifuge a 1,000 RCF na tsawon mintuna 3 don ba da damar ɗaure DNA na farko sannan a sanya centrifuge a 12,000 RCF na tsawon mintuna 1 don cire ruwan da ya rage. An canza ginshiƙin samfurin zuwa sabon bututun tattarawa sannan a wanke sau biyu. Don wankewa ta farko, a ƙara 500 µL na wankin buffer zuwa kowane bututu. A juya bututun sau 3-5 sannan a centrifuge a 12,000 RCF na tsawon minti 1. A jefar da ruwan daga bututun tattarawa sannan a mayar da harsashin tacewa cikin bututun tattarawa ɗaya. Don wankewa ta biyu, a ƙara 500 µL na wankin buffer a cikin matatar ba tare da an juya ba. An sanya samfuran a centrifuge a 12,000 RCF na tsawon minti 1. A mayar da matatar zuwa bututun LoBind® mai girman 1.5 mL sannan a ƙara 100 µL na ruwa mara ɗumi da aka riga aka ɗumama. An saka matatun a zafin ɗaki na minti 1 sannan a sanya su a centrifuge a 12,000 RCF na minti 1. An adana DNA ɗin da aka cire a -80°C.
An auna yawan DNA ta amfani da na'urar Qubit™ 4.0 Fluorometer. An shirya DNA ta amfani da Kit ɗin Qubit™ 1X dsDNA High Sensitivity Kit (Cat. No. Q33231) bisa ga umarnin masana'anta. An auna tsawon rabe-raben DNA ta amfani da TapeStation na Aglient™ 4150 ko 4200. An shirya DNA ta amfani da Agilent™ Genomic DNA Reagents (Cat. No. 5067-5366) da Genomic DNA ScreenTape (Cat. No. 5067-5365). An gudanar da shirye-shiryen ɗakin karatu ta amfani da Kit ɗin Barcoding na Oxford Nanopore Technologies™ (ONT) Rapid PCR (SQK-RPB004) bisa ga umarnin masana'anta. An tsara DNA ta amfani da sequencer na ONT GriDION™ Mk1 tare da ƙwayar kwarara ta Min106D (R 9.4.1). Saitunan jeri sune: babban daidaiton kiran tushe, mafi ƙarancin ƙimar q na 9, saitin barcode, da kuma yanke barcode. An yi jerin samfurori na tsawon awanni 72, bayan haka aka gabatar da bayanan kiran asali don ƙarin sarrafawa da bincike.
An gudanar da aikin sarrafa Bioinformatics ta amfani da hanyoyin da aka bayyana a baya (Greenman et al., 2024). An raba fayilolin FASTQ da aka samu daga jerin bayanai zuwa kundin adireshi ga kowane samfurin. Kafin nazarin bioinformatics, an sarrafa bayanan ta amfani da bututun mai zuwa: da farko, an haɗa fayilolin FASTQ na samfuran zuwa fayil ɗin FASTQ guda ɗaya. Sannan, an tace karatuttukan da suka fi guntu fiye da 1000 bp ta amfani da Filtlong v. 0.2.1, tare da sigar da aka canza ita ce –min_length 1000 (Wick, 2024). Kafin ƙarin tacewa, an sarrafa ingancin karantawa ta amfani da NanoPlot v. 1.41.3 tare da sigogi masu zuwa: –fastq –plots dot –N50 -o
Don rarrabuwar taxonomic, an rarraba karantawa da abubuwan da aka haɗa ta amfani da Kraken2 v. 2.1.2 (Wood et al., 2019). Samar da rahotanni da fitarwa fayiloli don karantawa da haɗuwa, bi da bi. Yi amfani da zaɓin sunayen-amfani don nazarin karantawa da haɗuwa. Zaɓuɓɓukan-matse-gzip da aka haɗa an ƙayyade su don sassan karantawa. An kiyasta yawan taxa a cikin metagenomes ta amfani da Bracken v. 2.8 (Lu et al., 2017). Da farko mun ƙirƙiri wani rumbun bayanai na kmer wanda ke ɗauke da tushe 1000 ta amfani da ginin bracken tare da sigogi masu zuwa: -d
An yi bayanin kwayoyin halitta da kuma kimanta yawan da ke tsakanin kwayoyin halitta ta amfani da sigar da aka gyara ta hanyar Maranga et al. (Maranga et al., 2023). Da farko, an cire guntun da suka fi guntu fiye da 500 bp daga dukkan taruka ta amfani da SeqKit v. 2.5.1 (Shen et al., 2016). Sannan aka haɗa taruka da aka zaɓa zuwa pan-metagenome. An gano taruka masu buɗewa (ORFs) ta amfani da Prodigal v. 1.0.1 (sigar Prodigal v. 2.6.3 mai layi ɗaya) tare da waɗannan sigogi: -d
An fara haɗa kwayoyin halitta bisa ga Kyoto Encyclopedia of Genes and Genomes (KEGG) ortholog (KO) da aka sanya ta eggNOG don kwatanta yawan hanyoyin kwayoyin halitta. An cire kwayoyin halitta ba tare da knockouts ko kwayoyin halitta masu knockouts da yawa ba kafin a yi nazari. Daga nan aka ƙididdige matsakaicin yawan kowanne KO a kowane samfuri kuma aka yi nazarin ƙididdiga. An ayyana kwayoyin halittar metabolism na PPA a matsayin kowace kwayar halitta da aka sanya a jere ko00640 a cikin ginshiƙin KEGG_Pathway, yana nuna rawar da ke takawa a cikin metabolism na propionate bisa ga KEGG. Kwayoyin halittar da aka gano suna da alaƙa da samar da PPA an jera su a cikin Ƙarin Tebur 1 (Reichardt et al., 2014; Yang et al., 2017). An gudanar da gwaje-gwajen permutation don gano metabolism na PPA da kwayoyin halittar samarwa waɗanda suka fi yawa a kowane nau'in samfurin. An yi permutations dubu ɗaya ga kowane kwayar halitta da aka bincika. An yi amfani da ƙimar p na 0.05 a matsayin yankewa don tantance mahimmancin ƙididdiga. An sanya bayanan aiki ga kwayoyin halitta daban-daban a cikin rukuni bisa ga bayanan kwayoyin halitta masu wakiltar a cikin rukuni. Ana iya gano Taxa da ke da alaƙa da metabolism na PPA da/ko samar da PPA ta hanyar daidaita ID na contig a cikin fayilolin fitarwa na Kraken2 tare da ID na contig iri ɗaya da aka riƙe yayin bayanin aiki ta amfani da eggNOG. An gudanar da gwajin mahimmanci ta amfani da gwajin Mann-Whitney U da aka bayyana a baya. An yi gyara don gwaji da yawa ta amfani da hanyar Benjamini-Hochberg. An yi amfani da ƙimar p na ≤ 0.05 azaman yankewa don tantance mahimmancin ƙididdiga.
An tantance bambancin ƙwayoyin cuta na hanji na bera ta amfani da ma'aunin bambancin Simpson. Ba a lura da wani bambanci mai mahimmanci tsakanin samfuran sarrafawa da PPA ba dangane da bambancin nau'in halittu da nau'ikan halittu (darajar p ga nau'in halittu: 0.18, ƙimar p ga nau'in halittu: 0.16) (Hoto na 1). Daga nan aka kwatanta abubuwan da ke cikin ƙwayoyin cuta ta amfani da babban nazarin abubuwan da ke cikin sinadaran (PCA). Hoto na 2 ya nuna tarin samfuran ta hanyar phyla ɗinsu, yana nuna cewa akwai bambance-bambance a cikin abun da ke cikin ƙwayoyin cuta tsakanin samfuran PPA da na sarrafawa. Wannan tarin bai bayyana sosai a matakin kwayoyin halitta ba, yana nuna cewa PPA yana shafar wasu ƙwayoyin cuta (Hoto na 1 na Ƙarin Bayani).
Siffa ta 1. Bambancin nau'in Alpha da kuma nau'in halittar ƙwayoyin cuta na hanjin beraye. Tsarin akwatunan da ke nuna alamun bambancin Simpson na nau'in (A) da nau'in (B) a cikin samfuran PPA da sarrafawa. An ƙayyade mahimmanci ta amfani da gwajin Mann-Whitney U, kuma an yi gyare-gyare da yawa ta amfani da hanyar Benjamini-Hochberg. ns, ƙimar p ba ta da mahimmanci (p>0.05).
Siffa ta 2. Sakamakon babban nazarin sassan da ke cikin ƙwayoyin cuta na hanjin linzamin kwamfuta a matakin nau'in. Babban taswirar nazarin sassan yana nuna rarrabawar samfuran a cikin manyan sassan su guda biyu na farko. Launuka suna nuna nau'in samfurin: Berayen da aka fallasa ga PPA shunayya ne kuma beraye masu sarrafawa rawaya ne. Babban sassan 1 da 2 an zana su a kan axis na x da y, bi da bi, kuma an bayyana su a matsayin rabon bambancin da aka bayyana.
Ta amfani da bayanan ƙidaya da aka canza na RLE, an lura da raguwa mai mahimmanci a cikin matsakaicin rabo na Bacteroidetes/Bacilli a cikin beraye masu sarrafawa da PPA (ikon sarrafawa: 9.66, PPA: 3.02; ƙimar p = 0.0011). Wannan bambanci ya faru ne saboda yawan Bacteroidetes a cikin beraye masu sarrafawa idan aka kwatanta da sarrafawa, kodayake bambancin bai kasance mai mahimmanci ba (matsayin sarrafawa CLR: 5.51, ma'anar PPA CLR: 6.62; ƙimar p = 0.054), yayin da yawan Bacteroidetes yayi kama (matsayin sarrafawa CLR: 7.76, ma'anar PPA CLR: 7.60; ƙimar p = 0.18).
Binciken yawan mambobi na ƙwayoyin cuta na hanji ya nuna cewa nau'in phylum 1 da nau'in 77 sun bambanta sosai tsakanin samfuran PPA da samfuran sarrafawa (Jadawali na Ƙarin 2). Yawancin nau'ikan PPA 59 ya fi na samfuran sarrafawa girma, yayin da yawan nau'ikan 16 kawai a cikin samfuran sarrafawa ya fi na samfuran PPA girma (Hoto na 3).
Siffa ta 3. Bambancin yawan taxa a cikin ƙwayoyin halittar ciki na PPA da beraye masu sarrafawa. Tsarin aman wuta yana nuna bambance-bambance a cikin yawan nau'ikan halitta (A) ko nau'in halitta (B) tsakanin samfuran PPA da na sarrafawa. Dige-dige masu launin toka ba su nuna wani bambanci mai mahimmanci a cikin yawan taxa ba. Dige-dige masu launi suna nuna bambance-bambance masu mahimmanci a cikin yawa (ƙimar p ≤ 0.05). Manyan dige-dige 20 tare da mafi girman bambance-bambance a cikin yawa tsakanin nau'ikan samfura an nuna su a cikin ja da shuɗi mai haske (samfuran sarrafawa da PPA), bi da bi. Dige-dige masu launin rawaya da shunayya sun kasance aƙalla sau 2.7 sun fi yawa a cikin samfuran sarrafawa ko na PPA fiye da na sarrafawa. Dige-dige baƙi suna wakiltar dige-dige masu yawa daban-daban, tare da matsakaicin bambance-bambancen CLR tsakanin -1 da 1. An ƙididdige ƙimar P ta amfani da gwajin Mann-Whitney U kuma an gyara su don gwaji da yawa ta amfani da hanyar Benjamini-Hochberg. Bambancin matsakaicin CLR mai ƙarfi yana nuna manyan bambance-bambance a cikin yawa.
Bayan mun yi nazarin abubuwan da ke cikin ƙwayoyin cuta na hanji, mun yi bayanin aiki na ƙwayoyin cuta. Bayan mun tace ƙwayoyin cuta marasa inganci, an gano jimillar ƙwayoyin cuta na musamman guda 378,355 a cikin dukkan samfuran. An yi amfani da yawan waɗannan ƙwayoyin cuta da aka canza don nazarin manyan abubuwan da ke cikin jiki (PCA), kuma sakamakon ya nuna babban matakin tattara nau'ikan samfuran bisa ga bayanan aikinsu (Hoto na 4).
Hoto na 4. Sakamakon PCA ta amfani da bayanin aiki na ƙwayoyin cuta na hanjin linzamin kwamfuta. Tsarin PCA yana nuna rarrabawar samfuran a cikin manyan abubuwan haɗinsu guda biyu na farko. Launuka suna nuna nau'in samfurin: beraye masu fallasa ga PPA suna da shunayya kuma beraye masu sarrafawa suna da rawaya. Babban sassan 1 da 2 an zana su akan axis na x da y, bi da bi, kuma an bayyana su azaman rabon bambancin da aka bayyana.
Mun sake duba yawan ƙwanƙwasa KEGG a cikin nau'ikan samfura daban-daban. An gano jimillar ƙwanƙwasa na musamman guda 3648, waɗanda 196 sun fi yawa a cikin samfuran sarrafawa kuma 106 sun fi yawa a cikin samfuran PPA (Hoto na 5). An gano jimillar kwayoyin halitta 145 a cikin samfuran sarrafawa da kwayoyin halitta 61 a cikin samfuran PPA, tare da yalwar abubuwa daban-daban. Hanyoyin da suka shafi metabolism na lipid da aminosugar sun fi wadata a cikin samfuran PPA (Tebur na Ƙarin 3). Hanyoyin da suka shafi metabolism na nitrogen da tsarin relay na sulfur sun fi wadata a cikin samfuran sarrafawa (Tebur na Ƙarin 3). Yawan kwayoyin halitta da suka shafi metabolism na aminosugar/nucleotide (ko:K21279) da metabolism na inositol phosphate (ko:K07291) sun fi girma a cikin samfuran PPA (Hoto na 5). Samfuran sarrafawa suna da ƙarin kwayoyin halitta da suka shafi metabolism na benzoate (ko:K22270), metabolism na nitrogen (ko:K00368), da glycolysis/gluconeogenesis (ko:K00131) (Hoto na 5).
Hoto na 5. Bambancin yawan KOs a cikin ƙwayoyin halittar ciki na PPA da beraye masu sarrafawa. Tsarin dutsen mai aman wuta yana nuna bambance-bambancen da ke cikin yawan ƙungiyoyi masu aiki (KOs). Digo-digo masu launin toka suna nuna KOs waɗanda yawansu bai bambanta sosai tsakanin nau'ikan samfura ba (ƙimar p > 0.05). Digo-digo masu launi suna nuna bambance-bambance masu mahimmanci a cikin yawa (ƙimar p ≤ 0.05). 20 KOs waɗanda ke da manyan bambance-bambance a cikin yawa tsakanin nau'ikan samfura an nuna su a ja da shuɗi mai haske, wanda ya dace da samfuran sarrafawa da PPA, bi da bi. Digo-digo masu launin rawaya da shunayya suna nuna KOs waɗanda suka kasance aƙalla ninki 2.7 fiye da samfuran sarrafawa da PPA, bi da bi. Digo-digo baƙi suna nuna KOs tare da yalwar yawa daban-daban, tare da matsakaicin bambance-bambancen CLR tsakanin -1 da 1. An ƙididdige ƙimar P ta amfani da gwajin Mann-Whitney U kuma an daidaita su don kwatantawa da yawa ta amfani da tsarin Benjamini-Hochberg. NaN yana nuna cewa KO ba ya cikin hanyar da ke cikin KEGG. Matsakaicin ƙimar bambancin CLR mai ƙarfi yana nuna manyan bambance-bambance a cikin yawa. Don ƙarin bayani game da hanyoyin da KOs ɗin da aka lissafa suka shiga, duba Ƙarin Tebur 3.
Daga cikin kwayoyin halittar da aka yi bayani dalla-dalla, kwayoyin halittar 1601 suna da yalwar da ta bambanta sosai tsakanin nau'ikan samfura (p ≤ 0.05), tare da kowace kwayar halitta ta kasance aƙalla ninki 2.7 fiye da haka. Daga cikin waɗannan kwayoyin halittar, kwayoyin halittar 4 sun fi yawa a cikin samfuran sarrafawa kuma kwayoyin halittar 1597 sun fi yawa a cikin samfuran PPA. Saboda PPA yana da kaddarorin ƙwayoyin cuta, mun bincika yawan metabolism na PPA da kwayoyin halittar samarwa tsakanin nau'ikan samfura. Daga cikin kwayoyin halittar da suka shafi metabolism na PPA 1332, kwayoyin halittar 27 sun fi yawa a cikin samfuran sarrafawa kuma kwayoyin halittar 12 sun fi yawa a cikin samfuran PPA. Daga cikin kwayoyin halittar da suka shafi samarwa na PPA 223, kwayar halittar 1 ta fi yawa a cikin samfuran PPA. Hoto na 6A ya kuma nuna yawan kwayoyin halittar da ke da hannu a metabolism na PPA, tare da yalwar da ta fi yawa a cikin samfuran sarrafawa da girman tasirin da ya fi girma, yayin da Hoto na 6B ya nuna kwayoyin halittar mutum ɗaya tare da yalwar da ta fi girma da aka lura a cikin samfuran PPA.
Hoto na 6. Bambancin yawan kwayoyin halitta masu alaƙa da PPA a cikin ƙwayoyin halittar hanji na linzamin kwamfuta. Tsarin aman wuta yana nuna bambance-bambancen yawan kwayoyin halitta da ke da alaƙa da metabolism na PPA (A) da samar da PPA (B). Digogi masu launin toka suna nuna kwayoyin halitta waɗanda yawansu bai bambanta sosai tsakanin nau'ikan samfura ba (ƙimar p > 0.05). Digogi masu launi suna nuna bambance-bambance masu mahimmanci a cikin yawa (ƙimar p ≤ 0.05). Kwayoyin halitta 20 waɗanda ke da manyan bambance-bambance a cikin yawa an nuna su a cikin ja da shuɗi mai haske (samfuran sarrafawa da PPA), bi da bi. Yawan digogi masu rawaya da shunayya ya fi aƙalla sau 2.7 girma a cikin samfuran sarrafawa da PPA fiye da samfuran sarrafawa. Digogi baƙi suna wakiltar kwayoyin halitta masu yawa daban-daban, tare da matsakaicin bambance-bambancen CLR tsakanin -1 da 1. An ƙididdige ƙimar P ta amfani da gwajin Mann-Whitney U kuma an gyara su don kwatantawa da yawa ta amfani da hanyar Benjamini-Hochberg. Kwayoyin halitta suna daidai da kwayoyin halitta masu wakilci a cikin kundin kwayar halitta mara yawan gaske. Sunayen kwayoyin halitta sun ƙunshi alamar KEGG wacce ke nuna kwayar KO. Bambancin CLR mai ƙarfi yana nuna yalwar da ta bambanta sosai. Alamar dash (-) tana nuna cewa babu wata alama ga kwayar halitta a cikin bayanan KEGG.
An gano nau'ikan kwayoyin halitta da suka shafi metabolism na PPA da/ko samarwa ta hanyar daidaita asalin kwayoyin halitta da ID na kwayar halitta. A matakin kwayoyin halitta, an gano nau'ikan kwayoyin halitta 130 suna da kwayoyin halitta da suka shafi metabolism na PPA kuma an gano nau'ikan kwayoyin halitta 61 suna da kwayoyin halitta da suka shafi samar da PPA (Karin Tebur 4). Duk da haka, babu wani nau'in halitta da ya nuna bambance-bambance masu mahimmanci a yawan (p > 0.05).
A matakin nau'in, an gano nau'ikan ƙwayoyin cuta 144 suna da kwayoyin halitta da ke da alaƙa da metabolism na PPA kuma an gano nau'ikan ƙwayoyin cuta 68 suna da kwayoyin halitta da ke da alaƙa da samar da PPA (Tebur na Ƙarin 5). Daga cikin masu metabolizing na PPA, ƙwayoyin cuta guda takwas sun nuna ƙaruwa mai yawa tsakanin nau'ikan samfura, kuma duk sun nuna manyan canje-canje a tasirinsu (Tebur na Ƙarin 6). Duk waɗanda aka gano masu metabolizing na PPA tare da manyan bambance-bambance a yalwa sun fi yawa a cikin samfuran PPA. Rarraba matakin nau'in ya nuna wakilan nau'ikan da ba su bambanta sosai tsakanin nau'ikan samfura ba, gami da nau'ikan Bacteroides da Ruminococcus da yawa, da kuma Duncania dubois, Myxobacterium enterica, Monococcus pectinolyticus, da Alcaligenes polymorpha. Daga cikin ƙwayoyin cuta masu samar da PPA, ƙwayoyin cuta guda huɗu sun nuna manyan bambance-bambance a yalwa tsakanin nau'ikan samfura. Nau'ikan da ke da manyan bambance-bambance a yalwa sun haɗa da Bacteroides novorossi, Duncania dubois, Myxobacterium enteritidis, da Ruminococcus bovis.
A cikin wannan binciken, mun bincika tasirin fallasa PPA akan ƙwayoyin cuta na hanji na beraye. PPA na iya haifar da martani daban-daban a cikin ƙwayoyin cuta saboda wasu nau'ikan halittu ne ke samar da shi, wasu nau'ikan halittu suna amfani da shi azaman tushen abinci, ko kuma yana da tasirin maganin rigakafi. Saboda haka, ƙarin sa ga yanayin hanji ta hanyar ƙarin abinci na iya samun sakamako daban-daban dangane da haƙuri, juriya, da ikon amfani da shi azaman tushen abinci. Ana iya kawar da nau'ikan ƙwayoyin cuta masu laushi kuma a maye gurbinsu da waɗanda suka fi juriya ga PPA ko kuma waɗanda za su iya amfani da shi azaman tushen abinci, wanda ke haifar da canje-canje a cikin abun da ke cikin ƙwayoyin cuta na hanji. Sakamakonmu ya nuna manyan bambance-bambance a cikin abun da ke cikin ƙwayoyin cuta amma babu wani tasiri akan bambancin ƙwayoyin cuta gabaɗaya. An lura da manyan tasirin a matakin nau'ikan, tare da fiye da nau'ikan 70 daban-daban sosai tsakanin PPA da samfuran sarrafawa (Jadawali na Ƙarin 2). Ƙarin kimantawa na abun da ke cikin samfuran da aka fallasa da PPA ya nuna babban bambancin nau'ikan ƙwayoyin cuta idan aka kwatanta da samfuran da ba a fallasa ba, yana nuna cewa PPA na iya haɓaka halayen haɓakar ƙwayoyin cuta da iyakance yawan ƙwayoyin cuta waɗanda za su iya rayuwa a cikin mahalli mai arzikin PPA. Don haka, PPA na iya haifar da canje-canje maimakon haifar da wargajewar bambancin ƙwayoyin cuta na hanji.
An riga an nuna cewa abubuwan kiyaye abinci kamar PPA suna canza yawan abubuwan da ke cikin hanji ba tare da shafar bambancin gaba ɗaya ba (Nagpal et al., 2021). A nan, mun lura da bambance-bambance mafi ban mamaki tsakanin nau'ikan Bacteroidetes a cikin phylum Bacteroidetes (wanda aka sani da Bacteroidetes a da), waɗanda aka wadatar da su sosai a cikin beraye da aka fallasa ga PPA. Ƙara yawan nau'ikan Bacteroides yana da alaƙa da ƙaruwar lalacewar majina, wanda zai iya ƙara haɗarin kamuwa da cuta da haɓaka kumburi (Cornick et al., 2015; Desai et al., 2016; Penzol et al., 2019). Wani bincike ya gano cewa beraye maza da aka haifa da aka yi wa magani da Bacteroides fragilis sun nuna halayen zamantakewa kamar na autism spectrum disorder (ASD) (Carmel et al., 2023), da sauran bincike sun nuna cewa nau'ikan Bacteroides na iya canza ayyukan garkuwar jiki da kuma haifar da cututtukan zuciya na autoimmune inflammatory cardiomyopathy (Gil-Cruz et al., 2019). Nau'ikan Ruminococcus, Prevotella, da Parabacteroides suma sun ƙaru sosai a cikin beraye da suka kamu da cutar PPA (Coretti et al., 2018). Wasu nau'ikan Ruminococcus suna da alaƙa da cututtuka kamar cutar Crohn ta hanyar samar da cytokines masu kumburi (Henke et al., 2019), yayin da nau'ikan Prevotella kamar Prevotella humani suna da alaƙa da cututtukan metabolism kamar hauhawar jini da kuma insulin sensitivity (Pedersen et al., 2016; Li et al., 2017). A ƙarshe, mun gano cewa rabon Bacteroidetes (wanda aka sani da Firmicutes a da) zuwa Bacteroidetes ya yi ƙasa sosai a cikin beraye da aka fallasa ga PPA fiye da beraye masu sarrafawa saboda yawan adadin Bacteroidetes. An riga an nuna cewa wannan rabo muhimmin alama ce ta yanayin gida na hanji, kuma rikice-rikice a cikin wannan rabon suna da alaƙa da yanayin cututtuka daban-daban (Turpin et al., 2016; Takezawa et al., 2021; An et al., 2023), gami da cututtukan kumburi na hanji (Stojanov et al., 2020). Gabaɗaya, nau'ikan phylum Bacteroidetes sun fi shafar yawan PPA na abinci. Wannan na iya zama saboda haƙurin PPA mai yawa ko ikon amfani da PPA a matsayin tushen kuzari, wanda aka nuna gaskiya ne ga aƙalla nau'in guda ɗaya, Hoylesella enocea (Hitch et al., 2022). A madadin haka, fallasa PPA na uwa na iya haɓaka ci gaban tayi ta hanyar sa hanjin 'ya'yan bera ya fi saurin kamuwa da mulkin mallaka na Bacteroidetes; duk da haka, ƙirar bincikenmu bai ba da damar irin wannan kimantawa ba.
Binciken abubuwan da ke cikin metagenomic ya nuna bambance-bambance masu yawa a cikin yawan kwayoyin halitta da ke da alaƙa da metabolism da samarwa na PPA, tare da beraye da aka fallasa ga PPA suna nuna yawan kwayoyin halitta da ke da alhakin samar da PPA, yayin da beraye da ba a fallasa ga PPA ba sun nuna yawan kwayoyin halitta da ke da alhakin metabolism na PAA (Hoto na 6). Waɗannan sakamakon sun nuna cewa tasirin PPA akan tsarin ƙwayoyin cuta bazai zama kawai saboda amfani da shi ba, in ba haka ba yawan kwayoyin halitta da ke da alaƙa da metabolism na PPA yakamata ya nuna yawan ƙwayoyin cuta a cikin ƙwayoyin cuta na beraye da aka fallasa ga PPA. Bayani ɗaya shine cewa PPA yana daidaita yawan ƙwayoyin cuta musamman ta hanyar tasirinsa na rigakafi maimakon ta hanyar amfani da ƙwayoyin cuta a matsayin abinci mai gina jiki. Nazarin da aka yi a baya sun nuna cewa PPA yana hana ci gaban Salmonella Typhimurium ta hanyar da ta dogara da kashi (Jacobson et al., 2018). Fuskantar yawan PPA na iya zaɓar ƙwayoyin cuta waɗanda ke da juriya ga kaddarorin ƙwayoyin cuta kuma ba lallai bane su iya metabolize ko samar da shi. Misali, nau'ikan Parabacteroides da yawa sun nuna yawan gaske a cikin samfuran PPA, amma ba a gano wani kwayar halitta da ke da alaƙa da metabolism ko samarwa na PPA ba (Teburin Ƙarin 2, 4, da 5). Bugu da ƙari, samar da PPA a matsayin samfurin fermentation yana yaɗuwa sosai tsakanin ƙwayoyin cuta daban-daban (Gonzalez-Garcia et al., 2017). Babban bambancin ƙwayoyin cuta na iya zama dalilin yawan ƙwayoyin halitta da ke da alaƙa da metabolism na PPA a cikin samfuran sarrafawa (Averina et al., 2020). Bugu da ƙari, an yi hasashen cewa kwayoyin halitta 27 (2.14%) kawai na kwayoyin halitta 1332 ne ke da alaƙa da metabolism na PPA kawai. Kwayoyin halitta da yawa da ke da alaƙa da metabolism na PPA suma suna da hannu a cikin wasu hanyoyin metabolism. Wannan ya ƙara nuna cewa yawan kwayoyin halitta da ke da hannu a metabolism na PPA ya fi yawa a cikin samfuran sarrafawa; waɗannan kwayoyin halitta na iya aiki a cikin hanyoyin da ba sa haifar da amfani da PPA ko samuwar su azaman samfurin esote. A wannan yanayin, kwayar halitta ɗaya kawai da ke da alaƙa da samar da PPA ta nuna bambance-bambance masu mahimmanci a cikin yawa tsakanin nau'ikan samfura. Sabanin kwayoyin halitta da ke da alaƙa da metabolism na PPA, an zaɓi kwayoyin halitta masu alama don samar da PPA saboda suna da hannu kai tsaye a cikin hanyar ƙwayoyin cuta don samar da PPA. A cikin beraye da aka fallasa ga PPA, an gano cewa dukkan nau'ikan halittu suna da ƙaruwar yalwa da ƙarfin samar da PPA sosai. Wannan yana goyan bayan hasashen cewa PPAs za su zaɓi masu samar da PPA don haka suna hasashen cewa ƙarfin samar da PPA zai ƙaru. Duk da haka, yawan kwayoyin halitta ba lallai bane ya yi daidai da bayyanar kwayoyin halitta; don haka, kodayake yawan kwayoyin halitta da ke da alaƙa da metabolism na PPA ya fi girma a cikin samfuran sarrafawa, ƙimar bayyanar na iya bambanta (Shi et al., 2014). Don tabbatar da alaƙar da ke tsakanin yawan kwayoyin halitta masu samar da PPA da samar da PPA, ana buƙatar nazarin bayyanar kwayoyin halitta da ke da hannu a samar da PPA.
Bayanin aiki na PPA da metagenomes na sarrafawa ya bayyana wasu bambance-bambance. Binciken PCA na abubuwan da ke cikin kwayar halitta ya nuna ƙungiyoyi daban-daban tsakanin PPA da samfuran sarrafawa (Hoto na 5). Tsarin tattarawa a cikin samfurin ya nuna cewa abubuwan da ke cikin kwayar halitta na sarrafawa sun fi bambanta, yayin da samfuran PPA suka haɗu. Rarraba ta hanyar abubuwan da ke cikin kwayar halitta yayi daidai da rarraba ta hanyar nau'ikan halittu. Don haka, bambance-bambance a cikin yawan hanyoyin hanya sun yi daidai da canje-canje a cikin yawan takamaiman nau'ikan da nau'ikan da ke cikinsu. A cikin samfuran PPA, hanyoyi biyu masu yawan gaske suna da alaƙa da metabolism na aminosugar/nucleotide sugar (ko:K21279) da hanyoyin metabolism na lipid da yawa (ko:K00647, ko:K03801; Tebur na Ƙarin 3). An san cewa kwayoyin halitta da ke da alaƙa da ko:K21279 suna da alaƙa da halittar Bacteroides, ɗaya daga cikin nau'ikan da ke da yawan nau'ikan da ke cikin samfuran PPA. Wannan enzyme na iya guje wa amsawar garkuwar jiki ta hanyar bayyana polysaccharides na capsular (Wang et al., 2008). Wannan na iya zama sanadin karuwar Bacteroidetes da aka lura a cikin beraye da aka fallasa PPA. Wannan ya cika yawan sinadarin kitse da aka gani a cikin kwayar halittar PPA. Kwayoyin cuta suna amfani da hanyar FASIIko:K00647 (fabB) don samar da kitse mai yawa, wanda zai iya yin tasiri ga hanyoyin metabolism na masu masaukin baki (Yao da Rock, 2015; Johnson et al., 2020), kuma canje-canje a cikin metabolism na lipid na iya taka rawa a cikin ci gaban jijiyoyi (Yu et al., 2020). Wata hanyar da ke nuna karuwar yawan samfuran PPA ita ce biosynthesis na hormone steroid (ko:K12343). Akwai shaida da ke ƙaruwa cewa akwai alaƙar da ke tsakanin ikon ƙwayoyin cuta na hanji don yin tasiri ga matakan hormones da kuma tasirin hormones, ta yadda matakan steroid masu yawa na iya haifar da sakamako mai kyau ga lafiya (Tetel et al., 2018).
Wannan binciken ba shi da iyaka da la'akari. Wani muhimmin bambanci shi ne cewa ba mu yi kimantawar ilimin halittar dabbobi ba. Saboda haka, ba zai yiwu a kammala kai tsaye ko canje-canje a cikin ƙwayoyin cuta suna da alaƙa da kowace cuta ba. Wani abin la'akari kuma shi ne cewa beraye a cikin wannan binciken an ciyar da su abinci iri ɗaya da uwayensu. Nazarin da za a yi nan gaba na iya tantance ko canzawa daga abinci mai wadataccen PPA zuwa abinci mara PPA yana inganta tasirinsa akan ƙwayoyin cuta. Wani iyakance na bincikenmu, kamar sauran mutane da yawa, shine ƙarancin girman samfurin. Kodayake ana iya zana sakamako mai inganci, babban girman samfurin zai samar da ƙarin ƙarfin ƙididdiga lokacin nazarin sakamakon. Hakanan muna yin taka tsantsan game da zana ƙarshe game da alaƙa tsakanin canje-canje a cikin ƙwayoyin cuta na hanji da kowace cuta (Yap et al., 2021). Abubuwan da suka rikice ciki har da shekaru, jinsi, da abinci na iya yin tasiri sosai ga tsarin ƙwayoyin cuta. Waɗannan abubuwan na iya bayyana rashin daidaito da aka lura a cikin wallafe-wallafen game da alaƙar ƙwayoyin cuta na hanji da cututtuka masu rikitarwa (Johnson et al., 2019; Lagod da Naser, 2023). Misali, an nuna cewa membobin dangin Bacteroidetes suna ƙaruwa ko raguwa a cikin dabbobi da mutane masu fama da ASD (Angelis et al., 2013; Kushak et al., 2017). Hakazalika, nazarin da aka yi kan tsarin hanji a cikin marasa lafiya da ke fama da cututtukan hanji mai kumburi ya gano ƙaruwa da raguwa a cikin nau'in taxa ɗaya (Walters et al., 2014; Forbes et al., 2018; Upadhyay et al., 2023). Don iyakance tasirin son zuciya na jinsi, mun yi ƙoƙarin tabbatar da daidaiton wakilcin jinsi don haka bambance-bambancen sun fi faruwa ne ta hanyar abinci. Ɗaya daga cikin ƙalubalen bayanin aiki shine cire jerin kwayoyin halitta masu yawa. Hanyar tattara kwayoyin halittarmu tana buƙatar asalin jerin 95% da kamanceceniya na tsawon 85%, da kuma ɗaukar nauyin daidaitawa 90% don kawar da tarin ƙarya. Duk da haka, a wasu lokuta, mun lura da COGs tare da bayanin iri ɗaya (misali, MUT) (Hoto na 6). Ana buƙatar ƙarin bincike don tantance ko waɗannan orthologs ɗin sun bambanta, suna da alaƙa da takamaiman nau'in halitta, ko kuma ko wannan iyakance ne ga hanyar haɗa kwayoyin halitta. Wani iyakancewar bayanin aiki shine yiwuwar rashin rarraba su; kwayar halittar ƙwayoyin cuta mmdA sanannen enzyme ne da ke da hannu a cikin haɗakar propionate, amma KEGG ba ta haɗa shi da hanyar metabolism na propionate ba. Sabanin haka, orthologs ɗin scpB da mmcD suna da alaƙa. Yawan kwayoyin halitta ba tare da takamaiman knockouts na iya haifar da rashin iya gano kwayoyin halittar da ke da alaƙa da PPA lokacin tantance yawan kwayoyin halitta. Nazarin nan gaba zai amfana daga nazarin metatranscriptome, wanda zai iya samar da zurfin fahimtar halayen aiki na microbiota na hanji da kuma haɗa bayyanar kwayoyin halitta da yuwuwar tasirin da ke ƙasa. Don nazarin da ya shafi takamaiman cututtukan ci gaban jijiyoyi ko cututtukan hanji mai kumburi, ana buƙatar kimantawa na jiki da ɗabi'a na dabbobi don haɗa canje-canje a cikin abun da ke cikin microbiome da waɗannan rikice-rikice. Ƙarin nazarin dasa microbiome na hanji zuwa beraye marasa ƙwayoyin cuta suma zasu zama da amfani don tantance ko microbiome shine abin da ke haifar da cuta.
A taƙaice, mun nuna cewa PPA na abinci yana aiki a matsayin wani abu da ke canza tsarin ƙwayoyin cuta na hanji. PPA wani abu ne da FDA ta amince da shi wanda ke kiyaye abinci iri-iri wanda, bayan an shafe shi na dogon lokaci, zai iya haifar da katsewar ƙwayoyin cuta na yau da kullun. Mun sami canje-canje a cikin yawan ƙwayoyin cuta da yawa, yana nuna cewa PPA na iya yin tasiri ga tsarin ƙwayoyin cuta na hanji. Canje-canje a cikin ƙwayoyin cuta na iya haifar da canje-canje a matakan wasu hanyoyin rayuwa, wanda zai iya haifar da canje-canje a cikin ilimin halittar jiki waɗanda suka dace da lafiyar mai masaukin baki. Ana buƙatar ƙarin bincike don tantance ko tasirin PPA na abinci akan tsarin ƙwayoyin cuta na iya haifar da dysbiosis ko wasu cututtuka. Wannan binciken ya shimfida harsashin karatu na gaba kan yadda tasirin PPA akan tsarin hanji zai iya shafar lafiyar ɗan adam.
Bayanan da aka gabatar a cikin wannan binciken suna samuwa a cikin ma'ajiyar bayanai ta yanar gizo. Sunan ma'ajiyar bayanai da lambar shiga sune: https://www.ncbi.nlm.nih.gov/, PRJNA1092431.
Kwamitin Kula da Dabbobi da Amfani da su na Jami'ar Central Florida (UCF-IACUC) ya amince da wannan binciken dabbobi (Lambar Izinin Amfani da Dabbobi: PROTO202000002). Wannan binciken ya yi daidai da dokoki, ƙa'idodi, da buƙatun hukumomi na gida.
NG: Ra'ayi, Shirya Bayanai, Nazarin Aiki, Bincike, Hanyar Aiki, Software, Ganin Hoto, Rubutu (asali daftarin aiki), Rubutu (bita & gyarawa). LA: Ra'ayi, Shirya Bayanai, Hanyar Aiki, Albarkatu, Rubutu (bita & gyarawa). SH: Bincike na yau da kullun, Software, Rubutu (bita & gyarawa). SA: Bincike, Rubutu (bita & gyarawa). Babban Alkali: Bincike, Rubutu (bita & gyarawa). SN: Ra'ayi, Gudanar da Ayyuka, Albarkatu, Kulawa, Rubutu (bita & gyarawa). TA: Ra'ayi, Gudanar da Ayyuka, Kulawa, Rubutu (bita & gyarawa).
Marubutan sun bayyana cewa ba su sami tallafin kuɗi ba don bincike, marubuta, da/ko buga wannan labarin.
Marubutan sun bayyana cewa an gudanar da binciken ne ba tare da wata alaƙa ta kasuwanci ko ta kuɗi da za a iya fassara ta a matsayin wata matsala ta sha'awa ba.
Duk ra'ayoyin da aka bayyana a cikin wannan labarin na marubuta ne kawai kuma ba lallai bane su nuna ra'ayoyin cibiyoyinsu, masu buga littattafai, editoci, ko masu bita ba. Duk wani samfuri da aka tantance a cikin wannan labarin, ko duk wani ikirari da masana'antun su suka yi, ba a ba da garantin ko amincewa da shi daga mai wallafawa ba.
Ana iya samun ƙarin kayan aikin wannan labarin akan layi: https://www.frontiersin.org/articles/10.3389/frmbi.2024.1451735/full#supplementary-material
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