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Impukane yejoni elimnyama (Hermetia illucens, L. 1758) sisinambuzane esitshabalalisayo esinamandla aphezulu wokusebenzisa icarbohydrate-rich organic by-products. Phakathi kweecarbohydrates, ijoni elimnyama libhabha lithembele kwiswekile enyibilikayo ekukhuleni kunye ne-lipid synthesis. Injongo yolu phononongo yayikukuvavanya iziphumo zeeswekile ezinyibilikayo eziqhelekileyo kuphuhliso, ukusinda, kunye neprofayili ye-asidi enamafutha yeempukane zejoni elimnyama. Yongeza ukutya kwenkukhu kunye ne-monosaccharides kunye ne-disaccharides ngokwahlukeneyo. I-cellulose yayisetyenziswa njengolawulo. Imibungu eyondla i-glucose, i-fructose, i-sucrose, kunye ne-maltose yakhula ngokukhawuleza kunokulawula imibungu. Ngokwahlukileyo koko, i-lactose yayinempembelelo eqhubekayo kwimibungu, inciphisa ukukhula kunye nokunciphisa ubunzima bokugqibela bomzimba womntu. Nangona kunjalo, zonke iiswekile ezinyibilikayo zenze imibungu yatyeba ngakumbi kunezo zondla ukutya kokulawula. Ngokucacileyo, iiswekile ezivavanyiweyo zenze iprofayili ye-asidi enamafutha. I-Maltose kunye ne-sucrose zonyuse umxholo we-asidi enamafutha xa kuthelekiswa neselulosi. Ngokwahlukileyo, i-lactose yonyusa i-bioaccumulation ye-dietary unsaturated fatty acids. Olu phononongo lolokuqala ukubonisa isiphumo seswekile enyibilikayo kukwakheka kweasidi enamafutha embungu yempukane yejoni elimnyama. Iziphumo zethu zibonisa ukuba i-carbohydrates evavanyiweyo inefuthe elibalulekileyo ekubunjweni kwe-asidi enamafutha eembungu zeempukane zejoni elimnyama kwaye ngoko ke zinokumisela isicelo sazo sokugqibela.
Imfuno yehlabathi yamandla kunye neprotheyini yezilwanyana iyaqhubeka nokunyuka1. Kumxholo wokufudumala kwehlabathi, kunyanzelekile ukufumana ezinye iindlela eziluhlaza kumandla efosili kunye neendlela zemveli zokuvelisa ukutya ngelixa kusandiswa imveliso. Izinambuzane zithembisa abagqatswa ukuba bayayilungisa le miba ngenxa yokwakheka kwazo kweekhemikhali ezisezantsi kunye nefuthe lokusingqongileyo xa kuthelekiswa nofuyo lwemveli2. Phakathi kwezinambuzane, umgqatswa ogqwesileyo wokujongana nale miba yimpukane yejoni elimnyama (BSF), iHermetia illucens (L. 1758), i-detritivorous species ekwazi ukutyisa kwiintlobo ezahlukeneyo ze-organic substrates3. Ke ngoko, ukunika amandla ezi substrates ngokufuya kwe-BSF kunokudala umthombo omtsha wemathiriyeli ekrwada ukuhlangabezana neemfuno zamashishini ahlukeneyo.
Imibungu ye-BSF (BSFL) inokutya kwimveliso yezolimo kunye nemveliso yemveliso yezolimo efana nengqolowa yabavundisi, iintsalela zemifuno, i-pulp yeziqhamo kunye nesonka esidala, ezilungele kakhulu ukukhula kwe-BSFL ngenxa ye-carbohydrate ephezulu (CH) 4,5, 6 umxholo. Imveliso enkulu ye-BSFL iphumela ekubunjweni kweemveliso ezimbini: i-feces, umxube weentsalela ze-substrate kunye neendle ezinokuthi zisetyenziswe njengesichumisi sokulima kwezityalo7, kunye nemibungu, eyona nto yenziwe ngamaprotheni, i-lipids kunye ne-chitin. Iiprotheyini kunye ne-lipids zisetyenziswa kakhulu ekufuyeni imfuyo, i-biofuel kunye ne-cosmetics8,9. Ngokuphathelele i-chitin, le biopolymer ifumana izicelo kwi-agri-food sector, biotechnology kunye nokhathalelo lwempilo10.
I-BSF sisinambuzane esizimelayo se-holometabolous, okuthetha ukuba i-metamorphosis yayo kunye nokuveliswa kwakhona, ngakumbi amanqanaba asebenzisa amandla obomi besinambuzane, anokuxhaswa ngokupheleleyo ngoovimba bezondlo abaveliswa ngexesha lokukhula kombungu11. Ngokukodwa, iprotheyini kunye ne-lipid synthesis ikhokelela ekuphuhlisweni komzimba wamanqatha, i-organ ebalulekileyo yokugcina ekhupha amandla ngexesha le-BSF engondliyo: i-prepupa (oko kukuthi, isigaba sokugqibela sombungu apho i-BSF ijika ibe mnyama ngelixa isondla kwaye ifuna. ukwenzela imeko-bume elungele ukuguquguquka), iipupa (oko kukuthi, inqanaba le-non-motile apho isinambuzane singena kwi-metamorphosis), kunye nabantu abadala12,13. I-CH ngowona mthombo wamandla ophambili ekutyeni kwe-BSF14. Phakathi kwezi zondlo, i-CH ye-fibrous efana ne-hemicellulose, i-cellulose kunye ne-lignin, ngokungafani ne-disaccharides kunye ne-polysaccharides (efana nesitashi), ayikwazi ukugaywa yi-BSFL15,16. Ukugaya kwe-CH kuyinyathelo lokuqala elibalulekileyo lokufunxa i-carbohydrates, ekugqibeleni i-hydrolyzed kwiishukela ezilula emathunjini16. Iiswekile ezilula zinokufunxwa (oko kukuthi, nge-intestinal peritrophic membrane) kunye ne-metabolized ukuvelisa amandla17. Njengoko kukhankanyiwe ngasentla, imibungu igcina amandla amaninzi njenge lipids kumzimba onamafutha12,18. I-lipids yokugcina iqulethe i-triglycerides (i-lipids engathathi hlangothi eyenziwe kwi-molecule ye-glycerol enye kunye ne-fatty acids ezintathu) ezidityaniswe yimibungu evela kwiishukela ezilula zokutya. Ezi CH zibonelela nge-acetyl-CoA substrates ezifunekayo kwi-fatty acid (FA) biosynthesis ngokusebenzisa i-fatty acid synthase kunye neendlela ze-thioesterase19. Iprofayili ye-fatty acid ye-H. illucens lipids ngokwemvelo ilawulwa yi-saturated fatty acids (SFA) kunye nenani eliphezulu le-lauric acid (C12: 0) 19,20. Ngoko ke, umxholo ophezulu we-lipid kunye nokwakheka kwe-asidi enamafutha ngokukhawuleza iba yimida yokusetyenziswa kwemibungu epheleleyo kwisondlo sezilwanyana, ngakumbi kwi-aquaculture apho i-polyunsaturated fatty acids (PUFA) iyadingeka21.
Ukusukela oko kufunyaniswe amandla e-BSFL okunciphisa inkunkuma yezinto eziphilayo, izifundo ngexabiso leemveliso ezahlukeneyo zibonise ukuba ukubunjwa kweBSFL ngokuyinxenye kulawulwa kukutya kwayo. Okwangoku, ukulawulwa kweprofayili ye-FA ye-H. illucens iyaqhubeka nokuphucula. Ikhono le-BSFL kwi-bioaccumulate PUFA libonakaliswe kwii-substrates ezizityebi ze-PUFA ezifana ne-algae, inkunkuma yeentlanzi, okanye izidlo ezinjenge-flaxseed, ezibonelela ngeprofayili ephezulu ye-FA kwisondlo sezilwanyana19,22,23. Ngokwahlukileyo, ngenxa yeemveliso ezingaxutywanga kwi-PUFA, akusoloko kukho ulungelelwaniso phakathi kweeprofayili ze-FA yokutya kunye ne-FA larval, ebonisa impembelelo yezinye izondlo24,25. Enyanisweni, isiphumo sokugaywa kwe-CH kwiiprofayili ze-FA zihlala zingaqondwa kakuhle kwaye ziphandwa ngaphantsi24,25,26,27.
Ngokolwazi lwethu, nangona i-monosaccharides epheleleyo kunye ne-disaccharides ininzi ekutyeni kwe-H. illucens, indima yabo yesondlo ihlala ingaqondwa kakuhle kwi-H. ilucens isondlo. Injongo yolu phononongo yayikukucacisa iziphumo zabo kwisondlo se-BSFL kunye nokubunjwa kwe-lipid. Siza kuvavanya ukukhula, ukusinda, kunye nemveliso yombungu phantsi kweemeko ezahlukeneyo zesondlo. Emva koko, siya kuchaza umxholo we-lipid kunye neprofayili ye-fatty acid yokutya ngalunye ukugqamisa iziphumo ze-CH kumgangatho wezondlo ze-BSFL.
Sacinga ukuba uhlobo lwe-CH oluvavanyiweyo luya kuchaphazela (1) ukukhula kombungu, (2) amanqanaba e-lipid ewonke, kunye (3) nokulungelelanisa iphrofayili ye-FA. I-Monosaccharides inokufunxwa ngokuthe ngqo, ngelixa i-disaccharides kufuneka ifakwe i-hydrolyzed. I-Monosaccharides ke ngoko ifumaneka ngakumbi njengemithombo yamandla ethe ngqo okanye izandulela ze-lipogenesis nge-FA synthase kunye neendlela ze-thioesterase, ngaloo ndlela ziphucula ukukhula kombungu we-H. illucens kunye nokukhuthaza ukuqokelelwa kwe-lipids yokugcina (ingakumbi i-lauric acid).
Uvavanyo lwe-CH luchaphazele umyinge wobunzima bombungu ngexesha lokukhula (umzobo 1). I-FRU, i-GLU, i-SUC kunye ne-MAL yandisa ubunzima bomzimba we-larval ngokufanayo kwi-control diet (CEL). Ngokuchaseneyo, i-LAC kunye ne-GAL zibonakale zilubambezela uphuhliso lombungu. Ngokucacileyo, i-LAC ibe nefuthe elibi kakhulu ekukhuleni kombungu xa kuthelekiswa ne-SUC kulo lonke ixesha lokukhula: 9.16 ± 1.10 mg ngokuchasene ne-15.00 ± 1.01 mg ngosuku lwe-3 (F6,21 = 12.77, p <0.001; Umzobo 1), 125.11 ± 125.16 mg kunye ne-211.79 ± 14.93 mg, ngokulandelanayo, ngosuku lwe-17 (F6,21 = 38.57, p <0.001; Umzobo 1).
Ukusebenzisa i-monosaccharides eyahlukeneyo (i-fructose (FRU), i-galactose (GAL), i-glucose (GLU)), i-disaccharides (i-lactose (LAC), i-maltose (MAL), i-sucrose (SUC)) kunye ne-cellulose (CEL) njengokulawula. Ukukhula kombungu owondliwa yimibungu yeempukane zejoni elimnyama. Inqaku ngalinye kwi-curve limele ubunzima bomntu ngamnye (mg) obalwe ngokulinganisa i-20 ye-larvae ekhethiweyo ekhethiweyo ukusuka kuluntu lwe-100 (n = 4). Iibar zemposiso zibonisa i-SD.
Ukutya kwe-CEL kubonelele ngokubalaseleyo ukusinda kombungu wama-95.5 ± 3.8%. Ngaphezu koko, ukusinda kwe-H. illucens ukutya okunokutya okune-soluble CH kwancitshiswa (GLM: χ = 107.13, df = 21, p <0.001), eyabangelwa yi-MAL kunye ne-SUC (disaccharides) kwi-CH efundwayo. Ukufa kwakungaphantsi kwe-GLU, FRU, GAL (monosaccharide), kunye ne-LAC (EMM: p <0.001, Umfanekiso 2).
Ibhokisi yebhokisi yokusinda imbungu yejoni elimnyama ephathwa nge-monosaccharides eyahlukeneyo (i-fructose, i-galactose, i-glucose), i-disaccharides (i-lactose, i-maltose, i-sucrose) kunye ne-cellulose njengolawulo. Unyango olunobumba olufanayo alufani kakhulu omnye komnye (EMM, p> 0.05).
Zonke izidlo ezivavanyiweyo ziye zavumela imibungu ukuba ifikelele kwinqanaba le-prepupal. Nangona kunjalo, ii-CHs ezivavanyiweyo zithande ukwandisa uphuhliso lombungu (F6,21=9.60, p<0.001; Itheyibhile 1). Ngokukodwa, imibungu eyondla i-GAL kunye ne-LAC ithathe ixesha elide ukufikelela kwisigaba se-prepupal xa kuthelekiswa nemibungu ekhuliswe kwi-CEL (CEL-GAL: p<0.001; CEL-LAC: p<0.001; Itheyibhile 1).
I-CH evavanyiweyo nayo yaba nemiphumo eyahlukileyo kubunzima bomzimba we-larva, kunye nobunzima bomzimba we-larvae wondla ukutya kwe-CEL ukufikelela kwi-180.19 ± 11.35 mg (F6,21 = 16.86, p <0.001; Umzobo 3). I-FRU, i-GLU, i-MAL kunye ne-SUC ibangele umlinganiselo wokugqibela wobunzima bomzimba obungaphezu kwe-200 mg, owawuphezulu kakhulu kune-CEL (p <0.05). Ngokwahlukileyo, izibungu ezondla i-GAL kunye ne-LAC zinesisindo somzimba esisezantsi, i-avareji ye-177.64 ± 4.23 mg kunye ne-156.30 ± 2.59 mg, ngokulandelanayo (p <0.05). Esi siphumo sasibonakaliswe ngakumbi nge-LAC, apho ubunzima bomzimba bokugqibela buphantsi kunokutya kokulawula (CEL-LAC: umehluko = 23.89 mg; p = 0.03; Umfanekiso 3).
Ithetha ubunzima bokugqibela bombungu ngamnye obonakaliswa njengamabala ombungu (mg) kunye neempukane zejoni elimnyama ezichazwe njenge-histogram (g) zondla i-monosaccharides eyahlukeneyo (i-fructose, i-galactose, i-glucose), i-disaccharides (i-lactose, i-maltose, i-sucrose) kunye ne-cellulose (njengolawulo). Oonobumba bekholomu bamele amaqela ahluke kakhulu kubunzima bombungu bubonke (p <0.001). Oonobumba abanxulunyaniswa namabala ombungu bamele amaqela anobunzima bombungu obahluke kakhulu (p <0.001). Iibar zemposiso zibonisa i-SD.
Ubukhulu bobunzima bomntu ngamnye babuzimeleyo kubunzima bokugqibela bekholoni yombungu. Enyanisweni, ukutya okuqukethe i-FRU, i-GLU, i-MAL, kunye ne-SUC ayizange ikhulise ubunzima be-larval obuveliswe kwitanki xa kuthelekiswa ne-CEL (Umfanekiso 3). Nangona kunjalo, i-LAC iyancipha kakhulu ubunzima obupheleleyo (CEL-LAC: umehluko = 9.14 g; p <0.001; Umfanekiso 3).
Uluhlu loku-1 lubonisa isivuno (imibungu/imini). Okubangel 'umdla kukuba, izivuno ezifanelekileyo ze-CEL, i-MAL kunye ne-SUC zazifana (iThebhile 1). Ngokwahlukileyo, i-FRU, i-GAL, i-GLU kunye ne-LAC yanciphisa isivuno xa kuthelekiswa ne-CEL (Itheyibhile 1). I-GAL kunye ne-LAC yenze eyona nto imbi kakhulu: isivuno sasiyisiqingatha saya kuphela kwi-0.51 ± 0.09 g imibungu / ngosuku kunye ne-0.48 ± 0.06 g imibungu / ngosuku, ngokulandelanayo (Itheyibhile 1).
I-Monosaccharides kunye ne-disaccharides yandisa umxholo we-lipid we-CF larvae (iThebhile 1). Kwisidlo se-CLE, iimbungu ezinomxholo we-lipid we-23.19 ± 0.70% womxholo we-DM zifunyenwe. Ukuthelekisa, umxholo we-lipid ophakathi kwimibungu eyondliwe ngeswekile enyibilikayo yayingaphezulu kwe-30% (Itheyibhile 1). Nangona kunjalo, ii-CHs ezivavanyiweyo zonyusa umxholo wabo wamafutha kwinqanaba elifanayo.
Njengoko bekulindelekile, izifundo zeCG zichaphazele iphrofayili ye-FA yeembungu ukuya kumaqondo ahlukeneyo (umzobo 4). Umxholo we-SFA wawuphezulu kuzo zonke izidlo kwaye ufikelele ngaphezu kwe-60%. I-MAL kunye ne-SUC ayilingani iprofayili ye-FA, eyakhokelela ekwandeni komxholo we-SFA. Kwimeko ye-MAL, ngakolunye uhlangothi, oku kungalingani kwakhokelela kakhulu ekunciphiseni kumxholo we-monounsaturated fatty acids (MUFA) (F6,21 = 7.47; p <0.001; Umzobo 4). Kwelinye icala, kwi-SUC, ukuhla bekuyiyunifomu ngakumbi phakathi kwe-MUFA kunye ne-PUFA. I-LAC kunye ne-MAL zineziphumo ezichaseneyo kwi-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). Umlinganiselo osezantsi we-SFA kwi-LAC-wondliwe imibungu ibonakala inyusa umxholo we-MUFA. Ngokukodwa, amanqanaba e-MUFA ayephezulu kwi-LAC-fed larvae xa kuthelekiswa nezinye iishukela ezinyibilikayo ngaphandle kwe-GAL (F6,21 = 7.47; p <0.001; Umfanekiso 4).
Ukusebenzisa i-monosaccharides eyahlukeneyo (i-fructose (i-FRU), i-galactose (i-GAL), i-glucose (GLU)), i-disaccharides (i-lactose (i-LAC), i-maltose (MAL), i-sucrose (SUC)) kunye ne-cellulose (CEL) njengokulawula, ibhokisi yebhokisi ye-fatty acid. ukwakheka okondla ijoni elimnyama imibungu impukane. Iziphumo zichazwa njengepesenti yeFAME iyonke. Unyango oluphawulwe ngoonobumba abahlukeneyo luhluke kakhulu (p <0.001). (a) Umlinganiselo weeasidi ezinamafutha; (b) Ii-monounsaturated fatty acids; (c) I-Polyunsaturated fatty acids.
Phakathi kwe-fatty acids echongiweyo, i-lauric acid (i-C12: i-0) yayiyeyona nto iphambili kuyo yonke i-spectra ebonwayo (ngaphezu kwe-40%). Ezinye ii-SFA ezikhoyo ziyi-palmitic acid (C16: 0) (ngaphantsi kwe-10%), i-stearic acid (C18: 0) (ngaphantsi kwe-2.5%) kunye ne-capric acid (C10: 0) (ngaphantsi kwe-1.5%). I-MUFAs ikakhulu imelwe yi-oleic acid (C18: 1n9) (ngaphantsi kwe-9.5%), ngelixa i-PUFAs yayiyi-linoleic acid (C18: 2n6) (ngaphantsi kwe-13.0%) (jonga i-Supplementary Table S1). Ukongeza, inxalenye encinci yeekhompawundi ayikwazanga ukuchongwa, ngakumbi kwi-spectra ye-CEL larvae, apho inani elingaziwayo le-compound 9 (UND9) libalelwa kumyinge we-2.46 ± 0.52% (jonga i-Supplementary Table S1). Uhlalutyo lwe-GC × GC-FID lucebise ukuba ingaba yi-20-carbon fatty acid ene-five or six bond double bonds (jonga i-Supplementary Figure S5).
Uhlalutyo lwe-PERMANOVA lubonakalise amaqela amathathu ahlukeneyo asekelwe kwiiprofayili ze-fatty acid (F6,21 = 7.79, p <0.001; Umfanekiso 5). Uhlalutyo lwamacandelo enqununu (PCA) lwe-spectrum ye-TBC lubonisa oku kwaye luchazwa ngamacandelo amabini (umzobo 5). Amacandelo anqununu achaza i-57.9% yokuhluka kwaye ibandakanya, ngokokubaluleka, i-lauric acid (C12: 0), i-oleic acid (C18: 1n9), i-palmitic acid (C16: 0), i-stearic acid (C18: 0), kunye i-linolenic acid (C18: 3n3) (jonga umfanekiso we-S4). Icandelo lesibini lichaze i-26.3% yokuhluka kwaye ibandakanya, ngokokubaluleka, i-decanoic acid (C10: 0) kunye ne-linoleic acid (C18: 2n6 cis) (jonga i-Supplementary Figure S4). Iiprofayili zokutya eziqukethe iishukela ezilula (i-FRU, i-GAL kunye ne-GLU) zibonise iimpawu ezifanayo. Ngokwahlukileyo, i-disaccharides ivelise iiprofayili ezahlukeneyo: i-MAL kunye ne-SUC kwelinye icala kunye ne-LAC kwelinye. Ngokukodwa, i-MAL yayikuphela kweswekile eyatshintsha iprofayili ye-FA xa kuthelekiswa ne-CEL. Ukongeza, iprofayili ye-MAL yayihluke kakhulu kwiiprofayili ze-FRU kunye ne-GLU. Ngokukodwa, iprofayili ye-MAL ibonise umlinganiselo ophezulu we-C12: 0 (54.59 ± 2.17%), okwenza ukuba ifaniswe ne-CEL (43.10 ± 5.01%), i-LAC (43.35 ± 1.31%), i-FRU (48.90 ± 1.97%) kunye I-GLU (48.38 ± 2.17%) iiprofayili (jonga i-Supplementary Uluhlu S1). I-spectrum ye-MAL ikwabonise owona mxholo usezantsi we-C18:1n9 (9.52 ± 0.50%), owayahlula ngakumbi kwi-LAC (12.86 ± 0.52%) kunye ne-CEL (12.40 ± 1.31%). Umkhwa ofanayo wabonwa kwi-C16: 0. Kwicandelo lesibini, i-spectrum ye-LAC ibonise umxholo ophezulu we-C18: 2n6 (17.22 ± 0.46%), ngelixa i-MAL ibonise ephantsi (12.58 ± 0.67%). I-C18: 2n6 nayo yahlula i-LAC kulawulo (CEL), ebonisa amanqanaba aphantsi (13.41 ± 2.48%) (jonga i-Supplementary Table S1).
PCA iyelenqe iprofayili ye-fatty acid yejoni elimnyama libhabha imibungu kunye ne-monosaccharides eyahlukeneyo (i-fructose, i-galactose, i-glucose), i-disaccharides (i-lactose, i-maltose, i-sucrose) kunye neselulosi njengolawulo.
Ukufunda iziphumo zesondlo zeeshukela ezinyibilikayo kwi-H. illucens larvae, i-cellulose (CEL) kwisondlo senkukhu yatshintshwa nge-glucose (GLU), i-fructose (FRU), i-galactose (GAL), i-maltose (MAL), i-sucrose (SUC), kunye lactose (LAC). Nangona kunjalo, i-monosaccharides kunye ne-disaccharides zineziphumo ezahlukeneyo kuphuhliso, ukusinda, kunye nokwakhiwa kwemibungu ye-HF. Ngokomzekelo, i-GLU, i-FRU, kunye neefom zabo ze-disaccharide (MAL kunye ne-SUC) zenze iziphumo ezixhasayo ezixhasayo ekukhuleni kombungu, okubavumela ukuba bafezekise ubunzima bomzimba bokugqibela obuphezulu kune-CEL. Ngokungafani ne-CEL engenakudliwa, i-GLU, i-FRU, kunye ne-SUC inokudlula umqobo wamathumbu kwaye isebenze njengemithombo ebalulekileyo yezondlo kwizidlo ezicwangcisiweyo16,28. I-MAL ayinabo abathuthi bezilwanyana ezithile kwaye kucingelwa ukuba i-hydrolyzed ukuya kwi-molecule ezimbini ze-glucose ngaphambi kokuba i-assimilation15. Ezi molekyuli zigcinwa kumzimba wezinambuzane njengomthombo wamandla ngokuthe ngqo okanye njenge-lipids18. Okokuqala, ngokumalunga nokugqibela, ezinye zeeyantlukwano ze-intramodal eziqatshelweyo zinokubakho ngenxa yeyantlukwano encinci kwireyishini yesini. Ngokwenene, kwi-H. illucens, ukuzala kusenokuzenzekela: iimazi ezindala ngokwendalo zinendawo ezaneleyo zokubeka amaqanda kwaye zinzima kunamadoda29. Nangona kunjalo, ukuqokelelwa kwe-lipid kwi-BSFL idibanisa kunye nokutya okunyibilikayo kwe-CH2, njengoko kubonwe ngaphambili kwi-GLU kunye ne-xylose26,30. Ngokomzekelo, u-Li et al.30 waphawula ukuba xa i-8% ye-GLU yongezwa kwi-larval diet, umxholo we-lipid we-BSF izibungu zanda nge-7.78% xa kuthelekiswa nokulawula. Iziphumo zethu zihambelana nolu qwalaselo, lubonisa ukuba umxholo we-fat ku-larvae wondla iswekile e-soluble wawuphezulu kune-larvae ezondla ukutya kwe-CEL, xa kuthelekiswa nokunyuka kwe-8.57% kunye ne-GLU supplementation. Okumangalisayo kukuba, iziphumo ezifanayo zabonwa kwiimbungu ezondliwe i-GAL kunye ne-LAC, ngaphandle kwemiphumo emibi ekukhuleni kombungu, ubunzima bomzimba bokugqibela, kunye nokusinda. Imibungu eyondla i-LAC yayincinci kakhulu kunezo zondla ukutya kwe-CEL, kodwa umxholo wabo wamafutha wawuthelekiseka nemibungu eyondla ezinye iiswekile ezinyibilikayo. Ezi ziphumo zibonisa iziphumo eziqhubekayo ze-lactose kwi-BSFL. Okokuqala, ukutya kuqulethe inani elikhulu le-CH. Iinkqubo zokufunxa kunye ne-hydrolysis ye-monosaccharides kunye ne-disaccharides, ngokulandelanayo, inokufikelela kwi-saturation, ibangele iibhotile kwinkqubo yokulinganisa. Ngokuphathelele i-hydrolysis, iqhutyelwa yi-α- kunye ne-β-glucosidases 31. Ezi enzymes ziye zakhetha i-substrates ngokuxhomekeke kubukhulu bazo kunye neebhondi zekhemikhali (α okanye i-β linkages) phakathi kwe-monosaccharides yazo eyi-15. I-Hydrolysis ye-LAC kwi-GLU kunye ne-GAL iqhutyelwa yi-β-galactosidase, i-enzyme umsebenzi wayo ubonakaliswe kumathumbu e-BSF 32. Nangona kunjalo, ukubonakaliswa kwayo kunokungonelanga xa kuthelekiswa nomthamo we-LAC osetyenziswe yimibungu. Ngokwahlukileyo, i-α-glucosidase maltase kunye ne-sucrase 15, eyaziwayo ukuba ibonakaliswe kakhulu kwizinambuzane, ziyakwazi ukuphula izixa ezikhulu ze-MAL kunye ne-sucrose SUC, ngaloo ndlela zinciphisa le mpembelelo yokwanelisa. Okwesibini, iziphumo eziqhubekayo zinokuba ngenxa yokunciphisa ukuvuselela komsebenzi we-amylase yamathumbu ezinambuzane kunye nokucotha kwendlela yokuziphatha yokutya xa kuthelekiswa nolunye unyango. Enyanisweni, i-sugars e-soluble ziye zachongwa njengento evuselela umsebenzi we-enzyme ebalulekileyo kwi-insect digestion, njenge-amylase, kwaye njengezinto ezibangela ukusabela kokutya33,34,35. Iqondo lokuvuselela liyahluka ngokuxhomekeke kwisakhiwo semolekyuli yeswekile. Ngapha koko, i-disaccharides ifuna i-hydrolysis ngaphambi kokufunxwa kwaye ithande ukuvuselela i-amylase ngaphezu kwe-monosaccharides34 yabo. Ngokwahlukileyo, i-LAC inefuthe elincinci kwaye ifunyenwe ingakwazi ukuxhasa ukukhula kwezinambuzane kwiintlobo ezahlukeneyo33,35. Umzekelo, kwi-pest Spodoptera exigua (Boddie 1850), akukho msebenzi we-hydrolytic we-LAC ufunyenwe kwiincindi ze-caterpillar midgut enzymes36.
Ngokumalunga ne-FA spectrum, iziphumo zethu zibonisa iziphumo ezibalulekileyo zokumodareyitha kwe-CH evavanyiweyo. Ngokucacileyo, nangona i-lauric acid (i-C12: 0) ibalwa ngaphantsi kwe-1% ye-FA iyonke ekudleni, yayilawula kuzo zonke iiprofayili (jonga i-Supplementary Table S1). Oku kuhambelana nedatha yangaphambili yokuba i-lauric acid idityaniswe kwi-CH yokutya kwi-H. iillucens ngendlela ebandakanya i-acetyl-CoA carboxylase kunye ne-FA synthase19,27,37. Iziphumo zethu ziqinisekisa ukuba i-CEL ininzi i-indigestible kwaye isebenza "njenge-agent agent" kwi-BSF yokulawula ukutya, njengoko kuxoxwe kwizifundo ezininzi ze-BSFL38,39,40. Ukutshintsha i-CEL nge-monosaccharides kunye ne-disaccharides ngaphandle kwe-LAC kwandisa umlinganiselo we-C12: i-0, ebonisa ukunyuka kwe-CH ngokunyuka kwezibungu. Okubangela umdla kukuba, i-disaccharides ye-MAL kunye ne-SUC ikhuthaza ukuhlanganiswa kwe-lauric acid ngokufanelekileyo ngakumbi kune-monosaccharides yabo, iphakamisa ukuba nangona iqondo eliphezulu le-polymerization ye-GLU kunye ne-FRU, kwaye ekubeni i-Drosophila kuphela kwe-sucrose transporter echongiwe kwiintlobo zeprotheyini yezilwanyana, abathuthi be-disaccharide. Isenokungabikho kumathumbu e-H. illucens larvae15, ukusetyenziswa kwe- I-GLU kunye ne-FRU yanda. Nangona kunjalo, nangona i-GLU kunye ne-FRU ngokwethiyori zenziwe ngokulula ngakumbi nge-metabolism yi-BSF, zikwaye zenziwa ngokulula ngakumbi nge-substrates kunye ne-gut microorganisms, ezinokubangela ukuthotywa kwazo ngokukhawuleza kunye nokuncipha kokusetyenziswa yimibungu xa kuthelekiswa needisaccharides.
Ekuqaleni, umxholo we-lipid we-larvae ezondliwe i-LAC kunye ne-MAL yayithelekiseka, ebonisa ukufumaneka okufanayo kwe-bioavailability kwezi swekile. Nangona kunjalo, ngokumangalisayo, iphrofayili ye-FA ye-LAC yayityebile kwi-SFA, ngakumbi nge-C12 ephantsi: umxholo we-0, xa kuthelekiswa ne-MAL. Enye ingcamango yokuchaza lo mahluko kukuba i-LAC inokuvuselela i-bioaccumulation ye-FA yokutya nge-acetyl-CoA FA synthase. Ukuxhasa le ngcamango, imibungu ye-LAC yayinomlinganiselo ophantsi we-decanoate (C10: 0) (0.77 ± 0.13%) kunokutya kwe-CEL (1.27 ± 0.16%), ebonisa ukunciphisa i-FA synthase kunye nemisebenzi ye-thioesterase19. Okwesibini, i-acids ye-fatty acids ithathwa njengento ephambili echaphazela ukubunjwa kwe-SFA ye-H. illucens27. Kwiimvavanyo zethu, i-linoleic acid (i-C18: i-2n6) ibalwa kwi-54.81% ye-fatty acids yokutya, kunye nomlinganiselo we-LAC izibungu ziyi-17.22 ± 0.46% kunye ne-MAL 12.58 ± 0.67%. I-Oleic acid (cis + trans C18: 1n9) (23.22% ekudleni) ibonise indlela efanayo. Umlinganiselo we-α-linolenic acid (C18: 3n3) iphinde ixhase i-hypothesis ye-bioaccumulation. Le asidi enamafutha iyaziwa ukuba iqokelele kwi-BSFL phezu kokutyebisa kwe-substrate, njengokongezwa kwekhekhe le-flaxseed, ukuya kuthi ga kwi-6-9% ye-fatty acids iyonke kwi-larvae19. Kwizidlo ezityetyisiweyo, i-C18: i-3n3 inokuthi ifikelele kwi-35% ye-fatty acids acids. Nangona kunjalo, kwisifundo sethu, i-C18: i-3n3 ibalwa kuphela i-2.51% yeprofayili ye-fatty acid. Nangona umlinganiselo ofunyenwe kwindalo wawungaphantsi kwimibungu yethu, lo mlinganiso wawuphezulu kwi-LAC (0.87 ± 0.02%) kune-MAL (0.49 ± 0.04%) (p <0.001; jonga i-Supplementary Table S1). Ukutya kwe-CEL kunomlinganiselo ophakathi we-0.72 ± 0.18%. Ekugqibeleni, i-acid palmitic (C16: 0) kwi-CF larvae ibonisa igalelo leendlela zokwenziwa kunye nokutya kwe-FA19. Hoc et al. I-19 yaqaphela ukuba i-C16: i-0 synthesis yancitshiswa xa ukutya kwakutyetyiswe ngesidlo se-flaxseed, okwakubangelwa ukuncipha kokufumaneka kwe-acetyl-CoA substrate ngenxa yokunciphisa umlinganiselo we-CH. Okumangalisayo kukuba, nangona izidlo zombini zinomxholo ofanayo we-CH kwaye i-MAL ibonisa i-bioavailability ephezulu, izibungu ze-MAL zibonise umlinganiselo ophantsi we-C16: 0 (10.46 ± 0.77%), kanti i-LAC ibonise umlinganiselo ophezulu, i-accounting ye-12.85 ± 0.27% (p <0.05; bona ITheyibhile eyoNgezelelweyo S1). Ezi ziphumo zibonisa impembelelo enzima yezondlo kwi-BSFL yokugaya kunye nemetabolism. Okwangoku, uphando ngesi sihloko lucokiseke ngakumbi kwiLepidoptera kuneDiptera. Kwimibungu, i-LAC ichongiwe njengento evuselelayo ebuthathaka yokuziphatha kokutya xa kuthelekiswa nezinye iishukela ezinyibilikayo ezifana ne-SUC kunye ne-FRU34,35. Ngokukodwa, kwi-Spodopteralittoralis (i-Boisduval 1833), ukusetyenziswa kwe-MAL kuvuselele umsebenzi we-amylolytic emathunjini ukuya kumlinganiselo omkhulu kune-LAC34. Imiphumo efanayo kwi-BSFL inokuchaza ukuvuselela okuphuculweyo kwe-C12: indlela ye-0 yokwenziwa kwi-larvae ye-MAL, ehambelana nokunyuka kwamathumbu e-CH, ukondla ixesha elide, kunye nesenzo se-amylase yamathumbu. Ukuvuselela okuncinci kwesingqisho sokutyisa phambi kwe-LAC kunokuchaza ukukhula okucothayo kwemibungu ye-LAC. Ngaphezu koko, uLiu Yanxia et al. I-27 yaqaphela ukuba ubomi beshelufu ye-lipids kwi-H. illucens substrates yayinde kune-CH. Ke ngoko, imibungu ye-LAC inokuthembela ngakumbi kwi-lipids yokutya ukugqiba uphuhliso lwabo, olunokunyusa umxholo wabo wokugqibela we-lipid kunye nokulungelelanisa iprofayili yabo ye-asidi enamafutha.
Ngokolwazi lwethu, zizifundo ezimbalwa kuphela eziye zavavanya iziphumo ze-monosaccharide kunye ne-disaccharide eyongeziweyo kukutya kwe-BSF kwiiprofayili zabo ze-FA. Okokuqala, uLi et al. I-30 ivavanye imiphumo ye-GLU kunye ne-xylose kwaye yabona amanqanaba e-lipid afana neyethu kwi-8% yokongeza. Iprofayile ye-FA yayingacaciswanga kwaye iqulathe ikakhulu i-SFA, kodwa akukho mahluko ufunyenwe phakathi kweeswekile ezimbini okanye xa zaziswa ngaxeshanye30. Ngapha koko, uCohn et al. I-41 ayizange ibonise umphumo we-20% ye-GLU, i-SUC, i-FRU kunye ne-GAL ukongeza kwisondlo senkukhu kwiiprofayili ze-FA ezifanelekileyo. Le mifanekiso ifunyenwe kwi-technical kunokuba i-biological replicates, leyo, njengoko ichazwe ngababhali, inokunciphisa uhlalutyo lwamanani. Ngaphezu koko, ukungabikho kolawulo lwe-iso-sugar (ukusebenzisa i-CEL) kunciphisa ukuchazwa kweziphumo. Kutshanje, izifundo ezimbini zikaNugroho RA et al. ubonise izinto ezingaqhelekanga kwi-FA spectra42,43. Kwisifundo sokuqala, uNugroho RA et al. I-43 ivavanye umphumo wokongeza i-FRU kwisidlo se-palm kernel ebilisiweyo. Iprofayili ye-FA yeembungu ezibangelwayo zibonise amanqanaba aphezulu ngokungaqhelekanga e-PUFA, ngaphezu kwe-90% ephuma kukutya okuqukethe i-10% FRU (efana nokufunda kwethu). Nangona oku kutya kuqulethe iipellet zentlanzi ezityebileyo ze-PUFA, ixabiso leprofayili ye-FA exeliweyo ye-larvae ekulawuleni ukutya okubandakanya i-100% ye-PCM evundisiweyo yayingahambelani nayo nayiphi na iprofayili exelwe ngaphambili, ngakumbi inqanaba elingaqhelekanga le-C18: 3n3 ye-17.77 ± 1.67% kunye ne-26.08 ± 0.20% ye-conjugated linoleic acid (C18: 2n6t), i-isomer enqabileyo ye-linoleic acid. Uphononongo lwesibini lubonise iziphumo ezifanayo ezibandakanya i-FRU, i-GLU, i-MAL kunye ne-SUC42 kwi-fermented palm kernel meal. Ezi zifundo, njengezethu, ziqaqambisa ubunzima obukhulu ekuthelekiseni iziphumo ezivela kulingo lokutya kombungu we-BSF, njengokhetho lolawulo, ukusebenzisana neminye imithombo yezondlo, kunye neendlela zohlalutyo lwe-FA.
Ngexesha lovavanyo, siqaphele ukuba umbala kunye nevumba le-substrate yahluka ngokuxhomekeke kukutya okusetyenzisiweyo. Oku kuphakamisa ukuba i-microorganisms inokudlala indima kwiziphumo ezibonwe kwi-substrate kunye nenkqubo yokugaya i-larvae. Ngapha koko, i-monosaccharides kunye ne-disaccharides zixutywa ngokulula nge-colonizing microorganisms. Ukusetyenziswa ngokukhawuleza kweswekile enyibilikayo ngama microorganisms kunokubangela ukukhutshwa kweemveliso ezininzi ze-microbial metabolic ezifana ne-ethanol, i-lactic acid, i-short-chain fatty acids (umz. i-acetic acid, i-propionic acid, i-butyric acid) kunye ne-carbon dioxide44. Ezinye zezi zixhobo zinokuba noxanduva lweziphumo ezinobuthi ezinobuthi kwimibungu ebonwa nguCohn et al.41 phantsi kweemeko ezifanayo zophuhliso. Umzekelo, i-ethanol iyingozi kwizinambuzane45. Izixa ezikhulu zokukhutshwa kwekharbon diokside kunokubangela ukuqokelelana kwayo emazantsi etanki, nto leyo enokuthi ivimbe umoya we-oxygen ukuba ukujikeleza komoya akuvumeli ukukhululwa kwayo. Ngokumalunga ne-SCFAs, iimpembelelo zabo kwiinambuzane, ngakumbi i-H. illucens, zihlala zingaqondwa kakuhle, nangona i-lactic acid, i-propionic acid, kunye ne-butyric acid ibonakaliswe ukuba iyabulala kwiCallosobruchus maculatus (Fabricius 1775)46. Kwi-Drosophila melanogaster Meigen 1830, ezi SCFAs ziziphawuli ezivumbayo ezikhokela amabhinqa kwiindawo zokubeka i-oviposition, ecebisa indima enenzuzo kuphuhliso lombungu47. Nangona kunjalo, i-acetic acid ihlelwa njengento enobungozi kwaye inokuthintela ngokubonakalayo ukukhula kombungu47. Ngokwahlukileyo koko, i-lactate ephuma kwi-microbially isandula kufunyaniswa inefuthe lokukhusela kwi-invasive gut microbes kwi-Drosophila48. Ngaphezu koko, i-microorganisms kwinkqubo yokugaya idlala indima kwi-CH digestion kwizinambuzane49. Imiphumo ye-Physiological ye-SCFAs kwi-gut microbiota, efana nesantya sokutya kunye ne-gene expression, ichazwe kwii-vertebrates ze-50. Basenokuba nefuthe le-trophic kwi-H. illucens larvae kwaye banokuba negalelo ngokuyinxenye kulawulo lweeprofayili ze-FA. Izifundo malunga nemiphumo yesondlo yale mveliso ye-microbial fermentation iya kucacisa imiphumo yazo kwi-H. ilucens isondlo kwaye inike isiseko sezifundo zexesha elizayo kwii-microorganisms eziluncedo okanye ezinobungozi malunga nophuhliso lwabo kunye nexabiso le-FA-rich substrates. Kule nkalo, indima ye-microorganisms kwiinkqubo zokugaya izinambuzane ezifuywa ngobuninzi zikhula ngokuqhubekayo. Izinambuzane ziqala ukujongwa njenge-bioreactors, zibonelela nge-pH kunye neemeko ze-oxygenation eziququzelela ukuphuhliswa kwe-microorganisms ezikhethekileyo ekuthotyweni okanye ukuchithwa kwezondlo ezinzima kwiinambuzane zokugaya i-51. Kungekudala, i-Xiang et al.52 ibonise ukuba, ngokomzekelo, i-inoculation yenkunkuma yezinto eziphilayo kunye nomxube webhaktheriya ivumela i-CF ukutsala iibhaktheriya ezikhethekileyo kwi-lignocellulose degradation, ukuphucula ukuthotywa kwayo kwi-substrate xa kuthelekiswa ne-substrates ngaphandle kwemibungu.
Ekugqibeleni, malunga nokusetyenziswa okuzuzisayo kwenkunkuma yezinto eziphilayo ngu-H. illucens, i-CEL kunye ne-SUC yokutya ivelise inani eliphezulu leembungu ngosuku. Oku kuthetha ukuba ngaphandle kobunzima bokugqibela obusezantsi bomntu ngamnye, ubunzima bubonke bombungu obuveliswe kwi-substrate equkethe i-CH engagayekiyo buthelekiseka naleyo ifunyenwe kwi-homosaccharide yokutya equlethe i-monosaccharides kunye ne-disaccharides. Kuphononongo lwethu, kubalulekile ukuba uqaphele ukuba amanqanaba ezinye izondlo zanele ukuxhasa ukukhula kombungu kunye nokuba ukongezwa kwe-CEL kufuneka kukhawulelwe. Nangona kunjalo, ukubunjwa kokugqibela kwemibungu kuyahluka, kugxininisa ukubaluleka kokukhetha isicwangciso esifanelekileyo sokuqhayisa izinambuzane. Izibungu ze-CEL ezondliwa ngokutya okupheleleyo zifaneleke ngakumbi ukusetyenziswa njengesondlo sezilwanyana ngenxa yokuqukethwe kwamafutha aphantsi kunye namanqanaba aphantsi e-lauric acid, kanti i-larvae ezondliwa nge-SUC okanye i-MAL yokutya ifuna ukuchithwa ngokucinezela ukunyusa ixabiso leoli, ngakumbi kwi-biofuel. icandelo. I-LAC ifumaneka kwimveliso yobisi kwimveliso yobisi efana ne-whey evela kwimveliso yesonka samasi. Kutshanje, ukusetyenziswa kwayo (3.5% lactose) kuphuculwe ubunzima bomzimba bombungu bokugqibela53. Nangona kunjalo, ukutya okulawulwayo kwesi sifundo kuqulethe isiqingatha se-lipid content. Ngoko ke, iziphumo eziqhubekayo ze-LAC zinokuthi zichaswe yi-larval bioaccumulation ye-lipids yokutya.
Njengoko kubonisiwe kwizifundo zangaphambili, iipropathi ze-monosaccharides kunye ne-disaccharides zichaphazela kakhulu ukukhula kwe-BSFL kwaye zimodareyitha iprofayili yayo ye-FA. Ngokukodwa, i-LAC ibonakala idlala indima eqhubekayo ngexesha lokuphuhliswa kombungu ngokunciphisa ukufumaneka kwe-CH ye-dietary lipid absorption, ngaloo ndlela ikhuthaza i-UFA bioaccumulation. Kulo mongo, kuya kuba nomdla ukwenza i-bioassays usebenzisa izidlo ezidibanisa i-PUFA kunye ne-LAC. Ngaphaya koko, indima yee-microorganisms, ngakumbi indima ye-microbial metabolites (efana ne-SCFAs) ephuma kwiinkqubo zokuvutshelwa kweswekile, ihlala isisihloko sophando esifanele ukuphandwa.
Izinambuzane zifunyenwe kwikholoni ye-BSF yeLabhoratri ye-Functional and Evolutionary Entomology esekwe kwi-2017 kwi-Agro-Bio Tech, i-Gembloux, eBelgium (ukufumana iinkcukacha ezingaphezulu malunga neendlela zokukhulisa, jonga i-Hoc et al. 19). Kulingo lovavanyo, i-2.0 g yamaqanda e-BSF aqokelelwa ngokungenamkhethe yonke imihla ukusuka kwiikheji zokuzala kwaye afukanywa kwi-2.0 kg ye-70% yokutya okumanzi kwenkukhu (Aveve, Leuven, Belgium). Kwiintsuku ezintlanu emva kokuqanduselwa, imibungu yahlulwa kwi-substrate kwaye ibalwe ngesandla ngeenjongo zokulinga. Ubunzima bokuqala bebhetshi nganye bulinganisiwe. Umyinge wobunzima bomntu ngamnye wawuyi-7.125 ± 0.41 mg, kwaye umyinge wonyango ngalunye uboniswa kwiTheyibhile eyoNgezelelweyo ye-S2.
Ukuqulunqwa kokutya kwaguqulwa kwisifundo sikaBarragan-Fonseca et al. 38 . Ngokufutshane, i-compromise ifunyenwe phakathi komgangatho ofanayo wokutya kwiinkukhu ze-larval, umxholo ofanayo owomileyo (DM) umxholo, i-CH ephezulu (i-10% esekelwe kwisidlo esitsha) kunye nokuthungwa, ekubeni i-sugars elula kunye ne-disaccharides ayinayo impahla yesicatshulwa. Ngokutsho kolwazi lomenzi (i-Chicken Feed, i-AVEVE, i-Leuven, eBelgium), i-CH evavanyiweyo (okt iswekile e-soluble) yongezwa ngokwahlukileyo njengesisombululo se-autoclaved aqueous (15.9%) kwi-diet equkethe i-16.0% yeprotheyini, i-5.0% ye-lipids iyonke, I-11.9% yokutya kwenkukhu egayiweyo equka uthuthu kunye ne-4.8% yefayibha. Kwisitya ngasinye se-750 ml (17.20 × 11.50 × 6.00 cm, AVA, Tempsee, Belgium), 101.9 g ye-autoclaved CH isisombululo ixutywe kunye ne-37.8 g yokutya kwenkukhu. Kwisidlo ngasinye, umxholo owomileyo wawuyi-37.0%, kubandakanywa iprotheni e-homogeneous (11.7%), i-lipids e-homogeneous (3.7%) kunye ne-sugar homogeneous (26.9% ye-CH eyongeziweyo). Uvavanyo lwe-CH lwaluyi-glucose (GLU), i-fructose (FRU), i-galactose (GAL), i-maltose (MAL), i-sucrose (SUC) kunye ne-lactose (LAC). Ukutya okulawulwayo kwakuquka i-cellulose (CEL), ethathwa njengento engavumelekanga kwi-H. illucens larvae 38. Iimbungu ezilikhulu zeentsuku ezi-5 zifakwe kwi-tray efakwe isiciko esinomngxuma oyi-1 cm phakathi kwaye igqunywe ngeplastiki ye-mosquito net. Ukutya ngakunye kwaphindwa izihlandlo ezine.
Ubunzima bombungu bulinganiswe kwiintsuku ezintathu emva kokuqala kovavanyo. Ngomlinganiselo ngamnye, i-larvae ye-20 yasuswa kwi-substrate isebenzisa amanzi afudumele angenazintsholongwane kunye ne-forceps, yomisiwe, kwaye ilinganiswe (STX223, Ohaus Scout, Parsippany, USA). Emva kokulinganisa, imibungu ibuyiselwa kumbindi we-substrate. Imilinganiselo yayithathwa rhoqo kathathu ngeveki de kwavela i-prepupa yokuqala. Kweli nqanaba, qokelela, ubale, kwaye ulinganise yonke imibungu njengoko kuchaziwe ngaphambili. Inqanaba elahlukileyo lesi-6 lemibungu (okt, imibungu emhlophe ehambelana nenqanaba lombungu elandulela inqanaba lokukhula) kunye ne-prepupae (okt, inqanaba lokugqibela lombungu apho imibungu ye-BSF ijika ibe mnyama, iyeke ukutyisa, kwaye ifune indawo elungele ukuguquguquka kwemibungu) kwaye igcine kwindawo - I-18 ° C yokuhlalutya ukwakheka. Isivuno sasibalwe njengomlinganiselo wobunzima obupheleleyo bezinambuzane (imibungu kunye ne-prepupae yesigaba sesi-6) efunyenwe kwisitya ngasinye (g) kwixesha lophuhliso (d). Onke amaxabiso aphakathi kwisicatshulwa abonakaliswe ngolu hlobo: ithetha ± SD.
Onke amanyathelo alandelayo asebenzisa i-solvents (i-hexane (i-Hex), i-chloroform (CHCl3), i-methanol (i-MeOH)) yenziwa phantsi kwe-fume hood kwaye ifuna ukunxiba iiglavu ze-nitrile, ii-apron kunye neziglasi zokhuseleko.
Iimbungu ezimhlophe zomiswa kwi-FreeZone6 i-freeze dryer (i-Labconco Corp., i-Kansas City, i-MO, i-USA) i-72 h kwaye yaphantsi (IKA A10, e-Staufen, eJamani). I-lipids iyonke yatsalwa kwi-± 1 g yomgubo kusetyenziswa indlela ye-Folch 54. Isiqulatho sokufuma esishiyekileyo sesampulu nganye ene-lyophilized yamiselwa ngokuphindwe kabini kusetyenziswa isihlalutyi sokufuma (MA 150, Sartorius, Göttiggen, Germany) ukulungisa i-lipids iyonke.
Iilipids zizonke ziye zaguqulwa phantsi kweemeko ze-acidic ukufumana i-fatty acid methyl esters. Ngokufutshane, malunga ne-10 mg lipids/100 µl isisombululo se-CHCl3 (100 µl) satshatyalaliswa ngenitrogen kwityhubhu eyi-8 ml yePyrex© (SciLabware – DWK Life Sciences, London, UK). I-tube ifakwe kwi-Hex (0.5 ml) (PESTINORM®SUPRTRACE n-Hexane> 95% yohlalutyo lwe-organic trace, i-VWR Chemicals, Radnor, PA, USA) kunye ne-Hex / MeOH / BF3 (20/25/55) isisombululo (0.5) ml) kwindawo yokuhlambela amanzi kwi-70 °C nge-90 min. Emva kokupholisa, i-10% isisombululo se-H2SO4 samanzi (0.2 ml) kunye nesisombululo esigcweleyo se-NaCl (0.5 ml) songezwa. Xuba ityhubhu kwaye ugcwalise umxube ngeHex ecocekileyo (8.0 mL). Inxalenye yesigaba esiphezulu idluliselwe kwi-vial kwaye ihlalutywe nge-chromatography yegesi kunye ne-flame ionization detector (GC-FID). Iisampulu zahlaziywa kusetyenziswa i-Trace GC Ultra (i-Thermo Scientific, Waltham, MA, USA) exhotywe nge-injector ekwahlulayo / engenakuqhekeka (240 ° C) kwimodi yokwahlula (i-split flow: 10 mL / min), ikholomu ye-Stabilwax®-DA ( 30 m, 0.25 mm id, 0.25 μm, Restek Corp., Bellefonte, PA, USA) kunye ne-FID (250 °C). Inkqubo yokushisa ibekwe ngolu hlobo lulandelayo: i-50 ° C kwi-1 min, inyuka ukuya kwi-150 ° C kwi-30 ° C / min, inyuka ukuya kwi-240 ° C kwi-4 ° C / min kwaye iqhubeke kwi-240 ° C kwi-5 min. I-Hex isetyenziswe njengento engenanto kunye nomgangatho wesalathiso oqukethe i-37 fatty acid methyl esters (i-Supelco 37-component FAMEmix, Sigma-Aldrich, Overijse, Belgium) yayisetyenziselwa ukuchongwa. Ukuchongwa kwe-unsaturated fatty acids (UFAs) kwaqinisekiswa yi-GC ebanzi ye-dimensional GC (GC × GC-FID) kunye nobukho be-isomers buchongwe ngokuchanekileyo ngokuguqulwa okuncinci kwendlela ye-Ferrara et al. 55. Iinkcukacha zesixhobo zifumaneka kwi-Supplementary Table S3 kunye neziphumo kwi-Supplementary Figure S5.
Idatha inikezelwe kwifomati yespredishithi ye-Excel (Microsoft Corporation, Redmond, WA, USA). Uhlalutyo lwamanani lwenziwa kusetyenziswa i-R Studio (inguqulo 2023.12.1 + 402, Boston, USA) 56. Idatha ngesisindo sobubungu, ixesha lophuhliso kunye nemveliso yaqikelelwa kusetyenziswa imodeli yomgca (LM) (umyalelo "lm", i-R package "stats" 56) njengoko ihambelana nokusabalalisa kweGaussian. Amazinga okusinda usebenzisa uhlalutyo lwemodeli ye-binomial aqikelelwa kusetyenziswa imodeli yomgca jikelele (GLM) (i-command "glm", iphakheji ye-R "lme4" 57). Isiqhelo kunye ne-homoscedasticity yaqinisekiswa ngokusebenzisa uvavanyo lwe-Shapiro (umyalelo "shapiro.test", iphakheji ye-R "izibalo" ze-56) kunye nohlalutyo lokuhluka kwedatha (umyalelo we-betadisper, iphakheji ye-R "vegan" 58). Emva kokuhlalutya ngokubini kwee-p-values ezibalulekileyo (p <0.05) ukusuka kuvavanyo lwe-LM okanye lwe-GLM, ukungafani okukhulu phakathi kwamaqela kufunyenwe kusetyenziswa uvavanyo lwe-EMM (umyalelo "emmeans", iphakheji ye-R "emmeans" 59).
Umboniso opheleleyo we-FA uthelekiswe kusetyenziswa uhlalutyo lwemvume ye-multivariate yomahluko (okt permMANOVA; umyalelo “adonis2”, iphakheji ye-R “vegan” 58) kusetyenziswa umgama we-Euclidean matrix kunye neemvume ezingama-999. Oku kunceda ukuchonga i-acids ezinamafutha aphenjelelwa yindalo ye-carbohydrates yokutya. Umahluko omkhulu kwiiprofayili ze-FA zahlaziywa ngakumbi kusetyenziswa uthelekiso lwesibini. Emva koko idatha yabonwa kusetyenziswa uhlalutyo lwecandelo eliphambili (PCA) (umyalelo "PCA", iphakheji ye-R "FactoMineR" 60). I-FA enoxanduva lwale mahluko ichongiwe ngokutolika izangqa ezihambelanayo. Aba baviwa baqinisekiswa ngokusebenzisa uhlalutyo lwendlela enye ye-variance (ANOVA) (i-command "aov", iphakheji ye-R "izibalo" 56 ) ilandelwa luvavanyo lwe-post hoc ka-Tukey (umyalelo we-TukeyHSD, iphakheji ye-R "izibalo" ze-56). Ngaphambi kokuhlalutya, ukuqheleka kwavavanywa kusetyenziswa uvavanyo lwe-Shapiro-Wilk, i-homoscedasticity ihlolwe ngokusebenzisa uvavanyo lwe-Bartlett (i-command "bartlett.test", iphakheji ye-R "izibalo" ze-56), kunye nendlela ye-nonparametric isetyenziswe ukuba akukho nanye yeengcamango ezimbini. . Ucazululo luthelekiswe (umyalelo "kruskal.test", iphakheji ye-R "izibalo" 56 ), kwaye emva koko iimvavanyo ze-Dunn ze-post hoc zasetyenziswa (command dunn.test, iphakheji ye-R "dunn.test" 56).
Inguqulelo yokugqibela yombhalo-ngqangi yahlolwa kusetyenziswa uMhleli weGrammarly njengomhloli wesiNgesi wesiNgesi ( Grammarly Inc., San Francisco, California, USA) 61 .
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Ixesha lokuposa: Dec-25-2024