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Impukane yesosha elimnyama (Hermetia illucens, L. 1758) iyisinambuzane esibulalayo esinamandla amakhulu okusebenzisa imikhiqizo ephilayo ecebile nge-carbohydrate. Phakathi kwama-carbohydrate, izimpukane zamasosha amnyama zithembele kushukela oncibilikayo ukuze zikhule kanye nokuhlanganiswa kwe-lipid. Inhloso yalolu cwaningo kwakuwukuhlola imiphumela kashukela oncibilikayo ovamile ekuthuthukisweni, ukuphila, kanye nephrofayili ye-acid enamafutha ezimpukane zamasosha amnyama. Faka ukudla kwenkukhu ngama-monosaccharides nama-disaccharides ngokuhlukana. I-Cellulose yayisetshenziswa njengokulawula. Izibungu ezifunza i-glucose, i-fructose, i-sucrose, ne-maltose zikhule ngokushesha kunokulawula izibungu. Ngokuphambene, i-lactose yaba nomthelela oqhubekayo ezibungwini, yehlisa ukukhula futhi yehlisa isisindo somzimba sokugcina somuntu ngamunye. Kodwa-ke, wonke ushukela oncibilikayo wenza izibungu zikhuluphale kunalezo ezidla ukudla okulawulayo. Ngokuphawulekayo, ushukela ohloliwe wakha iphrofayili ye-fatty acid. I-Maltose ne-sucrose kwenyusa okuqukethwe kwe-asidi egcwele amafutha uma kuqhathaniswa ne-cellulose. Ngokuphambene nalokho, i-lactose yandisa i-bioaccumulation ye-dietary unsaturated fatty acids. Lolu cwaningo lungokokuqala ukukhombisa umphumela kashukela oncibilikayo ekwakhiweni kwe-acid enamafutha ezibungu zempukane yamasosha amnyama. Imiphumela yethu ibonisa ukuthi ama-carbohydrate ahloliwe anomphumela obalulekile ekwakhekeni kwe-fatty acid yezibungu zempukane yamasosha amnyama futhi ngenxa yalokho angase anqume ukuthi zisebenza kanjani ekugcineni.
Isidingo somhlaba wonke samandla kanye namaprotheni ezilwane siyaqhubeka nokukhula1. Esimweni sokufudumala kwembulunga yonke, kubalulekile ukuthola ezinye izindlela eziluhlaza esikhundleni samandla ezinto ezimbiwa phansi kanye nezindlela zokukhiqiza ukudla zendabuko ngenkathi kukhula umkhiqizo. Izinambuzane zithembisa abantu abazobhekana nalezi zinkinga ngenxa yokuncipha kwamakhemikhali azo kanye nomthelela emvelweni uma kuqhathaniswa nokufuywa kwendabuko2. Phakathi kwezinambuzane, ikhandidethi elihle kakhulu lokubhekana nalezi zinkinga yimpukane yesosha elimnyama (BSF), i-Hermetia illucens (L. 1758), uhlobo oluyingozi olukwazi ukuzondla ngezinhlobonhlobo ze-organic substrates3. Ngakho-ke, ukuqhakambisa lawa ma-substrates ngokuzalaniswa kwe-BSF kungakha umthombo omusha wezinto ezingavuthiwe ukuhlangabezana nezidingo zezimboni ezihlukahlukene.
Izibungu ze-BSF (BSFL) zingadla emikhiqizweni yezolimo neyezimboni zezolimo ezifana nokusanhlamvu kwabaphisi botshwala, izinsalela zemifino, ukhuni lwesithelo nesinkwa esidala, ezifanele ikakhulukazi ukukhula kwe-BSFL ngenxa ye-carbohydrate ephezulu (CH)4,5, 6 okuqukethwe. Ukukhiqizwa okukhulu kwe-BSFL kubangela ukwakheka kwemikhiqizo emibili: indle, ingxube yezinsalela ze-substrate nendle engasetshenziswa njengomanyolo wokutshala izitshalo7, nezibungu, ezakhiwe ngokuyinhloko ngamaprotheni, lipids kanye ne-chitin. Amaprotheni nama-lipids asetshenziswa kakhulu ekufuyweni kwemfuyo, i-biofuel kanye nezimonyo8,9. Ngokuqondene ne-chitin, le biopolymer ithola izinhlelo zokusebenza emkhakheni we-agri-food, biotechnology kanye nokunakekelwa kwezempilo10.
I-BSF iyisilokazana esizimelayo se-holometabolous, okusho ukuthi ukushintshashintsha kwaso nokukhiqizwa kabusha, ikakhulukazi izigaba ezidla amandla omjikelezo wokuphila kwalesi sinambuzane, kungasekelwa ngokuphelele izinqolobane zomsoco ezikhiqizwa ngesikhathi sokukhula kwezibungu11. Ngokuqondile, ukuhlanganiswa kwamaprotheni kanye ne-lipid kuholela ekuthuthukisweni komzimba wamafutha, isitho esibalulekile sokugcina esikhipha amandla ngesikhathi se-BSF yokungadli: i-prepupa (okungukuthi, isigaba sokugcina sezibungu lapho izibungu ze-BSF ziba mnyama ngenkathi zidla futhi zicinga. endaweni elungele ukuguquguquka kwesimo sezulu), iziphungumangathi (okungukuthi, isigaba esingaguquguquki lapho isinambuzane sishintshashintsha), kanye nabantu abadala12,13. I-CH ingumthombo wamandla oyinhloko ekudleni kwe-BSF14. Phakathi kwalezi zondlo, i-fibrous CH efana ne-hemicellulose, i-cellulose ne-lignin, ngokungafani nama-disaccharides nama-polysaccharides (njengesitashi), ayikwazi ukugaywa yi-BSFL15,16. Ukugaya kwe-CH kuyisinyathelo sokuqala esibalulekile sokumuncwa ama-carbohydrate, agcina e-hydrolyzed abe ushukela olula emathunjini16. Ushukela olula ungabe usumuncwa (okungukuthi, ngolwelwesi lwamathumbu) bese ugaywa ukuze ukhiqize amandla17. Njengoba kushiwo ngenhla, izibungu zigcina amandla engeziwe njengama-lipids emzimbeni wamafutha12,18. I-lipids yokugcina iqukethe ama-triglycerides (ama-lipids angathathi hlangothi akhiwe nge-molecule eyodwa ye-glycerol namafutha amathathu) ahlanganiswe izibungu ezivela kushukela olula wokudla. Lezi CH zihlinzeka ngama-acetyl-CoA substrates adingekayo ku-fatty acid (FA) biosynthesis ngokusebenzisa i-fatty acid synthase kanye nezindlela ze-thioesterase19. Iphrofayili ye-fatty acid ye-H. illucens lipids ilawulwa ngokwemvelo ngama-saturated fatty acids (SFA) anenani eliphezulu le-lauric acid (C12:0)19,20. Ngakho-ke, okuqukethwe kwe-lipid ephezulu kanye nokwakheka kwe-acid enamafutha kukhula ngokushesha kuba yizici ezikhawulela ukusetshenziswa kwezibungu eziphelele ekudleni kwezilwane, ikakhulukazi emfuleni wasemanzini lapho kudingeka khona ama-polyunsaturated fatty acids (PUFA)21.
Selokhu kwatholakala amandla e-BSFL okunciphisa udoti wezinto eziphilayo, izifundo ngenani lemikhiqizo ehlukahlukene ekhiqizwayo ziye zabonisa ukuthi ukwakheka kwe-BSFL ngokwengxenye kulawulwa ukudla kwayo. Njengamanje, ukulawulwa kwephrofayili ye-FA ye-H. illucens iyaqhubeka nokuba ngcono. Ikhono le-BSFL lokuhlanganisa i-PUFA ye-bioaccumulate liye laboniswa kuma-substrates acebile e-PUFA afana ne-algae, imfucumfucu yezinhlanzi, noma ukudla okufana ne-flaxseed, okunikeza iphrofayili ye-FA yekhwalithi ephezulu yokudla kwezilwane19,22,23. Ngokuphambene, ngemikhiqizo engacetshiswanga ku-PUFA, akukhona ngaso sonke isikhathi ukuhlobana phakathi kwamaphrofayili e-FA yokudla kanye ne-FA larval, okubonisa ithonya lezinye izakhi24,25. Eqinisweni, umthelela we-CH egayekayo kumaphrofayela we-FA uhlala ungaqondakali kahle futhi ucwaningwa kancane24,25,26,27.
Ngokwazi kwethu konke, nakuba ama-monosaccharides aphelele nama-disaccharides eningi ekudleni kwe-H. illucens, indima yabo yokudla okunomsoco ihlala ingaqondakali kahle ku-H. illucens umsoco. Inhloso yalolu cwaningo kwakuwukucacisa imiphumela yabo ekudleni kwe-BSFL kanye nokwakheka kwe-lipid. Sizohlola ukukhula, ukuphila, kanye nokukhiqiza kwezibungu ngaphansi kwezimo ezihlukahlukene zokudla okunomsoco. Khona-ke, sizochaza okuqukethwe kwe-lipid kanye nephrofayili ye-fatty acid yokudla ngakunye ukugqamisa imiphumela ye-CH kukhwalithi yomsoco ye-BSFL.
Sacabanga ukuthi imvelo ye-CH ehloliwe izothinta (1) ukukhula kwezibungu, (2) amazinga e-lipid esephelele, futhi (3) ilungise iphrofayela ye-FA. Ama-monosaccharides angamuncwa ngokuqondile, kanti ama-disaccharides kufanele afakwe nge-hydrolyzed. Ngakho-ke ama-monosaccharides atholakala kakhulu njengemithombo yamandla eqondile noma izandulela ze-lipogenesis ngokusebenzisa izindlela ze-FA synthase kanye ne-thioesterase, ngaleyo ndlela kuthuthukiswe ukukhula kwezibungu ze-H. illucens futhi kugqugquzelwe ukunqwabelana kwama-reserve lipids (ikakhulukazi i-lauric acid).
I-CH ehloliwe yathinta isilinganiso sesisindo somzimba sezibungu ngesikhathi sokukhula (Umfanekiso 1). I-FRU, i-GLU, i-SUC kanye ne-MAL yandisa isisindo somzimba wezibungu ngokufana nokudla kokulawula (CEL). Ngokuphambene, i-LAC ne-GAL zibonakale zibambezela ukukhula kwezibungu. Ngokuphawulekayo, i-LAC ibe nomthelela omubi omkhulu ekukhuleni kwezibungu uma kuqhathaniswa ne-SUC phakathi nesikhathi sokukhula: 9.16 ± 1.10 mg ngokumelene ne-15.00 ± 1.01 mg ngosuku lwe-3 (F6,21 = 12.77, p <0.001; Fig. 1), 125.11 ± mg kanye ne-211.79 ± 14.93 mg, ngokulandelana, ngosuku lwe-17 (F6,21 = 38.57, p <0.001; Umfanekiso 1).
Kusetshenziswa ama-monosaccharides ahlukene (i-fructose (FRU), i-galactose (GAL), i-glucose (GLU)), i-disaccharides (lactose (LAC), i-maltose (MAL), i-sucrose (SUC)) kanye ne-cellulose (CEL) njengezilawuli. Ukukhula kwezibungu ezifunzwa izibungu ezimpukane zamasosha amnyama. Iphuzu ngalinye ejikeni limelela isisindo somuntu ngamunye esimaphakathi (mg) esibalwa ngokukala izibungu ezikhethwe ngokungahleliwe ezingama-20 esibalweni sezibungu eziyi-100 (n = 4). Amabha wephutha amele i-SD.
Ukudla kwe-CEL kunikeze ukuphila kwezibungu okuhle kakhulu okungama-95.5 ± 3.8%. Ngaphezu kwalokho, ukusinda kwe-H. illucens ukudla okuphakelwayo okuqukethe i-CH encibilikayo kwehlisiwe (GLM: χ = 107.13, df = 21, p <0.001), okubangelwa i-MAL ne-SUC (ama-disaccharides) ku-CH efundwayo. Ukufa kwakungaphansi kwalokho kwe-GLU, FRU, GAL (monosaccharide), kanye ne-LAC (EMM: p <0.001, Umfanekiso 2).
Ibhokisi lokusinda kwezibungu ezimpukane zamasosha amnyama eziphathwe ngama-monosaccharides ahlukahlukene (i-fructose, i-galactose, i-glucose), ama-disaccharides (i-lactose, i-maltose, i-sucrose) kanye ne-cellulose njengezilawuli. Ukwelashwa ngohlamvu olufanayo akuhlukile kakhulu komunye nomunye (EMM, p > 0.05).
Konke ukudla okuhloliwe kwavumela izibungu ukuthi zifinyelele esigabeni sokuqala. Kodwa-ke, ama-CH ahloliwe avame ukwandisa ukukhula kwezibungu (F6,21=9.60, p<0.001; Ithebula 1). Ikakhulukazi, izibungu ezifunza i-GAL ne-LAC zithathe isikhathi eside ukufinyelela esigabeni sangaphambi kokuphuza uma kuqhathaniswa nezibungu ezikhuliswe ku-CEL (CEL-GAL: p<0.001; CEL-LAC: p<0.001; Ithebula 1).
I-CH ehloliwe nayo yaba nemiphumela ehlukene ngesisindo somzimba wezibungu, isisindo somzimba wezibungu esondla ukudla kwe-CEL sifinyelela ku-180.19 ± 11.35 mg (F6,21 = 16.86, p <0.001; Umfanekiso 3). I-FRU, i-GLU, i-MAL ne-SUC ibangele isisindo somzimba sokugcina sezibungu esingaphezu kuka-200 mg, esasiphezulu kakhulu kune-CEL (p <0.05). Ngokuphambene, izibungu eziphakela i-GAL ne-LAC zinesisindo somzimba esiphansi, isilinganiso esingu-177.64 ± 4.23 mg kanye ne-156.30 ± 2.59 mg, ngokulandelanayo (p <0.05). Lo mphumela wabonakala kakhulu nge-LAC, lapho isisindo somzimba sokugcina sasiphansi kunokudla kokulawula (CEL-LAC: umehluko = 23.89 mg; p = 0.03; Umfanekiso 3).
Isisindo sokugcina sezibungu ngasinye esivezwa njengamachashaza ezibungu (mg) nezimpukane zesosha elimnyama ezivezwa njenge-histogram (g) eziphakela ama-monosaccharides ahlukene (i-fructose, i-galactose, i-glucose), ama-disaccharides (i-lactose, i-maltose, i-sucrose) kanye ne-cellulose (njengokulawula). Izinhlamvu zekholomu zimelela amaqembu ahluke kakhulu esisindweni sezibungu esiphelele (p <0.001). Izinhlamvu ezihlotshaniswa namachashaza wezibungu zimelela amaqembu anesisindo sobungu esihluke kakhulu (p <0.001). Amabha wephutha amele i-SD.
Ubukhulu besisindo somuntu ngamunye bebuzimele esilinganisweni sokugcina sesisindo sekoloni lezibungu. Eqinisweni, ukudla okuqukethe i-FRU, i-GLU, i-MAL, ne-SUC akuzange kukhulise isisindo sezibungu esiphelele esikhiqizwa ethangini uma kuqhathaniswa ne-CEL (Umfanekiso 3). Kodwa-ke, i-LAC yehle kakhulu isisindo esiphelele (CEL-LAC: umehluko = 9.14 g; p <0.001; Umfanekiso 3).
Ithebula 1 likhombisa isivuno (izibungu/usuku). Kuyathakazelisa ukuthi izivuno ezifanele ze-CEL, i-MAL ne-SUC zazifana (Ithebula 1). Ngokuphambene, i-FRU, i-GAL, i-GLU ne-LAC inciphise isivuno uma iqhathaniswa ne-CEL (Ithebula 1). I-GAL ne-LAC yenze okubi kakhulu: isivuno sehliswe ngesigamu saba ngu-0.51 ± 0.09 g kuphela izibungu/ngosuku kanye nezibungu ezingu-0.48 ± 0.06 g/ngosuku, ngokulandelana (Ithebula 1).
Ama-monosaccharides nama-disaccharides andise okuqukethwe kwe-lipid kwezibungu ze-CF (Ithebula 1). Ekudleni kwe-CLE, kutholwe izibungu ezinokuqukethwe kwe-lipid engu-23.19 ± 0.70% ye-DM. Uma kuqhathaniswa, isilinganiso se-lipid sokuqukethwe kwezibungu eziphakelwa ushukela oncibilikayo sasingaphezu kwama-30% (Ithebula 1). Kodwa-ke, ama-CH ahlolwe andise okuqukethwe kwawo kwamafutha ngezinga elifanayo.
Njengoba kulindelekile, izihloko ze-CG zithinte iphrofayili ye-FA yezibungu ngamadigri ahlukahlukene (Fig. 4). Okuqukethwe kwe-SFA bekuphezulu kukho konke ukudla futhi kufinyelele ngaphezu kwama-60%. I-MAL ne-SUC ayizange ilinganisele iphrofayela ye-FA, okuholele ekwenyukeni kokuqukethwe kwe-SFA. Endabeni ye-MAL, ngakolunye uhlangothi, lokhu kungalingani kwaholela ngokuyinhloko ekunciphiseni kokuqukethwe kwe-monounsaturated fatty acids (MUFA) (F6,21 = 7.47; p <0.001; Umfanekiso 4). Ngakolunye uhlangothi, ku-SUC, ukwehla bekufana kakhulu phakathi kwe-MUFA ne-PUFA. I-LAC ne-MAL ibe nemiphumela ephambene ku-FA spectrum (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; Umfanekiso 4). Ingxenye ephansi ye-SFA kuzibungu eziphakelwa i-LAC ibonakala inyusa okuqukethwe kwe-MUFA. Ikakhulukazi, amazinga e-MUFA ayephezulu kuzibungu ezifunzwa i-LAC uma kuqhathaniswa nabanye ushukela oncibilikayo ngaphandle kwe-GAL (F6,21 = 7.47; p <0.001; Umfanekiso 4).
Ukusebenzisa i-monosaccharides ehlukene (i-fructose (FRU), i-galactose (GAL), i-glucose (GLU)), i-disaccharides (lactose (LAC), i-maltose (MAL), i-sucrose (SUC)) kanye ne-cellulose (CEL) njengezilawuli, ibhokisi lebhokisi lama-fatty acid. ukwakheka okuphakelwa isosha elimnyama izibungu zempukane. Imiphumela ivezwa njengephesenti lengqikithi ye-FAME. Ukwelashwa okumakwe ngezinhlamvu ezihlukene kuhluke kakhulu (p <0.001). (a) Ingxenye yama-saturated fatty acids; (b) ama-monounsaturated fatty acids; (c) Amafutha e-Polyunsaturated.
Phakathi kwama-fatty acids ahlonziwe, i-lauric acid (C12: 0) yayibusa kuzo zonke izibukeli ezibonwayo (ngaphezu kwama-40%). Amanye ama-SFA akhona kwakuyi-palmitic acid (C16:0) (ngaphansi kuka-10%), i-stearic acid (C18:0) (ngaphansi kuka-2.5%) kanye ne-capric acid (C10:0) (ngaphansi kuka-1.5%). Ama-MUFA amelwe ngokuyinhloko i-oleic acid (C18:1n9) (ngaphansi kuka-9.5%), kuyilapho ama-PUFA ayenziwe ngokuyinhloko nge-linoleic acid (C18:2n6) (ngaphansi kuka-13.0%) (bona Ithebula Le-Supplementary S1). Ukwengeza, ingxenye encane yezinhlanganisela ayikwazanga ukukhonjwa, ikakhulukazi ku-spectra yezibungu ze-CEL, lapho inombolo yekhompiyutha engaziwa engu-9 (UND9) ibalwa ngokwesilinganiso esingu-2.46 ± 0.52% (bheka Ithebula Lokwengeza S1). Ukuhlaziywa kwe-GC×GC-FID kuphakamise ukuthi kungase kube i-asidi yamafutha ekhabhoni engu-20 enamabhondi aphindwe kabili amahlanu noma ayisithupha (bona Umfanekiso Owengeziwe S5).
Ukuhlaziywa kwe-PERMANOVA kwembule amaqembu amathathu ahlukene asekelwe kumaphrofayili e-fatty acid (F6,21 = 7.79, p <0.001; Umfanekiso 5). Ukuhlaziywa kwengxenye eyinhloko (PCA) ye-spectrum ye-TBC kubonisa lokhu futhi kuchazwa izingxenye ezimbili (Umfanekiso 5). Izingxenye eziyinhloko zichaze i-57.9% yokuhluka futhi zifake, ngokulandelana kokubaluleka, i-lauric acid (C12: 0), i-oleic acid (C18: 1n9), i-palmitic acid (C16: 0), i-stearic acid (C18: 0), kanye i-linolenic acid (C18: 3n3) (bheka Umfanekiso S4). Ingxenye yesibili ichaze i-26.3% yokuhluka futhi ihlanganisa, ngokulandelana kokubaluleka, i-decanoic acid (C10: 0) ne-linoleic acid (C18: 2n6 cis) (bheka Umfanekiso Ongeziwe S4). Amaphrofayili okudla aqukethe ushukela olula (i-FRU, i-GAL ne-GLU) abonise izici ezifanayo. Ngokuphambene, ama-disaccharides akhiqize amaphrofayili ahlukene: i-MAL ne-SUC ngakolunye uhlangothi kanye ne-LAC ngakolunye. Ikakhulukazi, i-MAL ukuphela kukashukela owashintsha iphrofayili ye-FA uma kuqhathaniswa ne-CEL. Ngaphezu kwalokho, iphrofayili ye-MAL yayihluke kakhulu kumaphrofayili we-FRU kanye ne-GLU. Ikakhulukazi, iphrofayela ye-MAL ibonise ingxenye ephezulu kakhulu ye-C12:0 (54.59 ± 2.17%), okwenza iqhathaniswe ne-CEL (43.10 ± 5.01%), i-LAC (43.35 ± 1.31%), i-FRU (48.90 ± 1.97%) kanye GLU (48.38 ± 2.17%) amaphrofayili (bona Okungeziwe Ithebula S1). I-spectrum ye-MAL iphinde yabonisa okuqukethwe okuphansi kwe-C18:1n9 (9.52 ± 0.50%), okuphinde kwayihlukanisa ne-LAC (12.86 ± 0.52%) kanye ne-CEL (12.40 ± 1.31%). Umkhuba ofanayo ubonwe ku-C16:0. Engxenyeni yesibili, i-spectrum ye-LAC ibonise okuqukethwe okuphezulu kwe-C18:2n6 (17.22 ± 0.46%), kuyilapho i-MAL ibonise okuphansi kakhulu (12.58 ± 0.67%). I-C18: I-2n6 iphinde ihlukanise i-LAC ekulawuleni (CEL), ebonise amazinga aphansi (13.41 ± 2.48%) (bheka I-Supplementary Table S1).
Isiqephu se-PCA sephrofayili ye-fatty acid ye-black soldier fly izibungu ezine-monosaccharides ehlukene (i-fructose, i-galactose, i-glucose), i-disaccharides (i-lactose, i-maltose, i-sucrose) ne-cellulose njengokulawula.
Ukutadisha imiphumela yokudla okunomsoco kashukela oncibilikayo kuzibungu ze-H. illucens, i-cellulose (CEL) ekudleni kwenkukhu kwathathelwa indawo i-glucose (GLU), i-fructose (FRU), i-galactose (GAL), i-maltose (MAL), i-sucrose (SUC), kanye i-lactose (LAC). Kodwa-ke, ama-monosaccharides nama-disaccharides abe nemiphumela ehlukile ekuthuthukisweni, ekusindeni, nasekubunjweni kwezibungu ze-HF. Isibonelo, i-GLU, FRU, kanye namafomu azo ama-disaccharide (i-MAL ne-SUC) abe nemiphumela emihle esekelayo ekukhuleni kwezibungu, okubavumela ukuba bazuze izisindo zokugcina eziphezulu kune-CEL. Ngokungafani ne-CEL engagayeki, i-GLU, i-FRU, ne-SUC ingadlula umgoqo wamathumbu futhi isebenze njengemithombo ebalulekile yezakhi ekudleni okuhlanganisiwe16,28. I-MAL ayinazo izithuthi zezilwane ezithile futhi kucatshangwa ukuthi i-hydrolyzed ibe ama-molecule amabili e-glucose ngaphambi kwe-assimilation15. Lawa ma-molecule agcinwa emzimbeni wezinambuzane njengomthombo wamandla oqondile noma njengama-lipids18. Okokuqala, ngokuphathelene nalokhu kwakamuva, okunye ukuhluka okubonwayo kwe-intramodal kungase kube ngenxa yokuhluka okuncane kwezilinganiso zobulili. Ngempela, ku-H. illucens, ukuzala kungase kwenzeke ngokuzenzakalelayo: abesifazane abadala ngokwemvelo banezindawo ezanele zokubekela amaqanda futhi basinda kunabesilisa29. Kodwa-ke, ukuqoqwa kwe-lipid ku-BSFL kuhlotshaniswa nokudla okuncibilikayo kwe-CH2, njengoba kuphawulwe ngaphambilini ku-GLU ne-xylose26,30. Isibonelo, u-Li et al.30 waphawula ukuthi lapho i-8% GLU yengezwa ekudleni kwezibungu, okuqukethwe kwe-lipid kwezibungu ze-BSF kukhuphuke ngo-7.78% uma kuqhathaniswa nezilawuli. Imiphumela yethu ihambisana nalokhu okubhekwayo, okubonisa ukuthi okuqukethwe kwamafutha ezibungwini ezidla ushukela oncibilikayo kwakuphakeme kunezibungu ezidla ukudla kwe-CEL, uma kuqhathaniswa nokunyuka kwe-8.57% nge-GLU supplementation. Ngokumangalisayo, imiphumela efanayo yabonwa kuzibungu ezifunzwa i-GAL ne-LAC, naphezu kwemiphumela emibi ekukhuleni kwezibungu, isisindo somzimba sokugcina, nokuphila. Izibungu ezaziphakelwa i-LAC zazizincane kakhulu kunalezo ezaziphakelwa ukudla kwe-CEL, kodwa amafutha azo ayeqhathaniswa nezibungu eziphakela omunye ushukela oncibilikayo. Le miphumela igqamisa imiphumela eqhubekayo ye-lactose ku-BSFL. Okokuqala, ukudla kuqukethe inani elikhulu le-CH. Amasistimu wokumunca kanye ne-hydrolysis yama-monosaccharides nama-disaccharides, ngokulandelana, angafinyelela ekugcwaleni, okubangele amabhodlela enqubweni yokufanisa. Ngokuqondene ne-hydrolysis, yenziwa yi-α- kanye ne-β-glucosidases 31. Lawa ma-enzyme aye akhetha ama-substrates kuye ngobukhulu bawo kanye nezibopho zamakhemikhali (izixhumanisi ze-α noma ze-β) phakathi kwama-monosaccharides awo angu-15. I-Hydrolysis ye-LAC kuya ku-GLU ne-GAL yenziwa yi-β-galactosidase, i-enzyme umsebenzi wayo oboniswe emathunjini e-BSF 32. Kodwa-ke, ukusho kwayo kungase kunganele uma kuqhathaniswa nenani le-LAC elidliwe izibungu. Ngokuphambene, i-α-glucosidase maltase kanye ne-sucrase 15, ezaziwa ngokuvezwa kakhulu ezinambuzaneni, ziyakwazi ukudiliza inani elikhulu le-MAL ne-sucrose SUC, ngaleyo ndlela kukhawulelwe lo mphumela wokusuthisa. Okwesibili, imiphumela eqhubekayo ingase ibe ngenxa yokuncipha kokukhuthazwa komsebenzi wezinambuzane wamathumbu e-amylase kanye nokuncipha kokuziphatha kokudla uma kuqhathaniswa nezinye izindlela zokwelapha. Ngempela, ushukela oncibilikayo ubonwe njengezikhuthazo zomsebenzi we-enzyme obalulekile ekugayweni kwezinambuzane, njenge-amylase, futhi njengezinto ezibangela impendulo yokudla33,34,35. Izinga lokuvuselela liyahlukahluka kuye ngesakhiwo samangqamuzana kashukela. Eqinisweni, ama-disaccharides adinga i-hydrolysis ngaphambi kokumuncwa futhi athambekele ekuvuseleleni i-amylase ngaphezu kwama-monosaccharides awo ayingxenye34. Ngokuphambene, i-LAC inomphumela omncane futhi itholakale ingakwazi ukusekela ukukhula kwezinambuzane ezinhlobonhlobo zezinhlobo33,35. Isibonelo, ku-spodoptera exigua ewuhlupho (Boddie 1850), awukho umsebenzi we-hydrolytic we-LAC otholwe ekukhishweni kwama-enzymes e-caterpillar midgut36.
Mayelana ne-spectrum ye-FA, imiphumela yethu ikhombisa imiphumela ebalulekile yokushintshashintsha kwe-CH ehloliwe. Ngokuphawulekayo, nakuba i-lauric acid (C12:0) ibalwa ngaphansi kuka-1% wesamba se-FA ekudleni, yayibusa kuwo wonke amaphrofayili (bona Ithebula Le-Supplementary S1). Lokhu kuhambisana nedatha yangaphambilini yokuthi i-lauric acid ihlanganiswa kusuka ekudleni kwe-CH ku-H. illucens ngendlela ehlanganisa i-acetyl-CoA carboxylase ne-FA synthase19,27,37. Imiphumela yethu iqinisekisa ukuthi i-CEL ayigayeki kakhulu futhi isebenza “njenge-ejenti eningi” ekudleni okulawula i-BSF, njengoba kuxoxiwe ezifundweni ezimbalwa ze-BSFL38,39,40. Ukushintsha i-CEL ngama-monosaccharides nama-disaccharides ngaphandle kwe-LAC kwenyusa isilinganiso se-C12:0, okubonisa ukwanda kokutholwa kwe-CH yizibungu. Kuyathakazelisa ukuthi ama-disaccharides i-MAL ne-SUC akhuthaza ukuhlanganiswa kwe-lauric acid ngokuphumelelayo kune-monosaccharides yawo ehlanganisiwe, ephakamisa ukuthi naphezu kwezinga eliphezulu le-polymerization ye-GLU ne-FRU, futhi njengoba i-Drosophila kuwukuphela kwe-sucrose transporter ekhonjwe ezinhlobonhlobo zamaprotheni ezilwane, abathuthi be-disaccharide. kungenzeka ingabi khona emathunjini e-H. illucens larvae15, ukusetshenziswa I-GLU ne-FRU iyanda. Kodwa-ke, nakuba i-GLU ne-FRU ngokwethiyori igaywa kalula yi-BSF, ibuye igaywe kalula ngama-substrates nama-gut microorganisms, okungase kubangele ukuwohloka kwawo okushesha kakhulu kanye nokuncipha kokusetshenziswa kwezibungu uma kuqhathaniswa nama-disaccharides.
Uma uthi nhlá, okuqukethwe kwe-lipid kwezibungu ezifunzwa i-LAC ne-MAL kwakuqhathaniswa, okubonisa ukutholakala okufanayo kwe-bioavailability kwalawa ushukela. Kodwa-ke, ngokumangalisayo, iphrofayela ye-FA ye-LAC yayicebile ku-SFA, ikakhulukazi ngokuqukethwe okuphansi kwe-C12:0, uma kuqhathaniswa ne-MAL. Enye inkolelo-mbono yokuchaza lo mehluko ukuthi i-LAC ingase ikhuthaze ukuqoqwa kwe-bioaccumulation ye-FA yokudla nge-acetyl-CoA FA synthase. Isekela le nkolelo-mbono, izibungu ze-LAC zinesilinganiso esiphansi se-decanoate (C10:0) (0.77 ± 0.13%) kunokudla kwe-CEL (1.27 ± 0.16%), okubonisa ukunciphisa kwe-FA synthase kanye nemisebenzi ye-thioesterase19. Okwesibili, ama-fatty acids okudla abhekwa njengesici esiyinhloko esithonya ukwakheka kwe-SFA kwe-H. illucens27. Ekuhloleni kwethu, i-linoleic acid (C18:2n6) ibalwa ku-54.81% wamafutha e-dietary fatty acids, kanti ingxenye yezibungu ze-LAC ingu-17.22 ± 0.46% kanye ne-MAL 12.58 ± 0.67%. I-Oleic acid (cis + trans C18: 1n9) (23.22% ekudleni) ibonise umkhuba ofanayo. Isilinganiso se-α-linolenic acid (C18: 3n3) siphinde sisekele i-bioaccumulation hypothesis. Le asidi enamafutha yaziwa ukuthi inqwabelana ku-BSFL lapho kunothisa i-substrate, njengokwengezwa kwekhekhe le-flaxseed, kuze kufike ku-6-9% wawo wonke ama-acids anamafutha ezibungu19. Ekudleni okunothisiwe, i-C18:3n3 ingafinyelela ku-35% wesamba samafutha acid okudla. Kodwa-ke, esifundweni sethu, i-C18: 3n3 ibalwa kuphela i-2.51% yephrofayili ye-fatty acid. Nakuba ingxenye etholakala emvelweni yayiphansi ezibungwini zethu, le ngxenye yayiphezulu kuzibungu ze-LAC (0.87 ± 0.02%) kune-MAL (0.49 ± 0.04%) (p <0.001; bheka Ithebula Lokwengeza S1). Ukudla kwe-CEL bekunengxenye emaphakathi engu-0.72 ± 0.18%. Ekugcineni, isilinganiso se-palmitic acid (C16:0) kuzibungu ze-CF sibonisa umnikelo wezindlela zokwenziwa kanye nokudla kwe-FA19. Hoc et al. I-19 ibone ukuthi i-C16: i-0 synthesis yancishiswa lapho ukudla kunothiswa ngesidlo se-flaxseed, okwakubangelwa ukwehla kokutholakala kwe-acetyl-CoA substrate ngenxa yokwehla kwesilinganiso se-CH. Ngokumangazayo, nakuba kokubili ukudla kunokuqukethwe kwe-CH okufanayo futhi i-MAL ibonise i-bioavailability ephakeme, izibungu ze-MAL zibonise isilinganiso esiphansi se-C16: 0 (10.46 ± 0.77%), kanti i-LAC ibonise ingxenye ephezulu, i-accounting ye-12.85 ± 0.27% (p <0.05; bheka; bheka Ithebula Lokwengeza S1). Le miphumela iqokomisa ithonya eliyinkimbinkimbi lezakhamzimba ekugayweni kwe-BSFL kanye ne-metabolism. Njengamanje, ucwaningo ngalesi sihloko lucophelela kakhulu ku-Lepidoptera kunase-Diptera. Kumacimbi, i-LAC yabonakala njengesikhuthazi esibuthakathaka sokuziphatha kokuphakelayo uma kuqhathaniswa nabanye ushukela oncibilikayo njenge-SUC ne-FRU34,35. Ikakhulukazi, ku-Spodopteralittoralis (Boisduval 1833), ukusetshenziswa kwe-MAL kukhuthaze umsebenzi we-amylolytic emathunjini ngezinga elikhulu kune-LAC34. Imiphumela efanayo ku-BSFL ingase ichaze ukugqugquzelwa okuthuthukisiwe kwe-C12:0 indlela yokwenziwa kwezibungu ze-MAL, ehlotshaniswa nokwanda kwe-CH emuncwa emathunjini, ukondla isikhathi eside, kanye nesenzo se-amylase yamathumbu. Ukuncipha kokukhuthazwa kwesigqi sokudla phambi kwe-LAC kungase kuchaze nokukhula kancane kwezibungu ze-LAC. Ngaphezu kwalokho, uLiu Yanxia et al. 27 yaphawula ukuthi impilo yeshalofu ye-lipids ku-H. illucens substrates yayinde kunaleyo ye-CH. Ngakho-ke, izibungu ze-LAC zingase zithembele kakhulu kuma-lipids okudla ukuze ziqedele ukuthuthukiswa kwazo, okungase kwandise okuqukethwe kwazo kwe-lipid yokugcina futhi kuguqule iphrofayili yazo ye-fatty acid.
Ngokwazi kwethu konke, izifundo ezimbalwa kuphela ezihlole imiphumela ye-monosaccharide kanye ne-disaccharide eyengeziwe ekudleni kwe-BSF kumaphrofayili abo e-FA. Okokuqala, uLi et al. I-30 ihlole imiphumela ye-GLU ne-xylose futhi yabona amazinga e-lipid afana nawethu ngesilinganiso sokungezwa esingu-8%. Iphrofayili ye-FA ayizange inemininingwane futhi iqukethe ikakhulukazi i-SFA, kodwa akukho mehluko owatholakala phakathi koshukela ababili noma lapho bethulwa kanyekanye30. Ngaphezu kwalokho, uCohn et al. I-41 ayizange ibonise umphumela we-20% we-GLU, SUC, FRU kanye ne-GAL okungeziwe ekudleni kwenkukhu kumaphrofayela e-FA afanele. Lezi zibukeli zitholwe kusukela ekuphindaphindweni kwezobuchwepheshe kunezinto eziphilayo, lokho, njengoba kuchazwe ababhali, kungase kukhawulele ukuhlaziywa kwezibalo. Ngaphezu kwalokho, ukuntuleka kokulawulwa kwe-iso-ushukela (ukusebenzisa i-CEL) kunciphisa ukuchazwa kwemiphumela. Muva nje, izifundo ezimbili zikaNugroho RA et al. wabonisa okudidayo ku-FA spectra42,43. Esifundweni sokuqala, uNugroho RA et al. 43 ihlole umphumela wokwengeza i-FRU ekudleni okuvutshiwe kwe-palm kernel. Iphrofayili ye-FA yezibungu eziwumphumela ibonise amazinga aphezulu ngokungavamile e-PUFA, angaphezu kwama-90% awo athathwe ekudleni okuqukethe i-10% FRU (efana nocwaningo lwethu). Nakuba lokhu kudla kwakuqukethe ama-pellets ezinhlanzi anothile nge-PUFA, amanani ephrofayili ye-FA abikiwe ezibungu ekudleni okulawulwayo okuhlanganisa i-100% ye-PCM evutshiwe ayengahambelani nanoma iyiphi iphrofayili ebikwe ngaphambilini, ikakhulukazi izinga elingavamile le-C18: 3n3 ye-17.77 ± 1.67% kanye no-26.08 ± 0.20% we-conjugated linoleic acid (C18:2n6t), i-isomer engavamile ye-linoleic acid. Ucwaningo lwesibili lubonise imiphumela efanayo ehlanganisa i-FRU, i-GLU, i-MAL ne-SUC42 ekudleni okuvutshiwe kwe-palm kernel. Lezi zifundo, njengolwethu, zigqamisa ubunzima obungathi sína ekuqhathaniseni imiphumela evela ocwaningweni lokudla kwezibungu lwe-BSF, olufana nokukhetha kokulawula, ukusebenzisana neminye imithombo yezakhi, nezindlela zokuhlaziya i-FA.
Ngesikhathi sokuhlolwa, sabona ukuthi umbala nephunga le-substrate kuyahlukahluka kuye ngokudla okusetshenzisiwe. Lokhu kusikisela ukuthi ama-microorganisms angase adlale indima emiphumeleni ebonwa ku-substrate kanye nesistimu yokugaya izibungu. Eqinisweni, ama-monosaccharides nama-disaccharides agaywa kalula nge-colonizing microorganisms. Ukusetshenziswa ngokushesha kukashukela oncibilikayo ngama-microorganisms kungase kuphumele ekukhishweni kwenani elikhulu lemikhiqizo ye-microbial metabolic efana ne-ethanol, i-lactic acid, i-short-chain fatty acids (isb. i-acetic acid, i-propionic acid, i-butyric acid) kanye ne-carbon dioxide44. Ezinye zalezi zinhlanganisela zingase zibe nesibopho semiphumela enobuthi ebulalayo ezibungwini ezibuye zibonwe nguCohn et al.41 ngaphansi kwezimo ezifanayo zokuthuthuka. Isibonelo, i-ethanol iyingozi ezinambuzaneni45. Inani elikhulu lokukhishwa kwe-carbon dioxide lingase libangele ukunqwabelana kwayo phansi ethangini, okungase kuncishe umoya-mpilo osemkhathini uma ukuzungeza komoya kungavumeli ukukhululwa kwawo. Ngokuphathelene nama-SCFA, imiphumela yawo ezinambuzaneni, ikakhulukazi ama-H. illucens, ahlala engaqondakali kahle, nakuba i-lactic acid, i-propionic acid, ne-butyric acid iboniswe ukuthi iyabulala ku-Callosobruchus maculatus (Fabricius 1775)46. Ku-Drosophila melanogaster Meigen 1830, lawa ma-SCFA angamaka okuhogela aqondisa abantu besifazane kuzindawo zokuphuma kwamaqanda, okuphakamisa indima enenzuzo ekuthuthukisweni kwezibungu47. Nokho, i-acetic acid ihlukaniswa njengento eyingozi futhi ingavimbela kakhulu ukukhula kwezibungu47. Ngokuphambene, i-lactate etholwe yi-microbially isanda kutholakala inomphumela wokuvikela kumagciwane ahlasela amathumbu e-Drosophila48. Ngaphezu kwalokho, ama-microorganisms ohlelweni lokugaya adlala indima ekugayweni kwe-CH ezinambuzaneni49. Imiphumela ye-physiological ye-SCFAs ku-gut microbiota, njengezinga lokudla kanye ne-gene expression, ichazwe kuma-vertebrates 50. Zingase futhi zibe nomthelela omubi kuzibungu ze-H. illucens futhi zingafaka isandla ngokwengxenye ekulawuleni amaphrofayili e-FA. Izifundo ngemiphumela yomsoco yale mikhiqizo yokuvutshelwa kwe-microbial izocacisa imiphumela yayo ku-H. illucens umsoco futhi inikeze isisekelo sezifundo zesikhathi esizayo mayelana nama-microorganisms azuzisayo noma ayingozi ngokuya ngokukhula kwawo kanye nenani lama-substrates ane-FA-rich. Mayelana nalokhu, indima yama-microorganisms ezinqubweni zokugaya izinambuzane ezifuywa ngobuningi iya ngokuya kufundwa. Izinambuzane seziqala ukubhekwa njengama-bioreactors, zihlinzeka nge-pH nezimo ze-oxygenation ezenza kube lula ukuthuthukiswa kwama-microorganisms akhethekile ekucekeleni phansi noma ekuqothulweni kwemisoco okunzima ukuthi izinambuzane zigaye i-51. Muva nje, u-Xiang et al.52 ubonise ukuthi, isibonelo, ukujova kwemfucuza yezinto eziphilayo ngenhlanganisela yebhaktheriya kuvumela i-CF ukuheha amabhaktheriya akhethekile e-lignocellulose degradation, ukuthuthukisa ukuwohloka kwayo ku-substrate uma kuqhathaniswa nama-substrates ngaphandle kwezibungu.
Okokugcina, mayelana nokusetshenziswa okunenzuzo kwemfucuza yezinto eziphilayo ngu-H. illucens, ukudla kwe-CEL kanye ne-SUC kwakhiqiza inani eliphakeme kakhulu lezibungu ngosuku. Lokhu kusho ukuthi naphezu kwesisindo sokugcina esiphansi somuntu ngamunye, isisindo sesibungu esiphelele esikhiqizwa engxenyeni engaphansi ehlanganisa i-CH engagayeki siqhathaniswa naleyo etholakala ekudleni okune-homosaccharide okuqukethe ama-monosaccharides nama-disaccharides. Esifundweni sethu, kubalulekile ukuqaphela ukuthi amazinga ezinye izakhamzimba anele ukusekela ukukhula kwezibungu nokuthi ukungezwa kwe-CEL kufanele kube nomkhawulo. Kodwa-ke, ukwakheka kokugcina kwezibungu kuyahluka, okugqamisa ukubaluleka kokukhetha isu elifanele lokuqhakambisa izinambuzane. Izibungu ze-CEL eziphakelwa ukudla okuphelele zifaneleka kakhulu ukusetshenziswa njengokudla kwezilwane ngenxa yokuqukethwe kwazo okunamafutha aphansi kanye namazinga aphansi e-lauric acid, kuyilapho izibungu ezifunzwa nge-SUC noma i-MAL yokudla zidinga ukwehlisa amafutha ngokucindezela ukukhulisa inani lamafutha, ikakhulukazi ku-biofuel. umkhakha. I-LAC itholakala embonini yobisi ngemikhiqizo efana ne-whey evela ekukhiqizeni ushizi. Muva nje, ukusetshenziswa kwayo (i-3.5% lactose) kuthuthukise isisindo somzimba wezibungu sokugcina53. Kodwa-ke, ukudla okulawulwayo kulolu cwaningo kwakuqukethe ingxenye ye-lipid. Ngakho-ke, imiphumela eqhubekayo ye-LAC kungenzeka ukuthi iphikisana ne-larval bioaccumulation yama-lipids okudla.
Njengoba kukhonjisiwe ocwaningweni lwangaphambilini, izakhiwo zama-monosaccharides nama-disaccharides zithinta kakhulu ukukhula kwe-BSFL futhi zimodela iphrofayili yayo ye-FA. Ikakhulukazi, i-LAC ibonakala idlala indima eqhubekayo phakathi nokuthuthukiswa kwezibungu ngokunciphisa ukutholakala kwe-CH yokumuncwa kwe-lipid yokudla, ngaleyo ndlela ikhuthaza ukuqoqwa kwe-UFA bioaccumulation. Kulo mongo, kungathakazelisa ukwenza ama-bioassays kusetshenziswa ukudla okuhlanganisa i-PUFA ne-LAC. Ngaphezu kwalokho, indima yama-microorganisms, ikakhulukazi indima yama-microbial metabolites (njengama-SCFA) atholakala ezinqubweni zokuvutshelwa kukashukela, isalokhu iyisihloko socwaningo esifanele ukuphenywa.
Izinambuzane zitholwe kukholoni ye-BSF ye-Laboratory of Functional and Evolutionary Entomology eyasungulwa ngo-2017 e-Agro-Bio Tech, Gembloux, Belgium (ukuthola imininingwane eyengeziwe mayelana nezindlela zokukhulisa, bheka i-Hoc et al. 19). Ocwaningweni lokuhlola, u-2.0 g wamaqanda e-BSF aqoqwa ngokungahleliwe nsuku zonke emakhejini okuzalanisa futhi afakwa ku-2.0 kg wokudla kwenkukhu okumanzi okungu-70% (Aveve, Leuven, Belgium). Ezinsukwini ezinhlanu ngemva kokuchanyuselwa, izibungu zazihlukaniswa ku-substrate futhi zibalwe mathupha ngezinjongo zokuhlola. Isisindo sokuqala seqoqo ngalinye silinganisiwe. Isilinganiso sesisindo somuntu siqu sasingu-7.125 ± 0.41 mg, futhi isilinganiso sokwelashwa ngakunye sikhonjiswe Kuthebula Lokwengeza S2.
Ukwakhiwa kokudla kwaguqulwa kusukela ocwaningweni lukaBarragan-Fonseca et al. 38 . Kafushane, kutholwe ukuyekethisa phakathi kwekhwalithi efanayo yokudla kwezinkukhu zezibungu, okuqukethwe kwento eyomile efanayo (DM), i-CH ephezulu (10% esekelwe ekudleni okusha) kanye nokuthungwa, njengoba ushukela olula nama-disaccharides awanazo izinto zombhalo. Ngokusho kolwazi lomkhiqizi (I-Chicken Feed, AVEVE, Leuven, Belgium), i-CH ehloliwe (okungukuthi ushukela oncibilikayo) yengezwe ngokuhlukana njengesixazululo se-aqueous autoclaved (15.9%) ekudleni okuhlanganisa amaprotheni angu-16.0%, ama-lipids angu-5.0%. I-11.9% yokudla kwenkukhu egayiwe ehlanganisa umlotha kanye ne-fibre engu-4.8%. Embizeni ngayinye engu-750 ml (17.20 × 11.50 × 6.00 cm, AVA, Tempsee, Belgium), 101.9 g we-autoclaved CH solution yayixutshwe ne-37.8 g yenkukhu yokudla. Ekudleni ngakunye, okuqukethwe kwento eyomile kwakungu-37.0%, okuhlanganisa amaprotheni afanayo (11.7%), ama-lipids a-homogeneous (3.7%) kanye noshukela we-homogeneous (26.9% we-CH eyengeziwe). I-CH ihlolwe i-glucose (GLU), i-fructose (FRU), i-galactose (GAL), i-maltose (MAL), i-sucrose (SUC) ne-lactose (LAC). Ukudla okulawulwayo kwakuqukethe i-cellulose (CEL), ebhekwa njengengenakudliwa yizibungu ze-H. illucens 38. Izibungu eziyikhulu ezinezinsuku eziyi-5 zafakwa ethreyini efakwe isivalo esinembobo engu-1 cm ububanzi phakathi futhi embozwe ngenethi yomiyane yepulasitiki. Ukudla ngakunye kwaphindwa izikhathi ezine.
Izisindo zezibungu zikalwa ezinsukwini ezintathu ngemva kokuqala kokuhlolwa. Esilinganisweni ngasinye, izibungu ezingu-20 zakhishwa ku-substrate kusetshenziswa amanzi afudumele oyinyumba nama-forceps, omisiwe, futhi kukalwa (STX223, Ohaus Scout, Parsippany, USA). Ngemva kokukala, izibungu zazibuyiselwa enkabeni ye-substrate. Izilinganiso zazithathwa njalo kathathu ngesonto kuze kube yilapho kuvela i-prepupa yokuqala. Kuleli qophelo, qoqa, ubale, futhi ukale zonke izibungu njengoba kuchaziwe ngaphambili. Izibungu ezihlukene zesigaba 6 (okungukuthi, izibungu ezimhlophe ezihambisana nesigaba sezibungu esandulela isigaba sokuqala) kanye ne-prepupae (okungukuthi, isigaba sokugcina sezibungu lapho izibungu ze-BSF ziba mnyama, ziyeke ukudla, futhi zifune indawo efanelekile yokushintshashintsha) futhi zigcine - 18°C ukuze kuhlaziywe ukwakheka. Isivuno sibalwa njengesilinganiso sesamba sesamba sezinambuzane (izibungu nama-prepupae esiteji sesi-6) esitholwe ngesitsha ngasinye (g) kuya esikhathini sokuthuthukiswa (d). Wonke amanani asho embhalweni avezwa njengalokhu: kusho ± SD.
Zonke izinyathelo ezalandela kusetshenziswa izincibilikisi (i-hexane (Hex), i-chloroform (CHCl3), i-methanol (i-MeOH)) zenziwa ngaphansi kwe-fume hood futhi kwakudingeka kugqokwe amagilavu e-nitrile, amaphinifa kanye nezibuko zokuphepha.
Izibungu ezimhlophe zomiswa endaweni yokomisa i-FreeZone6 (Labconco Corp., Kansas City, MO, USA) amahora angu-72 bese zagaywa phansi (IKA A10, Staufen, Germany). Isamba se-lipids sakhishwa ku-±1 g wempushana kusetshenziswa indlela ye-Folch 54. Umswakama osele wesampula ngayinye ene-lyophilized kwanqunywa ngokuphindwe kabili kusetshenziswa ukuhlaziya umswakama (MA 150, Sartorius, Göttiggen, Germany) ukuze kulungiswe inani lama-lipids.
Isamba se-lipids sashintshwa ngaphansi kwezimo ze-asidi ukuze kutholwe i-fatty acid methyl esters. Kafushane, cishe i-10 mg lipids/100 µl isixazululo se-CHCl3 (100 µl) sahwamuka nge-nitrogen ku-8 ml Pyrex© tube (SciLabware – DWK Life Sciences, London, UK). Ishubhu lafakwa ku-Hex (0.5 ml) (PESTINORM®SUPRTRACE n-Hexane > 95% ukuze kuhlaziywe i-organic trace analysis, i-VWR Chemicals, Radnor, PA, USA) kanye nesixazululo se-Hex/MeOH/BF3 (20/25/55) (0.5) ml) kubhavu wamanzi ku-70 °C imizuzu engama-90. Ngemuva kokupholisa, isisombululo se-H2SO4 esinamanzi esingu-10% (0.2 ml) nesisombululo se-NaCl esigcwele (0.5 ml) sengeziwe. Hlanganisa ishubhu bese ugcwalisa ingxube nge-Hex ehlanzekile (8.0 mL). Ingxenye yesigaba esiphezulu yadluliselwa ku-vial futhi yahlaziywa nge-chromatography yegesi ngomtshina we-ionization yomlilo (GC-FID). Amasampuli ahlaziywa kusetshenziswa i-Trace GC Ultra (Thermo Scientific, Waltham, MA, USA) efakwe umjovo ohlukanisayo/ongahlukani (240 °C) kumodi yokuhlukanisa (ukugeleza kokuhlukana: 10 mL/min), ikholomu ye-Stabilwax®-DA ( 30 m, 0.25 mm id, 0.25 μm, Restek Corp., Bellefonte, PA, USA) kanye ne-FID (250 °C). Uhlelo lokushisa lwahlelwa kanje: 50 °C iminithi elingu-1, lukhula lube ngu-150 °C ku-30 °C/min, lukhule lube ngu-240 °C ku-4 °C/min futhi luqhubeke ku-240 °C imizuzu emi-5. I-Hex isetshenziswe njengento engenalutho kanye nezinga lereferensi eliqukethe ama-37 fatty acid methyl esters (i-Supelco-component ye-37-FAMEmix, Sigma-Aldrich, Overijse, Belgium) yasetshenziselwa ukuhlonza. Ukuhlonzwa kwama-unsaturated fatty acids (UFAs) kwaqinisekiswa yi-GC enezinhlangothi ezimbili ezibanzi (GC × GC-FID) futhi ukuba khona kwama-isomers kwanqunywa ngokunembile ukujwayela kancane kwendlela ye-Ferrara et al. 55. Imininingwane yezinsimbi ingatholakala kokuthi Ithebula Lokwengeza S3 kanye nemiphumela ku-Supplementary Figure S5.
Idatha yethulwa ngefomethi yesipredishithi se-Excel (Microsoft Corporation, Redmond, WA, USA). Ukuhlaziywa kwezibalo kwenziwe kusetshenziswa i-R Studio (inguqulo 2023.12.1+402, Boston, USA) 56. Idatha yesisindo sezibungu, isikhathi sokuthuthukiswa kanye nokukhiqiza kulinganiselwa kusetshenziswa imodeli yomugqa (LM) (umyalo "lm", iphakheji ye-R "izibalo" 56 ) njengoba ilingana nokusatshalaliswa kwe-Gaussian. Amazinga okusinda kusetshenziswa ukuhlaziya imodeli ye-binomial alinganiselwa kusetshenziswa imodeli yomugqa ovamile (i-GLM) (umyalo “glm”, iphakheji le-R elithi “lme4” 57). Ukujwayelekile kanye ne-homoscedasticity kwaqinisekiswa kusetshenziswa ukuhlolwa kwe-Shapiro (umyalo "shapiro.test", iphakheji ye-R "izibalo" ze-56) nokuhlaziywa kokuhlukahluka kwedatha (i-command betadisper, iphakheji ye-R "vegan" 58). Ngemva kokuhlaziywa okubili kwamanani e-p abalulekile (p <0.05) kusuka kuhlolo lwe-LM noma lwe-GLM, umehluko omkhulu phakathi kwamaqembu utholwe kusetshenziswa ukuhlolwa kwe-EMM (iyala okuthi “emmeans”, iphakheji ye-R “emeans” 59).
Ukubuka okuphelele kwe-FA kuqhathaniswe kusetshenziswa ukuhlaziya kwemvume ye-multivariate kokuhluka (okungukuthi permMANOVA; umyalo othi “adonis2”, iphakethe le-R elithi “vegan” 58) kusetshenziswa i-matrix yebanga le-Euclidean nezimvume ezingu-999. Lokhu kusiza ukuhlonza ama-fatty acids athonywa uhlobo lwama-carbohydrates okudla. Umehluko omkhulu kumaphrofayili we-FA wabuye wahlaziywa kusetshenziswa ukuqhathanisa okubili. Idatha yabe isibonwa kusetshenziswa ukuhlaziywa kwengxenye eyinhloko (PCA) (umyalo "PCA", iphakheji ye-R "FactoMineR" 60). I-FA ebophezele kulo mehluko ikhonjwe ngokuhumusha imibuthano yokuxhumanisa. Laba makhandidethi baqinisekiswa kusetshenziswa ukuhlaziya kwendlela eyodwa yokuhluka (ANOVA) (umyalo “aov”, iphakethe le-R “izibalo” 56 ) kulandelwe ukuhlolwa kwe-post hoc ka-Tukey ( umyalo TukeyHSD, “izibalo” zephakheji le-R 56 ). Ngaphambi kokuhlaziywa, ukujwayelekile kwahlolwa kusetshenziswa ukuhlolwa kwe-Shapiro-Wilk, i-homoscedasticity yahlolwa kusetshenziswa ukuhlolwa kwe-Bartlett (i-command “bartlett.test”, iphakethe le-R “izibalo” 56), futhi kwasetshenziswa indlela ye-nonparametric uma kungekho kokucatshangelwa okubili okufinyelelwe. . Ukuhlaziya kwaqhathaniswa (i-command “kruskal.test”, iphakheji ye-R “izibalo” 56 ), kwase kusetshenziswa ukuhlolwa kwe-post hoc ka-Dunn (command dunn.test, R package “dunn.test” 56 ).
Inguqulo yokugcina yombhalo wesandla yahlolwa kusetshenziswa i-Grammarly Editor njengomhloli wokuhlola wesiNgisi (Grammarly Inc., San Francisco, California, USA) 61 .
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Isikhathi sokuthumela: Dec-25-2024