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Kuphatikiza mankhwala ophera tizilombo ochokera ku zomera kungasonyeze kuyanjana kwabwino kapena kotsutsana ndi tizilombo. Popeza matenda omwe amafalitsidwa ndi udzudzu wa Aedes akuchulukirachulukira komanso kukana kwa udzudzu wa Aedes ku mankhwala ophera tizilombo, kuphatikiza kwa mankhwala a terpene makumi awiri mphambu asanu ndi atatu ochokera ku mafuta ofunikira a zomera kunapangidwa ndikuyesedwa motsutsana ndi magawo a mphutsi ndi akuluakulu a Aedes aegypti. Mafuta asanu ofunikira a zomera (EOs) poyamba adayesedwa kuti agwire bwino ntchito popha larvicidal komanso kugwiritsa ntchito akuluakulu, ndipo mankhwala awiri akuluakulu adapezeka mu EO iliyonse kutengera zotsatira za GC-MS. Mankhwala akuluakulu omwe adadziwika adagulidwa, omwe ndi diallyl disulfide, diallyl trisulfide, carvone, limonene, eugenol, methyl eugenol, eucalyptol, eudesmol ndi udzudzu alpha-pinene. Kuphatikiza kwa mankhwala awa kunakonzedwa pogwiritsa ntchito mlingo wochepa ndipo zotsatira zake zogwirizana komanso zotsutsana zidayesedwa ndikudziwika. Mankhwala abwino kwambiri ophera zilonda amapezeka posakaniza limonene ndi diallyl disulfide, ndipo mankhwala abwino kwambiri ophera zilonda amapezeka posakaniza carvone ndi limonene. Mankhwala opangidwa ndi Temphos omwe amagwiritsidwa ntchito m'malonda ndi mankhwala a akuluakulu a Malathion adayesedwa padera komanso m'magulu awiri ndi terpenoids. Zotsatira zake zidawonetsa kuti kuphatikiza kwa temephos ndi diallyl disulfide ndi malathion ndi eudesmol kunali kothandiza kwambiri. Mankhwala amphamvu awa ndi omwe angagwiritsidwe ntchito polimbana ndi Aedes aegypti.
Mafuta ofunikira a zomera (EOs) ndi ma metabolites achiwiri omwe ali ndi mankhwala osiyanasiyana opatsa thanzi ndipo akukhala ofunikira kwambiri m'malo mwa mankhwala ophera tizilombo. Sikuti ndi ochezeka komanso osavuta kugwiritsa ntchito, komanso ndi osakaniza a mankhwala osiyanasiyana opatsa thanzi, zomwe zimachepetsanso mwayi wokhala ndi kukana mankhwala1. Pogwiritsa ntchito ukadaulo wa GC-MS, ofufuza adafufuza zomwe zili m'mafuta ofunikira a zomera zosiyanasiyana ndipo adapeza mankhwala opitilira 3,000 ochokera ku zomera 17,500 zonunkhira2, zomwe zambiri zidayesedwa kuti zipeze mphamvu zophera tizilombo ndipo akuti zili ndi zotsatira zophera tizilombo3,4. Kafukufuku wina akuwonetsa kuti poizoni wa gawo lalikulu la mankhwalawo ndi wofanana kapena wokulirapo kuposa wa ethylene oxide yake yosaphika. Koma kugwiritsa ntchito mankhwala payokha kungapatsenso mwayi wopanga kukana, monga momwe zilili ndi mankhwala ophera tizilombo5,6. Chifukwa chake, cholinga chamakono chili pakukonzekera zosakaniza za mankhwala opangidwa ndi ethylene oxide kuti apititse patsogolo mphamvu yophera tizilombo ndikuchepetsa mwayi wokana tizilombo. Mankhwala ogwira ntchito omwe ali mu EOs amatha kuwonetsa zotsatira zogwirizana kapena zotsutsana mu kuphatikiza komwe kukuwonetsa ntchito yonse ya EO, mfundo yomwe yatsindikitsidwa bwino mu maphunziro omwe ofufuza akale adachita7,8. Pulogalamu yowongolera ma vector imaphatikizaponso EO ndi zigawo zake. Ntchito yowononga udzudzu ya mafuta ofunikira yaphunziridwa kwambiri pa udzudzu wa Culex ndi Anopheles. Maphunziro angapo ayesa kupanga mankhwala othandiza pophatikiza zomera zosiyanasiyana ndi mankhwala ophera tizilombo opangidwa kuti awonjezere poizoni wonse ndikuchepetsa zotsatira zoyipa9. Koma maphunziro a mankhwala oterewa motsutsana ndi Aedes aegypti akadali osowa. Kupita patsogolo kwa sayansi ya zamankhwala ndi chitukuko cha mankhwala ndi katemera kwathandiza kuthana ndi matenda ena ofalitsidwa ndi ma vector. Koma kukhalapo kwa ma serotypes osiyanasiyana a kachilomboka, komwe kamafalitsidwa ndi udzudzu wa Aedes aegypti, kwapangitsa kuti mapulogalamu a katemera alephereke. Chifukwa chake, matenda otere akachitika, mapulogalamu owongolera ma vector ndiye njira yokhayo yopewera kufalikira kwa matendawa. Pakadali pano, kuwongolera Aedes aegypti ndikofunikira kwambiri chifukwa ndi njira yofunika kwambiri yopezera mavairasi osiyanasiyana ndi ma serotype awo omwe amayambitsa malungo a dengue, Zika, malungo a dengue hemorrhagic, malungo achikasu, ndi zina zotero. Chinthu chofunika kwambiri ndichakuti chiwerengero cha matenda pafupifupi onse ofalitsidwa ndi Aedes chikuwonjezeka chaka chilichonse ku Egypt ndipo chikuwonjezeka padziko lonse lapansi. Chifukwa chake, pankhaniyi, pakufunika kwambiri kupanga njira zowongolera zachilengedwe komanso zothandiza za Aedes aegypti. Omwe angakhalepo pankhaniyi ndi EOs, mankhwala omwe ali nawo, ndi kuphatikiza kwawo. Chifukwa chake, kafukufukuyu anayesa kupeza kuphatikiza kogwira mtima kwa mankhwala ofunikira a EO ochokera ku zomera zisanu zomwe zili ndi mphamvu zophera tizilombo (monga, timbewu ta ...
Ma EO onse osankhidwa adawonetsa mphamvu yopha ma larvicidal motsutsana ndi Aedes aegypti ndi 24-hour LC50 kuyambira 0.42 mpaka 163.65 ppm. Ntchito yopha ma larvicidal yapamwamba kwambiri idalembedwa pa peppermint (Mp) EO yokhala ndi LC50 ya 0.42 ppm pa maola 24, kutsatiridwa ndi adyo (As) yokhala ndi LC50 ya 16.19 ppm pa maola 24 (Table 1).
Kupatula Ocimum Sainttum, Os EO, ma EO ena onse anayi omwe adawunikidwa adawonetsa zotsatira zodziwikiratu za allercidal, ndi ma LC50 values oyambira 23.37 mpaka 120.16 ppm panthawi yomwe adawonetsedwa maola 24. Thymophilus striata (Cl) EO inali yothandiza kwambiri kupha akuluakulu omwe anali ndi LC50 value ya 23.37 ppm mkati mwa maola 24 atawonetsedwa, kutsatiridwa ndi Eucalyptus maculata (Em) yomwe inali ndi LC50 value ya 101.91 ppm (Table 1). Kumbali inayi, mtengo wa LC50 wa Os sunadziwikebe chifukwa chiŵerengero chachikulu cha imfa cha 53% chidalembedwa pa mlingo wapamwamba kwambiri (Chithunzi Chowonjezera 3).
Ma compound awiri akuluakulu omwe ali mu EO iliyonse adadziwika ndikusankhidwa kutengera zotsatira za database ya NIST laibulale, kuchuluka kwa dera la GC chromatogram, ndi zotsatira za MS spectra (Table 2). Pa EO As, ma compound akuluakulu omwe adadziwika anali diallyl disulfide ndi diallyl trisulfide; pa EO Mp ma compound akuluakulu omwe adadziwika anali carvone ndi limonene, pa EO Em ma compound akuluakulu omwe adadziwika anali eudesmol ndi eucalyptol; Pa EO Os, ma compound akuluakulu omwe adadziwika anali eugenol ndi methyl eugenol, ndipo pa EO Cl, ma compound akuluakulu omwe adadziwika anali eugenol ndi α-pinene (Chithunzi 1, Zithunzi Zowonjezera 5–8, Table Yowonjezera 1–5).
Zotsatira za mass spectrometry ya main terpenoids a mafuta osankhidwa ofunikira (A-diallyl disulfide; B-diallyl trisulfide; C-eugenol; D-methyl eugenol; E-limonene; F-aromatic ceperone; G-α-pinene; H-cineole; R-eudamol).
Mankhwala onse asanu ndi anayi (diallyl disulfide, diallyl trisulfide, eugenol, methyl eugenol, carvone, limonene, eucalyptol, eudesmol, α-pinene) adapezeka kuti ndi mankhwala othandiza omwe ndi zigawo zazikulu za EO ndipo adayesedwa payekhapayekha motsutsana ndi Aedes aegypti pamlingo wa mphutsi. . Eudesmol ya compound inali ndi ntchito yayikulu kwambiri yopha mphutsi yokhala ndi LC50 ya 2.25 ppm pambuyo pa maola 24 atakhudzidwa. Mankhwalawa a diallyl disulfide ndi diallyl trisulfide nawonso apezeka kuti ali ndi zotsatirapo zopha mphutsi, ndi milingo yapakati yoopsa pakati pa 10–20 ppm. Ntchito yopha mphutsi yapakati idawonedwanso pa mankhwala a eugenol, limonene ndi eucalyptol okhala ndi LC50 values ya 63.35 ppm, 139.29 ppm. ndi 181.33 ppm patatha maola 24, motsatana (Table 3). Komabe, palibe mphamvu yayikulu yopha larvicidal ya methyl eugenol ndi carvone yomwe idapezeka ngakhale pamlingo wapamwamba kwambiri, kotero ma LC50 values sanawerengedwe (Table 3). Larvicide yopangidwa ndi Temephos inali ndi kuchuluka kwakupha kwa 0.43 ppm motsutsana ndi Aedes aegypti pa maola 24 atakhudzidwa (Table 3, Table Supplementary 6).
Mankhwala asanu ndi awiri (diallyl disulfide, diallyl trisulfide, eucalyptol, α-pinene, eudesmol, limonene ndi carvone) adadziwika kuti ndi mankhwala akuluakulu a EO ndipo adayesedwa payekhapayekha motsutsana ndi udzudzu wa Aedes waku Egypt. Malinga ndi kusanthula kwa Probit regression, Eudesmol idapezeka kuti ili ndi mphamvu yayikulu yokhala ndi LC50 ya 1.82 ppm, kutsatiridwa ndi Eucalyptol yokhala ndi LC50 ya 17.60 ppm pa nthawi yowonekera maola 24. Mankhwala asanu otsala omwe adayesedwa anali owopsa pang'ono kwa akuluakulu omwe ali ndi LC50 kuyambira 140.79 mpaka 737.01 ppm (Table 3). Malathion yopangidwa ndi organophosphorus inali yochepa mphamvu kuposa eudesmol ndipo inali yokwera kuposa mankhwala ena asanu ndi limodzi, yokhala ndi LC50 ya 5.44 ppm pa nthawi yowonekera maola 24 (Table 3, Table Yowonjezera 6).
Mankhwala asanu ndi awiri amphamvu a lead ndi organophosphorus tamephosate adasankhidwa kuti apange ma binary mixtures a LC50 doses awo mu chiŵerengero cha 1:1. Ma binary mixture 28 onse adakonzedwa ndikuyesedwa kuti awone mphamvu yawo yopha larvicidal motsutsana ndi Aedes aegypti. Ma binary mixture asanu ndi anayi adapezeka kuti ndi othandizana, ma binary mixture 14 anali otsutsana, ndipo ma binary mixture asanu sanali othandizana ndi larvicidal. Pakati pa ma binary mixture, ma binary mixture a diallyl disulfide ndi temofol anali othandiza kwambiri, ndipo 100% ya imfa idawonedwa patatha maola 24 (Table 4). Mofananamo, ma binary mixture a limonene ndi diallyl disulfide ndi eugenol ndi thymetphos adawonetsa kuthekera kwabwino ndi 98.3% ya imfa ya mphutsi (Table 5). Kuphatikiza 4 kotsalako, komwe ndi eudesmol kuphatikiza eucalyptol, eudesmol kuphatikiza limonene, eucalyptol kuphatikiza alpha-pinene, alpha-pinene kuphatikiza temephos, kunawonetsanso mphamvu yopha zilonda, ndipo chiwerengero cha imfa chomwe chikuyembekezeka chikuposa 90%. Chiwerengero cha imfa chomwe chikuyembekezeka chili pafupi ndi 60-75%. (Table 4). Komabe, kuphatikiza kwa limonene ndi α-pinene kapena eucalyptus kunawonetsa zotsatira zotsutsana. Momwemonso, zosakaniza za Temephos ndi eugenol kapena eucalyptus kapena eudesmol kapena diallyl trisulfide zapezeka kuti zili ndi zotsatira zotsutsana. Momwemonso, kuphatikiza kwa diallyl disulfide ndi diallyl trisulfide ndi kuphatikiza kwa chilichonse mwa izi ndi eudesmol kapena eugenol ndizotsutsana mu zochita zawo zopha zilonda. Kutsutsana kwanenedwanso ndi kuphatikiza kwa eudesmol ndi eugenol kapena α-pinene.
Mwa zosakaniza zonse 28 za binary zomwe zinayesedwa kuti ziwonetse mphamvu ya asidi ya akuluakulu, zosakaniza 7 zinali zogwirizana, 6 sizinagwire ntchito, ndipo 15 zinali zotsutsana. Zosakaniza za eudesmol ndi eucalyptus ndi limonene ndi carvone zinapezeka kuti ndizothandiza kwambiri kuposa zosakaniza zina zogwirizana, ndipo chiwerengero cha imfa pa maola 24 cha 76% ndi 100%, motsatana (Table 5). Malathion yawonedwa kuti ikuwonetsa zotsatira zogwirizana ndi zosakaniza zonse za mankhwala kupatula limonene ndi diallyl trisulfide. Kumbali ina, kutsutsa kwapezeka pakati pa diallyl disulfide ndi diallyl trisulfide ndi kuphatikiza kwa chilichonse mwa izo ndi eucalyptus, kapena eucalyptol, kapena carvone, kapena limonene. Mofananamo, kuphatikiza kwa α-pinene ndi eudesmol kapena limonene, eucalyptol ndi carvone kapena limonene, ndi limonene ndi eudesmol kapena malathion kunawonetsa zotsatira zotsutsana ndi larvicidal. Pa zosakaniza zisanu ndi chimodzi zotsalazo, panalibe kusiyana kwakukulu pakati pa imfa zomwe zimayembekezeredwa ndi zomwe zimawonedwa (Table 5).
Kutengera zotsatira zogwirizana ndi mlingo wochepa wa mankhwala, poizoni wawo wopha tizilombo toyambitsa matenda motsutsana ndi udzudzu wambiri wa Aedes aegypti unasankhidwa ndipo unayesedwanso. Zotsatira zake zinasonyeza kuti imfa ya tizilombo toyambitsa matenda pogwiritsa ntchito njira ziwiri zophatikizana za eugenol-limonene, diallyl disulfide-limonene ndi diallyl disulfide-timephos inali 100%, pomwe imfa yomwe imayembekezeredwa ya tizilombo toyambitsa matenda inali 76.48%, 72.16% ndi 63.4%, motsatana (Table 6). . Kuphatikiza kwa limonene ndi eudesmol sikunali kogwira ntchito kwenikweni, ndipo 88% ya imfa ya tizilombo toyambitsa matenda inawonedwa mkati mwa maola 24 (Table 6). Mwachidule, njira zinayi zosakanikirana za tizilombo toyambitsa matenda zinawonetsanso zotsatira za tizilombo toyambitsa matenda motsutsana ndi Aedes aegypti zikagwiritsidwa ntchito pamlingo waukulu (Table 6).
Kuphatikiza katatu kogwirizana kunasankhidwa kuti kuyesedwe kwa anthu akuluakulu kuti kulamulire kuchuluka kwa Aedes aegypti akuluakulu. Pofuna kusankha kuphatikiza koyesera pa magulu akuluakulu a tizilombo, choyamba tinayang'ana kwambiri pa kuphatikiza kwabwino kwambiri kwa terpene kogwirizana, komwe ndi carvone plus limonene ndi eucalyptol plus eudesmol. Kachiwiri, kuphatikiza kwabwino kwambiri kogwirizana kunasankhidwa kuchokera ku kuphatikiza kwa organophosphate malathion yopangidwa ndi terpenoids. Tikukhulupirira kuti kuphatikiza kwa malathion ndi eudesmol ndiko kuphatikiza kwabwino kwambiri koyesera pa magulu akuluakulu a tizilombo chifukwa cha kufa kwakukulu komwe kumawonedwa komanso kuchuluka kochepa kwambiri kwa LC50 kwa zosakaniza zomwe zikufunika. Malathion imasonyeza mgwirizano wogwirizana ndi α-pinene, diallyl disulfide, eucalyptus, carvone ndi eudesmol. Koma ngati tiyang'ana pa kuchuluka kwa LC50, Eudesmol ili ndi mtengo wotsika kwambiri (2.25 ppm). Ma LC50 omwe anawerengedwa a malathion, α-pinene, diallyl disulfide, eucalyptol ndi carvone anali 5.4, 716.55, 166.02, 17.6 ndi 140.79 ppm. motsatana. Ma LC50 awa akusonyeza kuti kuphatikiza kwa malathion ndi eudesmol ndiye kuphatikiza koyenera kwambiri pankhani ya mlingo. Zotsatira zake zasonyeza kuti kuphatikiza kwa carvone plus limonene ndi eudesmol plus malathion kunali ndi imfa yowonedwa 100% poyerekeza ndi imfa yoyembekezeredwa ya 61% mpaka 65%. Kuphatikiza kwina, eudesmol plus eucalyptol, kunawonetsa kuchuluka kwa imfa ya 78.66% pambuyo pa maola 24 atagwiritsidwa ntchito, poyerekeza ndi kuchuluka kwa imfa yoyembekezeredwa ya 60%. Kuphatikiza konseko katatu komwe kunasankhidwa kunawonetsa zotsatira zogwirizana ngakhale kugwiritsidwa ntchito pamlingo waukulu motsutsana ndi Aedes aegypti wamkulu (Table 6).
Mu kafukufukuyu, zomera zosankhidwa monga Mp, As, Os, Em ndi Cl zinawonetsa zotsatira zabwino zakupha pa mphutsi ndi za akuluakulu a Aedes aegypti. Mp EO inali ndi mphamvu yowononga mphutsi kwambiri yokhala ndi LC50 ya 0.42 ppm, kutsatiridwa ndi As, Os ndi Em EO yokhala ndi LC50 ya pansi pa 50 ppm patatha maola 24. Zotsatirazi zikugwirizana ndi kafukufuku wakale wa udzudzu ndi ntchentche zina zotuluka m'madzi10,11,12,13,14. Ngakhale mphamvu yowononga mphutsi ya Cl ndi yotsika kuposa mafuta ena ofunikira, yokhala ndi LC50 ya 163.65 ppm patatha maola 24, mphamvu yake yowononga mphutsi ndi yapamwamba kwambiri yokhala ndi LC50 ya 23.37 ppm patatha maola 24. Mp, As ndi Em EOs adawonetsanso mphamvu yabwino yolimbana ndi allercidal yokhala ndi ma LC50 values omwe ali pakati pa 100–120 ppm pa maola 24 atakhudzidwa, koma anali otsika poyerekeza ndi mphamvu yawo yopha tizilombo toyambitsa matenda. Kumbali ina, EO Os idawonetsa mphamvu yochepa yolimbana ndi allercidal ngakhale pa mlingo wapamwamba kwambiri wochiritsira. Chifukwa chake, zotsatira zake zikuwonetsa kuti poizoni wa ethylene oxide ku zomera amatha kusiyana kutengera gawo la chitukuko cha udzudzu15. Zimatengeranso kuchuluka kwa kulowa kwa EOs m'thupi la tizilombo, kuyanjana kwawo ndi ma enzyme enaake, komanso mphamvu ya udzudzu pa gawo lililonse la chitukuko16. Kafukufuku wambiri wasonyeza kuti chigawo chachikulu cha mankhwala ndi chinthu chofunikira kwambiri pa ntchito yachilengedwe ya ethylene oxide, popeza ndiye amachititsa kuti pakhale mankhwala ambiri 3,12,17,18. Chifukwa chake, tidaganizira za mankhwala awiri akuluakulu mu EO iliyonse. Kutengera zotsatira za GC-MS, diallyl disulfide ndi diallyl trisulfide zinadziwika ngati mankhwala akuluakulu a EO As, zomwe zikugwirizana ndi malipoti am'mbuyomu19,20,21. Ngakhale malipoti am'mbuyomu adawonetsa kuti menthol inali imodzi mwa mankhwala ake akuluakulu, carvone ndi limonene zinadziwikanso ngati mankhwala akuluakulu a Mp EO22,23. Kapangidwe ka Os EO kanawonetsa kuti eugenol ndi methyl eugenol ndiye mankhwala akuluakulu, zomwe zikufanana ndi zomwe ofufuza am'mbuyomu16,24 adapeza. Eucalyptol ndi eucalyptol zanenedwa kuti ndi mankhwala akuluakulu omwe amapezeka mu mafuta a masamba a Em, zomwe zikugwirizana ndi zomwe ofufuza ena adapeza25,26 koma mosiyana ndi zomwe Olalade et al.27 adapeza. Kuchuluka kwa cineole ndi α-pinene kunawonedwa mu mafuta ofunikira a melaleuca, zomwe zikufanana ndi maphunziro am'mbuyomu28,29. Kusiyana kwapadera kwa kapangidwe ndi kuchuluka kwa mafuta ofunikira otengedwa kuchokera ku mitundu yofanana ya zomera m'malo osiyanasiyana kwanenedwa ndipo kwawonedwanso mu kafukufukuyu, komwe kumakhudzidwa ndi momwe zomera zimakulira, nthawi yokolola, gawo la chitukuko, kapena zaka za zomera. Kuwonekera kwa mitundu ya chemotypes, ndi zina zotero.22,30,31,32. Kenako mankhwala ofunikira omwe adapezeka adagulidwa ndikuyesedwa kuti awone zotsatira zake zopha mphutsi ndi zotsatira zake pa udzudzu wa Aedes aegypti wamkulu. Zotsatira zake zidawonetsa kuti ntchito yopha mphutsi ya diallyl disulfide inali yofanana ndi ya EO As yopanda kanthu. Koma ntchito ya diallyl trisulfide ndi yokwera kuposa EO As. Zotsatirazi ndizofanana ndi zomwe Kimbaris et al. 33 adapeza pa Culex philippines. Komabe, mankhwala awiriwa sanawonetse ntchito yabwino yopha mphutsi motsutsana ndi udzudzu womwe ukufuna, zomwe zikugwirizana ndi zotsatira za Plata-Rueda et al 34 pa Tenebrio molitor. Os EO imagwira ntchito motsutsana ndi gawo la mphutsi la Aedes aegypti, koma osati motsutsana ndi gawo la akuluakulu. Zatsimikiziridwa kuti ntchito yopha zilonda za mankhwala akuluakulu ndi yotsika kuposa ya crude Os EO. Izi zikutanthauza kuti ntchito ya mankhwala ena ndi momwe amachitira zinthu mu crude ethylene oxide. Methyl eugenol yokha ili ndi ntchito yochepa, pomwe eugenol yokha ili ndi ntchito yopha zilonda zapakati. Izi zikutsimikizira, kumbali imodzi, 35,36, ndipo kumbali ina, zikutsutsana ndi zomwe ofufuza akale adaganiza 37,38. Kusiyana kwa magulu ogwira ntchito a eugenol ndi methyleugenol kungayambitse poizoni wosiyanasiyana ku tizilombo tomwe tikufuna 39. Limonene idapezeka kuti ili ndi ntchito yopha zilonda zapakati, pomwe zotsatira za carvone sizinali zazikulu. Mofananamo, poizoni wochepa wa limonene kwa tizilombo tating'onoting'ono ta akuluakulu komanso poizoni wambiri wa carvone zimathandizira zotsatira za maphunziro ena am'mbuyomu 40 koma zikutsutsana ndi zina 41. Kupezeka kwa ma bond awiri pa malo onse a intracyclic ndi exocyclic kungawonjezere ubwino wa mankhwala awa monga larvicides3,41, pomwe carvone, yomwe ndi ketone yokhala ndi ma alpha ndi beta carbon osakhuta, ikhoza kuwonetsa kuthekera kwakukulu kwa poizoni mwa akuluakulu42. Komabe, makhalidwe a limonene ndi carvone ndi otsika kwambiri kuposa EO yonse Mp (Table 1, Table 3). Pakati pa terpenoids omwe adayesedwa, eudesmol idapezeka kuti ili ndi ntchito yayikulu kwambiri yopha larvicidal komanso ya akuluakulu yokhala ndi LC50 value pansi pa 2.5 ppm, zomwe zimapangitsa kuti ikhale mankhwala abwino kwambiri oletsa udzudzu wa Aedes. Kuchita kwake bwino kuposa kwa EO Em yonse, ngakhale izi sizikugwirizana ndi zomwe Cheng et al.40 adapeza. Eudesmol ndi sesquiterpene yokhala ndi mayunitsi awiri a isoprene omwe sasinthasintha kwambiri kuposa ma monoterpenes okhala ndi mpweya monga eucalyptus ndipo motero ali ndi kuthekera kwakukulu ngati mankhwala ophera tizilombo. Eucalyptol yokha ili ndi mphamvu yowononga mabakiteriya akuluakulu kuposa larvicidal, ndipo zotsatira za maphunziro akale zimathandizira ndikutsutsa izi37,43,44. Ntchito yokhayo ndi yofanana ndi ya EO Cl yonse. Monoterpene ina ya bicyclic, α-pinene, ili ndi mphamvu yochepa pa Aedes aegypti kuposa larvicidal effect, zomwe ndizosiyana ndi zotsatira za EO Cl yonse. Ntchito yonse yopha tizilombo ya terpenoids imakhudzidwa ndi lipophilicity yawo, kusasinthasintha, nthambi ya kaboni, malo owonetsera, malo ogwirira ntchito, magulu ogwira ntchito ndi malo awo45,46. Mankhwalawa amatha kugwira ntchito powononga kusonkhanitsa kwa maselo, kuletsa ntchito yopuma, kusokoneza kufalikira kwa mitsempha, ndi zina zotero.47 Organophosphate yopangidwa ndi Temephos idapezeka kuti ili ndi mphamvu yopha mabakiteriya apamwamba kwambiri yokhala ndi LC50 ya 0.43 ppm, zomwe zikugwirizana ndi deta ya Lek -Utala48. Ntchito ya akuluakulu ya synthetic organophosphorus malathion idanenedwa pa 5.44 ppm. Ngakhale kuti ma organophosphate awiriwa asonyeza kuti achita bwino polimbana ndi mitundu ya Aedes aegypti ya m'ma laboratories, kukana kwa udzudzu ku mankhwala amenewa kwanenedwa m'madera osiyanasiyana padziko lonse lapansi49. Komabe, palibe malipoti ofanana ndi amenewa okhudza kukula kwa kukana mankhwala azitsamba omwe apezeka50. Chifukwa chake, zomera zimaonedwa ngati njira zina zomwe zingatheke m'malo mwa mankhwala ophera tizilombo m'mapulogalamu owongolera tizilombo.
Mphamvu yopha ma larvicidal inayesedwa pa ma binary mixtures 28 (1:1) opangidwa kuchokera ku terpenoids amphamvu ndi terpenoids okhala ndi thymetphos, ndipo ma synergistics 9 anapezeka kuti ndi othandizana, 14 otsutsana ndi 5 otsutsana. Palibe zotsatira. Kumbali ina, mu bioassay ya mphamvu ya akuluakulu, ma synergistics 7 anapezeka kuti ndi othandizana, ma synergistics 15 anali otsutsana, ndipo ma synergistics 6 ananenedwa kuti alibe zotsatira. Chifukwa chomwe ma synergistics ena amapangira mphamvu yogwirizana chingakhale chifukwa cha ma conducts omwe amalumikizana nthawi imodzi m'njira zosiyanasiyana zofunika, kapena kuletsa motsatizana kwa ma enzymes ofunikira osiyanasiyana a njira inayake yachilengedwe51. Kuphatikiza kwa limonene ndi diallyl disulfide, eucalyptus kapena eugenol kunapezeka kuti kumathandizirana m'magwiritsidwe ang'onoang'ono ndi akuluakulu (Table 6), pomwe kuphatikiza kwake ndi eucalyptus kapena α-pinene kunapezeka kuti kumakhudza mphutsi. Pa avareji, limonene imawoneka kuti ndi synergist yabwino, mwina chifukwa cha kukhalapo kwa magulu a methyl, kulowa bwino mu stratum corneum, ndi njira yosiyana ya kagwiridwe ka ntchito52,53. Zanenedwa kale kuti limonene ingayambitse zotsatira zoopsa polowa m'matumbo a tizilombo (poizoni wokhudzana ndi kukhudzana), kukhudza dongosolo la m'mimba (antifeedant), kapena kukhudza dongosolo la kupuma (fumigation activity), 54 pomwe phenylpropanoids monga eugenol ingakhudze ma enzymes a kagayidwe kachakudya 55. Chifukwa chake, kuphatikiza kwa mankhwala okhala ndi njira zosiyanasiyana zogwirira ntchito kungapangitse kuti chisakanizocho chikhale ndi zotsatira zoopsa kwambiri. Eucalyptol idapezeka kuti imagwirizana ndi diallyl disulfide, eucalyptus kapena α-pinene, koma kuphatikiza kwina ndi mankhwala ena sikunali koopsa kapena kotsutsana. Kafukufuku woyambirira adawonetsa kuti eucalyptol ili ndi ntchito yoletsa acetylcholinesterase (AChE), komanso octamine ndi GABA receptors56. Popeza ma cyclic monoterpenes, eucalyptol, eugenol, ndi zina zotero zingakhale ndi njira yofanana ndi ntchito yawo yowononga mitsempha, 57 motero zimachepetsa zotsatira zawo zonse kudzera mu kuletsana. Momwemonso, kuphatikiza kwa Temephos ndi diallyl disulfide, α-pinene ndi limonene kunapezeka kuti kumagwirizana, kuthandizira malipoti am'mbuyomu a mphamvu yogwirizana pakati pa mankhwala azitsamba ndi organophosphates zopangidwa58.
Kuphatikiza kwa eudesmol ndi eucalyptol kunapezeka kuti kumakhudza mgwirizano pa magawo a mphutsi ndi akuluakulu a Aedes aegypti, mwina chifukwa cha njira zawo zosiyanasiyana zogwirira ntchito chifukwa cha kapangidwe kake kosiyanasiyana ka mankhwala. Eudesmol (sesquiterpene) ingakhudze dongosolo lopumira 59 ndipo eucalyptol (monoterpene) ingakhudze acetylcholinesterase 60. Kuphatikizika kwa zosakaniza pamalo awiri kapena kuposerapo kungapangitse kuti kuphatikizako kukhale koopsa. Mu kafukufuku wa zinthu za munthu wamkulu, malathion inapezeka kuti imagwirizana ndi carvone kapena eucalyptol kapena eucalyptol kapena diallyl disulfide kapena α-pinene, zomwe zikusonyeza kuti imagwirizana ndi kuwonjezera kwa limonene ndi di. Ndi mankhwala abwino opha tizilombo toyambitsa matenda omwe amagwiritsidwa ntchito pa ntchito yonse ya mankhwala a terpene, kupatula allyl trisulfide. Thangam ndi Kathiresan61 adanenanso za zotsatira zofanana za mphamvu ya malathion yogwirizana ndi zotulutsa zitsamba. Yankho la mgwirizanowu likhoza kukhala chifukwa cha zotsatira za poizoni za malathion ndi phytochemicals pa ma enzymes ochotsa tizilombo. Ma Organophosphates monga malathion nthawi zambiri amagwira ntchito poletsa ma cytochrome P450 esterases ndi monooxygenases62,63,64. Chifukwa chake, kuphatikiza malathion ndi njira izi zogwirira ntchito ndi ma terpenes ndi njira zosiyanasiyana zogwirira ntchito kungapangitse kuti pakhale zotsatira zoopsa pa udzudzu.
Kumbali ina, kutsutsa kumasonyeza kuti mankhwala osankhidwawo sagwira ntchito kwambiri pophatikizana kuposa mankhwala aliwonse okha. Chifukwa cha kutsutsa m'maphatikizidwe ena chingakhale chakuti mankhwala ena amasintha khalidwe la mankhwala ena mwa kusintha liwiro la kuyamwa, kugawa, kagayidwe kachakudya, kapena kutulutsa. Ofufuza oyambirira ankaona kuti izi ndi zomwe zimayambitsa kutsutsa m'maphatikizidwe a mankhwala. Mamolekyulu Njira yomwe ingakhalepo 65. Mofananamo, zomwe zimayambitsa kutsutsa zitha kukhala zokhudzana ndi njira zofanana zogwirira ntchito, mpikisano wa mankhwala omwe ali m'gulu la receptor kapena malo omwe akufunidwa. Nthawi zina, kuletsa kosapikisana kwa mapuloteni omwe akufunidwa kungachitikenso. Mu kafukufukuyu, mankhwala awiri a organosulfur, diallyl disulfide ndi diallyl trisulfide, adawonetsa zotsatira zotsutsana, mwina chifukwa cha kupikisana ndi malo omwewo omwe akufunidwa. Momwemonso, mankhwala awiriwa a sulfure adawonetsa zotsatira zotsutsana ndipo sanakhudzepo akaphatikizidwa ndi eudesmol ndi α-pinene. Eudesmol ndi alpha-pinene ndi ozungulira, pomwe diallyl disulfide ndi diallyl trisulfide ndi aliphatic m'chilengedwe. Kutengera kapangidwe ka mankhwala, kuphatikiza kwa mankhwala awa kuyenera kuwonjezera mphamvu yakupha yonse popeza malo omwe amafunidwa nthawi zambiri amakhala osiyana34,47, koma mwa kuyesa tinapeza kuti kudana, komwe kungakhale chifukwa cha ntchito ya mankhwala awa m'zinthu zina zosadziwika zomwe zimachitika m'thupi chifukwa cha kuyanjana. Mofananamo, kuphatikiza kwa cineole ndi α-pinene kunapanga mayankho otsutsana, ngakhale ofufuza kale adanena kuti mankhwala awiriwa ali ndi zolinga zosiyana za ntchito47,60. Popeza mankhwala onsewa ndi a cyclic monoterpenes, pakhoza kukhala malo ena ofanana omwe angapikisane kuti amange ndikukhudza poizoni wonse wa awiriawiri ophatikizana omwe adaphunziridwa.
Kutengera ndi kuchuluka kwa LC50 ndi kufa komwe kunawonedwa, mitundu iwiri yabwino kwambiri ya terpene yogwirizana idasankhidwa, yomwe ndi iwiri ya carvone + limonene ndi eucalyptol + eudesmol, komanso organophosphorus malathion yopangidwa ndi terpenes. Kuphatikiza koyenera kwambiri kwa mankhwala a malathion + Eudesmol kunayesedwa mu bioassay ya tizilombo ta akuluakulu. Yang'anani magulu akuluakulu a tizilombo kuti mutsimikizire ngati kuphatikiza kogwira mtima kumeneku kungagwire ntchito motsutsana ndi anthu ambiri m'malo akuluakulu owonekera. Kuphatikiza konseku kukuwonetsa mphamvu yogwirizana motsutsana ndi magulu akuluakulu a tizilombo. Zotsatira zofananazi zidapezeka pakuphatikiza koyenera kwambiri kwa mankhwala opha tizilombo omwe adayesedwa motsutsana ndi magulu akuluakulu a mphutsi za Aedes aegypti. Chifukwa chake, tinganene kuti kuphatikiza kogwira mtima kwa mankhwala opha tizilombo komanso opha tizilombo a zomera za EO ndi njira yabwino yolimbana ndi mankhwala omwe alipo ndipo angagwiritsidwe ntchito kwambiri poletsa kuchuluka kwa Aedes aegypti. Momwemonso, kuphatikiza kogwira mtima kwa mankhwala ophera larvicides kapena mankhwala ophera achikulire ndi terpenes kungagwiritsidwenso ntchito kuchepetsa mlingo wa thymetphos kapena malathion woperekedwa kwa udzudzu. Kuphatikiza kwamphamvu kumeneku kungapereke mayankho a kafukufuku wamtsogolo pa kusintha kwa kukana mankhwala mu udzudzu wa Aedes.
Mazira a Aedes aegypti adasonkhanitsidwa kuchokera ku Regional Medical Research Centre, Dibrugarh, Indian Council of Medical Research ndipo adasungidwa kutentha koyenera (28 ± 1 °C) ndi chinyezi (85 ± 5%) mu Dipatimenti ya Zoology, Gauhati University motsatira izi: Arivoli adafotokozedwa ndi ena. Pambuyo pobereka, mphutsi zidapatsidwa chakudya cha mphutsi (ufa wa mabisiketi agalu ndi yisiti mu chiŵerengero cha 3:1) ndipo akuluakulu adapatsidwa 10% glucose solution. Kuyambira tsiku lachitatu kuchokera pamene adatuluka, udzudzu wamkazi wamkulu adaloledwa kuyamwa magazi a makoswe a albino. Zilowerereni pepala losefera m'madzi mu galasi ndikuliyika mu khola loikira mazira.
Zitsanzo za zomera zomwe zinasankhidwa monga masamba a eucalyptus (Myrtaceae), basil woyera (Lamiaceae), timbewu ta mint (Lamiaceae), melaleuca (Myrtaceae) ndi mababu a allium (Amaryllidaceae). Zinasonkhanitsidwa kuchokera ku Guwahati ndipo zinapezeka ndi Dipatimenti ya Botany, Gauhati University. Zitsanzo za zomera zomwe zinasonkhanitsidwa (500 g) zinaphwanyidwa ndi madzi pogwiritsa ntchito chipangizo cha Clevenger kwa maola 6. EO yochotsedwayo inasonkhanitsidwa m'mabotolo oyera agalasi ndikusungidwa pa 4°C kuti iphunzire zambiri.
Kuopsa kwa tizilombo toyambitsa matenda m'matupi kunaphunziridwa pogwiritsa ntchito njira zosinthidwa pang'ono za World Health Organization 67. Gwiritsani ntchito DMSO ngati emulsifier. Kuchuluka kwa EO kulikonse kunayesedwa poyamba pa 100 ndi 1000 ppm, kuwonetsa mphutsi 20 pa kubwerezabwereza kulikonse. Kutengera zotsatira zake, kuchuluka kwa tizilombo kunagwiritsidwa ntchito ndipo imfa zinalembedwa kuyambira ola limodzi mpaka maola 6 (pa nthawi ya ola limodzi), komanso pa maola 24, maola 48 ndi maola 72 mutalandira chithandizo. Kuchuluka kwa tizilombo toyambitsa matenda (LC50) kunapezeka pambuyo pa maola 24, 48 ndi 72 mutapezeka. Kuchuluka kulikonse kunayesedwa katatu pamodzi ndi kulamulira koipa (madzi okha) ndi kulamulira koyenera kamodzi (madzi okonzedwa ndi DMSO). Ngati kufalikira kwa tizilombo kumachitika ndipo mphutsi zoposa 10% za gulu lolamulira zimafa, kuyesaku kumabwerezedwanso. Ngati kuchuluka kwa imfa m'gulu lolamulira kuli pakati pa 5-10%, gwiritsani ntchito njira yowongolera ya Abbott 68.
Njira yomwe Ramar et al. 69 adafotokoza idagwiritsidwa ntchito poyesa matenda a munthu wamkulu motsutsana ndi Aedes aegypti pogwiritsa ntchito acetone ngati chosungunulira. EO iliyonse idayesedwa poyamba motsutsana ndi udzudzu wa Aedes aegypti wachikulire pamlingo wa 100 ndi 1000 ppm. Ikani 2 ml ya yankho lililonse lokonzedwa ku nambala ya Whatman. Chidutswa chimodzi cha pepala losefera (kukula 12 x 15 cm2) ndikusiya acetone isungunuke kwa mphindi 10. Pepala losefera lokhala ndi acetone 2 ml yokha linagwiritsidwa ntchito ngati chowongolera. Acetone ikatha, pepala losefera lokhala ndi chowongolera ndi pepala lowongolera limayikidwa mu chubu chozungulira (10 cm kuya). Udzudzu khumi wa masiku 3 mpaka 4 wosadya magazi unasamutsidwa kukhala katatu pamlingo uliwonse. Kutengera zotsatira za mayeso oyamba, kuchuluka kosiyanasiyana kwa mafuta osankhidwa kunayesedwa. Imfa zinalembedwa pa ola limodzi, maola awiri, maola atatu, maola anayi, maola asanu, maola asanu ndi limodzi, maola makumi awiri ndi anayi, maola makumi asanu ndi limodzi ndi maola makumi asanu ndi awiri ndi awiri ndi maola makumi asanu ndi awiri ndi awiri (72) udzudzu utatuluka. Werengani kuchuluka kwa LC50 kuti mudziwe nthawi yomwe udzudzu unakumana nayo pa maola 24, maola makumi anayi ndi asanu ndi atatu ndi maola asanu ndi awiri (72). Ngati chiwerengero cha imfa cha gulu loyang'anira chapitirira 20%, bwerezani mayeso onse. Mofananamo, ngati chiwerengero cha imfa m'gulu loyang'anira chapitirira 5%, sinthani zotsatira za zitsanzo zomwe zachiritsidwa pogwiritsa ntchito fomula ya Abbott68.
Kuyeza kwa mpweya (Agilent 7890A) ndi mass spectrometry (Accu TOF GCv, Jeol) kunachitidwa kuti afufuze zinthu zomwe zili mu mafuta ofunikira osankhidwa. GC inali ndi chowunikira cha FID ndi capillary column (HP5-MS). Mpweya wonyamula unali helium, ndipo kuchuluka kwa madzi kunali 1 ml/min. Pulogalamu ya GC imayika Allium sativum pa 10:80-1M-8-220-5M-8-270-9M ndi Ocimum Sainttum pa 10:80-3M-8-200-3M-10-275-1M-5 – 280, ya timbewu ta ...
Ma compound akuluakulu a EO iliyonse adazindikirika kutengera kuchuluka kwa dera komwe kudawerengedwa kuchokera ku zotsatira za GC chromatogram ndi mass spectrometry (zotchulidwa ku database ya NIST 70 standards).
Mankhwala awiri akuluakulu mu EO iliyonse adasankhidwa kutengera zotsatira za GC-MS ndipo adagulidwa kuchokera ku Sigma-Aldrich pa 98–99% yoyera kuti apeze zotsatira zina za bioassays. Mankhwalawa adayesedwa kuti awone ngati ali ndi mphamvu yopha ma larvicides komanso mphamvu ya akuluakulu motsutsana ndi Aedes aegypti monga tafotokozera pamwambapa. Mankhwala opangidwa ndi larvicides tamephosate (Sigma Aldrich) ndi mankhwala a akuluakulu a malathion (Sigma Aldrich) adawunikidwa kuti ayerekeze kugwira ntchito kwawo ndi mankhwala osankhidwa a EO, potsatira njira yomweyi.
Zosakaniza za binary za mankhwala osankhidwa a terpene ndi mankhwala a terpene kuphatikiza ma organophosphates ogulitsa (tilephos ndi malathion) zinakonzedwa mwa kusakaniza mlingo wa LC50 wa mankhwala aliwonse oyenerera mu chiŵerengero cha 1:1. Zosakaniza zokonzedwazo zinayesedwa pa magawo a mphutsi ndi akuluakulu a Aedes aegypti monga tafotokozera pamwambapa. Kuyesa kulikonse kwa bioassay kunachitika katatu pa kuphatikiza kulikonse komanso katatu pa mankhwala aliwonse omwe alipo pakuphatikiza kulikonse. Imfa ya tizilombo tomwe tikufunira inalembedwa patatha maola 24. Werengani kuchuluka kwa imfa komwe kumayembekezeredwa pa kusakaniza kwa binary pogwiritsa ntchito njira yotsatirayi.
kumene E = chiŵerengero cha imfa chomwe chikuyembekezeka cha udzudzu wa Aedes aegypti chifukwa cha kuphatikizana kwa binary, mwachitsanzo kulumikizana (A + B).
Zotsatira za kusakaniza kulikonse kwa binary zidalembedwa kuti ndi zogwirizana, zotsutsana, kapena zopanda zotsatira kutengera mtengo wa χ2 wowerengedwa ndi njira yomwe Pavla52 adafotokoza. Werengani mtengo wa χ2 wa kuphatikiza kulikonse pogwiritsa ntchito fomula yotsatirayi.
Zotsatira za kuphatikizana zinatanthauzidwa ngati mgwirizano pamene mtengo wowerengedwa wa χ2 unali waukulu kuposa mtengo wa tebulo wa madigiri ofanana a ufulu (95% nthawi yodzidalira) ndipo ngati imfa yowonedwa inapezeka kuti yapitirira imfa yoyembekezeredwa. Mofananamo, ngati mtengo wowerengedwa wa χ2 wa kuphatikiza kulikonse unapitirira mtengo wa tebulo ndi madigiri ena a ufulu, koma imfa yowonedwa ndi yotsika kuposa imfa yoyembekezeredwa, chithandizocho chimaonedwa kuti ndi chotsutsana. Ndipo ngati mu kuphatikiza kulikonse mtengo wowerengedwa wa χ2 uli wochepera kuposa mtengo wa tebulo m'madigiri ofanana a ufulu, kuphatikizako kumaonedwa kuti kulibe mphamvu.
Kuphatikiza katatu kapena kanayi komwe kungatheke kugwirizana (mphutsi 100 ndi tizilombo 50 topha tizilombo toyambitsa matenda komanso akuluakulu) kunasankhidwa kuti kuyesedwe motsutsana ndi tizilombo tochuluka. Akuluakulu) amapitilira monga momwe tafotokozera pamwambapa. Pamodzi ndi zosakaniza, mankhwala omwe alipo m'zosakaniza zomwe zasankhidwa adayesedwanso pa kuchuluka kofanana kwa mphutsi za Aedes aegypti ndi akuluakulu. Chiŵerengero chophatikizana ndi gawo limodzi la mlingo wa LC50 wa mankhwala amodzi oyenera ndi gawo la mlingo wa LC50 wa mankhwala ena omwe ali m'gululi. Mu kafukufuku wa zochita za akuluakulu, mankhwala osankhidwa adasungunuka mu acetone yosungunulira ndikugwiritsidwa ntchito papepala losefera lomwe lakulungidwa mu chidebe cha pulasitiki cha silinda cha 1300 cm3. Acetone idasanduka nthunzi kwa mphindi 10 ndipo akuluakulu adatulutsidwa. Mofananamo, mu kafukufuku wa zochita za larvicidal, mlingo wa mankhwala ofanana a LC50 poyamba unasungunuka m'mavoliyumu ofanana a DMSO kenako nkusakanizidwa ndi lita imodzi ya madzi yosungidwa m'zidebe za pulasitiki za 1300 cc, ndipo mphutsi zidatulutsidwa.
Kusanthula kwadzidzidzi kwa deta 71 ya imfa yolembedwa kunachitika pogwiritsa ntchito SPSS (mtundu 16) ndi pulogalamu ya Minitab kuti awerengere kuchuluka kwa LC50.
Nthawi yotumizira: Julayi-01-2024