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Ntchentche ya msilikali wakuda (Hermetia illucens, L. 1758) ndi tizilombo toononga kwambiri tomwe timatha kugwiritsa ntchito zinthu zopangidwa ndi carbohydrate. Pakati pazakudya, ntchentche zankhondo zakuda zimadalira shuga wosungunuka kuti akule komanso kaphatikizidwe ka lipid. Cholinga cha kafukufukuyu chinali kuwunika zotsatira za shuga wosungunuka wamba pakukula, kukhala ndi moyo, komanso mbiri yamafuta amtundu wa ntchentche zankhondo zakuda. Onjezerani chakudya cha nkhuku ndi monosaccharides ndi ma disaccharides mosiyana. Cellulose idagwiritsidwa ntchito ngati chowongolera. Mphutsi zodyetsa shuga, fructose, sucrose, ndi maltose zidakula mwachangu kuposa mphutsi zowongolera. Mosiyana ndi zimenezi, lactose inali ndi zotsatira zopitirizabe pa mphutsi, kuchepetsa kukula ndi kuchepetsa kulemera kwa thupi. Komabe, mashuga onse osungunuka amapangitsa mphutsi kukhala zonenepa kuposa zomwe zimadyetsa chakudya chowongolera. Makamaka, mashuga oyesedwa adapanga mawonekedwe amafuta acid. Maltose ndi sucrose zimachulukitsa kuchuluka kwamafuta acid poyerekeza ndi cellulose. Mosiyana ndi zimenezi, lactose inachulukitsa bioaccumulation ya zakudya unsaturated mafuta zidulo. Kafukufukuyu ndi woyamba kusonyeza zotsatira za shuga wosungunuka pamafuta a asidi amtundu wa mphutsi za ntchentche za msilikali wakuda. Zotsatira zathu zikuwonetsa kuti ma carbohydrate oyesedwa amakhudza kwambiri kuchuluka kwa mafuta amtundu wa mphutsi zamtundu wakuda wankhondo ndipo atha kudziwa momwe angagwiritsire ntchito pomaliza.
Kufunika kwapadziko lonse kwa mphamvu ndi mapuloteni a nyama kukupitilira 1. Pankhani ya kutentha kwa dziko, ndikofunikira kupeza njira zina zobiriwira m'malo mwa mphamvu zamafuta ndi njira zachikhalidwe zopangira chakudya ndikuwonjezera kupanga. Tizilombo tikulonjeza ofuna kuthana ndi mavutowa chifukwa cha kuchepa kwa mankhwala komanso kuwonongeka kwa chilengedwe poyerekeza ndi ulimi wa ziweto zakale2. Pakati pa tizilombo, munthu wabwino kwambiri kuti athetse vutoli ndi ntchentche yakuda ya msilikali (BSF), Hermetia illucens (L. 1758), mitundu yowononga yomwe imatha kudyetsa mitundu yosiyanasiyana ya organic substrates3. Chifukwa chake, kulemekeza magawo awa kudzera mu kuswana kwa BSF kumatha kupanga gwero latsopano la zida zopangira kuti zikwaniritse zosowa zamafakitale osiyanasiyana.
Mphutsi za BSF (BSFL) zimatha kudya zinthu zaulimi ndi zaulimi monga tirigu, zotsalira za masamba, zamkati za zipatso ndi mkate wakale, zomwe ndizofunikira kwambiri pakukula kwa BSFL chifukwa cha kuchuluka kwa carbohydrate (CH) 4,5, 6 zomwe. Kupanga kwakukulu kwa BSFL kumapangitsa kupanga zinthu ziwiri: ndowe, chisakanizo cha zotsalira za gawo lapansi ndi ndowe zomwe zingagwiritsidwe ntchito ngati feteleza wa kulima mbewu7, ndi mphutsi, zomwe zimapangidwa makamaka ndi mapuloteni, lipids ndi chitin. Mapuloteni ndi lipids amagwiritsidwa ntchito makamaka paulimi wa ziweto, biofuel ndi zodzoladzola8,9. Ponena za chitin, biopolymer iyi imapeza ntchito mu gawo lazakudya, biotechnology ndi chisamaliro chaumoyo10.
BSF ndi tizilombo tomwe timakhala tomwe timapanga holometabolous, kutanthauza kuti kusintha kwake ndi kuberekana, makamaka magawo owononga mphamvu pa moyo wa tizilombo, amatha kuthandizidwa ndi nkhokwe zosungirako zakudya zomwe zimapangidwa panthawi ya kukula kwa mphutsi11. Makamaka, kaphatikizidwe ka mapuloteni ndi lipid kumabweretsa kukula kwa thupi lamafuta, chiwalo chofunikira chosungiramo mphamvu chomwe chimatulutsa mphamvu panthawi yopanda chakudya cha BSF: prepupa (ie, gawo lomaliza la mphutsi pomwe mphutsi za BSF zimasanduka zakuda podyetsa ndikusaka. kwa malo oyenera kusintha kusintha kwa zinthu), pupae (ie, siteji yosasunthika pamene tizilombo timapanga masinthidwe), ndi akuluakulu12,13. CH ndiye gwero lalikulu lamphamvu muzakudya za BSF14. Pakati pa zakudya izi, fibrous CH monga hemicellulose, cellulose ndi lignin, mosiyana ndi ma disaccharides ndi polysaccharides (monga wowuma), sangathe kudyetsedwa ndi BSFL15,16. Kugaya kwa CH ndi gawo lofunikira poyambira kuyamwa kwa ma carbohydrate, omwe pamapeto pake amapangidwa ndi hydrolyzed kukhala shuga wosavuta m'matumbo16. Mashuga osavuta amatha kuyamwa (mwachitsanzo, kudzera m'matumbo a m'mimba) ndikusinthidwa kuti apange mphamvu17. Monga tafotokozera pamwambapa, mphutsi zimasunga mphamvu zochulukirapo monga lipids m'thupi lamafuta12,18. Ma lipids osungira amakhala ndi ma triglycerides (ma lipids osalowerera omwe amapangidwa kuchokera ku molekyulu imodzi ya glycerol ndi mafuta atatu amafuta acid) opangidwa ndi mphutsi kuchokera ku shuga wosavuta wazakudya. CH izi zimapereka magawo a acetyl-CoA ofunikira kuti mafuta a asidi (FA) biosynthesis apangidwe kudzera mumafuta acid synthase ndi thioesterase pathways19. Mafuta amtundu wa H. illucens lipids mwachibadwa amakhala ndi mafuta odzaza mafuta (SFA) omwe ali ndi lauric acid (C12: 0) 19,20. Chifukwa chake, kuchuluka kwa lipids komanso mafuta a asidi amafuta akuchulukirachulukira kukhala zinthu zomwe zimalepheretsa kugwiritsa ntchito mphutsi pazakudya zanyama, makamaka m'zamoyo zam'madzi momwe ma polyunsaturated fatty acids (PUFA) amafunikira21.
Chiyambireni kupezeka kwa kuthekera kwa BSFL kuchepetsa zinyalala za organic, kafukufuku wokhudzana ndi mtengo wazinthu zosiyanasiyana zapakhomo awonetsa kuti kapangidwe ka BSFL kumayendetsedwa ndi zakudya zake. Pakalipano, malamulo a FA mbiri ya H. illucens akupitirizabe kusintha. Kuthekera kwa BSFL kupanga bioaccumulate PUFA kwawonetsedwa pazigawo zolemera za PUFA monga algae, zinyalala za nsomba, kapena zakudya monga flaxseed, zomwe zimapereka mbiri yapamwamba ya FA pazakudya zanyama19,22,23. Mosiyana ndi izi, pazogulitsa zomwe sizinalemeredwe mu PUFA, sinthawi zonse pali kulumikizana pakati pazakudya za FA ndi FA larval, zomwe zikuwonetsa mphamvu ya zakudya zina24,25. M'malo mwake, zotsatira za digestible CH pazambiri za FA zimakhalabe zomveka bwino komanso zosakanika24,25,26,27.
Monga momwe tikudziwira, ngakhale kuti ma monosaccharides ndi ma disaccharides ochuluka ali ochuluka muzakudya za H. illucens, gawo lawo lazakudya silidziwika bwino mu H. ilucens zakudya. Cholinga cha kafukufukuyu chinali kufotokozera zotsatira zake pazakudya za BSFL komanso kapangidwe ka lipid. Tidzawunikanso kukula, kupulumuka, ndi kutulutsa kwa mphutsi pansi pazakudya zosiyanasiyana. Kenako, tidzafotokozera zamafuta amafuta ndi mbiri yamafuta amafuta pazakudya zilizonse kuti tiwonetse zotsatira za CH pazakudya za BSFL.
Tinkaganiza kuti mtundu wa CH woyesedwa ungakhudze (1) kukula kwa mphutsi, (2) kuchuluka kwa lipids, ndi (3) kusintha mbiri ya FA. Ma monosaccharides amatha kuyamwa mwachindunji, pomwe ma disaccharides ayenera kukhala ndi hydrolyzed. Monosaccharides motero amapezeka ngati magwero achindunji amphamvu kapena zotsogola za lipogenesis kudzera mu njira za FA synthase ndi thioesterase, potero zimakulitsa kukula kwa mphutsi za H. illucens ndikulimbikitsa kudzikundikira kwa lipids (makamaka lauric acid).
CH yoyesedwa inakhudza kulemera kwa thupi la mphutsi panthawi ya kukula (mkuyu 1). FRU, GLU, SUC ndi MAL anawonjezera kulemera kwa mphutsi mofanana ndi zakudya zowongolera (CEL). Mosiyana ndi izi, LAC ndi GAL zimawoneka zolepheretsa kukula kwa mphutsi. Makamaka, LAC inali ndi vuto lalikulu pa kukula kwa mphutsi poyerekeza ndi SUC nthawi yonse ya kukula: 9.16 ± 1.10 mg motsutsana ndi 15.00 ± 1.01 mg pa tsiku 3 (F6,21 = 12.77, p <0.001; Mkuyu 1), 125.12 ± 125.11 mg mg ndi 211.79 ± 14.93 mg, motero, pa tsiku 17 (F6,21 = 38.57, p <0.001; Mkuyu 1).
Kugwiritsa ntchito monosaccharides osiyanasiyana (fructose (FRU), galactose (GAL), shuga (GLU)), disaccharides (lactose (LAC), maltose (MAL), sucrose (SUC)) ndi mapadi (CEL) monga amawongolera. Kukula kwa mphutsi zomwe zimadyetsedwa ndi mphutsi zakuda zankhondo. Mfundo iliyonse pamphepete imayimira kulemera kwake kwa munthu (mg) wowerengedwa poyeza mphutsi za 20 zosankhidwa mwachisawawa kuchokera ku gulu la mphutsi za 100 (n = 4). Zolemba zolakwika zikuyimira SD.
Chakudya cha CEL chinapereka moyo wabwino kwambiri wa mphutsi wa 95.5 ± 3.8%. Komanso, kupulumuka kwa H. illucens kudyetsedwa zakudya zomwe zili ndi soluble CH kunachepetsedwa (GLM: χ = 107.13, df = 21, p <0.001), zomwe zinayambitsidwa ndi MAL ndi SUC (disaccharides) mu CH yophunzira. Kufa kunali kochepa kuposa GLU, FRU, GAL (monosaccharide), ndi LAC (EMM: p <0.001, Chithunzi 2).
Boxplot ya kupulumuka kwa mphutsi zankhondo zakuda zomwe zimathandizidwa ndi ma monosaccharides osiyanasiyana (fructose, galactose, glucose), ma disaccharides (lactose, maltose, sucrose) ndi mapadi monga zowongolera. Chithandizo chokhala ndi chilembo chomwecho sizosiyana kwambiri ndi wina ndi mnzake (EMM, p> 0.05).
Zakudya zonse zoyesedwa zimalola kuti mphutsi zifike pamlingo wa prepupal. Komabe, ma CH omwe adayesedwa amakulitsa kukula kwa mphutsi (F6,21=9.60, p<0.001; Gulu 1). Makamaka, mphutsi zodyetsera GAL ndi LAC zinatenga nthawi yaitali kuti zifike pa siteji ya prepupal poyerekeza ndi mphutsi zomwe zinaleredwa pa CEL (CEL-GAL: p<0.001; CEL-LAC: p<0.001; Table 1).
CH yoyesedwa inalinso ndi zotsatira zosiyana pa kulemera kwa mphutsi, ndi kulemera kwa thupi la mphutsi kudyetsa zakudya za CEL kufika ku 180.19 ± 11.35 mg (F6,21 = 16.86, p <0.001; Chithunzi 3). FRU, GLU, MAL ndi SUC zinapangitsa kuti thupi likhale lolemera kwambiri kuposa 200 mg, lomwe linali lalikulu kwambiri kuposa la CEL (p <0.05). Mosiyana ndi zimenezi, mphutsi zodyetsera GAL ndi LAC zinali ndi zolemera zochepa za thupi, pafupifupi 177.64 ± 4.23 mg ndi 156.30 ± 2.59 mg, motsatira (p <0.05). Izi zidadziwika kwambiri ndi LAC, pomwe kulemera kwa thupi komaliza kunali kocheperako poyerekeza ndi zakudya zowongolera (CEL-LAC: kusiyana = 23.89 mg; p = 0.03; Chithunzi 3).
Kutanthauza kulemera komaliza kwa mphutsi zamtundu uliwonse zomwe zimawonetsedwa ngati mawanga a mphutsi (mg) ndi ntchentche zakuda zakuda zomwe zimafotokozedwa ngati histogram (g) zimadyetsa ma monosaccharides osiyanasiyana (fructose, galactose, glucose), ma disaccharides (lactose, maltose, sucrose) ndi cellulose (monga ulamuliro). Zilembo zamagulu zimayimira magulu osiyana kwambiri ndi kulemera kwa mphutsi (p <0.001). Malembo okhudzana ndi mawanga a mphutsi amaimira magulu omwe ali ndi zolemera zosiyana kwambiri za mphutsi (p <0.001). Zolemba zolakwika zikuyimira SD.
Kulemera kwapayekha kunali kosiyana ndi kulemera komaliza kwa mphutsi. M'malo mwake, zakudya zomwe zimakhala ndi FRU, GLU, MAL, ndi SUC sizinawonjezere kulemera kwa mphutsi yopangidwa mu thanki poyerekeza ndi CEL (Chithunzi 3). Komabe, LAC inachepetsa kwambiri kulemera konse (CEL-LAC: kusiyana = 9.14 g; p <0.001; Chithunzi 3).
Gulu 1 likuwonetsa zokolola (mphutsi/tsiku). Chochititsa chidwi, zokolola zabwino za CEL, MAL ndi SUC zinali zofanana (Table 1). Mosiyana, FRU, GAL, GLU ndi LAC adachepetsa zokolola poyerekeza ndi CEL (Table 1). GAL ndi LAC zidachita zoyipa kwambiri: zokolola zidachepetsedwa kukhala 0.51 ± 0.09 g mphutsi / tsiku ndi 0.48 ± 0.06 g / tsiku, motsatana (Table 1).
Ma monosaccharides ndi ma disaccharides amawonjezera kuchuluka kwa lipid mu mphutsi za CF (Table 1). Pazakudya za CLE, mphutsi zomwe zili ndi lipids za 23.19 ± 0.70% za DM zinapezedwa. Poyerekeza, kuchuluka kwa lipid mu mphutsi zomwe zimadyetsedwa ndi shuga wosungunuka zinali zoposa 30% (Table 1). Komabe, ma CH oyesedwa adawonjezera mafuta awo pamlingo womwewo.
Monga kuyembekezera, maphunziro a CG adakhudza mbiri ya FA ya mphutsi ku madigiri osiyanasiyana (mkuyu 4). Zomwe zili mu SFA zinali zambiri muzakudya zonse ndipo zidafika kupitilira 60%. MAL ndi SUC sanagwirizane ndi mbiri ya FA, zomwe zinapangitsa kuti SFA ichuluke. Pankhani ya MAL, kumbali imodzi, kusalinganika kumeneku kunayambitsa makamaka kuchepa kwa zomwe zili mu monounsaturated fatty acids (MUFA) (F6,21 = 7.47; p <0.001; Chithunzi 4). Kumbali ina, kwa SUC, kuchepa kunali kofanana pakati pa MUFA ndi PUFA. LAC ndi MAL zinali ndi zotsatira zosiyana pa mawonekedwe a 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; Chithunzi 4). Kuchepa kwa SFA mu mphutsi zodyetsedwa ndi LAC zikuwoneka kuti zikuwonjezera zomwe zili mu MUFA. Makamaka, milingo ya MUFA inali yayikulu mu mphutsi zodyetsedwa ndi LAC poyerekeza ndi shuga wina wosungunuka kupatula GAL (F6,21 = 7.47; p <0.001; Chithunzi 4).
Kugwiritsa ntchito ma monosaccharides osiyanasiyana (fructose (FRU), galactose (GAL), shuga (GLU)), disaccharides (lactose (LAC), maltose (MAL), sucrose (SUC)) ndi mapadi (CEL) monga zowongolera, bokosi lamafuta acid. Zomwe zimadyetsedwa ndi mphutsi za msilikali wakuda. Zotsatira zimawonetsedwa ngati kuchuluka kwa FAME. Zochizira zolembedwa ndi zilembo zosiyanasiyana ndizosiyana kwambiri (p <0.001). (a) Chigawo chamafuta amafuta acids; (b) Mafuta a monounsaturated mafuta acids; (c) Mafuta a polyunsaturated mafuta acids.
Pakati pa mafuta acids odziwika, lauric acid (C12: 0) anali wamkulu m'mawonekedwe onse (kuposa 40%). Ma SFA ena omwe analipo anali palmitic acid (C16: 0) (osakwana 10%), stearic acid (C18: 0) (osakwana 2.5%) ndi capric acid (C10: 0) (osakwana 1.5%). Ma MUFA makamaka ankayimiridwa ndi oleic acid (C18: 1n9) (osakwana 9.5%), pamene PUFAs makamaka ankapangidwa ndi linoleic acid (C18: 2n6) (osakwana 13.0%) (onani Supplementary Table S1). Kuonjezera apo, kagawo kakang'ono ka mankhwala sikunadziwike, makamaka mu mawonekedwe a mphutsi za CEL, kumene osadziwika pawiri nambala 9 (UND9) amawerengera pafupifupi 2.46 ± 0.52% (onani Supplementary Table S1). Kusanthula kwa GC × GC-FID kunawonetsa kuti ikhoza kukhala 20-carbon fatty acid yokhala ndi ma bond asanu kapena asanu ndi limodzi (onani Supplementary Figure S5).
Kusanthula kwa PERMANOVA kunavumbulutsa magulu atatu osiyana kutengera mbiri yamafuta acid (F6,21 = 7.79, p <0.001; Chithunzi 5). Principal component analysis (PCA) ya TBC spectrum ikuwonetsera izi ndipo ikufotokozedwa ndi zigawo ziwiri (Chithunzi 5). Zigawo zazikuluzikulu zinafotokozera 57.9% ya kusiyana kwake ndipo zinaphatikizapo, monga kufunikira, lauric acid (C12: 0), oleic acid (C18: 1n9), palmitic acid (C16: 0), stearic acid (C18: 0), ndi linolenic acid (C18: 3n3) (onani Chithunzi S4). Chigawo chachiwiri chinafotokozera 26.3% ya kusiyana kwake ndipo chinaphatikizapo, mwadongosolo, decanoic acid (C10: 0) ndi linoleic acid (C18: 2n6 cis) (onani Supplementary Figure S4). Mbiri yazakudya zomwe zimakhala ndi shuga wosavuta (FRU, GAL ndi GLU) zidawonetsanso zofanana. Mosiyana ndi izi, ma disaccharides adapereka mbiri zosiyanasiyana: MAL ndi SUC mbali imodzi ndi LAC mbali inayo. Makamaka, MAL ndiye shuga yekhayo yemwe adasintha mbiri ya FA poyerekeza ndi CEL. Kuphatikiza apo, mbiri ya MAL inali yosiyana kwambiri ndi mbiri ya FRU ndi GLU. Makamaka, mbiri ya MAL idawonetsa gawo lalikulu kwambiri la C12: 0 (54.59 ± 2.17%), kupangitsa kuti ifanane ndi CEL (43.10 ± 5.01%), LAC (43.35 ± 1.31%), FRU (48.90 ± 1.97%) ndi Mbiri ya GLU (48.38 ± 2.17%) (onani Zowonjezera Gulu S1). Mawonekedwe a MAL adawonetsanso zotsika kwambiri za C18:1n9 (9.52 ± 0.50%), zomwe zimasiyanitsanso ndi LAC (12.86 ± 0.52%) ndi CEL (12.40 ± 1.31%). Mchitidwe wofananawo udawonedwa ku C16: 0. Mu gawo lachiwiri, mawonekedwe a LAC adawonetsa C18: 2n6 yapamwamba kwambiri (17.22 ± 0.46%), pomwe MAL adawonetsa otsika kwambiri (12.58 ± 0.67%). C18: 2n6 inasiyanitsanso LAC kuchokera ku ulamuliro (CEL), yomwe inasonyeza milingo yotsika (13.41 ± 2.48%) (onani Supplementary Table S1).
PCA chiwembu chamafuta acid mbiri ya msilikali wakuda ntchentche mphutsi ndi monosaccharides osiyana (fructose, galactose, shuga), disaccharides (lactose, maltose, sucrose) ndi mapadi monga ulamuliro.
Kuti muphunzire za thanzi la shuga wosungunuka pa mphutsi za H. illucens, mapadi (CEL) mu chakudya cha nkhuku adasinthidwa ndi shuga (GLU), fructose (FRU), galactose (GAL), maltose (MAL), sucrose (SUC), ndi lactose (LAC). Komabe, ma monosaccharides ndi ma disaccharides anali ndi zotsatira zosiyana pakukula, kupulumuka, komanso mapangidwe a mphutsi za HF. Mwachitsanzo, GLU, FRU, ndi mawonekedwe awo a disaccharide (MAL ndi SUC) ali ndi zotsatira zabwino zothandizira pakukula kwa mphutsi, zomwe zimawathandiza kuti akwaniritse kulemera kwakukulu kwa thupi kuposa CEL. Mosiyana ndi indigestible CEL, GLU, FRU, ndi SUC imatha kudutsa chotchinga chamatumbo ndikukhala ngati magwero ofunikira pazakudya zopangidwa16,28. MAL ilibe zonyamulira nyama zenizeni ndipo imaganiziridwa kuti imapangidwa ndi hydrolyzed kukhala mamolekyu awiri a shuga isanachitike15. Mamolekyuwa amasungidwa m'thupi la tizilombo ngati gwero lamphamvu kapena ngati lipids18. Choyamba, ponena za zotsirizirazi, zina mwazosiyana zomwe zimawonedwa mu intramodal zingakhale chifukwa cha kusiyana kwakung'ono kwa chiwerengero cha kugonana. Zoonadi, mu H. illucens, kubereka kungakhale kwachisawawa: zazikazi zazikulu mwachibadwa zimakhala ndi mazira okwanira ndipo zimakhala zolemera kuposa amuna29. Komabe, kudzikundikira kwa lipid mu BSFL kumagwirizana ndi kudya kosungunuka kwa CH2, monga momwe tawonera kale kwa GLU ndi xylose26,30. Mwachitsanzo, Li et al.30 adawona kuti pamene 8% GLU idawonjezeredwa ku chakudya cha mphutsi, mphutsi za lipid za BSF zidakwera ndi 7.78% poyerekeza ndi zowongolera. Zotsatira zathu zimagwirizana ndi zomwe taziwonazi, zomwe zikuwonetsa kuti mafuta omwe ali mu mphutsi omwe amadyetsa shuga wosungunuka anali apamwamba kuposa a mphutsi omwe amadyetsa zakudya za CEL, poyerekeza ndi kuwonjezeka kwa 8.57% ndi GLU supplementation. Chodabwitsa n'chakuti zotsatira zofananazi zinawonedwa mu mphutsi zomwe zimadyetsedwa GAL ndi LAC, ngakhale kuti zotsatira zake zimakhala zovuta pakukula kwa mphutsi, kulemera kwa thupi lomaliza, ndi kupulumuka. Mphutsi zodyetsera LAC zinali zazing'ono kwambiri kuposa zomwe zimadyetsedwa ndi CEL, koma mafuta awo anali ofanana ndi mphutsi zomwe zimadyetsa shuga wina wosungunuka. Zotsatirazi zikuwonetsa zotsatira za kupitiliza kwa lactose pa BSFL. Choyamba, zakudya zimakhala ndi kuchuluka kwa CH. Mayamwidwe ndi ma hydrolysis machitidwe a monosaccharides ndi ma disaccharides, motsatana, amatha kufikira machulukitsidwe, zomwe zimapangitsa kuti pakhale zopinga munjira yofananira. Ponena za hydrolysis, imachitika ndi α- ndi β-glucosidases 31. Ma enzymes awa asankha magawo ang'onoang'ono malinga ndi kukula kwake ndi zomangira zamagulu (α kapena β linkages) pakati pa ma monosaccharides awo 15. Hydrolysis ya LAC ku GLU ndi GAL imachitika ndi β-galactosidase, enzyme yomwe ntchito yake yasonyezedwa m'matumbo a BSF 32. Komabe, mawu ake angakhale osakwanira poyerekeza ndi kuchuluka kwa LAC yomwe imadyedwa ndi mphutsi. Mosiyana ndi zimenezi, α-glucosidase maltase ndi sucrase 15, zomwe zimadziwika kuti zimafotokozedwa mochuluka mu tizilombo, zimatha kuwononga MAL ndi sucrose SUC yambiri, motero kuchepetsa zotsatira zokhutiritsa izi. Kachiwiri, zotsatira zotsalira zimatha kukhala chifukwa cha kuchepa kwamphamvu kwa ntchito ya m'matumbo amylase komanso kuchepa kwa kadyedwe poyerekeza ndi mankhwala ena. Zoonadi, mashuga osungunuka adziwika kuti ndi olimbikitsa ntchito ya enzyme yofunikira kuti tizilombo toyambitsa matenda, monga amylase, ndi zomwe zimayambitsa kuyankha kwa chakudya33,34,35. Mlingo wa kukondoweza kumasiyanasiyana malinga ndi kapangidwe ka maselo a shuga. M'malo mwake, ma disaccharides amafunikira hydrolysis asanayambe kuyamwa ndipo amakonda kulimbikitsa amylase kuposa ma monosaccharides34 awo. Mosiyana ndi zimenezi, LAC ili ndi mphamvu yochepa kwambiri ndipo yapezeka kuti sichitha kuthandizira kukula kwa tizilombo mu mitundu yosiyanasiyana33,35. Mwachitsanzo, mu tizilombo Sodoptera exigua (Boddie 1850), palibe ntchito ya hydrolytic ya LAC yomwe idapezeka muzotulutsa za ma enzymes a caterpillar midgut36.
Ponena za mawonekedwe a FA, zotsatira zathu zikuwonetsa zotsatira zazikulu za CH yoyesedwa. Makamaka, ngakhale kuti lauric acid (C12: 0) inali yocheperapo 1% ya FA yonse muzakudya, imayang'anira mbiri yonse (onani Supplementary Table S1). Izi zikugwirizana ndi deta yapitayi kuti lauric acid imapangidwa kuchokera ku zakudya za CH mu H. illucens kudzera munjira yokhudzana ndi acetyl-CoA carboxylase ndi FA synthase19,27,37. Zotsatira zathu zimatsimikizira kuti CEL ndi yosagawika kwambiri ndipo imagwira ntchito ngati "bulking agent" muzakudya zowongolera za BSF, monga momwe tafotokozera m'maphunziro angapo a BSFL38,39,40. Kusintha CEL ndi ma monosaccharides ndi ma disaccharides ena osati LAC kunachulukitsa chiŵerengero cha C12: 0, kusonyeza kuwonjezeka kwa CH ndi mphutsi. Chosangalatsa ndichakuti, ma disaccharides MAL ndi SUC amalimbikitsa kaphatikizidwe ka lauric acid mogwira mtima kuposa ma monosaccharides awo, kutanthauza kuti ngakhale kuchuluka kwa polymerization ya GLU ndi FRU, ndipo popeza Drosophila ndiye yekhayo wonyamula sucrose yemwe wadziwika mumitundu yama protein anyama, onyamula ma disaccharide. sangakhalepo m'matumbo a H. illucens larvae15, kugwiritsa ntchito GLU ndi FRU zikuwonjezeka. Komabe, ngakhale GLU ndi FRU zimasinthidwa mosavuta ndi BSF, zimasinthidwanso mosavuta ndi tizirombo tating'onoting'ono tating'onoting'ono, zomwe zingayambitse kuwonongeka kwawo mwachangu komanso kuchepa kwa kugwiritsidwa ntchito ndi mphutsi poyerekeza ndi ma disaccharides.
Poyamba, lipid zomwe zili mu mphutsi zodyetsedwa ndi LAC ndi MAL zinali zofanana, kusonyeza bioavailability ofanana a shugawa. Komabe, chodabwitsa, mbiri ya FA ya LAC inali yolemera mu SFA, makamaka ndi C12: 0 yotsika, poyerekeza ndi MAL. Lingaliro limodzi lofotokozera kusiyana kumeneku ndikuti LAC ikhoza kuyambitsa bioaccumulation ya zakudya za FA kudzera pa acetyl-CoA FA synthase. Kuthandizira lingaliro ili, mphutsi za LAC zinali ndi chiŵerengero chotsika kwambiri cha decanoate (C10: 0) (0.77 ± 0.13%) kuposa zakudya za CEL (1.27 ± 0.16%), kusonyeza kuchepa kwa FA synthase ndi thioesterase zochita19. Chachiwiri, zakudya zamafuta acids zimaonedwa kuti ndizofunikira kwambiri zomwe zimapangitsa kuti SFA ikhale ndi H. illucens27. Muzoyesera zathu, linoleic acid (C18: 2n6) inali 54.81% ya zakudya zamafuta acids, ndipo gawo la LAC mphutsi ndi 17.22 ± 0.46% ndi MAL 12.58 ± 0.67%. Oleic acid (cis + trans C18: 1n9) (23.22% muzakudya) adawonetsa zofanana. Chiŵerengero cha α-linolenic acid (C18: 3n3) chimathandizanso kuti bioaccumulation hypothesis. Asidi wamafutawa amadziwika kuti amawunjikana mu BSFL pakukula kwa gawo lapansi, monga kuwonjezera keke ya flaxseed, mpaka 6-9% yamafuta onse amafuta mu mphutsi19. Muzakudya zopatsa thanzi, C18: 3n3 imatha kuwerengera mpaka 35% yazakudya zonse zamafuta acid. Komabe, mu phunziro lathu, C18: 3n3 inali ndi 2.51% yokha ya mafuta a asidi. Ngakhale kuti chiwerengero chopezeka m'chilengedwe chinali chochepa mu mphutsi zathu, chiwerengerochi chinali chachikulu mu LAC mphutsi (0.87 ± 0.02%) kuposa MAL (0.49 ± 0.04%) (p <0.001; onani Supplementary Table S1). Zakudya za CEL zinali ndi gawo lapakati la 0.72 ± 0.18%. Pomaliza, chiŵerengero cha palmitic acid (C16:0) mu mphutsi za CF chikuwonetsa kuthandizira kwa njira zopangira komanso zakudya za FA19. Hoc et al. 19 idawona kuti kaphatikizidwe ka C16: 0 idachepetsedwa pomwe zakudya zidakulitsidwa ndi ufa wa flaxseed, zomwe zidanenedwa ndi kuchepa kwa kupezeka kwa gawo lapansi la acetyl-CoA chifukwa cha kuchepa kwa chiŵerengero cha CH. Chodabwitsa n'chakuti, ngakhale kuti zakudya zonse ziwirizi zinali ndi CH zofanana ndipo MAL inawonetsa bioavailability yapamwamba, mphutsi za MAL zimasonyeza C16: 0 chiwerengero chochepa kwambiri (10.46 ± 0.77%), pamene LAC inasonyeza kuchuluka kwakukulu, kuwerengera 12.85 ± 0.27% (p <0.05; onani Zowonjezera Table S1). Zotsatirazi zikuwonetsa zovuta zazakudya pa BSFL digestion ndi metabolism. Pakadali pano, kafukufuku pamutuwu ndiwokwanira ku Lepidoptera kuposa ku Diptera. Mu mbozi, LAC idadziwika ngati cholimbikitsa chofooka cha kudya poyerekeza ndi mashuga ena osungunuka monga SUC ndi FRU34,35. Makamaka, mu Spodopteralittoralis (Boisduval 1833), kumwa kwa MAL kunalimbikitsa ntchito ya amylolytic m'matumbo kwambiri kuposa LAC34. Zotsatira zofananira mu BSFL zitha kufotokozera kukondoweza kowonjezereka kwa C12: njira yopangira 0 mu mphutsi za MAL, zomwe zimalumikizidwa ndi kuchulukitsidwa kwamatumbo a CH, kudyetsa kwanthawi yayitali, ndi matumbo amylase. Kukondoweza pang'ono kwa nyimbo yodyetsera pamaso pa LAC kungafotokozerenso kukula kwapang'onopang'ono kwa mphutsi za LAC. Komanso, Liu Yanxia et al. 27 adawona kuti alumali moyo wa lipids mu magawo a H. illucens unali wautali kuposa wa CH. Chifukwa chake, mphutsi za LAC zitha kudalira kwambiri ma lipids azakudya kuti amalize kukula kwawo, zomwe zitha kuwonjezera kuchuluka kwamafuta awo omaliza ndikuwongolera mawonekedwe awo amafuta acid.
Monga momwe tikudziwira, ndi maphunziro ochepa okha omwe ayesa zotsatira za monosaccharide ndi ma disaccharide owonjezera pazakudya za BSF pa mbiri yawo ya FA. Choyamba, Li et al. 30 idayesa zotsatira za GLU ndi xylose ndikuwona milingo ya lipid yofanana ndi yathu pamlingo wowonjezera wa 8%. Mbiri ya FA sinafotokozedwe mwatsatanetsatane ndipo idapangidwa makamaka ndi SFA, koma palibe kusiyana komwe kunapezeka pakati pa mashuga awiriwa kapena ataperekedwa nthawi imodzi30. Komanso, Cohn et al. 41 sinawonetse zotsatira za 20% GLU, SUC, FRU ndi GAL kuwonjezera pazakudya za nkhuku pambiri za FA. Zowonera izi zidapezedwa kuchokera kuukadaulo m'malo motengera zamoyo, zomwe, monga tafotokozera olemba, zitha kuchepetsa kusanthula kwa ziwerengero. Kuphatikiza apo, kusowa kwa iso-sugar control (pogwiritsa ntchito CEL) kumachepetsa kutanthauzira kwazotsatira. Posachedwapa, maphunziro awiri a Nugroho RA et al. adawonetsa zolakwika mu mawonekedwe a FA42,43. Mu phunziro loyamba, Nugroho RA et al. 43 adayesa momwe angawonjezere FRU ku chakudya cha kanjedza chofufumitsa. Mbiri ya FA ya mphutsi zomwe zinayambitsa mphutsi zinawonetsa PUFA yapamwamba kwambiri, yoposa 90% yomwe inachokera ku zakudya zomwe zili ndi 10% FRU (zofanana ndi phunziro lathu). Ngakhale zakudyazi zinali ndi ma pellets a nsomba olemera a PUFA, mbiri ya FA ya mphutsi pazakudya zowongolera zomwe zimakhala ndi 100% fermented PCM sizinagwirizane ndi mbiri iliyonse yomwe inanenedwa kale, makamaka mlingo wa C18: 3n3 wa 17.77 ± 1.67% ndi 26.08 ± 0.20% pa conjugated linoleic acid (C18: 2n6t), isomer yosowa ya linoleic acid. Kafukufuku wachiwiri adawonetsa zotsatira zofananira kuphatikiza FRU, GLU, MAL ndi SUC42 muzakudya za kanjedza zofufumitsa. Maphunzirowa, monga athu, amawunikira zovuta zazikulu pakuyerekeza zotsatira za mayeso a zakudya za mphutsi za BSF, monga kusankha kowongolera, kuyanjana ndi magwero ena azakudya, ndi njira zowunikira za FA.
Poyesera, tidawona kuti mtundu ndi fungo la gawo lapansi limasiyanasiyana malinga ndi zakudya zomwe zimagwiritsidwa ntchito. Izi zikusonyeza kuti tizilombo tating'onoting'ono titha kutenga nawo gawo pazotsatira zomwe zimawonedwa mu gawo lapansi komanso m'mimba ya mphutsi. M'malo mwake, ma monosaccharides ndi ma disaccharides amapangidwa mosavuta popanga tizilombo toyambitsa matenda. Kumwa mwachangu kwa shuga wosungunuka ndi tizilombo kungayambitse kutulutsa kwazinthu zambiri zamagetsi zamagetsi monga ethanol, lactic acid, mafuta afupiafupi (monga acetic acid, propionic acid, butyric acid) ndi carbon dioxide44. Zina mwazinthuzi zimatha kukhala ndi vuto lakupha kwa mphutsi zomwe zimawonedwanso ndi Cohn et al.41 pamikhalidwe yofananira yachitukuko. Mwachitsanzo, Mowa amawononga tizilombo45. Kuchuluka kwa mpweya woipa wa carbon dioxide ukhoza kuchititsa kuti pansi pa thanki iwunjike, zomwe zingawononge mpweya wa mpweya ngati mpweya sulola kuti utuluke. Ponena za ma SCFAs, zotsatira zawo pa tizilombo, makamaka H. illucens, zimakhalabe zomveka bwino, ngakhale kuti lactic acid, propionic acid, ndi butyric acid awonetsedwa kuti ndi oopsa mu Callosobruchus maculatus (Fabricius 1775)46. Mu Drosophila melanogaster Meigen 1830, ma SCFA awa ndi zolembera zomwe zimatsogolera azimayi kumalo opangira mazira, zomwe zikuwonetsa gawo lopindulitsa pakukula kwa mphutsi47. Komabe, asidi acetic amatchulidwa ngati chinthu chowopsa ndipo amatha kulepheretsa kukula kwa mphutsi47. Mosiyana ndi izi, lactate yopangidwa ndi tizilombo tating'onoting'ono tapezeka kuti ili ndi chitetezo ku tizilombo toyambitsa matenda mu Drosophila48. Kuphatikiza apo, tizilombo tating'onoting'ono m'matumbo a m'mimba timakhalanso ndi gawo la CH digestion mu tizilombo49. Zotsatira zakuthupi za SCFAs pamatumbo a microbiota, monga kuchuluka kwa chakudya ndi ma jini, zafotokozedwa m'magulu amtundu wa 50. Zitha kukhalanso ndi zotsatira zoyipa pa mphutsi za H. illucens ndipo zitha kuthandizira pang'ono pakuwongolera mbiri ya FA. Maphunziro okhudzana ndi zakudya zamagulu a tizilombo toyambitsa matenda a tizilombo toyambitsa matenda adzafotokozera zotsatira zake pa zakudya za H. illucens ndikupereka maziko a maphunziro amtsogolo pa tizilombo toyambitsa matenda opindulitsa kapena owononga pokhudzana ndi chitukuko chawo komanso phindu la magawo olemera a FA. Pachifukwa ichi, ntchito ya tizilombo tating'onoting'ono m'matumbo a tizilombo tomwe timalimidwa kwambiri tikuphunziridwa kwambiri. Tizilombo timayamba kuonedwa ngati bioreactors, kupereka pH ndi oxygenation zinthu zomwe zimathandizira kuti tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda tizilombo toyambitsa matenda. Posachedwapa, Xiang et al.52 adawonetsa kuti, mwachitsanzo, kulowetsedwa kwa zinyalala za organic ndi kusakaniza kwa bakiteriya kumapangitsa CF kukopa mabakiteriya omwe amadziwika kwambiri ndi kuwonongeka kwa lignocellulose, kupititsa patsogolo kuwonongeka kwake mu gawo lapansi poyerekeza ndi magawo opanda mphutsi.
Potsirizira pake, ponena za kugwiritsidwa ntchito kopindulitsa kwa zinyalala za organic ndi H. illucens, zakudya za CEL ndi SUC zinapanga chiwerengero chachikulu cha mphutsi patsiku. Izi zikutanthauza kuti ngakhale kulemera kochepa komaliza kwa munthu payekhapayekha, kulemera kwa mphutsi yonse yomwe imapangidwa pa gawo lapansi lopangidwa ndi indigestible CH ndi yofanana ndi yomwe imapezeka pazakudya za homosaccharide zomwe zimakhala ndi monosaccharides ndi ma disaccharides. Mu phunziro lathu, nkofunika kuzindikira kuti milingo ya zakudya zina ndizokwanira kuthandizira kukula kwa mphutsi komanso kuti kuwonjezera kwa CEL kuyenera kukhala kochepa. Komabe, mapangidwe omaliza a mphutsi amasiyana, kuwonetsa kufunikira kosankha njira yoyenera yopangira tizilombo. Mphutsi za CEL zodyetsedwa ndi chakudya chathunthu ndizoyenera kugwiritsidwa ntchito ngati chakudya cha ziweto chifukwa chamafuta ochepa komanso kuchuluka kwa asidi a lauric acid, pomwe mphutsi zodyetsedwa ndi zakudya za SUC kapena MAL zimafunikira kuwononga mafuta pokakamiza kuti mafuta achuluke, makamaka mu biofuel. gawo. LAC imapezeka muzakudya zamkaka monga whey kuchokera kupanga tchizi. Posachedwapa, kugwiritsa ntchito kwake (3.5% lactose) kunasintha kulemera kwa thupi la mphutsi53. Komabe, zakudya zowongolera mu kafukufukuyu zinali ndi theka la zomwe zili mu lipid. Chifukwa chake, zotsatira zopitilira muyeso za LAC zitha kukhala zotsutsidwa ndi mphutsi ya bioaccumulation ya lipids yazakudya.
Monga momwe zasonyezedwera ndi maphunziro apitalo, katundu wa monosaccharides ndi ma disaccharides amakhudza kwambiri kukula kwa BSFL ndikusintha mbiri yake ya FA. Makamaka, LAC ikuwoneka kuti ikuchitapo kanthu panthawi ya kukula kwa mphutsi mwa kuchepetsa kupezeka kwa CH kwa mayamwidwe a lipid m'zakudya, motero kumalimbikitsa UFA bioaccumulation. Munkhaniyi, zingakhale zosangalatsa kuchita mayeso a bioassay pogwiritsa ntchito zakudya zophatikiza PUFA ndi LAC. Kuphatikiza apo, gawo la tizilombo tating'onoting'ono, makamaka gawo la ma metabolites ang'onoang'ono (monga ma SCFAs) ochokera kunjira zowotchera shuga, umakhalabe mutu wofufuza woyenera kufufuzidwa.
Tizilombo tinapezedwa kuchokera ku gulu la BSF la Laboratory of Functional and Evolutionary Entomology yomwe inakhazikitsidwa mu 2017 ku Agro-Bio Tech, Gembloux, Belgium (kuti mudziwe zambiri za njira zolerera, onani Hoc et al. 19). Pamayesero oyesera, 2.0 g ya mazira a BSF adasonkhanitsidwa mwachisawawa tsiku lililonse kuchokera ku makola oswana ndikuyikidwa mu 2.0 kg ya 70% chakudya cha nkhuku chonyowa (Aveve, Leuven, Belgium). Patatha masiku asanu kuswa, mphutsi zinalekanitsidwa ndi gawo lapansi ndikuwerengera pamanja kuti ziyesedwe. Kulemera koyamba kwa batchi iliyonse kumayesedwa. Kulemera kwapakati pamunthu kunali 7.125 ± 0.41 mg, ndipo pafupifupi pamankhwala aliwonse akuwonetsedwa mu Supplementary Table S2.
Mapangidwe a zakudya adasinthidwa kuchokera ku kafukufuku wa Barragan-Fonseca et al. 38 . Mwachidule, kusagwirizana kunapezeka pakati pa chakudya chofanana cha nkhuku za mphutsi, zomwe zili zowuma (DM), CH (10% yochokera ku zakudya zatsopano) ndi kapangidwe kake, popeza shuga wosavuta ndi ma disaccharides alibe malemba. Malinga ndi chidziwitso cha wopanga (Chicken Feed, AVEVE, Leuven, Belgium), CH yoyesedwa (ie shuga wosungunuka) idawonjezedwa padera ngati yankho lamadzimadzi la autoclaved (15.9%) pazakudya zomwe zimakhala ndi 16.0% mapuloteni, 5.0% okwana lipids, 11.9% ya chakudya chankhuku chokhala ndi phulusa ndi 4.8% ya ulusi. Mumtsuko uliwonse wa 750 ml (17.20 × 11.50 × 6.00 cm, AVA, Tempsee, Belgium), 101.9 g ya autoclaved CH njira inasakanizidwa ndi 37.8 g ya chakudya cha nkhuku. Pazakudya zilizonse, zouma zouma zinali 37.0%, kuphatikiza mapuloteni osakanikirana (11.7%), lipids osakanikirana (3.7%) ndi shuga wofanana (26.9% wa CH wowonjezera). CH adayesedwa anali glucose (GLU), fructose (FRU), galactose (GAL), maltose (MAL), sucrose (SUC) ndi lactose (LAC). Chakudya chowongolera chinali ndi cellulose (CEL), yomwe imatengedwa kuti ndi yosavomerezeka kwa H. illucens mphutsi 38. Mphutsi zana zamasiku 5 zidayikidwa mu thireyi yokhala ndi chivindikiro chokhala ndi dzenje la 1 cm pakati ndikukutidwa ndi ukonde wa udzudzu wa pulasitiki. Chakudya chilichonse chinabwerezedwa kanayi.
Kulemera kwa larval kunayesedwa patatha masiku atatu chiyambi cha kuyesera. Pa muyeso uliwonse, mphutsi 20 zimachotsedwa ku gawo lapansi pogwiritsa ntchito madzi otentha osabala ndi mphamvu, zouma, ndi zolemera (STX223, Ohaus Scout, Parsippany, USA). Pambuyo kuyeza, mphutsi zinabwezedwa pakati pa gawo lapansi. Miyezo idatengedwa nthawi zonse katatu pa sabata mpaka prepupa yoyamba itawonekera. Panthawiyi, sonkhanitsani, werengani, ndi kuyeza mphutsi zonse monga tafotokozera kale. Osiyana siteji 6 mphutsi (ie, mphutsi zoyera zogwirizana ndi mphutsi siteji ya prepupal) ndi prepupae (ie, gawo lomaliza la mphutsi pamene mphutsi za BSF zimasanduka zakuda, zimasiya kudya, ndi kufunafuna malo oyenera kusintha) ndi kusunga pa - 18 ° C kuti mufufuze zolemba. Zokolola zinawerengedwa ngati chiŵerengero cha kuchuluka kwa tizilombo (mphutsi ndi prepupae za siteji 6) zomwe zimapezeka pa mbale (g) mpaka nthawi yachitukuko (d). Zolinga zonse zomwe zili m'malembawo zimafotokozedwa motere: zikutanthauza ± SD.
Masitepe onse ogwiritsira ntchito zosungunulira (hexane (Hex), chloroform (CHCl3), methanol (MeOH)) adachitidwa pansi pa fume hood ndipo amafunikira kuvala magolovesi a nitrile, ma apuloni ndi magalasi otetezera.
Mphutsi zoyera zinaumitsidwa mu chowumitsira chowumitsa cha FreeZone6 (Labconco Corp., Kansas City, MO, USA) kwa maola 72 kenako n’kugwa (IKA A10, Staufen, Germany). Ma lipids onse adatengedwa kuchokera ku ± 1 g ya ufa pogwiritsa ntchito njira ya Folch 54. Chinyezi chotsalira cha chitsanzo chilichonse chokhala ndi lyophilized chinatsimikiziridwa mobwerezabwereza pogwiritsa ntchito makina opangira chinyezi (MA 150, Sartorius, Göttiggen, Germany) kuti akonze lipids yonse.
Ma lipids onse adasinthidwa pansi pamikhalidwe ya acidic kuti apeze mafuta acid methyl esters. Mwachidule, pafupifupi 10 mg lipids/100 µl CHCl3 solution (100 µl) inasinthidwa ndi nayitrogeni mu chubu cha 8 ml Pyrex© (SciLabware – DWK Life Sciences, London, UK). Chubucho chinayikidwa mu Hex (0.5 ml) (PESTINORM®SUPRTRACE n-Hexane> 95% ya organic trace analysis, VWR Chemicals, Radnor, PA, USA) ndi Hex/MeOH/BF3 (20/25/55) yankho (0.5) ml) mu osamba madzi pa 70 °C kwa 90 min. Pambuyo pozizira, 10% yamadzimadzi amadzimadzi a H2SO4 (0.2 ml) ndi njira yodzaza NaCl (0.5 ml) inawonjezeredwa. Sakanizani chubu ndikudzaza kusakaniza ndi Hex yoyera (8.0 mL). Gawo lina lapamwamba linasamutsidwa ku vial ndikuwunikidwa ndi gas chromatography yokhala ndi chowunikira chamoto cha ionization (GC-FID). Zitsanzo zinasanthulidwa pogwiritsa ntchito Trace GC Ultra (Thermo Scientific, Waltham, MA, USA) yokhala ndi jekeseni wogawanika / wosagawanika (240 ° C) mumsewu wogawanika (kugawanika kwa 10 mL / min), ndime ya Stabilwax®-DA ( 30 m, 0.25 mm id, 0.25 μm, Restek Corp., Bellefonte, PA, USA) ndi FID (250 ° C). Pulogalamu ya kutentha inakhazikitsidwa motere: 50 ° C kwa 1 min, kuwonjezeka mpaka 150 ° C pa 30 ° C / min, kuwonjezeka mpaka 240 ° C pa 4 ° C / min ndikupitiriza 240 ° C kwa 5 min. Hex idagwiritsidwa ntchito ngati yopanda kanthu komanso muyezo womwe uli ndi 37 fatty acid methyl esters (Supelco 37-component FAMEmix, Sigma-Aldrich, Overijse, Belgium) idagwiritsidwa ntchito pozindikiritsa. Kuzindikiritsa kwa unsaturated mafuta acids (UFAs) kunatsimikiziridwa ndi GC (GC × GC-FID) yokwanira iwiri (GC × GC-FID) ndipo kukhalapo kwa ma isomer kunatsimikiziridwa molondola ndi kusintha pang'ono kwa njira ya Ferrara et al. 55. Zambiri za zida zitha kupezeka mu Supplementary Table S3 ndi zotsatira mu Supplementary Figure S5.
Zambiri zimaperekedwa mumtundu wa spreadsheet wa Excel (Microsoft Corporation, Redmond, WA, USA). Kusanthula kwa chiwerengero kunachitika pogwiritsa ntchito R Studio (mtundu 2023.12.1+402, Boston, USA) 56. Deta pa kulemera kwa mphutsi, nthawi yachitukuko ndi zokolola zimayesedwa pogwiritsa ntchito chitsanzo cha mzere (LM) (command "lm", R phukusi "stats" 56) monga momwe akufunira kugawa kwa Gaussian. Kupulumuka pogwiritsa ntchito kusanthula kwachitsanzo cha binomial kunayesedwa pogwiritsa ntchito chitsanzo cha mzere (GLM) (command "glm", R phukusi "lme4" 57). Chizoloŵezi chokhazikika ndi homoscedasticity chinatsimikiziridwa pogwiritsa ntchito mayeso a Shapiro (command "shapiro.test", R phukusi "stats" 56) ndi kusanthula kusiyana kwa deta (command betadisper, R phukusi "vegan" 58). Pambuyo pakuwunika pawiri kwa p-values (p <0.05) kuchokera ku mayeso a LM kapena GLM, kusiyana kwakukulu pakati pamagulu kudazindikirika pogwiritsa ntchito mayeso a EMM (lamulirani "emmeans", phukusi la R "emmeans" 59).
Mawonekedwe athunthu a FA adafanizidwa pogwiritsa ntchito kusanthula kwamitundu yosiyanasiyana (ie permMANOVA; lamulo "adonis2", R phukusi "vegan" 58) pogwiritsa ntchito matrix amtunda wa Euclidean ndi zololeza 999. Izi zimathandiza kuzindikira mafuta acids omwe amakhudzidwa ndi chikhalidwe cha zakudya zama carbohydrate. Kusiyana kwakukulu m'mbiri za FA kudawunikidwanso pogwiritsa ntchito kufananitsa pawiri. Detayo idawonetsedwa pogwiritsa ntchito principal component analysis (PCA) (command "PCA", R phukusi "FactoMineR" 60). FA yomwe idayambitsa kusiyana kumeneku idadziwika potanthauzira mabwalo olumikizana. Otsatirawa adatsimikiziridwa pogwiritsa ntchito njira imodzi yowunikira kusiyana (ANOVA) (command "aov", R phukusi "stats" 56 ) kutsatiridwa ndi Tukey's post hoc test (command TukeyHSD, R phukusi "stats" 56 ). Asanayambe kusanthula, chizolowezi chinayesedwa pogwiritsa ntchito mayeso a Shapiro-Wilk, homoscedasticity idafufuzidwa pogwiritsa ntchito mayeso a Bartlett (command "bartlett.test", R phukusi "stats" 56), ndipo njira yopanda malire idagwiritsidwa ntchito ngati palibe malingaliro awiriwa adakwaniritsidwa. . Kusanthula kunafanizidwa (command "kruskal.test", R phukusi "stats" 56 ), ndiyeno mayesero a Dunn a post hoc anagwiritsidwa ntchito ( command dunn.test, R phukusi "dunn.test" 56 ).
Mtundu womaliza wa malembo apamanjawo unafufuzidwa pogwiritsa ntchito Grammarly Editor monga chowerengera cha Chingelezi (Grammarly Inc., San Francisco, California, USA) 61 .
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Nthawi yotumiza: Dec-25-2024