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Ntsintsi ea lesole le letšo (Hermetia illucens, L. 1758) ke kokoanyana e senyang e nang le bokhoni bo phahameng ba ho sebelisa lihlahisoa tsa tlhaho tse nang le lik'habohaedreite. Har'a lik'habohaedreite, lesole le letšo le fofa le itšetlehile ka tsoekere e qhibilihang bakeng sa kholo le lipid synthesis. Sepheo sa phuputso ena e ne e le ho hlahloba liphello tsa tsoekere e tloaelehileng e qhibilihang ho nts'etsopele, ho phela, le boemo ba asiti e mafura ea lintsintsi tsa masole a batho ba batšo. Eketsa phepo ea likhoho ka monosaccharides le disaccharides ka thoko. Cellulose e ne e sebelisoa e le taolo. Li-larvae tse fepileng tsoekere, fructose, sucrose le maltose li ile tsa hola ka potlako ho feta liboko tse laolang. Ka lehlakoreng le leng, lactose e bile le phello e tsoelang pele ho li-larvae, e fokotsa khōlo le ho fokotsa boima ba 'mele ba ho qetela. Leha ho le joalo, tsoekere eohle e qhibilihang e ile ea nontša li-larvae ho feta tse fepang lijo tsa taolo. Ka ho hlakileng, tsoekere e lekiloeng e ile ea bopa boemo ba acid e mafura. Maltose le sucrose li ekelitse mafura a mafura a mangata ha a bapisoa le selulose. Ka lehlakoreng le leng, lactose e ile ea eketsa bioaccumulation ea dietary unsaturated fatty acids. Thuto ena ke ea pele e bontšang phello ea tsoekere e qhibilihang ka har'a motsoako oa asiti e mafura ea li-larvae tsa lesole le letšo. Liphetho tsa rona li bonts'a hore lik'habohaedreite tse lekiloeng li na le phello e kholo mofuteng oa mafura a mafura a li-larvae tsa masole a matšo 'me ka hona li ka etsa qeto ea ts'ebeliso ea tsona ea ho qetela.
Tlhokahalo ea lefats'e ea matla le protheine ea liphoofolo e ntse e eketseha1. Boemong ba ho futhumala ha lefatše, hoa hlokahala ho fumana mekhoa e meng e tala ho feta matla a fossil le mekhoa ea setso ea tlhahiso ea lijo ha ho ntse ho eketsa tlhahiso. Likokoanyana li tšepisa bonkgetheng ho rarolla mathata ana ka lebaka la ho fokotseha ha tsona ka lik'hemik'hale le tšusumetso ea tikoloho ha li bapisoa le temo ea khale ea liphoofolo2. Har'a likokoanyana, mokhethoa ea hloahloa oa ho rarolla mathata ana ke "black soldier fly" (BSF), Hermetia illucens (L. 1758), mofuta o kotsi o khonang ho iphepa ka mefuta e fapaneng ea li-organic substrates3. Ka hona, ho matlafatsa li-substrates tsena ka ho ikatisa ha BSF ho ka theha mohloli o mocha oa thepa e tala ho fihlela litlhoko tsa liindasteri tse fapaneng.
BSF larvae (BSFL) e ka iphepa ka lihlahisoa tsa temo le tsa indasteri ea temo tse kang lijo-thollo tsa ba ritelo, masalla a meroho, makhasi a litholoana le bohobe ba khale, tse loketseng kholo ea BSFL ka lebaka la lik'habohaedreite tse ngata (CH)4,5, 6 litaba. Tlhahiso e kholo ea BSFL e fella ka ho thehoa ha lihlahisoa tse peli: mantle, motsoako oa masala a substrate le mantle a ka sebelisoang e le manyolo bakeng sa temo ea limela7, le li-larvae, tse entsoeng haholo-holo ka liprotheine, lipids le chitin. Liprotheine le lipids li sebelisoa haholo temong ea liphoofolo, biofuel le litlolo8,9. Ha e le chitin, biopolymer ena e fumana lits'ebetso lekaleng la agri-food, biotechnology le tlhokomelo ea bophelo bo botle10.
BSF ke kokoanyana ea tlhaho ea holometabolous, ho bolelang hore metamorphosis ea eona le ho ikatisa, haholo-holo mekhahlelo e jang matla ea nako ea bophelo ba kokoanyana, e ka tšehetsoa ka ho feletseng ke mehloli ea limatlafatsi e hlahisoang nakong ea kholo ea leruane11. Haholo-holo, protheine le lipid synthesis li lebisa ho nts'etsopele ea 'mele oa mafura, setho sa bohlokoa sa polokelo se ntšang matla nakong ea mekha ea ho se fepe ea BSF: prepupa (ke hore, mohato oa ho qetela oa larvae nakong eo li-larvae tsa BSF li fetohang batsho ha li ntse li fepa le ho batla. bakeng sa tikoloho e loketseng metamorphosis), li-pupae (ke hore, mohato o sa sisinyeheng oo kokoanyana e bang le metamorphosis), le batho ba baholo12,13. CH ke mohloli o ka sehloohong oa matla lijong tsa BSF14. Har'a limatlafatsi tsena, fibrous CH e kang hemicellulose, cellulose le lignin, ho fapana le li-disaccharides le polysaccharides (tse kang starch), li ke ke tsa cheka ke BSFL15,16. Tšimoloho ea CH ke mohato oa bohlokoa oa pele oa ho monya lik'habohaedreite, tseo qetellong li nang le hydrolyzed ho tsoekere e bonolo ka maleng16. Tsoekere e bonolo joale e ka monngoa (ke hore, ka lera la mala a mala) ebe e sebelisoa ho hlahisa matla17. Joalokaha ho boletsoe ka holimo, li-larvae li boloka matla a feteletseng joaloka lipids 'meleng oa mafura12,18. Lipids tsa polokelo li na le li-triglycerides (li-lipids tse sa nke lehlakore tse entsoeng ka molek'hule e le 'ngoe ea glycerol le li-acid tse tharo tse mafura) tse entsoeng ke li-larvae tse tsoang ho tsoekere e bonolo ea lijo. Li-CH tsena li fana ka li-substrates tsa acetyl-CoA tse hlokahalang bakeng sa biosynthesis ea mafura a acid (FA) ka mafura a acid synthase le litsela tsa thioesterase19. Mafura a mafura a H. illucens lipids ka tlhaho a laoloa ke saturated fatty acids (SFA) e nang le karolo e phahameng ea lauric acid (C12: 0) 19,20. Ka hona, litaba tse phahameng tsa lipid le sebopeho sa asiti e mafura li fetoha lintlha tse fokotsang ts'ebeliso ea li-larvae kaofela lijong tsa liphoofolo, haholo-holo temong ea metsing moo ho hlokahalang polyunsaturated fatty acids (PUFA)21.
Ho tloha ha ho sibolloa bokhoni ba BSFL ba ho fokotsa litšila tsa tlhaho, liphuputso tse mabapi le boleng ba lihlahisoa tse fapa-fapaneng li bontšitse hore sebopeho sa BSFL se laoloa ka mokhoa o itseng ke lijo tsa eona. Hajoale, taolo ea profil ea FA ea H. illucens e ntse e ntlafala. Bokhoni ba BSFL ho bioaccumulate PUFA bo bontšitsoe ka li-substrates tse ruileng tsa PUFA tse kang algae, litšila tsa tlhapi, kapa lijo tse kang li-flaxseed, tse fanang ka boemo bo phahameng ba FA profil ea phepo ea liphoofolo19,22,23. Ka lehlakoreng le leng, bakeng sa lihlahisoa tse sa ntlafatsoang ho PUFA, ha ho kamehla ho nang le kamano pakeng tsa litlaleho tsa FA tsa lijo le FA larval, e bontšang tšusumetso ea limatlafatsi tse ling24,25. Haele hantle, phello ea CH e sa tsitsang ho li-profaele tsa FA e lula e sa utloisisoe hantle ebile e sa batlisisoe ka tlase24,25,26,27.
Ho ea ka tsebo ea rona, le hoja li-monosaccharides le li-disaccharides tse feletseng li le ngata lijong tsa H. illucens, karolo ea bona ea phepo e ntse e sa utloisisoe hantle ho H. illucens phepo e nepahetseng. Sepheo sa thuto ena e ne e le ho hlakisa litlamorao tsa bona ho phepo ea BSFL le sebopeho sa lipid. Re tla hlahloba kholo, ho phela, le tlhahiso ea li-larvae tlas'a maemo a fapaneng a phepo e nepahetseng. Joale, re tla hlalosa litaba tsa lipid le profil ea acid e mafura ea lijo ka 'ngoe ho totobatsa litlamorao tsa CH ho boleng ba phepo ea BSFL.
Re ile ra nahana hore mofuta oa CH o lekiloeng o tla ama (1) kholo ea larval, (2) maemo a lipid kaofela, le (3) ho fetola boemo ba FA. Li-monosaccharides li ka monngoa ka ho toba, athe li-disaccharides tsona li tlameha ho kenngoa ka hydrolyzed. Ka hona, li-monosaccharides li fumaneha haholoanyane e le mehloli e tobileng ea matla kapa li-precursors tsa lipogenesis ka FA synthase le thioesterase pathways, ka tsela eo li matlafatsa H. illucens kgolo ea larval le ho khothalletsa ho bokellana ha lipids (haholo-holo lauric acid).
CH e lekiloeng e ile ea ama boima ba 'mele bo tloaelehileng ba li-larvae nakong ea khōlo (setšoantšo sa 1). FRU, GLU, SUC le MAL li ile tsa eketsa boima ba 'mele oa larval ka mokhoa o tšoanang le oa ho laola lijo (CEL). Ka lehlakoreng le leng, LAC le GAL li ne li bonahala li sitisa tsoelo-pele ea li-larval. Ka ho hlakileng, LAC e ne e e-na le phello e mpe e kholo ho kholo ea larval ha e bapisoa le SUC ho pholletsa le nako ea kgolo: 9.16 ± 1.10 mg khahlanong le 15.00 ± 1.01 mg ka letsatsi la 3 (F6,21 = 12.77, p <0.001; Setšoantšo sa 1), 125.12 ± 125.11 mg mg le 211.79 ± 14.93 mg, ka ho latellana, ka letsatsi la 17 (F6,21 = 38.57, p <0.001; Setšoantšo sa 1).
Ho sebelisa li-monosaccharides tse fapaneng (fructose (FRU), galactose (GAL), tsoekere (GLU)), disaccharides (lactose (LAC), maltose (MAL), sucrose (SUC)) le cellulose (CEL) e le taolo. Ho hola ha li-larvae tse feptjoang ke lesole le letšo. Ntlha e 'ngoe le e 'ngoe e mothinya e emela boima ba motho ka mong (mg) bo baloang ka boima ba li-larvae tse 20 tse khethiloeng ka mokhoa o sa reroang ho tsoa ho baahi ba 100 larvae (n = 4). Li-barbar tsa liphoso li emela SD.
Lijo tsa CEL li fane ka bophelo bo botle ba li-larval tsa 95.5 ± 3.8%. Ho feta moo, ho phela ha H. illucens ho fepa lijo tse nang le CH e qhibilihang ho fokotsoe (GLM: χ = 107.13, df = 21, p <0.001), e neng e bakoa ke MAL le SUC (disaccharides) ho CH e ithutoang. Lefu le ne le le tlase ho feta la GLU, FRU, GAL (monosaccharide), le LAC (EMM: p <0.001, Setšoantšo sa 2).
Lebokose la ho pholoha ha li-larvae tse fofang tsa masole a batšo tse tšoaroang ka li-monosaccharides tse fapaneng (fructose, galactose, glucose), li-disaccharides (lactose, maltose, sucrose) le selulose e le taolo. Liphekolo tse nang le lengolo le tšoanang ha li fapane haholo ho tse ling (EMM, p> 0.05).
Lijo tsohle tse lekiloeng li ile tsa lumella li-larvae hore li fihle boemong ba prepupal. Leha ho le joalo, li-CH tse lekiloeng li ne li atisa ho lelefatsa tsoelo-pele ea larval (F6,21 = 9.60, p<0.001; Lethathamo la 1). Haholo-holo, li-larvae tse fepa GAL le LAC li ile tsa nka nako e telele ho fihla sethaleng sa prepupal ha li bapisoa le li-larvae tse hōlisitsoeng ho CEL (CEL-GAL: p<0.001; CEL-LAC: p<0.001; Letlapa la 1).
CH e lekiloeng e ile ea boela ea e-ba le liphello tse fapaneng ho boima ba 'mele oa larva, ka boima ba' mele ba li-larvae bo fepa lijo tsa CEL ho fihla ho 180.19 ± 11.35 mg (F6,21 = 16.86, p <0.001; Setšoantšo sa 3). FRU, GLU, MAL le SUC li hlahisitse boima ba 'mele oa ho qetela oa larval oa ho feta 200 mg, o neng o phahame haholo ho feta oa CEL (p <0.05). Ka lehlakoreng le leng, li-larvae tse fepa GAL le LAC li ne li e-na le boima ba 'mele bo tlaase, ka karolelano 177.64 ± 4.23 mg le 156.30 ± 2.59 mg, ka ho latellana (p <0.05). Phello ena e ne e bonahala haholo ka LAC, moo boima ba 'mele ba ho qetela bo neng bo le tlase ho feta lijo tsa taolo (CEL-LAC: phapang = 23.89 mg; p = 0.03; Setšoantšo sa 3).
Bolela boima ba ho qetela ba li-larvae ka bomong tse hlahisoang e le matheba a larval (mg) le lintsintsi tse ntšo tsa sesole tse hlalositsoeng e le histogram (g) li fepa monosaccharides tse fapaneng (fructose, galactose, glucose), disaccharides (lactose, maltose, sucrose) le selulose (e le taolo). Litlhaku tsa li-columnar li emela lihlopha tse fapaneng haholo ka boima ba 'mele oa larval (p <0.001). Litlhaku tse amanang le matheba a li-larval li emela lihlopha tse nang le boima ba 'mele bo fapaneng haholo (p <0.001). Li-barbar tsa liphoso li emela SD.
Boima ba motho ka mong bo ne bo ikemetse ho boima ba ho qetela ba kolone ea larval. Ha e le hantle, lijo tse nang le FRU, GLU, MAL, le SUC ha lia ka tsa eketsa boima ba li-larval bo hlahisoang ka tanka ha bo bapisoa le CEL (Setšoantšo sa 3). Leha ho le joalo, LAC e theohile haholo boima ba boima ba 'mele (CEL-LAC: phapang = 9.14 g; p <0.001; Setšoantšo sa 3).
Letlapa la 1 le bonts'a chai (li-larvae/letsatsi). Hoa thahasellisa hore lihlahisoa tse nepahetseng tsa CEL, MAL le SUC li ne li tšoana (Letlapa la 1). Ka lehlakoreng le leng, FRU, GAL, GLU le LAC li fokolitse lihlahisoa ha li bapisoa le CEL (Letlapa la 1). GAL le LAC li entse hampe ka ho fetisisa: chai e ne e le halofo ho ea ho 0.51 ± 0.09 g larvae feela / letsatsi le 0.48 ± 0.06 g larvae / letsatsi, ka ho latellana (Letlapa la 1).
Li-monosaccharides le li-disaccharides li ile tsa eketsa litaba tsa lipid tsa CF larvae (Letlapa la 1). Lijong tsa CLE, ho ile ha fumanoa li-larvae tse nang le lipid content ea 23.19 ± 0.70% ea litaba tsa DM. Ha ho bapisoa, karolelano ea lipid ka har'a li-larvae tse feptjoang ka tsoekere e qhibilihang e ne e feta 30% (Letlapa la 1). Leha ho le joalo, li-CH tse lekiloeng li ile tsa eketsa mafura a tsona ka tekanyo e tšoanang.
Joalokaha ho lebeletsoe, lihlooho tsa CG li ile tsa ama boemo ba FA ea li-larvae ho ea ka likhato tse sa tšoaneng (setšoantšo sa 4). Likahare tsa SFA li ne li phahame lijong tsohle 'me li fihlile ho feta 60%. MAL le SUC li ne li sa leka-lekane boemo ba FA, e leng se lebisitseng ho eketseha ha litaba tsa SFA. Tabeng ea MAL, ka lehlakoreng le leng, ho se leka-lekane hona ho ile ha lebisa haholo-holo ho fokotseha ha litaba tsa monounsaturated fatty acids (MUFA) (F6,21 = 7.47; p <0.001; Setšoantšo sa 4). Ka lehlakoreng le leng, bakeng sa SUC, ho fokotseha ho ne ho tšoana haholo pakeng tsa MUFA le PUFA. LAC le MAL li bile le litlamorao tse fapaneng ho sebono sa FA (SFA: F6,21 = 8.74; p <0.001; MUFA: F6,21 = 7.47; p <0.001; PUFA: χ2 = 19.60; Df = 6; p <0.001; Setšoantšo 4). Karolo e tlase ea SFA ho larvae e feptjoang ke LAC e bonahala e eketsa litaba tsa MUFA. Haholo-holo, maemo a MUFA a ne a phahame ho li-larvae tse feptjoang ka LAC ha li bapisoa le tsoekere e meng e qhibilihang ntle le GAL (F6,21 = 7.47; p <0.001; Setšoantšo sa 4).
Ho sebelisa li-monosaccharides tse fapaneng (fructose (FRU), galactose (GAL), tsoekere (GLU)), disaccharides (lactose (LAC), maltose (MAL), sucrose (SUC)) le cellulose (CEL) e le taolo, lebokose la morero oa mafura a acid. sebopeho se feptjoang ke lesole le letšo le fofang larvae. Liphetho li hlahisoa e le liperesente tsa kakaretso ea FAME. Liphekolo tse tšoailoeng ka litlhaku tse fapaneng li fapane haholo (p <0.001). (a) Palo ea li- saturated fatty acids; (b) Li-monounsaturated fatty acids; (c) Polyunsaturated fatty acids.
Har'a li-fatty acids tse khethiloeng, lauric acid (C12: 0) e ne e le eona e ka sehloohong ho bohle ba boneng (ho feta 40%). Li-SFA tse ling tse teng e ne e le palmitic acid (C16: 0) (tlase ho 10%), stearic acid (C18: 0) (tlase ho 2.5%) le capric acid (C10: 0) (tlase ho 1.5%). Li-MUFA li ne li emeloa haholo ke oleic acid (C18: 1n9) (tlase ho 9.5%), ha PUFA e ne e entsoe haholo ka linoleic acid (C18: 2n6) (tlase ho 13.0%) (bona Supplementary Table S1). Ho phaella moo, karolo e nyenyane ea metsoako e ne e ke ke ea tsejoa, haholo-holo ka har'a li-larvae tsa CEL, moo palo e sa tsejoeng ea motsoako oa 9 (UND9) e neng e le ka karolelano ea 2.46 ± 0.52% (sheba Supplementary Table S1). Tlhahlobo ea GC × GC-FID e bontšitse hore e ka 'na ea e-ba 20-carbon fatty acid e nang le li-bond tse peli tse hlano kapa tse tšeletseng (sheba Supplementary Figure S5).
Tlhahlobo ea PERMANOVA e senotse lihlopha tse tharo tse fapaneng tse thehiloeng ho mafura a mafura a mafura (F6,21 = 7.79, p <0.001; Setšoantšo sa 5). Principal component analysis (PCA) ea TBC spectrum e bontša sena 'me e hlalosoa ke likarolo tse peli (Setšoantšo sa 5). Likarolo tse ka sehloohong li hlalositse 57.9% ea phapang 'me li kenyelelitse, ka tatellano ea bohlokoa, lauric acid (C12: 0), oleic acid (C18: 1n9), palmitic acid (C16: 0), stearic acid (C18: 0), le linolenic acid (C18: 3n3) (sheba Setšoantšo sa S4). Karolo ea bobeli e hlalositse 26.3% ea phapang mme e kenyelletsa, ka tatellano ea bohlokoa, decanoic acid (C10: 0) le linoleic acid (C18: 2n6 cis) (bona Supplementary Figure S4). Litlaleho tsa lijo tse nang le tsoekere e bonolo (FRU, GAL le GLU) li bontšitse litšobotsi tse tšoanang. Ka lehlakoreng le leng, li-disaccharides li ile tsa fana ka lintlha tse fapaneng: MAL le SUC ka lehlakoreng le leng le LAC ka ho le leng. Haholo-holo, MAL e ne e le eona feela tsoekere e fetotseng profil ea FA ha e bapisoa le CEL. Ntle le moo, boemo ba MAL bo ne bo fapane haholo le ba FRU le GLU. Haholo-holo, boemo ba MAL bo bontšitse karolo e phahameng ka ho fetisisa ea C12: 0 (54.59 ± 2.17%), e etsa hore e bapise le CEL (43.10 ± 5.01%), LAC (43.35 ± 1.31%), FRU (48.90 ± 1.97%) le GLU (48.38 ± 2.17%) profiles (bona Tlatsetso Lethathamo la S1). MAL spectrum e boetse e bonts'a C18: 1n9 e tlaase ka ho fetisisa (9.52 ± 0.50%), e ileng ea e arola hape ho LAC (12.86 ± 0.52%) le CEL (12.40 ± 1.31%) spectra. Mokhoa o tšoanang o ile oa hlokomeloa bakeng sa C16: 0. Karolong ea bobeli, sekhahla sa LAC se bonts'itse litaba tse phahameng ka ho fetisisa tsa C18: 2n6 (17.22 ± 0.46%), ha MAL e bonts'a e tlase (12.58 ± 0.67%). C18: 2n6 e boetse e khetholla LAC ho tloha taolong (CEL), e bontšitseng maemo a tlaase (13.41 ± 2.48%) (sheba Supplementary Table S1).
PCA morero oa profil ea mafura a mafura a lesole le letšo le fofa larvae le li-monosaccharides tse fapaneng (fructose, galactose, glucose), li-disaccharides (lactose, maltose, sucrose) le selulose e le taolo.
Ho ithuta litlamorao tsa phepo ea tsoekere e qhibilihang ho H. illucens larvae, cellulose (CEL) ho fepa ea likhoho e ile ea nkeloa sebaka ke tsoekere (GLU), fructose (FRU), galactose (GAL), maltose (MAL), sucrose (SUC), le lactose (LAC). Leha ho le joalo, li-monosaccharides le li-disaccharides li bile le litlamorao tse fapaneng ho nts'etsopele, ho phela, le sebopeho sa li-larvae tsa HF. Ka mohlala, GLU, FRU, le mefuta ea bona ea li-disaccharide (MAL le SUC) li bile le liphello tse ntle tse tšehetsang khōlo ea larval, e ba lumellang ho finyella boima ba 'mele bo phahameng ho feta CEL. Ho fapana le indigestible CEL, GLU, FRU, le SUC li ka feta mokoallo oa mala 'me tsa sebetsa e le mehloli ea bohlokoa ea limatlafatsi lijong tse entsoeng16,28. MAL e haelloa ke lipalangoang tse khethehileng tsa liphoofolo 'me ho nahanoa hore e na le hydrolyzed ho limolek'hule tse peli tsa tsoekere pele ho assimilation15. Limolek'hule tsena li bolokoa 'meleng oa likokoanyana e le mohloli o tobileng oa matla kapa e le lipids18. Ntlha ea pele, mabapi le ho qetela, tse ling tsa liphapang tsa intramodal tse hlokometsoeng li ka 'na tsa e-ba ka lebaka la phapang e nyenyane ea likarohano tsa thobalano. Ka 'nete, ho H. illucens, ho ikatisa ho ka 'na ha itlela feela: tse tšehali tse hōlileng ka tlhaho li na le mehloli e lekaneng ea ho behela mahe 'me li boima ho feta tse tona29. Leha ho le joalo, ho bokellana ha lipid ho BSFL ho tsamaisana le ho kenngoa ha CH2 ka har'a lijo, joalo ka ha ho hlokometsoe pele bakeng sa GLU le xylose26,30. Ka mohlala, Li et al.30 o hlokometse hore ha 8% GLU e eketsoa lijong tsa larval, lihlahisoa tsa lipid tsa BSF larvae li eketsehile ka 7.78% ha li bapisoa le taolo. Liphetho tsa rona li lumellana le litlhaloso tsena, tse bontšang hore mafura a mafura a li-larvae a fepa tsoekere e qhibilihang a ne a phahame ho feta a li-larvae a fepa lijo tsa CEL, ha a bapisoa le keketseho ea 8.57% le GLU supplementation. Ho makatsang ke hore liphello tse tšoanang li ile tsa hlokomeloa ho li-larvae tse feptjoang GAL le LAC, ho sa tsotellehe liphello tse mpe tsa kholo ea li-larva, boima ba 'mele oa ho qetela le ho phela. Li-larvae tse feptjoang ka LAC li ne li le nyane haholo ho feta tse feptjoang ka lijo tsa CEL, empa mafura a tsona a ne a tšoana le libolu tse fepa tsoekere e 'ngoe e qhibilihang. Liphetho tsena li totobatsa litlamorao tsa lactose ho BSFL. Ntlha ea pele, lijo li na le palo e kholo ea CH. Sistimi ea ho monya le ea hydrolysis ea li-monosaccharides le li-disaccharides, ka ho latellana, e ka fihla ho tlala, e bakang mathata ts'ebetsong ea assimilation. Ha e le hydrolysis, e etsoa ke α- le β-glucosidases 31 . Li-enzyme tsena li khethile li-substrates ho itšetlehile ka boholo ba tsona le lik'hemik'hale tsa lik'hemik'hale (α kapa β linkages) pakeng tsa li-monosaccharides tsa tsona tse 15. Hydrolysis ea LAC ho GLU le GAL e etsoa ke β-galactosidase, enzyme eo mosebetsi oa eona o bontšitsoeng ka maleng a BSF 32. Leha ho le joalo, polelo ea eona e kanna ea se lekane ha e bapisoa le palo ea LAC e jeoang ke li-larvae. Ka lehlakoreng le leng, α-glucosidase maltase le sucrase 15, tse tsejoang ka hore li hlahisoa haholo ke likokoanyana, li khona ho senya boholo ba MAL le sucrose SUC, kahoo li fokotsa phello ena e khotsofatsang. Taba ea bobeli, litlamorao tsa ts'ebetso e ka ba ka lebaka la phokotso ea ts'usumetso ea ts'ebetso ea amylase ea mala a likokoanyana le ho lieha ha boitšoaro ba ho fepa ha ho bapisoa le liphekolo tse ling. Ha e le hantle, tsoekere e qhibilihang e 'nile ea tsejoa e le li-stimulators tsa mosebetsi oa enzyme ea bohlokoa bakeng sa tšilo ea likokoanyana, tse kang amylase,' me e le lisosa tsa karabo ea phepo33,34,35. Tekanyo ea ho hlasimolla e fapana ho latela sebopeho sa limolek'hule tsa tsoekere. Ha e le hantle, li-disaccharides li hloka hydrolysis pele li kenngoa 'me li atisa ho susumetsa amylase ho feta monosaccharides ea tsona34. Ka lehlakoreng le leng, LAC e na le phello e bobebe 'me e fumanoe e sa khone ho tšehetsa kholo ea likokoanyana mefuteng e fapaneng33,35. Mohlala, sesenying sa Sodoptera exigua (Boddie 1850), ha ho ts'ebetso ea hydrolytic ea LAC e fumanoeng ho li-extracts tsa li-enzyme tsa popane36.
Mabapi le sekhahla sa FA, liphetho tsa rona li bonts'a litlamorao tse kholo tsa CH e lekiloeng. Ka ho hlakileng, le hoja lauric acid (C12: 0) e ne e le ka tlaase ho 1% ea kakaretso ea FA lijong, e ne e laola litlalehong tsohle (sheba Supplementary Table S1). Sena se lumellana le data e fetileng ea hore lauric acid e entsoe ho tloha ho lijo tsa CH ho H. illucens ka tsela e amanang le acetyl-CoA carboxylase le FA synthase19,27,37. Liphetho tsa rona li tiisa hore CEL ha e khone ho jeoa haholo 'me e sebetsa e le "motlakase" lijong tsa taolo ea BSF, joalo ka ha ho tšohloa lithutong tse' maloa tsa BSFL38,39,40. Ho fetola CEL ka li-monosaccharides le li-disaccharides tse ling ntle le LAC ho ile ha eketsa karolelano ea C12: 0, e bontšang ho eketseha ha CH ka li-larvae. Ho khahlisang, li-disaccharides MAL le SUC li khothaletsa motsoako oa lauric acid ka mokhoa o atlehileng ho feta monosaccharides ea tsona, e fana ka maikutlo a hore leha ho na le boemo bo phahameng ba polymerization ea GLU le FRU, mme kaha Drosophila ke eona feela sepalangoang sa sucrose se fumanoeng mefuteng ea liprotheine ea liphoofolo, ba tsamaisang li-disaccharide. e kanna ea se be teng maleng a H. illucens larvae15, tšebeliso ea GLU le FRU lia eketseha. Leha ho le joalo, leha GLU le FRU li sebelisoa habonolo ke BSF, li boetse li ts'oaroa habonolo ke li-substrates le li-microorganisms tsa mala, tse ka bakang ho senyeha ha tsona ka potlako le ho fokotseha ha tšebeliso ea li-larvae ha li bapisoa le li-disaccharides.
Ha u habanya feela, litaba tsa lipid tsa li-larvae tse feptjoang ka LAC le MAL li ne li bapisoa, li bonts'a bioavailability e tšoanang ea tsoekere ena. Leha ho le joalo, ho makatsang ke hore boemo ba FA ba LAC bo ne bo ruile ho SFA, haholo-holo ka litaba tse tlaase tsa C12: 0, ha li bapisoa le MAL. Khopolo e 'ngoe ea ho hlalosa phapang ena ke hore LAC e ka susumetsa bioaccumulation ea lijo tsa FA ka acetyl-CoA FA synthase. Ho tšehetsa khopolo-taba ena, li-larvae tsa LAC li ne li e-na le tekanyo e tlaase ka ho fetisisa ea decanoate (C10: 0) (0.77 ± 0.13%) ho feta lijo tsa CEL (1.27 ± 0.16%), tse bontšang ho fokotseha ha FA synthase le mesebetsi ea thioesterase19. Ea bobeli, mafura a mafura a lijo a nkoa e le eona ntho e ka sehloohong e susumetsang sebopeho sa SFA sa H. illucens27. Litekong tsa rona, linoleic acid (C18: 2n6) e ne e le 54.81% ea mafura a mafura a lijong, 'me karolo ea LAC larvae e le 17.22 ± 0.46% le MAL 12.58 ± 0.67%. Oleic acid (cis + trans C18: 1n9) (23.22% lijong) e bontšitse mokhoa o tšoanang. Karolelano ea α-linolenic acid (C18: 3n3) e boetse e tšehetsa khopolo ea bioaccumulation. Asiti ena e mafura e tsebahala ka ho ipokellela ho BSFL holim'a matlafatso ea substrate, joalo ka ho eketsoa ha kuku ea flaxseed, ho fihla ho 6-9% ea kakaretso ea mafura a acid ka har'a larvae19. Lijong tse matlafatsang, C18: 3n3 e ka ikarabella ho fihlela ho 35% ea kakaretso ea mafura a mafura a lijo. Leha ho le joalo, thutong ea rona, C18: 3n3 e ne e le 2.51% feela ea profil ea mafura a mafura. Le hoja karolo e fumanoang tlhahong e ne e le tlaase ho li-larvae tsa rona, karolo ena e ne e le holimo ho li-larvae tsa LAC (0.87 ± 0.02%) ho feta MAL (0.49 ± 0.04%) (p <0.001; sheba Tlatsetso Letlapa la S1). Lijo tsa CEL li ne li e-na le karolo e bohareng ea 0.72 ± 0.18%. Qetellong, karo-karolelano ea palmitic acid (C16: 0) ho larvae ea CF e bonahatsa tlatsetso ea litsela tsa maiketsetso le lijo tsa FA19. Hoc et al. 19 e hlokometse hore C16: 0 synthesis e fokotsehile ha lijo li ntse li ntlafatsoa ka lijo tsa flaxseed, tse neng li bakoa ke ho fokotseha ha ho fumaneha ha substrate ea acetyl-CoA ka lebaka la ho fokotseha ha karo-karolelano ea CH. Ho makatsang ke hore le hoja lijo tseo ka bobeli li ne li e-na le CH e tšoanang le MAL e bontšitse bioavailability e phahameng, MAL larvae e bontšitse tekanyo e tlaase ka ho fetisisa ea C16: 0 (10.46 ± 0.77%), athe LAC e bontšitse karolo e phahameng, e ikarabellang bakeng sa 12.85 ± 0.27% (p <0.05; bona; bona Tlatsetso Tafole S1). Liphetho tsena li totobatsa tšusumetso e rarahaneng ea limatlafatsi ho tšilo ea lijo ea BSFL le metabolism. Hajoale, lipatlisiso mabapi le taba ena li entsoe ka botlalo ho Lepidoptera ho feta ho Diptera. Ho popane, LAC e ile ea khetholloa e le mokhoa o fokolang oa ho fepa ha o bapisoa le tsoekere e meng e qhibilihang e kang SUC le FRU34,35. Haholo-holo, ho Spodopteralittoralis (Boisduval 1833), tšebeliso ea MAL e ile ea susumetsa mosebetsi oa amylolytic ka maleng ka tekanyo e kholo ho feta LAC34. Liphello tse tšoanang ho BSFL li ka hlalosa ts'ebetso e ntlafetseng ea C12: tsela ea maiketsetso ea 0 ho li-larvae tsa MAL, tse amanang le ho eketseha ha mala a CH, ho fepa nako e telele, le ketso ea mala amylase. Ho fokotseha ho fokolang ha morethetho oa phepo ka pel'a LAC ho ka boela ha hlalosa kholo e liehang ea li-larvae tsa LAC. Ho feta moo, Liu Yanxia et al. 27 e hlokometse hore nako ea bophelo ba lipids ho H. illucens substrates e ne e le telele ho feta ea CH. Ka hona, li-larvae tsa LAC li ka itšetleha haholo ka lipids tsa lijo ho phethela nts'etsopele ea tsona, tse ka eketsang litaba tsa tsona tsa ho qetela tsa lipid le ho fetola boemo ba tsona ba mafura a mafura.
Ho latela tsebo ea rona, ke liphuputso tse 'maloa feela tse lekileng litlamorao tsa tlatsetso ea monosaccharide le disaccharide lijong tsa BSF liprofinseng tsa bona tsa FA. Ea pele, Li et al. 30 e ile ea lekola litlamorao tsa GLU le xylose mme ea bona maemo a lipid a ts'oanang le a rona ka sekhahla sa tlatsetso ea 8%. Boemo ba FA bo ne bo sa qaqisoe mme bo ne bo entsoe haholo ka SFA, empa ha ho liphapang tse ileng tsa fumanoa lipakeng tsa tsoekere tse peli kapa ha li hlahisoa ka nako e le 'ngoe30. Ho feta moo, Cohn et al. 41 ha ea ka ea bontša phello ea 20% GLU, SUC, FRU le GAL tlatsetso ho fepa ea likhoho ho li-profiles tse fapaneng tsa FA. Lipontšo tsena li nkiloe ho tsoa ho litsebi tsa theknoloji ho e-na le likopi tsa likokoana-hloko, tseo, joalokaha li hlalositsoe ke bangoli, li ka fokotsang tlhahlobo ea lipalo-palo. Ho feta moo, ho haella ha taolo ea iso-sugar (ho sebelisa CEL) ho fokotsa tlhaloso ea liphetho. Haufinyane tjena, lithuto tse peli tsa Nugroho RA et al. e bontšitse liphapang ho FA spectra42,43. Thutong ea pele, Nugroho RA et al. 43 e ile ea leka phello ea ho kenyelletsa FRU lijong tsa kernel ea palema e lomositsoeng. Profaele ea FA ea li-larvae tse hlahisitsoeng li bonts'itse maemo a phahameng ka mokhoa o sa tloaelehang a PUFA, ho feta 90% ea eona e nkiloeng lijong tse nang le 10% FRU (e ts'oanang le thuto ea rona). Le hoja lijo tsena li ne li e-na le li-pellets tsa litlhapi tse ruileng tsa PUFA, litekanyetso tse tlalehiloeng tsa FA tsa li-larvae lijong tsa taolo tse nang le 100% PCM e lomositsoeng li ne li sa lumellane le boemo leha e le bofe bo tlalehiloeng pele, haholo-holo boemo bo sa tloaelehang ba C18: 3n3 ea 17.77 ± 1.67% le 26.08 ± 0.20% bakeng sa conjugated linoleic acid (C18: 2n6t), isomer e sa tloaelehang ea linoleic acid. Boithuto ba bobeli bo bontšitse liphetho tse ts'oanang ho kenyelletsa FRU, GLU, MAL le SUC42 lijong tsa kernel ea palema e lomositsoeng. Lithuto tsena, joalo ka tsa rona, li totobatsa mathata a tebileng a ho bapisa liphetho tsa liteko tsa lijo tsa larval tsa BSF, joalo ka khetho ea taolo, tšebelisano le mehloli e meng ea limatlafatsi, le mekhoa ea tlhahlobo ea FA.
Nakong ea liteko, re hlokometse hore 'mala le monko oa substrate li fapana ho latela lijo tse sebelisoang. Sena se fana ka maikutlo a hore likokoana-hloko li ka bapala karolo ho liphello tse hlokometsoeng ka substrate le tsamaiso ea tšilo ea lijo ea li-larvae. Ha e le hantle, li-monosaccharides le li-disaccharides li senyeha habonolo ka ho bokella likokoana-hloko. Tšebeliso e potlakileng ea tsoekere e qhibilihang ke likokoana-hloko e ka etsa hore ho lokolloe lihlahisoa tse ngata tsa microbial metabolism tse kang ethanol, lactic acid, short-chain fatty acids (mohlala, acetic acid, propionic acid, butyric acid) le carbon dioxide44. E meng ea metsoako ena e ka 'na ea ikarabella bakeng sa liphello tse kotsi tse chefo ho larvae hape e hlokometsoeng ke Cohn et al.41 tlas'a maemo a tšoanang a tsoelo-pele. Mohlala, ethanol e kotsi ho likokoanyana45. Mosi o mongata oa carbon dioxide o ka ’na oa fella ka ho bokellana ha oona ka tlaase ho tanka, e leng se ka ’nang sa tima sepakapaka sa oksijene haeba potoloho ea moea e sa lumelle hore e lokolloe. Mabapi le li-SCFA, liphello tsa tsona ho likokoanyana, haholo-holo H. illucens, li lula li sa utloisisoe hantle, le hoja lactic acid, propionic acid, le butyric acid e bontšitsoe hore e bolaea Callosobruchus maculatus (Fabricius 1775)46. Ho Drosophila melanogaster Meigen 1830, li-SCFA tsena ke matšoao a monko a tataisang basali ho ea libakeng tsa oviposition, tse fanang ka maikutlo a karolo e molemo ho nts'etsopele ea larval47. Leha ho le joalo, asiti ea acetic e khetholloa e le ntho e kotsi 'me e ka thibela haholo tsoelo-pele ea larval47. Ka lehlakoreng le leng, lactate e nkiloeng ka likokoana-hloko e sa tsoa fumanoa e na le ts'ireletso khahlanong le likokoana-hloko tse hlaselang ka maleng ho Drosophila48. Ho feta moo, likokoana-hloko tse tsamaisong ea tšilo ea lijo li boetse li phetha karolo ea CH tshilo ea likokoanyana49. Liphello tsa physiological tsa SCFAs ka gut microbiota, tse kang tekanyo ea ho fepa le liphatsa tsa lefutso, li 'nile tsa hlalosoa ka li-vertebrates 50. Li ka boela tsa ba le phello ea trophic ho li-larvae tsa H. illucens 'me li ka kenya letsoho ka mokhoa o itseng taolong ea litlaleho tsa FA. Liphuputso tse mabapi le liphello tsa phepo ea lihlahisoa tsena tsa likokoana-hloko li tla hlakisa liphello tsa tsona ho H. illucens phepo e nepahetseng le ho fana ka motheo oa lithuto tsa nakong e tlang mabapi le likokoana-hloko tse molemo kapa tse kotsi ho latela tsoelo-pele ea tsona le boleng ba li-substrates tse ruileng tsa FA. Tabeng ena, karolo ea likokoana-hloko ka har'a ts'ebetso ea tšilo ea lijo ea likokoanyana tse ruuoang ka bongata e ntse e tsoela pele ho ithutoa. Likokoanyana li qala ho nkoa e le li-bioreactors, tse fanang ka pH le maemo a oksijene a thusang nts'etsopele ea likokoana-hloko tse khethehileng ka ho senyeha kapa ho senya limatlafatsi tseo ho leng thata hore likokoanyana li li sile 51. Morao tjena, Xiang et al.52 o bontšitse hore, ka mohlala, ho kenngoa ha litšila tsa tlhaho ka motsoako oa baktheria ho lumella CF ho hohela libaktheria tse khethehileng ho senyeha ha lignocellulose, ho ntlafatsa ho senyeha ha eona ka har'a substrate ha e bapisoa le substrates ntle le li-larvae.
Qetellong, mabapi le tšebeliso e ntle ea litšila tsa tlhaho ka H. illucens, lijo tsa CEL le SUC li hlahisitse palo e phahameng ka ho fetisisa ea li-larvae ka letsatsi. Sena se bolela hore ho sa tsotellehe boima bo tlaase ba ho qetela ba batho ka bomong, boima bohle ba larval bo hlahisoang ka substrate e nang le indigestible CH bo bapisoa le bo fumanoang lijong tsa homosaccharide tse nang le monosaccharides le disaccharides. Thutong ea rona, ke habohlokoa ho hlokomela hore litekanyetso tsa limatlafatsi tse ling li lekane ho tšehetsa kholo ea palo ea li-larval le hore ho eketsoa ha CEL ho lokela ho fokotsoa. Leha ho le joalo, sebopeho sa ho qetela sa li-larvae se fapane, se totobatsa bohlokoa ba ho khetha leano le nepahetseng bakeng sa ho matlafatsa likokoanyana. Li-larvae tsa CEL tse feptjoang ka lijo tse feletseng li loketse ho sebelisoa e le lijo tsa liphoofolo ka lebaka la mafura a tsona a tlaase le maemo a tlaase a lauric acid, athe li-larvae tse feptjoang ka lijo tsa SUC kapa MAL li hloka ho senyeha ka ho hatella ho eketsa boleng ba oli, haholo-holo ka biofuel. lefapha. LAC e fumanoa lihlahisoa tsa indasteri ea lebese tse kang whey e tsoang tlhahiso ea chisi. Haufinyane tjena, tšebeliso ea eona (3.5% lactose) e ntlafalitse boima ba 'mele oa ho qetela oa larval53. Leha ho le joalo, lijo tsa taolo thutong ena li ne li na le halofo ea litaba tsa lipid. Ka hona, liphello tse tsoelang pele tsa LAC e ka 'na eaba li ile tsa hanyetsanoa ke larval bioaccumulation ea lipids ea lijo.
Joalo ka ha ho bonts'itsoe ke lithuto tse fetileng, thepa ea monosaccharides le disaccharides e ama kholo ea BSFL haholo mme e hlophisa boemo ba eona ba FA. Haholo-holo, LAC e bonahala e bapala karolo ea tsoelo-pele nakong ea tsoelo-pele ea larval ka ho fokotsa ho fumaneha ha CH bakeng sa ho monya lipid, ka tsela eo ho khothalletsa UFA bioaccumulation. Boemong bona, ho ka ba monate ho etsa liteko tsa bioloji ho sebelisa lijo tse kopanyang PUFA le LAC. Ho feta moo, karolo ea likokoana-hloko, haholo-holo karolo ea metabolites ea likokoana-hloko (joalo ka li-SCFA) tse nkiloeng mekhoeng ea ho belisoa ha tsoekere, e ntse e le sehlooho sa lipatlisiso se lokelang ho etsoa lipatlisiso.
Likokoanyana li ile tsa fumanoa kolone ea BSF ea Laboratory of Functional and Evolutionary Entomology e thehiloeng ka 2017 Agro-Bio Tech, Gembloux, Belgium (bakeng sa lintlha tse ling mabapi le mekhoa ea ho holisa, bona Hoc et al. 19). Bakeng sa liteko tsa liteko, 2.0 g ea mahe a BSF a ne a bokelloa ka mokhoa o sa reroang letsatsi le leng le le leng ho tloha ka har'a mekotla ea ho ikatisa 'me a kenngoa ka 2.0 kg ea 70% ea lijo tsa likhoho tse metsi (Aveve, Leuven, Belgium). Matsatsi a mahlano ka mor'a ho qhotsoa, li-larvae li ile tsa aroloa ho tloha karolong e ka tlaase 'me li baloa ka letsoho bakeng sa liteko. Boima ba pele ba beche ka 'ngoe bo ile ba lekanngoa. Boima bo tloaelehileng ba motho ka mong e ne e le 7.125 ± 0.41 mg, 'me karolelano ea phekolo e' ngoe le e 'ngoe e bontšoa ho Supplementary Table S2.
Tlhahiso ea lijo e ile ea ikamahanya le thuto ea Barragan-Fonseca et al. 38 . Ka bokhuts'oanyane, ho ile ha fumanoa ho sekisetsa pakeng tsa boleng bo tšoanang ba phepelo bakeng sa likhoho tsa li-larval, litaba tse tšoanang tse omeletseng (DM), tse phahameng CH (10% ho latela lijo tse ncha) le sebopeho, kaha tsoekere e bonolo le li-disaccharides ha li na thepa ea mongolo. Ho latela tlhaiso-leseling ea moetsi (Chicken Feed, AVEVE, Leuven, Belgium), CH e lekiloeng (e leng tsoekere e qhibilihang) e ile ea eketsoa ka thoko e le tharollo ea metsi a autoclaved (15.9%) ho lijo tse nang le protheine ea 16.0%, 5.0% kakaretso ea lipids, 11.9% ea lijo tsa likhoho tsa fatše tse nang le molora le 4.8% ea fiber. Ka nkho e 'ngoe le e' ngoe ea 750 ml (17.20 × 11.50 × 6.00 cm, AVA, Tempsee, Belgium), 101.9 g ea tharollo ea autoclaved CH e kopantsoe le 37.8 g ea lijo tsa likhoho. Bakeng sa lijo ka 'ngoe, litaba tse omeletseng e ne e le 37.0%, ho kenyelletsa le protheine e ts'oanang (11.7%), lipids ea homogeneous (3.7%) le tsoekere e ts'oanang (26.9% ea CH e ekelitsoeng). CH e lekiloeng e ne e le tsoekere (GLU), fructose (FRU), galactose (GAL), maltose (MAL), sucrose (SUC) le lactose (LAC). Lijo tsa ho laola li ne li e-na le cellulose (CEL), e nkoang e sa khonehe bakeng sa H. illucens larvae 38 . Li-larvae tse lekholo tsa matsatsi a 5 li ne li kenngoa ka har'a terei e kentsoeng sekoahelo se nang le lesoba la bophara ba 1 cm bohareng 'me se koahetsoe ka letlooa la menoang ea polasetiki. Lijo ka 'ngoe li ile tsa phetoa ka makhetlo a mane.
Boima ba larval bo ile ba lekanngoa matsatsi a mararo ka mor'a hore teko e qale. Bakeng sa tekanyo e 'ngoe le e' ngoe, li-larvae tse 20 li ile tsa tlosoa substrate ho sebelisoa metsi a futhumetseng a sa hloekang le li-forceps, tse omisitsoeng le tse lekantsoeng (STX223, Ohaus Scout, Parsippany, USA). Ka mor'a ho lekanya, li-larvae li ne li khutlisetsoa bohareng ba substrate. Litekanyo li ne li nkoa khafetsa ka makhetlo a mararo ka beke ho fihlela prepupa ea pele e hlaha. Mothating ona, bokella, bala, 'me u lekanye libokoana tsohle joalokaha ho hlalositsoe pejana. Li-larvae tse arohaneng tsa mohato oa 6 (ke hore, li-larvae tse tšoeu tse tsamaellanang le sethala se etellang pele sethaleng sa prepupal) le prepupae (ke hore, mohato oa ho qetela oa larvae nakong eo li-larvae tsa BSF li fetohang li ntšo, li khaotsa ho fepa, 'me li batla tikoloho e loketseng metamorphosis) le ho boloka ho - 18°C bakeng sa tlhahlobo ea motsoako. Kotulo e ne e baloa e le karo-karolelano ea boima ba kakaretso ea likokoanyana (li-larvae le li-prepupae tsa sethaleng sa 6) tse fumanoang ka sejana (g) ho isa nakong ea tsoelo-pele (d). Litekanyetso tsohle tsa sengoloa li hlahisoa e le: bolela ± SD.
Mehato eohle e ileng ea latela ka ho sebelisa solvents (hexane (Hex), chloroform (CHCl3), methanol (MeOH)) e ne e etsoa tlas'a mosi oa mosi 'me ho hlokahala hore ho roaloe liatlana tsa nitrile, li-aprone le likhalase tsa tšireletso.
Li-larvae tse tšoeu li ile tsa omisoa ka har'a mochini o omisang sehatsetsi oa FreeZone6 (Labconco Corp., Kansas City, MO, USA) ka lihora tse 72 'me tsa theoha (IKA A10, Staufen, Jeremane). Kakaretso ea lipids e ne e ntšitsoe ho ± 1 g ea phofo ho sebelisa mokhoa oa Folch 54. Mongobo o setseng oa sampole e 'ngoe le e' ngoe e nang le lyophilized e ile ea khethoa ka bobeli ho sebelisa mochine oa ho hlahloba mongobo (MA 150, Sartorius, Göttiggen, Jeremane) ho lokisa lipids kaofela.
Kakaretso ea lipids e ile ea fetoloa tlas'a maemo a acidic ho fumana mafura a methyl esters. Ka bokhutšoanyane, hoo e ka bang 10 mg lipids/100 µl CHCl3 solution (100 µl) e ile ea fetoloa mouoane ka naetrojene ka har'a tube ea 8 ml Pyrex© (SciLabware - DWK Life Sciences, London, UK). The tube e ile ea kenngoa ka Hex (0.5 ml) (PESTINORM®SUPRTRACE n-Hexane> 95% bakeng sa tlhahlobo ea organic trace analysis, VWR Chemicals, Radnor, PA, USA) le Hex/MeOH/BF3 (20/25/55) tharollo (0.5) ml) ka bateng ea metsi ka 70 ° C bakeng sa 90 min. Ka mor'a ho pholile, tharollo ea 10% ea metsi a H2SO4 (0.2 ml) le tharollo ea NaCl e tletseng (0.5 ml) e ile ea kenngoa. Kopanya tube 'me u tlatse motsoako ka Hex e hloekileng (8.0 mL). Karolo e 'ngoe ea karolo e ka holimo e ile ea fetisetsoa ho vial' me ea hlahlojoa ke chromatography ea khase e nang le mochine oa ionization oa mollo (GC-FID). Mehlala e ile ea hlahlojoa ho sebelisoa Trace GC Ultra (Thermo Scientific, Waltham, MA, USA) e nang le ente e arohaneng / e sa arohaneng (240 ° C) ka mokhoa oa ho arohana (phallo e arohaneng: 10 mL/min), kholomo ea Stabilwax®-DA ( 30 m, 0.25 mm id, 0.25 μm, Restek Corp., Bellefonte, PA, USA) le FID (250 °C). Lenaneo la mocheso le behiloe ka tsela e latelang: 50 ° C bakeng sa 1 min, ho eketseha ho 150 ° C ho 30 ° C / min, ho eketseha ho 240 ° C ho 4 ° C / min le ho tsoela pele ho 240 ° C bakeng sa 5 min. Hex e ne e sebelisoa e le sekheo se se nang letho le tekanyetso ea litšupiso e nang le 37 fatty acid methyl esters (Supelco 37-component FAMEmix, Sigma-Aldrich, Overijse, Belgium) e sebelisitsoe bakeng sa boitsebiso. Ho tsebahatsoa ha li-unsaturated fatty acids (UFAs) ho tiisitsoe ke GC e pharaletseng ea mahlakore a mabeli (GC × GC-FID) 'me boteng ba li-isomer bo ile ba khethoa ka nepo ka ho ikamahanya hanyenyane ha mokhoa oa Ferrara et al. 55. Lintlha tsa lisebelisoa li ka fumanoa ho Supplementary Table S3 le liphetho ho Supplementary Figure S5.
Lintlha li hlahisoa ka mokhoa oa Excel spreadsheet (Microsoft Corporation, Redmond, WA, USA). Tlhahlobo ea lipalo-palo e ile ea etsoa ho sebelisoa R Studio (version 2023.12.1+402, Boston, USA) 56 . Lintlha tse mabapi le boima ba 'mele, nako ea tsoelo-pele le tlhahiso li ne li hakanngoa ho sebelisoa mohlala oa linear (LM) (taelo "lm", R package "stats" 56) kaha li lumellana le kabo ea Gaussian. Litefiso tsa ho pholoha ho sebelisoa tlhahlobo ea li-binomial li ne li hakanngoa ho sebelisoa mokhoa o tloaelehileng oa linear (GLM) (taelo "glm", sephutheloana sa R "lme4" 57). Tloaelo le homoscedasticity li tiisitsoe ho sebelisoa tlhahlobo ea Shapiro (taelo "shapiro.test", R package "stats" 56) le tlhahlobo ea phapang ea data (taelo ea betadisper, R package "vegan" 58). Kamora ho hlahlojoa ka bobeli ka litekanyetso tsa bohlokoa tsa p (p <0.05) ho tsoa tlhahlobong ea LM kapa GLM, ho ile ha bonoa phapang e kholo lipakeng tsa lihlopha ho sebelisoa tlhahlobo ea EMM (taelo "emmeans", sephutheloana sa R "emmeans" 59).
Ponahalo e felletseng ea FA e bapisitsoe ho sebelisoa tlhahlobo ea tumello ea mefuta e mengata (ke hore permMANOVA; taelo "adonis2", R package "vegan" 58) ho sebelisoa matrix a sebaka sa Euclidean le litumello tse 999. Sena se thusa ho khetholla li-acid tse mafura tse susumetsoang ke mofuta oa lik'habohaedreite tsa lijo. Liphapang tse kholo litabeng tsa FA li ile tsa hlahlobjoa ho ea pele ho sebelisoa papiso ea mahlakore ka bobeli. Lintlha li ile tsa bonoa ho sebelisoa tlhahlobo ea likarolo tse ka sehloohong (PCA) (taelo "PCA", sephutheloana sa R "FactoMineR" 60). FA e ikarabellang bakeng sa liphapang tsena e ile ea khetholloa ka ho toloka li-circles tsa khokahano. Bakhethoa bana ba tiisitsoe ho sebelisoa tlhahlobo ea tsela e le 'ngoe ea phapang (ANOVA) (taelo "aov", R package "stats" 56 ) e lateloa ke tlhahlobo ea post hoc ea Tukey (taelo TukeyHSD, R package "stats" 56 ). Pele ho hlahlojoa, ho tloaeleha ho ile ha hlahlojoa ho sebelisoa tlhahlobo ea Shapiro-Wilk, homoscedasticity e ile ea hlahlojoa ho sebelisoa tlhahlobo ea Bartlett (taelo "bartlett.test", "pakete ea R" lipalo-palo" 56), 'me mokhoa o se nang parametric o ne o sebelisoa haeba leha e le efe ea likhopolo tse peli e ile ea finyelloa. . Litlhahlobo li ile tsa bapisoa (taelo "kruskal.test", R package "stats" 56 ), 'me joale liteko tsa post hoc tsa Dunn li ile tsa sebelisoa (command dunn.test, R package "dunn.test" 56 ).
Phetolelo ea ho qetela ea buka e ngotsoeng ka letsoho e ile ea hlahlojoa ho sebelisoa Grammarly Editor e le sebali sa Senyesemane sa ho hlahloba liphoso ( Grammarly Inc., San Francisco, California, USA) 61 .
Lintlha tse hlahisitsoeng le tse hlahlobiloeng nakong ea thuto ea hajoale li fumaneha ho tsoa ho sengoli se tsamaisanang le sona ka kopo e utloahalang.
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Nako ea poso: Dec-25-2024