Kuti zitheke bwinoletsa udzudzundi kuchepetsa kufalikira kwa matenda omwe ali nawo, njira zina zodalirika, zokhazikika komanso zosawononga chilengedwe m'malo mwa mankhwala ophera tizilombo ndizofunikira. Tinayesa chakudya cha mbewu kuchokera ku Brassicaceae (banja la Brassica) ngati gwero la isothiocyanates zochokera ku zomera zomwe zimapangidwa ndi enzymatic hydrolysis ya glucosinolates yosagwira ntchito m'thupi kuti igwiritsidwe ntchito polamulira Egyptian Aedes (L., 1762). Ufa wa mbewu zisanu zochotsedwa mafuta (Brassica juncea (L) Czern., 1859, Lepidium sativum L., 1753, Sinapis alba L., 1753, Thlaspi arvense L., 1753 ndi Thlaspi arvense - mitundu itatu yayikulu ya kutentha ndi kuwonongeka kwa enzymatic. Mankhwala achilengedwe. Kuti mudziwe poizoni (LC50) wa allyl isothiocyanate, benzyl isothiocyanate ndi 4-hydroxybenzylisothiocyanate ku mphutsi za Aedes aegypti pakadutsa maola 24 = 0.04 g/120 ml dH2O). LC50 values ya mpiru, mpiru woyera ndi mchira wa horsetail. Ufa wa mbewu unali 0.05, 0.08 ndi 0.05 motsatana poyerekeza ndi allyl isothiocyanate (LC50 = 19.35 ppm) ndipo 4. -Hydroxybenzylisothiocyanate (LC50 = 55.41 ppm) inali yoopsa kwambiri kwa mphutsi kudzera maola 24 pambuyo pa chithandizo kuposa 0.1 g/120 ml dH2O motsatana. Zotsatirazi zikugwirizana ndi kupanga ufa wa mbewu ya alfalfa. Kuchita bwino kwambiri kwa benzyl esters kumagwirizana ndi ma LC50 owerengedwa. Kugwiritsa ntchito ufa wa mbewu kungapereke njira yothandiza yowongolera udzudzu. Kugwira ntchito kwa ufa wa mbewu ya cruciferous ndi zigawo zake zazikulu za mankhwala motsutsana ndi mphutsi za udzudzu ndikuwonetsa momwe mankhwala achilengedwe omwe ali mu ufa wa mbewu ya cruciferous angagwirire ntchito ngati mankhwala ophera larvicide abwino kwambiri poletsa udzudzu.
Matenda opatsirana ndi tizilombo toyambitsa matenda omwe amayamba chifukwa cha udzudzu wa Aedes akadali vuto lalikulu padziko lonse lapansi. Kuchuluka kwa matenda opatsirana ndi udzudzu kumafalikira m'malo osiyanasiyana1,2,3 ndipo kumabwereranso, zomwe zimapangitsa kuti matenda oopsa ayambe kufalikira4,5,6,7. Kufalikira kwa matenda pakati pa anthu ndi nyama (monga chikungunya, dengue, Rift Valley fever, yellow fever ndi Zika virus) sikunachitikepo. Dengue fever yokha imaika anthu pafupifupi 3.6 biliyoni pachiwopsezo chotenga matenda m'madera otentha, ndipo matenda okwana 390 miliyoni amapezeka pachaka, zomwe zimapangitsa kuti anthu 6,100–24,300 afe pachaka8. Kubwereranso ndi kufalikira kwa kachilombo ka Zika ku South America kwakopa chidwi padziko lonse lapansi chifukwa cha kuwonongeka kwa ubongo komwe kumayambitsa mwa ana obadwa kwa akazi omwe ali ndi kachilomboka2. Kremer et al 3 akulosera kuti udzudzu wa Aedes udzapitirira kukula ndipo pofika chaka cha 2050, theka la anthu padziko lonse lapansi adzakhala pachiwopsezo chotenga matenda ndi ma arbovirus ofalitsidwa ndi udzudzu.
Kupatula katemera wa dengue ndi yellow fever wopangidwa posachedwapa, katemera wa matenda ambiri ofalitsidwa ndi udzudzu sanapangidwe9,10,11. Katemera akadalipo ochepa ndipo amagwiritsidwa ntchito kokha m'mayesero azachipatala. Kulamulira tizilombo toyambitsa udzudzu pogwiritsa ntchito mankhwala ophera tizilombo opangidwa kwakhala njira yofunika kwambiri yochepetsera kufalikira kwa matenda ofalitsidwa ndi udzudzu12,13. Ngakhale kuti mankhwala ophera tizilombo opangidwa ndi mankhwala othandiza kupha udzudzu, kugwiritsa ntchito mankhwala ophera tizilombo opangidwa ndi mankhwala kumakhudza kwambiri zamoyo zomwe sizili m'gulu la mankhwala ndipo kumaipitsa chilengedwe14,15,16. Choopsa kwambiri ndi chizolowezi chowonjezera kukana kwa udzudzu ku mankhwala ophera tizilombo17,18,19. Mavuto amenewa okhudzana ndi mankhwala ophera tizilombo afulumizitsa kufunafuna njira zina zothandiza komanso zoteteza chilengedwe kuti zisawononge tizilombo toyambitsa matenda.
Zomera zosiyanasiyana zapangidwa ngati magwero a mankhwala ophera tizilombo toyambitsa matenda kuti achepetse tizilombo20,21. Zinthu zomwe zili m'zomera nthawi zambiri zimakhala zotetezeka ku chilengedwe chifukwa zimatha kuwonongeka ndipo zimakhala ndi poizoni wochepa kapena wochepa kwa zamoyo zomwe sizili m'gulu la ziweto monga nyama zoyamwitsa, nsomba ndi amphibians20,22. Zokonzekera zitsamba zimadziwika kuti zimapanga mitundu yosiyanasiyana ya mankhwala ophera tizilombo okhala ndi njira zosiyanasiyana zogwirira ntchito kuti athetse bwino magawo osiyanasiyana a moyo wa udzudzu23,24,25,26. Mankhwala ochokera ku zomera monga mafuta ofunikira ndi zosakaniza zina zogwira ntchito za zomera zatchuka ndipo zapanga njira yopezera zida zatsopano zowongolera tizilombo toyambitsa udzudzu. Mafuta ofunikira, monoterpenes ndi sesquiterpenes amagwira ntchito ngati mankhwala othamangitsa, oletsa kudyetsa ndi ovicides27,28,29,30,31,32,33. Mafuta ambiri amasamba amachititsa imfa ya mphutsi za udzudzu, ma pupae ndi akuluakulu34,35,36, zomwe zimakhudza mitsempha, kupuma, endocrine ndi machitidwe ena ofunikira a tizilombo37.
Kafukufuku waposachedwapa wapereka chidziwitso pa momwe zomera za mpiru ndi mbewu zawo zingagwiritsire ntchito ngati gwero la mankhwala othandiza thupi. Chakudya cha mbewu za mpiru chayesedwa ngati mankhwala ophera tizilombo toyambitsa matenda38,39,40,41 ndipo chagwiritsidwa ntchito ngati njira yochepetsera udzu42,43,44 komanso kuwongolera tizilombo toyambitsa matenda tomwe timapezeka m'nthaka45,46,47,48,49,50, zakudya za zomera. nematodes 41,51, 52, 53, 54 ndi tizilombo 55, 56, 57, 58, 59, 60. Ntchito yopha bowa ya ufa wa mbewu izi imayambitsidwa ndi mankhwala oteteza zomera otchedwa isothiocyanates38,42,60. Mu zomera, mankhwala oteteza awa amasungidwa m'maselo a zomera monga ma glucosinolates osagwira ntchito thupi. Komabe, zomera zikawonongeka ndi kudya tizilombo kapena matenda opatsirana ndi tizilombo, ma glucosinolates amathiridwa ndi myrosinase kukhala isothiocyanates yogwira ntchito thupi55,61. Isothiocyanates ndi mankhwala osinthika omwe amadziwika kuti ali ndi mphamvu yolimbana ndi majeremusi ambiri komanso yopha tizilombo, ndipo kapangidwe kake, zochita zake zamoyo komanso kuchuluka kwake zimasiyana kwambiri pakati pa mitundu ya Brassicaceae42,59,62,63.
Ngakhale kuti isothiocyanates yochokera ku ufa wa mbewu ya mpiru imadziwika kuti imapha tizilombo, deta yokhudza ntchito ya zamoyo motsutsana ndi tizilombo toyambitsa matenda tofunikira pazachipatala siikupezeka. Kafukufuku wathu adafufuza momwe ufa wa mbewu zinayi zochotsedwa mafuta zimaphera tizilombo motsutsana ndi udzudzu wa Aedes. Mphutsi za Aedes aegypti zimaphera tizilombo toyambitsa matenda motsutsana ndi udzudzu wa Aedes. Cholinga cha kafukufukuyu chinali kuwunika momwe angagwiritsire ntchito ngati mankhwala ophera tizilombo omwe ndi abwino kwa chilengedwe poletsa udzudzu. Zigawo zitatu zazikulu za mankhwala a ufa wa mbewu, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC), ndi 4-hydroxybenzylisothiocyanate (4-HBITC) zinayesedwanso kuti ziyese ntchito ya zamoyo ya mankhwala awa pa mphutsi za udzudzu. Ili ndi lipoti loyamba loyesa momwe ufa wa mbewu zinayi za kabichi umagwirira ntchito komanso momwe zigawo zake zazikulu zimaphera tizilombo motsutsana ndi mphutsi za udzudzu.
Ma colonies a Aedes aegypti (mtundu wa Rockefeller) m'ma laboratories anasungidwa pa 26°C, 70% chinyezi (RH) ndi 10:14 h (L:D photoperiod). Akazi okwatiwa anaikidwa m'mabokosi apulasitiki (kutalika kwa 11 cm ndi m'mimba mwake 9.5 cm) ndipo anadyetsedwa kudzera mu botolo lodyetsera pogwiritsa ntchito magazi a ng'ombe opangidwa ndi citricated (HemoStat Laboratories Inc., Dixon, CA, USA). Kudyetsa magazi kunachitika mwachizolowezi pogwiritsa ntchito nembanemba multi-glass feeder (Chemglass, Life Sciences LLC, Vineland, NJ, USA) yolumikizidwa ku chubu chosambira chamadzi chozungulira (HAAKE S7, Thermo-Scientific, Waltham, MA, USA) ndi kutentha koyenera 37°C. Tambasulani filimu ya Parafilm M pansi pa chipinda chilichonse chodyetsera magalasi (dera la 154 mm2). Kenako chodyetsera chilichonse chinayikidwa pamwamba pa gridi yophimba khola lomwe linali ndi yaikazi yokwatiwa. Pafupifupi 350–400 μl ya magazi a ng'ombe anawonjezeredwa ku funnel yodyetsera galasi pogwiritsa ntchito Pasteur pipette (Fisherbrand, Fisher Scientific, Waltham, MA, USA) ndipo nyongolotsi zazikulu zinaloledwa kutuluka madzi kwa ola limodzi. Kenako akazi apakati anapatsidwa yankho la 10% la sucrose ndipo analoledwa kuikira mazira pa pepala lonyowa lophimbidwa ndi makapu a soufflé (1.25 fl oz kukula, Dart Container Corp., Mason, MI, USA). Ikani pepala lophikira lomwe lili ndi mazira m'thumba lotsekedwa (SC Johnsons, Racine, WI) ndikusunga pa 26°C. Mazirawo anaswedwa ndipo mphutsi pafupifupi 200–250 zinaleredwa m'mathireyi apulasitiki okhala ndi chisakanizo cha rabbit chow (ZuPreem, Premium Natural Products, Inc., Mission, KS, USA) ndi ufa wa chiwindi (MP Biomedicals, LLC, Solon, OH, USA), ndi fillet ya nsomba (TetraMin, Tetra GMPH, Meer, Germany) mu chiŵerengero cha 2:1:1. Mphutsi za m'mapeto achitatu zinagwiritsidwa ntchito mu kafukufuku wathu wa zamoyo.
Mbewu za zomera zomwe zagwiritsidwa ntchito mu kafukufukuyu zapezeka kuchokera ku magwero otsatirawa amalonda ndi aboma: Brassica juncea (brown mustard-Pacific Gold) ndi Brassica juncea (white mustard-Ida Gold) kuchokera ku Pacific Northwest Farmers' Cooperative, Washington State, USA; (Garden Cress) kuchokera ku Kelly Seed and Hardware Co., Peoria, IL, USA ndi Thlaspi arvense (Field Pennycress-Elisabeth) kuchokera ku USDA-ARS, Peoria, IL, USA; Palibe mbewu zomwe zagwiritsidwa ntchito mu kafukufukuyu zomwe zinapatsidwa mankhwala ophera tizilombo. Mbewu zonse za mbewu zinakonzedwa ndikugwiritsidwa ntchito mu kafukufukuyu mogwirizana ndi malamulo am'deralo ndi adziko lonse komanso mogwirizana ndi malamulo onse a m'deralo ndi adziko lonse. Kafukufukuyu sanayang'ane mitundu ya zomera zosinthidwa majini.
Mbewu za Brassica juncea (PG), Alfalfa (Ls), White mustard (IG), Thlaspi arvense (DFP) zinaphwanyidwa kukhala ufa wosalala pogwiritsa ntchito Retsch ZM200 ultracentrifugal mill (Retsch, Haan, Germany) yokhala ndi 0.75 mm mesh ndi Stainless steel rotor, mano 12, 10,000 rpm (Table 1). Ufa wa mbewu zophwanyidwa unasamutsidwa ku thimble ya pepala ndikuchotsedwa mafuta ndi hexane mu chipangizo cha Soxhlet kwa maola 24. Chitsanzo cha mpiru wochotsedwa mafuta chinatenthedwa pa 100 °C kwa ola limodzi kuti chichotse myrosinase ndikuletsa hydrolysis ya glucosinolates kuti apange isothiocyanates yogwira ntchito m'thupi. Ufa wa mbewu ya horsetail wochiritsidwa ndi kutentha (DFP-HT) unagwiritsidwa ntchito ngati choletsa kuipitsidwa mwa kuchepetsa kukula kwa myrosinase.
Kuchuluka kwa glucosinolate mu ufa wa mbewu zochotsedwa mafuta kunapezeka katatu pogwiritsa ntchito high-performance liquid chromatography (HPLC) malinga ndi protocol 64 yomwe idasindikizidwa kale. Mwachidule, 3 mL ya methanol inawonjezedwa ku chitsanzo cha 250 mg cha ufa wa mbewu zochotsedwa mafuta. Chitsanzo chilichonse chinayikidwa mu sonic bath kwa mphindi 30 ndikusiyidwa mumdima pa 23°C kwa maola 16. Kenako 1 mL aliquot ya organic layer idasefedwa kudzera mu fyuluta ya 0.45 μm kupita ku autosampler. Pogwira ntchito pa Shimadzu HPLC system (mapampu awiri a LC 20AD; SIL 20A autosampler; DGU 20As degasser; SPD-20A UV-VIS detector yowunikira pa 237 nm; ndi CBM-20A communication bus module), kuchuluka kwa glucosinolate mu ufa wa mbewu kunapezeka katatu. pogwiritsa ntchito Shimadzu LC Solution software version 1.25 (Shimadzu Corporation, Columbia, MD, USA). Mzerewu unali C18 Inertsil reverse phase column (250 mm × 4.6 mm; RP C-18, ODS-3, 5u; GL Sciences, Torrance, CA, USA). Mikhalidwe yoyambirira ya gawo loyenda idakhazikitsidwa pa 12% methanol/88% 0.01 M tetrabutylammonium hydroxide m'madzi (TBAH; Sigma-Aldrich, St. Louis, MO, USA) ndi kuchuluka kwa 1 mL/min. Pambuyo pobayira 15 μl ya chitsanzo, mikhalidwe yoyambirira idasungidwa kwa mphindi 20, kenako chiŵerengero cha solvent chidasinthidwa kukhala 100% methanol, ndi nthawi yonse yowunikira chitsanzo ya mphindi 65. Mzere wokhazikika (wochokera ku nM/mAb) udapangidwa ndi kuchepetsedwa kwa sinapine, glucosinolate ndi myrosin standards (Sigma-Aldrich, St. Louis, MO, USA) kuti ayese kuchuluka kwa sulfure mu ufa wa mbewu wochotsedwa mafuta. Kuchuluka kwa glucosinolate m'zitsanzo kunayesedwa pa Agilent 1100 HPLC (Agilent, Santa Clara, CA, USA) pogwiritsa ntchito OpenLAB CDS ChemStation version (C.01.07 SR2 [255]) yokhala ndi mzere womwewo komanso pogwiritsa ntchito njira yomwe yafotokozedwa kale. Kuchuluka kwa glucosinolate kunapezeka; kufanana pakati pa machitidwe a HPLC.
Allyl isothiocyanate (94%, yokhazikika) ndi benzyl isothiocyanate (98%) zinagulidwa ku Fisher Scientific (Thermo Fisher Scientific, Waltham, MA, USA). 4-Hydroxybenzylisothiocyanate inagulidwa ku ChemCruz (Santa Cruz Biotechnology, CA, USA). Pamene enzymatically hydrolyzed ndi myrosinase, glucosinolates, glucosinolates, ndi glucosinolates amapanga allyl isothiocyanate, benzyl isothiocyanate, ndi 4-hydroxybenzylisothiocyanate, motsatana.
Kuyesa kwa bioassay ku labotale kunachitika motsatira njira ya Muturi et al. 32 ndi zosintha. Zakudya zisanu za mbewu zopanda mafuta ambiri zinagwiritsidwa ntchito mu kafukufukuyu: DFP, DFP-HT, IG, PG ndi Ls. Mphutsi makumi awiri zinayikidwa mu beaker ya 400 mL yotayika ya njira zitatu (VWR International, LLC, Radnor, PA, USA) yokhala ndi madzi osungunuka a 120 mL (dH2O). Kuchuluka kwa mbewu zisanu ndi ziwiri kunayesedwa kuti ziwone ngati pali poizoni wa mphutsi za udzudzu: 0.01, 0.02, 0.04, 0.06, 0.08, 0.1 ndi 0.12 g ya chakudya cha mbewu/120 ml dH2O ya chakudya cha mbewu cha DFP, DFP-HT, IG ndi PG. Kuyesa koyambirira kumasonyeza kuti ufa wa mbewu ya Ls wochotsedwa mafuta ndi woopsa kwambiri kuposa ufa wina wa mbewu zinayi womwe unayesedwa. Chifukwa chake, tinasintha kuchuluka kwa mankhwala asanu ndi awiri a Ls seed food kuti kukhale kotsatira: 0.015, 0.025, 0.035, 0.045, 0.055, 0.065, ndi 0.075 g/120 mL dH2O.
Gulu loyang'anira lomwe silinalandire chithandizo (dH20, palibe chakudya chowonjezera cha mbewu) linaphatikizidwa kuti liwone ngati tizilombo tafa nthawi zonse pansi pa mayeso. Kuyesa kwa poizoni pa chakudya chilichonse cha mbewu kunaphatikizapo ma beakers atatu otsatizana atatu (mphutsi 20 zomaliza zachitatu pa beak iliyonse), kuti pakhale mabotolo 108. Zidebe zomwe zinachiritsidwa zinasungidwa kutentha kwa chipinda (20-21°C) ndipo kufa kwa mphutsi kunalembedwa mkati mwa maola 24 ndi 72 a nthawi zonse pamene mankhwala akugwiritsidwa ntchito. Ngati thupi la udzudzu ndi ziwalo zake sizikuyenda zikabowoledwa kapena kukhudzidwa ndi spatula yopyapyala yachitsulo chosapanga dzimbiri, mphutsi za udzudzu zimaonedwa kuti zafa. Mphutsi zakufa nthawi zambiri zimakhala zosayenda pansi pa chidebecho kapena pamwamba pa madzi. Kuyeseraku kunabwerezedwa katatu pa masiku osiyanasiyana pogwiritsa ntchito magulu osiyanasiyana a mphutsi, kuti pakhale mphutsi 180 zomwe zimawonetsedwa pa mankhwala aliwonse.
Kuopsa kwa AITC, BITC, ndi 4-HBITC ku mphutsi za udzudzu kunayesedwa pogwiritsa ntchito njira yomweyo yoyesera koma ndi mankhwala osiyanasiyana. Konzani mayankho a 100,000 ppm pa mankhwala aliwonse powonjezera 100 µL ya mankhwalawo ku 900 µL ya ethanol yokwanira mu chubu cha 2-mL centrifuge ndikugwedeza kwa masekondi 30 kuti musakanizike bwino. Kuchuluka kwa chithandizo kunatsimikiziridwa kutengera mayeso athu oyamba a bioassays, omwe adapeza kuti BITC ndi yoopsa kwambiri kuposa AITC ndi 4-HBITC. Kuti mudziwe poizoni, kuchuluka kwa BITC 5 (1, 3, 6, 9 ndi 12 ppm), kuchuluka kwa AITC 7 (5, 10, 15, 20, 25, 30 ndi 35 ppm) ndi kuchuluka kwa 6 kwa 4-HBITC (15, 15, 20, 25, 30 ndi 35 ppm). 30, 45, 60, 75 ndi 90 ppm). Chithandizo chowongolera chinaperekedwa ndi 108 μL ya absolute ethanol, yomwe ndi yofanana ndi kuchuluka kwakukulu kwa mankhwala. Kuyesa kwa bioanalysis kunabwerezedwa monga momwe tafotokozera pamwambapa, kuwonetsa mphutsi zonse 180 pa kuchuluka kwa mankhwala. Imfa ya mphutsi inalembedwa pa kuchuluka kulikonse kwa AITC, BITC, ndi 4-HBITC pambuyo pa maola 24 ogwiritsidwa ntchito mosalekeza.
Kusanthula kwa probit kwa deta 65 yokhudzana ndi imfa yokhudzana ndi mlingo kunachitika pogwiritsa ntchito pulogalamu ya Polo (Polo Plus, LeOra Software, mtundu 1.0) kuti awerengere kuchuluka kwa imfa 50% (LC50), kuchuluka kwa imfa 90% (LC90), malo otsetsereka, kuchuluka kwa imfa, ndi kuchuluka kwa imfa 95%. kutengera nthawi yodalirika ya kuchuluka kwa imfa pa kuchuluka kwa imfa komwe kumasinthidwa ndi log ndi ma curve a mlingo. Deta ya imfa imachokera pa deta yobwerezabwereza ya mphutsi 180 zomwe zimawonetsedwa pa kuchuluka kwa chithandizo chilichonse. Kusanthula kwadzidzidzi kunachitika padera pa chakudya chilichonse cha mbewu ndi gawo lililonse la mankhwala. Kutengera nthawi yodalirika ya 95% ya chiŵerengero cha imfa, poizoni wa ufa wa mbewu ndi mankhwala omwe amapezeka ku mphutsi za udzudzu ankaonedwa kuti ndi osiyana kwambiri, kotero nthawi yodalirika yokhala ndi mtengo wa 1 sinali yosiyana kwambiri, P = 0.0566.
Zotsatira za HPLC zodziwira ma glucosinolates akuluakulu mu ufa wa mbewu wochotsedwa mafuta DFP, IG, PG ndi Ls zalembedwa mu Gome 1. Ma glucosinolates akuluakulu mu ufa wa mbewu omwe adayesedwa adasiyana kupatula DFP ndi PG, zomwe zonse zinali ndi myrosinase glucosinolates. Kuchuluka kwa myrosinin mu PG kunali kokwera kuposa mu DFP, 33.3 ± 1.5 ndi 26.5 ± 0.9 mg/g, motsatana. Ufa wa mbewu wa Ls unali ndi 36.6 ± 1.2 mg/g glucoglycone, pomwe ufa wa mbewu wa IG unali ndi 38.0 ± 0.5 mg/g sinapine.
Mphutsi za udzudzu wa Ae. Aedes aegypti zinaphedwa pamene zinapatsidwa chakudya cha mbewu zochotsedwa mafuta, ngakhale kuti mphamvu ya mankhwalawa inasiyana malinga ndi mtundu wa zomera. DFP-NT yokha sinali poizoni kwa mphutsi za udzudzu pambuyo pa maola 24 ndi 72 atakhudzidwa (Table 2). Kuopsa kwa ufa wa mbewu yogwira ntchito kunawonjezeka ndi kuchuluka kwa ndende (Chithunzi 1A, B). Kuopsa kwa ufa wa mbewu kwa mphutsi za udzudzu kunasintha kwambiri kutengera 95% CI ya chiŵerengero cha mlingo woopsa wa LC50 pa mayeso a maola 24 ndi maola 72 (Table 3). Pambuyo pa maola 24, zotsatira za poizoni wa ufa wa mbewu za Ls zinali zazikulu kuposa mankhwala ena a mbewu, ndi ntchito yayikulu kwambiri komanso poizoni kwambiri kwa mphutsi (LC50 = 0.04 g/120 ml dH2O). Mphutsi sizinali zovuta ku DFP pa maola 24 poyerekeza ndi ufa wa mbewu za IG, Ls ndi PG, ndi LC50 values ya 0.115, 0.04 ndi 0.08 g/120 ml dH2O motsatana, zomwe zinali zapamwamba kwambiri kuposa mtengo wa LC50. 0.211 g/120 ml dH2O (Table 3). LC90 values ya DFP, IG, PG ndi Ls inali 0.376, 0.275, 0.137 ndi 0.074 g/120 ml dH2O, motsatana (Table 2). Kuchuluka kwakukulu kwa DPP kunali 0.12 g/120 ml dH2O. Pambuyo pa maola 24 owunikira, kufa kwapakati kwa mphutsi kunali 12% yokha, pomwe kufa kwapakati kwa mphutsi za IG ndi PG kunafika 51% ndi 82%, motsatana. Pambuyo pa maola 24 owunikira, imfa yapakati ya mphutsi chifukwa cha kuchuluka kwakukulu kwa Ls (0.075 g/120 ml dH2O) inali 99% (Chithunzi 1A).
Ma curve a imfa adayesedwa kuchokera ku yankho la mlingo (Probit) la mphutsi za Ae. Egypt (mphutsi zachitatu) mpaka kuchuluka kwa chakudya cha mbewu maola 24 (A) ndi maola 72 (B) pambuyo pa chithandizo. Mzere wolembedwa ndi dontho ukuyimira LC50 ya chithandizo cha chakudya cha mbewu. DFP Thlaspi arvense, DFP-HT Heat inactivated Thlaspi arvense, IG Sinapsis alba (Ida Gold), PG Brassica juncea (Pacific Gold), Ls Lepidium sativum.
Pakuwunika kwa maola 72, miyeso ya LC50 ya ufa wa mbewu wa DFP, IG ndi PG inali 0.111, 0.085 ndi 0.051 g/120 ml dH2O, motsatana. Pafupifupi mphutsi zonse zomwe zinapezeka ku ufa wa mbewu wa L zinafa pambuyo pa maola 72 zitapezeka, kotero deta ya imfa sinali yogwirizana ndi kusanthula kwa Probit. Poyerekeza ndi miyeso ina ya mbewu, mphutsi sizinali zokhudzidwa kwambiri ndi chithandizo cha ufa wa mbewu wa DFP ndipo zinali ndi miyeso yapamwamba ya LC50 (Matebulo 2 ndi 3). Patatha maola 72, miyeso ya LC50 ya chithandizo cha ufa wa mbewu wa DFP, IG ndi PG inayerekezeredwa kukhala 0.111, 0.085 ndi 0.05 g/120 ml dH2O, motsatana. Patatha maola 72 a kuwunikira, miyeso ya LC90 ya ufa wa mbewu wa DFP, IG ndi PG inali 0.215, 0.254 ndi 0.138 g/120 ml dH2O, motsatana. Pambuyo pa maola 72 owunikira, imfa yapakati ya mphutsi chifukwa cha chakudya cha DFP, IG ndi PG chomwe chimaperekedwa pamlingo wapamwamba wa 0.12 g/120 ml dH2O chinali 58%, 66% ndi 96% motsatana (Chithunzi 1B). Pambuyo pakuwunika kwa maola 72, chakudya cha PG chomwe chimaperekedwa chimapeza kuti ndi choopsa kwambiri kuposa chakudya cha IG ndi DFP.
Ma isothiocyanates opangidwa, allyl isothiocyanate (AITC), benzyl isothiocyanate (BITC) ndi 4-hydroxybenzylisothiocyanate (4-HBITC) amatha kupha mphutsi za udzudzu. Patatha maola 24 kuchokera pamene mankhwala aperekedwa, BITC inali yoopsa kwambiri kwa mphutsi ndi LC50 ya 5.29 ppm poyerekeza ndi 19.35 ppm ya AITC ndi 55.41 ppm ya 4-HBITC (Table 4). Poyerekeza ndi AITC ndi BITC, 4-HBITC ili ndi poizoni wochepa komanso LC50 yamtengo wapatali. Pali kusiyana kwakukulu pa poizoni wa mphutsi za udzudzu wa isothiocyanate ziwiri zazikulu (Ls ndi PG) mu chakudya champhamvu kwambiri cha mbewu. Kuopsa kwa poizoni kutengera chiŵerengero cha mlingo woopsa wa LC50 pakati pa AITC, BITC, ndi 4-HBITC kunawonetsa kusiyana kwa ziwerengero kotero kuti 95% CI ya chiŵerengero cha mlingo woopsa wa LC50 sinaphatikizepo mtengo wa 1 (P = 0.05, Table 4). Kuchuluka kwakukulu kwa BITC ndi AITC kunayerekezeredwa kuti kunapha 100% ya mphutsi zomwe zinayesedwa (Chithunzi 2).
Ma curve a imfa adayesedwa kuchokera ku yankho la mlingo (Probit) wa Ae. Patatha maola 24 kuchokera pamene chithandizo chinaperekedwa, mphutsi za ku Egypt (mphutsi zachitatu) zinafika pamlingo wa isothiocyanate wopangidwa. Mzere wolembedwa ndi madontho ukuyimira LC50 ya chithandizo cha isothiocyanate. Benzyl isothiocyanate BITC, allyl isothiocyanate AITC ndi 4-HBITC.
Kugwiritsa ntchito mankhwala ophera tizilombo toyambitsa matenda m'zomera monga mankhwala oletsa udzudzu kwakhala kuphunziridwa kwa nthawi yayitali. Zomera zambiri zimapanga mankhwala achilengedwe omwe ali ndi mphamvu yopha tizilombo37. Mankhwala awo ophera tizilombo amapereka njira ina yokongola m'malo mwa mankhwala ophera tizilombo opangidwa omwe ali ndi mphamvu yayikulu yoletsa tizilombo, kuphatikizapo udzudzu.
Zomera za mpiru zimalimidwa ngati mbewu za mbewu zawo, zomwe zimagwiritsidwa ntchito ngati zonunkhira komanso mafuta. Mafuta a mpiru akachotsedwa mu mbewu kapena mpiru ikachotsedwa kuti igwiritsidwe ntchito ngati biofuel, 69 chinthu chotsalacho chimakhala ufa wa mbewu wochotsedwa mafuta. Ufa wa mbewu uwu umasunga zinthu zambiri zachilengedwe za biochemical ndi ma enzymes a hydrolytic. Kuopsa kwa ufa wa mbewu uwu kumachitika chifukwa cha kupanga isothiocyanates55,60,61. Isothiocyanates zimapangidwa ndi hydrolysis ya glucosinolates ndi enzyme myrosinase panthawi ya madzi a ufa wa mbewu38,55,70 ndipo amadziwika kuti ali ndi zotsatira zopha bowa, mabakiteriya, nematicidal komanso tizilombo toyambitsa matenda, komanso zinthu zina kuphatikizapo zotsatira za mankhwala ndi mankhwala a chemotherapeutic61,62,70. Kafukufuku wambiri wasonyeza kuti zomera za mpiru ndi ufa wa mbewu zimagwira ntchito bwino ngati zofukiza motsutsana ndi nthaka ndi tizilombo tosungidwa chakudya57,59,71,72. Mu kafukufukuyu, tinayang'ana poizoni wa ufa wa mbewu zinayi ndi zinthu zake zitatu zogwira ntchito m'thupi monga AITC, BITC, ndi 4-HBITC ku mphutsi za udzudzu za Aedes. Aedes aegypti. Kuwonjezera ufa wa mbewu mwachindunji m'madzi okhala ndi mphutsi za udzudzu kukuyembekezeka kuyambitsa njira za enzymatic zomwe zimapanga isothiocyanates zomwe zimakhala zoopsa kwa mphutsi za udzudzu. Kusintha kwa biotransformation kumeneku kunawonetsedwa pang'ono ndi momwe ufa wa mbewu umagwirira ntchito komanso kutayika kwa mphamvu yopha tizilombo pamene ufa wa mbewu ya mpiru wa dwarf unakonzedwa kutentha musanagwiritse ntchito. Kuchiza kutentha kukuyembekezeka kuwononga ma enzymes a hydrolytic omwe amachititsa glucosinolates, potero kuletsa kupangika kwa isothiocyanates yogwira ntchito m'thupi. Uwu ndi kafukufuku woyamba kutsimikizira mphamvu yopha tizilombo ya ufa wa mbewu ya kabichi motsutsana ndi udzudzu m'malo a m'madzi.
Pakati pa ufa wa mbewu zomwe zinayesedwa, ufa wa mbewu za watercress (Ls) unali woopsa kwambiri, zomwe zinachititsa kuti Aedes albopictus afe kwambiri. Mphutsi za Aedes aegypti zinakonzedwa mosalekeza kwa maola 24. Ufa wa mbewu zitatu zotsalazo (PG, IG ndi DFP) unali ndi ntchito yochepa ndipo unapangitsa kuti imfa zambiri zichitike pambuyo pa maola 72 a chithandizo chosalekeza. Chakudya cha mbewu cha L chokha chinali ndi glucosinolates yochuluka, pomwe PG ndi DFP zinali ndi myrosinase ndipo IG inali ndi glucosinolate ngati glucosinolate yayikulu (Table 1). Glucotropaeolin imathiridwa ndi hydrolyzed ku BITC ndipo sinalbine imathiridwa ndi hydrolyzed ku 4-HBITC61,62. Zotsatira zathu za bioassay zikusonyeza kuti chakudya cha mbewu cha Ls ndi BITC yopangidwa ndi mankhwala ndi poizoni kwambiri ku mphutsi za udzudzu. Gawo lalikulu la chakudya cha mbewu cha PG ndi DFP ndi myrosinase glucosinolate, yomwe imathiridwa ndi hydrolyzed ku AITC. AITC ndi yothandiza kupha mphutsi za udzudzu ndi LC50 ya 19.35 ppm. Poyerekeza ndi AITC ndi BITC, 4-HBITC isothiocyanate ndi poizoni wochepa kwambiri kwa mphutsi. Ngakhale kuti AITC ndi poizoni wochepa kuposa BITC, LC50 yawo ndi yotsika poyerekeza ndi mafuta ambiri ofunikira omwe amayesedwa pa mphutsi za udzudzu32,73,74,75.
Ufa wa mbewu zathu zophimbidwa kuti zigwiritsidwe ntchito polimbana ndi mphutsi za udzudzu uli ndi glucosinolate imodzi yayikulu, yomwe imawerengera 98-99% ya glucosinolates yonse monga momwe HPLC imadziwira. Kuchuluka kwa ma glucosinolates ena kunapezeka, koma milingo yawo inali yochepera 0.3% ya glucosinolates yonse. Ufa wa mbewu za Watercress (L. sativum) uli ndi glucosinolates yachiwiri (sinigrin), koma gawo lawo ndi 1% ya glucosinolates yonse, ndipo kuchuluka kwawo sikuli kofunikira (pafupifupi 0.4 mg/g ufa wa mbewu). Ngakhale PG ndi DFP zili ndi glucosinolate yayikulu (myrosin) yomweyo, mphamvu ya larvicidal ya chakudya cha mbewu zawo imasiyana kwambiri chifukwa cha LC50 yawo. Zimasiyana poizoni wa powdery mildew. Kutuluka kwa mphutsi za Aedes aegypti kungakhale chifukwa cha kusiyana kwa ntchito ya myrosinase kapena kukhazikika pakati pa zakudya ziwiri za mbewu. Ntchito ya Myrosinase imagwira ntchito yofunika kwambiri pakupezeka kwa zinthu zopangidwa ndi hydrolysis monga isothiocyanates mu zomera za Brassicaceae76. Malipoti am'mbuyomu a Pocock et al.77 ndi Wilkinson et al.78 awonetsa kuti kusintha kwa ntchito ya myrosinase ndi kukhazikika kwake kungagwirizanenso ndi majini ndi zinthu zachilengedwe.
Kuchuluka kwa isothiocyanate komwe kumayembekezeredwa kuti kukhale ndi bioactive kunawerengedwa kutengera kuchuluka kwa LC50 pa chakudya chilichonse cha mbewu pa maola 24 ndi 72 (Table 5) poyerekeza ndi kugwiritsa ntchito mankhwala koyenera. Pambuyo pa maola 24, isothiocyanate mu chakudya cha mbewu inali yoopsa kwambiri kuposa mankhwala oyera. Ma LC50 omwe amawerengedwa kutengera magawo pa miliyoni (ppm) a mankhwala a mbewu za isothiocyanate anali otsika kuposa ma LC50 pakugwiritsa ntchito BITC, AITC, ndi 4-HBITC. Tinaona mphutsi zikudya ma pellets a mbewu (Chithunzi 3A). Chifukwa chake, mphutsi zimatha kukhudzidwa kwambiri ndi isothiocyanate ya poizoni mwa kudya ma pellets a mbewu. Izi zidawonekera kwambiri mu mankhwala a IG ndi PG a mbewu pa nthawi ya maola 24, pomwe kuchuluka kwa LC50 kunali kotsika ndi 75% ndi 72% kuposa mankhwala a AITC ndi 4-HBITC, motsatana. Mankhwala a L ndi DFP anali oopsa kwambiri kuposa isothiocyanate yoyera, ndipo LC50 values 24% ndi 41% otsika, motsatana. Mphutsi zomwe zinali mu chithandizo chowongolera zinayamba kukula bwino (Chithunzi 3B), pomwe mphutsi zambiri zomwe zinali mu chithandizo cha mbewu sizinaphuke ndipo kukula kwa mphutsi kunachedwa kwambiri (Chithunzi 3B,D). Mu Spodopteralitura, isothiocyanates zimagwirizanitsidwa ndi kuchedwa kukula ndi kuchedwa kukula79.
Mphutsi za udzudzu wa Ae. Aedes aegypti zinkakumana ndi ufa wa mbewu ya Brassica kwa maola 24-72. (A) Mphutsi zakufa zokhala ndi tinthu ta ufa wa mbewu mkamwa (zozunguliridwa); (B) Chithandizo chowongolera (dH20 popanda ufa wowonjezera wa mbewu) chimasonyeza kuti mphutsi zimakula bwino ndipo zimayamba kumera pambuyo pa maola 72 (C, D) Mphutsi zothandizidwa ndi ufa wa mbewu; ufa wa mbewu unasonyeza kusiyana kwa kukula ndipo sunamere.
Sitinaphunzire momwe isothiocyanates imakhudzira poizoni pa mphutsi za udzudzu. Komabe, kafukufuku wakale pa nyerere zofiira (Solenopsis invicta) wasonyeza kuti kuletsa glutathione S-transferase (GST) ndi esterase (EST) ndiye njira yayikulu yogwiritsira ntchito isothiocyanate bioactivity, ndipo AITC, ngakhale itakhala yochepa, ingalepheretsenso GST kugwira ntchito. nyerere zofiira zomwe zimatumizidwa kunja zimakhala zochepa. Mlingo wake ndi 0.5 µg/ml80. Mosiyana ndi zimenezi, AITC imaletsa acetylcholinesterase mu nkhupakupa zazikulu za chimanga (Sitophilus zeamais)81. Maphunziro ofanana ayenera kuchitika kuti afotokoze momwe isothiocyanate imagwirira ntchito mu mphutsi za udzudzu.
Timagwiritsa ntchito mankhwala a DFP osagwiritsidwa ntchito ndi kutentha kuti tithandizire lingaliro lakuti hydrolysis ya glucosinolates ya zomera kuti ipange isothiocyanates yogwira ntchito imagwira ntchito ngati njira yowongolera mphutsi za udzudzu pogwiritsa ntchito ufa wa mbewu ya mpiru. Ufa wa mbewu ya DFP-HT sunali woopsa pamlingo woyesedwa. Lafarga et al. 82 adanenanso kuti glucosinolates imakhudzidwa ndi kuwonongeka pa kutentha kwakukulu. Chithandizo cha kutentha chikuyembekezekanso kuwononga enzyme ya myrosinase mu ufa wa mbewu ndikuletsa hydrolysis ya glucosinolates kuti ipange isothiocyanates yogwira ntchito. Izi zidatsimikiziridwanso ndi Okunade et al. 75 adawonetsa kuti myrosinase imakhudzidwa ndi kutentha, zomwe zikusonyeza kuti ntchito ya myrosinase idalepheretsedwa kwathunthu pamene mbewu za mpiru, mpiru wakuda, ndi mizu yamagazi zidakumana ndi kutentha kopitilira 80°C. Njirazi zitha kupangitsa kuti ufa wa mbewu ya DFP wothiridwa ndi kutentha utayike.
Motero, ufa wa mbewu ya mpiru ndi ma isothiocyanate ake atatu akuluakulu ndi oopsa kwa mphutsi za udzudzu. Popeza pali kusiyana pakati pa ufa wa mbewu ndi mankhwala, kugwiritsa ntchito ufa wa mbewu kungakhale njira yothandiza yowongolera udzudzu. Pakufunika kupeza njira zoyenera komanso njira zoperekera zabwino kuti ziwongolere kugwiritsa ntchito ufa wa mbewu. Zotsatira zathu zikusonyeza kuti ufa wa mbewu ya mpiru ukhoza kugwiritsidwa ntchito m'malo mwa mankhwala ophera tizilombo. Ukadaulo uwu ukhoza kukhala chida chatsopano chowongolera tizilombo toyambitsa udzudzu. Chifukwa mphutsi za udzudzu zimakula bwino m'madzi ndipo ma glucosinolates a ufa wa mbewu amasinthidwa kukhala isothiocyanate yogwira ntchito akangolowa madzi, kugwiritsa ntchito ufa wa mbewu ya mpiru m'madzi okhala ndi udzudzu kumapereka mphamvu yayikulu yowongolera. Ngakhale kuti ntchito yopha ma larvicate ya isothiocyanate imasiyana (BITC > AITC > 4-HBITC), kafukufuku wowonjezereka akufunika kuti adziwe ngati kuphatikiza ufa wa mbewu ndi ma glucosinolates angapo kumawonjezera poizoni. Uwu ndi kafukufuku woyamba kuwonetsa zotsatira za ufa wa ufa wa mbewu wochotsedwa mafuta ndi ma isothiocyanate atatu ogwira ntchito pa udzudzu. Zotsatira za kafukufukuyu zawonetsa kuti ufa wa mbewu ya kabichi wochotsedwa mafuta, womwe umachokera ku mafuta ochokera ku mbewu, ukhoza kukhala mankhwala ophera nkhupakupa omwe angathandize kuletsa udzudzu. Izi zingathandize kwambiri kupeza mankhwala ophera nkhupakupa m'minda komanso kupanga mankhwala ophera nkhupakupa monga mankhwala otsika mtengo, othandiza, komanso oteteza chilengedwe.
Ma data omwe adapangidwa pa kafukufukuyu ndi kusanthula komwe kwapezeka akupezeka kwa wolemba woyenerera ngati pakufunika kutero. Pamapeto pa kafukufukuyu, zinthu zonse zomwe zidagwiritsidwa ntchito mu kafukufukuyu (tizilombo ndi ufa wa mbewu) zidawonongeka.
Nthawi yotumizira: Julayi-29-2024