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Hiki i ka hui ʻana o nā pūhui insecticidal i loaʻa i nā mea kanu ke hōʻike i ka synergistic a i ʻole nā mea kūʻē i nā mea ʻino.Ma muli o ka laha wikiwiki ʻana o nā maʻi i lawe ʻia e nā makika Aedes a me ka piʻi ʻana o ke kūʻē ʻana o ka heluna namu Aedes i nā lāʻau lāʻau kuʻuna, ua hoʻokumu ʻia a hoʻāʻo ʻia e kūʻē i nā pae larval a me nā pae makua o Aedes aegypti.ʻElima mea kanu ʻaila nui (EO) i loiloi mua ʻia no kā lākou larvicidal a me ka hoʻohana ʻana i ka pono o nā mākua, a ʻelua mau mea nui i ʻike ʻia i kēlā me kēia EO ma muli o nā hopena GC-MS.Ua kūʻai ʻia nā pūhui nui i ʻike ʻia, ʻo ia ka diallyl disulfide, diallyl trisulfide, carvone, limonene, eugenol, methyl eugenol, eucalyptol, eudesmol a me namu alpha-pinene.A laila hoʻomākaukau ʻia nā hui binary o kēia mau pūhui me ka hoʻohana ʻana i nā dosis sublethal a ua hoʻāʻo ʻia a hoʻoholo ʻia ko lākou hopena synergistic a antagonistic.Loaʻa nā haku mele larvicidal maikaʻi loa ma ka hui ʻana i ka limonene me ka diallyl disulfide, a loaʻa nā haku mele maikaʻi loa ma ka hui ʻana i ka carvone me ka limonene.Ua ho'āʻo kaʻawale ʻia ka larvicide synthetic ʻo Temphos a me ka lāʻau makua Malathion i hoʻokaʻawale ʻia a ma nā hui binary me nā terpenoids.Ua hōʻike nā hopena i ka hui pū ʻana o ka temephos a me diallyl disulfide a me ka malathion a me ka eudesmol ka hui maikaʻi loa.Hiki ke hoʻohana ʻia kēia mau hui pū me ka Aedes aegypti.
ʻO nā ʻaila koʻikoʻi o nā mea kanu (EO) nā metabolites lua i loaʻa nā pūhui bioactive like ʻole a ke lilo nei i mea nui ma ke ʻano he ʻokoʻa i nā pesticides synthetic.ʻAʻole wale lākou e pili ana i ke kaiapuni a me ka mea hoʻohana, akā he hui pū kekahi o nā pūhui bioactive like ʻole, e hōʻemi ana i ka hiki ke hoʻomohala i ka lāʻau kūʻē1.Ma ka hoʻohana ʻana i ka ʻenehana GC-MS, ua nānā nā mea noiʻi i nā mea i hoʻohui ʻia o nā ʻaila koʻikoʻi o nā mea kanu a ʻike ʻia ma mua o 3,000 pūhui mai 17,500 mea kanu ʻala2, ʻo ka hapa nui o ia mau mea i hoʻāʻo ʻia no nā waiwai insecticidal a ua hōʻike ʻia he hopena insecticidal3,4.Hōʻike kekahi mau haʻawina i ka like a i ʻole ka nui o ka ʻawaʻawa o ka mea nui o ka pūhui ma mua o kāna ethylene oxide.Akā ʻo ka hoʻohana ʻana i kēlā me kēia hui e waiho hou i kahi no ka hoʻomohala ʻana i ke kū'ē, e like me ke ʻano o nā insecticides kemika5,6.No laila, ʻo ka manaʻo nui o kēia manawa i ka hoʻomākaukau ʻana i nā hui ʻana o nā pūhui ethylene oxide e hoʻomaikaʻi i ka hopena insecticidal a hōʻemi i ka hiki ke kū i ka heluna kanaka pest.Hiki ke hōʻike ʻia nā pūhui hana hoʻokahi i loko o nā EO i nā hopena synergistic a i ʻole antagonistic i nā hui pū ʻana e hōʻike ana i ka hana holoʻokoʻa o ka EO, he ʻoiaʻiʻo i hoʻoikaika maikaʻi ʻia i nā haʻawina i hana ʻia e nā mea noiʻi mua7,8.Aia pū ka polokalamu hoʻokele vector i ka EO a me kāna mau ʻāpana.Ua aʻo nui ʻia ka hana namu o nā ʻaila koʻikoʻi ma nā namu Culex a me Anopheles.Ua ho'āʻo kekahi mau haʻawina e hoʻomohala i nā lāʻau pesticides maikaʻi ma o ka hoʻohui ʻana i nā mea kanu like ʻole me nā pesticides synthetic i hoʻohana ʻia ma ke kālepa e hoʻonui i ka ʻona a hoʻemi i nā hopena ʻaoʻao9.Akā ʻaʻole kakaikahi nā haʻawina o ia mau pūhui e kūʻē iā Aedes aegypti.Ua kōkua ka holomua ʻana i ka ʻepekema lapaʻau a me ka hoʻomohala ʻana i nā lāʻau lapaʻau a me nā lāʻau lapaʻau i ke kaua ʻana i kekahi mau maʻi i lawe ʻia e ka vector.Akā ʻo ka loaʻa ʻana o nā serotypes like ʻole o ka maʻi maʻi, i lawe ʻia e ka namu Aedes aegypti, ua alakaʻi i ka hāʻule ʻole o nā papahana lapaʻau.No laila, i ka wā e loaʻa ai ia mau maʻi, ʻo nā polokalamu hoʻokele vector wale nō ke koho e pale ai i ka laha ʻana o ka maʻi.I kēia manawa, he mea nui ka hoʻomalu ʻana i ka Aedes aegypti no ka mea, ʻo ia ke kumu nui o nā maʻi maʻi like ʻole a me kā lākou mau serotypes e hoʻoulu ai i ka dengue fever, Zika, dengue hemorrhagic fever, yellow fever, etc. Ke piʻi nei nā maʻi o nā maʻi Aedes-borne a pau i kēlā me kēia makahiki ma ʻAigupita a ke piʻi nei a puni ka honua.No laila, i loko o kēia pōʻaiapili, he mea koʻikoʻi ka hoʻomohala ʻana i nā hana hoʻomalu e pili ana i ke kaiapuni a me ka maikaʻi no ka lehulehu Aedes aegypti.ʻO nā moho kūpono e pili ana i kēia ʻano, ʻo ia nā EO, kā lākou hui pūhui, a me kā lākou hui pū ʻana.No laila, ua hoʻāʻo kēia haʻawina e ʻike i nā hui like synergistic kūpono o nā pūhui EO mea kanu nui mai nā mea kanu ʻelima me nā waiwai insecticidal (ʻo ia hoʻi, mint, basil hemolele, Eucalyptus spotted, Allium sulfur a me melaleuca) kūʻē iā Aedes aegypti.
Ua hōʻike nā EO i koho ʻia i ka hana larvicidal kūʻē iā Aedes aegypti me 24-h LC50 mai ka 0.42 a i ka 163.65 ppm.ʻO ka hana larvicidal kiʻekiʻe loa i hoʻopaʻa ʻia no ka peppermint (Mp) EO me kahi waiwai LC50 o 0.42 ppm ma 24 h, a ukali ʻia e ke kāleka (As) me kahi waiwai LC50 o 16.19 ppm ma 24 h (Table 1).
Me ka ʻokoʻa ʻo Ocimum Sainttum, Os EO, hōʻike nā EO ʻehā ʻē aʻe i nā hopena allercidal, me nā waiwai LC50 mai 23.37 a 120.16 ppm ma luna o ka manawa hōʻike 24-hola.ʻO Thymophilus striata (Cl) EO ka mea maikaʻi loa i ka pepehi kanaka makua me ka waiwai LC50 o 23.37 ppm i loko o 24 mau hola o ka hoʻolaha ʻana, a ukali ʻia e Eucalyptus maculata (Em) nona ka waiwai LC50 o 101.91 ppm (Table 1).Ma ka ʻaoʻao ʻē aʻe, ʻaʻole i hoʻoholo ʻia ka waiwai LC50 no Os ʻoiai ʻo ka helu make kiʻekiʻe loa o 53% i hoʻopaʻa ʻia ma ka helu kiʻekiʻe loa (Supplementary Figure 3).
Ua ʻike ʻia a koho ʻia nā pūhui mea nui ʻelua i kēlā me kēia EO ma muli o nā hopena waihona waihona waihona NIST, GC chromatogram area percentage, a me nā hualoaʻa MS spectra (Papa 2).No EO As, ʻo nā pūhui nui i ʻike ʻia ʻo ia ka diallyl disulfide a me ka diallyl trisulfide;no ka EO Mp na mea nui i ikeia he carvone a me ka limonene, no ka EO Em na mea nui i ikeia he eudesmol a me eucalyptol;No EO Os, ʻo nā pūhui nui i ʻike ʻia he eugenol a me ka methyl eugenol, a no ka EO Cl, ʻo nā pūhui nui i ʻike ʻia ʻo eugenol a me α-pinene (Figure 1, Nā Kiʻi Pākuʻi 5-8, Papa Pākuʻi 1-5).
Nā hualoaʻa o ka nui spectrometry o nā terpenoids nui o nā aila pono i koho ʻia (A-diallyl disulfide; B-diallyl trisulfide; C-eugenol; D-methyl eugenol; E-limonene; F-aromatic ceperone; G-α-pinene; H-cineole R-eudamol).
He ʻeiwa pūhui (diallyl disulfide, diallyl trisulfide, eugenol, methyl eugenol, carvone, limonene, eucalyptol, eudesmol, α-pinene) i ʻike ʻia he mau pūhui kūpono ʻo ia nā mea nui o ka EO a ua hoʻopaʻa ʻia i kēlā me kēia bioassayed kūʻē iā Aedes aegypti ma ka larval. nā pae..ʻO ka hui eudesmol ka hana larvicidal kiʻekiʻe loa me ka waiwai LC50 o 2.25 ppm ma hope o 24 mau hola o ka hoʻolaha.Ua ʻike pū ʻia nā pūhui diallyl disulfide a me diallyl trisulfide i nā hopena larvicidal hiki ke loaʻa, me ka mean sublethal doses ma ka laulā o 10-20 ppm.Ua ʻike hou ʻia ka hana larvicidal maʻalahi no nā pūhui eugenol, limonene a me eucalyptol me nā waiwai LC50 o 63.35 ppm, 139.29 ppm.a me 181.33 ppm ma hope o 24 mau hola (Papa 3).Eia nō naʻe, ʻaʻole i ʻike ʻia ka larvicidal koʻikoʻi o ka methyl eugenol a me ka carvone ma nā koina kiʻekiʻe loa, no laila ʻaʻole i helu ʻia nā koina LC50 (Table 3).Ua loa'a i ka larvicide synthetic Temephos ka nui o ka make o 0.43 ppm e kū'ē iā Aedes aegypti ma luna o 24 mau hola o ka ho'olaha (Papa 3, Papa Pāku'i 6).
ʻEhiku mau pūhui (diallyl disulfide, diallyl trisulfide, eucalyptol, α-pinene, eudesmol, limonene a me carvone) i ʻike ʻia ʻo ia nā pūhui nui o ka EO kūpono a ua hoʻāʻo ʻia i kēlā me kēia kūʻē i nā makika Aedes makua ʻAigupita.Wahi a Probit regression analysis, ʻike ʻia ʻo Eudesmol ka mea kiʻekiʻe loa me ka waiwai LC50 o 1.82 ppm, a ukali ʻia e Eucalyptol me kahi waiwai LC50 o 17.60 ppm i ka manawa hōʻike 24-hola.ʻO nā pūhui ʻelima i hoʻāʻo ʻia he mea pōʻino i nā pākeke me nā LC50 mai ka 140.79 a i ka 737.01 ppm (Table 3).ʻO ka synthetic organophosphorus malathion i emi iho ka ikaika ma mua o ka eudesmol a ʻoi aku ka kiʻekiʻe ma mua o nā pūhui ʻeono ʻē aʻe, me ka waiwai LC50 o 5.44 ppm ma luna o ka manawa hōʻike 24-hola (Table 3, Supplementary Table 6).
ʻEhiku mau pūhui alakaʻi ikaika a me ka organophosphorus tamephosate i koho ʻia no ka hoʻokumu ʻana i nā hui binary o kā lākou LC50 doses ma kahi ratio 1:1.Ua hoʻomākaukau ʻia a hoʻāʻo ʻia he 28 mau hui binary no kā lākou larvicidal efficacy e kūʻē iā Aedes aegypti.ʻEiwa hui pū ʻana i ʻike ʻia he synergistic, 14 hui pū ʻana he kūʻē, a ʻelima hui ʻaʻole larvicidal.Ma waena o nā hui synergistic, ʻo ka hui pū ʻana o diallyl disulfide a me temofol ka mea maikaʻi loa, me ka 100% make i ʻike ʻia ma hope o 24 mau hola (Table 4).Pēlā nō, ʻo ka hui ʻana o ka limonene me ka diallyl disulfide a me ka eugenol me thymetphos i hōʻike ʻia i ka maikaʻi maikaʻi me kahi make larval i ʻike ʻia he 98.3% (Papa 5).ʻO nā hui 4 i koe, ʻo ia hoʻi eudesmol plus eucalyptol, eudesmol plus limonene, eucalyptol plus alpha-pinene, alpha-pinene a me temephos, ua hōʻike pū kekahi i ka mana larvicidal koʻikoʻi, me nā helu make i ʻike ʻia ma mua o 90%.ʻO ka nui o ka make i manaʻo ʻia he kokoke i 60-75%.(Pakaukau 4).Eia naʻe, ʻo ka hui pū ʻana o ka limonene me α-pinene a i ʻole eucalyptus i hōʻike i nā hopena antagonistic.Pēlā nō, ʻike ʻia nā hui ʻana o Temephos me ka eugenol a i ʻole eucalyptus a i ʻole eudesmol a i ʻole diallyl trisulfide i loaʻa nā hopena antagonistic.Pēlā nō, ʻo ka hui pū ʻana o diallyl disulfide a me ka diallyl trisulfide a me ka hui pū ʻana o kekahi o kēia mau pūhui me ka eudesmol a i ʻole eugenol he mea kūʻē i kā lākou hana larvicidal.Ua hōʻike pū ʻia ka antagonism me ka hui pū ʻana o ka eudesmol me ka eugenol a i ʻole α-pinene.
ʻO nā hui ʻelua 28 a pau i hoʻāʻo ʻia no ka hana ʻawaʻawa makua, 7 hui pū ʻana he synergistic, 6 ʻaʻohe hopena, a he 15 antagonistic.ʻO ka hui pū ʻana o ka eudesmol me ka eucalyptus a me ka limonene me ka carvone i ʻike ʻia e ʻoi aku ka maikaʻi ma mua o nā hui synergistic ʻē aʻe, me nā helu make ma nā hola 24 o 76% a me 100%, kēlā me kēia (Papa 5).Ua ʻike ʻia ʻo Malathion e hōʻike i kahi hopena synergistic me nā hui pū ʻana a pau koe ka limonene a me ka diallyl trisulfide.Ma ka ʻaoʻao ʻē aʻe, ua loaʻa ka antagonism ma waena o ka diallyl disulfide a me ka diallyl trisulfide a me ka hui pū ʻana o kekahi o lākou me ka eucalyptus, a i ʻole eucalyptol, a i ʻole carvone, a i ʻole limonene.Pēlā nō, ʻo ka hui pū ʻana o α-pinene me ka eudesmol a i ʻole limonene, eucalyptol me ka carvone a i ʻole limonene, a me ka limonene me ka eudesmol a i ʻole malathion i hōʻike i nā hopena larvicidal antagonistic.No nā hui ʻeono i koe, ʻaʻohe ʻokoʻa koʻikoʻi ma waena o ka make i manaʻo ʻia a nānā ʻia (Table 5).
Ma muli o nā hopena synergistic a me nā ʻano sublethal, ua koho ʻia a hoʻāʻo hou ʻia kā lākou larvicidal toxicity e kūʻē i ka nui o nā makika Aedes aegypti.Ua hōʻike ʻia nā hopena he 100% ka make larval i ʻike ʻia me ka hoʻohana ʻana i nā hui binary eugenol-limonene, diallyl disulfide-limonene a me diallyl disulfide-timephos he 100%, ʻoiai ʻo ka make larval i manaʻo ʻia ʻo 76.48%, 72.16% a me 63.4% (Papa 6)..ʻO ka hui pū ʻana o ka limonene a me ka eudesmol ʻaʻole i ʻoi aku ka maikaʻi, me ka 88% o ka make larval i ʻike ʻia ma luna o ka manawa hōʻike 24-hola (Papa 6).Ma ka hōʻuluʻulu manaʻo, ua hōʻike pū nā hui binary ʻehā i nā hopena larvicidal synergistic e kūʻē iā Aedes aegypti ke hoʻohana ʻia ma ke ʻano nui (Papa 6).
Ua koho ʻia ʻekolu hui like ʻole no ka bioassay adultocidal e hoʻomalu i nā heluna nui o nā Aedes aegypti makua.No ke koho ʻana i nā hui e hoʻāʻo ai i nā kolo pepeke nui, ua nānā mua mākou i nā hui ʻelua synergistic terpene maikaʻi loa, ʻo ia ka carvone plus limonene a me eucalyptol plus eudesmol.ʻO ka lua, ua koho ʻia ka hui synergistic maikaʻi loa mai ka hui pū ʻana o organophosphate malathion a me terpenoids.Manaʻo mākou ʻo ka hui pū ʻana o ka malathion a me ka eudesmol ka hui maikaʻi loa no ka hoʻāʻo ʻana i nā kolo pepeke nui ma muli o ke kiʻekiʻe kiʻekiʻe o ka make a me ka haʻahaʻa haʻahaʻa LC50 o nā mea koho.Hōʻike ʻo Malathion i ka synergism i hui pū me ka α-pinene, diallyl disulfide, eucalyptus, carvone a me eudesmol.Akā inā mākou e nānā i nā waiwai LC50, ʻo Eudesmol ka waiwai haʻahaʻa loa (2.25 ppm).ʻO nā helu helu LC50 o malathion, α-pinene, diallyl disulfide, eucalyptol a me carvone he 5.4, 716.55, 166.02, 17.6 a me 140.79 ppm.pakahi.Hōʻike kēia mau waiwai i ka hui pū ʻana o ka malathion a me ka eudesmol ka hui maikaʻi loa i ka ʻōlelo o ka dosage.Ua hōʻike ʻia nā hopena i ka hui pū ʻana o carvone plus limonene a me eudesmol plus malathion i 100% i nānā ʻia ka make ma mua o ka make i manaʻo ʻia o 61% a 65%.ʻO kahi hui ʻē aʻe, eudesmol plus eucalyptol, i hōʻike i ka nui o ka make ʻana o 78.66% ma hope o 24 mau hola o ka hoʻolaha ʻana, i hoʻohālikelike ʻia me kahi helu make i manaʻo ʻia he 60%.ʻO nā hui ʻekolu i koho ʻia i hōʻike i nā hopena synergistic ʻoiai i hoʻohana ʻia ma ka pālākiō nui e kūʻē i ka Aedes aegypti makua (Papa 6).
I loko o kēia haʻawina, ua hōʻike ʻia nā EO mea kanu i koho ʻia e like me Mp, As, Os, Em a me Cl i nā hopena make i ka pae larval a me nā pae makua o Aedes aegypti.Loaʻa iā Mp EO ka hana larvicidal kiʻekiʻe loa me ka waiwai LC50 o 0.42 ppm, a ukali ʻia e As, Os a me Em EOs me ka waiwai LC50 o lalo o 50 ppm ma hope o 24 hola.Ua kūlike kēia mau hopena me nā noiʻi mua o nā makika a me nā nalo dipterous ʻē aʻe10,11,12,13,14.ʻOiai ʻoi aku ka liʻiliʻi o ka larvicidal potency o Cl ma mua o nā aila koʻikoʻi ʻē aʻe, me ka waiwai LC50 o 163.65 ppm ma hope o 24 mau hola, ʻoi aku ka kiʻekiʻe o kona hiki ke kanaka makua me ka waiwai LC50 o 23.37 ppm ma hope o 24 mau hola.Ua hōʻike pū ʻo Mp, As a me Em EOs i ka hiki ke allercidal maikaʻi me nā waiwai LC50 ma ka laulā o 100-120 ppm ma 24 mau hola o ka hoʻolaha ʻana, akā he haʻahaʻa haʻahaʻa ma mua o ko lākou larvicidal efficacy.Ma ka ʻaoʻao ʻē aʻe, ua hōʻike ʻo EO Os i kahi hopena allercidal haʻahaʻa ʻoiai ma ka nui o ka lāʻau lapaʻau kiʻekiʻe.No laila, hōʻike nā hualoaʻa he ʻokoʻa paha ka ʻawaʻawa o ka ethylene oxide i nā mea kanu ma muli o ke kūlana ulu o nā makika15.Ma muli o ka nui o ke komo ʻana o nā EO i loko o ke kino o ka pepeke, ko lākou launa pū ʻana me nā enzyme i hoʻopaʻa ʻia, a me ka hiki ke detoxification o ka makika i kēlā me kēia pae ulu16.Ua hōʻike ʻia kahi helu nui o nā haʻawina he mea nui ka mea hoʻohui kumu nui i ka hana olaola o ka ethylene oxide, ʻoiai ʻo ia ka hapa nui o nā pūhui āpau3,12,17,18.No laila, ua noʻonoʻo mākou i ʻelua hui nui i kēlā me kēia EO.Ma muli o nā hualoaʻa GC-MS, ua ʻike ʻia ka diallyl disulfide a me ka diallyl trisulfide ma ke ʻano he hui nui o EO As, i kūlike me nā hōʻike mua19,20,21.ʻOiai ua hōʻike ʻia nā hōʻike mua ʻo ka menthol kekahi o kāna mau pūhui nui, ua ʻike hou ʻia ka carvone a me ka limonene ma ke ʻano he pūhui nui o Mp EO22,23.Ua hōʻike ʻia ka moʻolelo haku mele o Os EO ʻo ka eugenol a me ka methyl eugenol nā pūhui nui, e like me nā ʻike o nā mea noiʻi mua16,24.Ua hōʻikeʻia ka Eucalyptol a me ka eucalyptol i nā mea nui i loaʻa i kaʻaila lau Em, i kūlike me nāʻike a kekahi poʻe noiʻi25,26 akā kū'ē i nā mea iʻikeʻia e Olalade et al.27.Uaʻikeʻia ka mana o ka cineole a me ka α-pinene i ka melaleuca hinu nui, e like me nā haʻawina mua28,29.Ua hōʻike ʻia nā ʻokoʻa ʻokoʻa i loko o ka haku mele ʻana a me ka neʻe ʻana o nā ʻaila koʻikoʻi i unuhi ʻia mai nā ʻano mea kanu like ma nā wahi like ʻole a ua ʻike pū ʻia i loko o kēia haʻawina, i hoʻopili ʻia e nā kūlana ulu o nā mea kanu, ka manawa o ka ʻohi ʻana, ka pae hoʻomohala, a i ʻole ka makahiki kanu.hiʻona o chemotypes, etc.22,30,31,32.Ua kūʻai ʻia a hoʻāʻo ʻia nā mea nui i ʻike ʻia no ka hopena larvicidal a me nā hopena i nā makika Aedes aegypti makua.Ua hōʻike ʻia nā hopena i hoʻohālikelike ʻia ka hana larvicidal o diallyl disulfide me ka hana o ka crude EO As.Akā ʻoi aku ka kiʻekiʻe o ka hana o diallyl trisulfide ma mua o EO As.Ua like kēia mau hopena me nā mea i loaʻa iā Kimbaris et al.33 ma Culex philippines.Eia naʻe, ʻaʻole i hōʻike kēia mau pūhui ʻelua i ka hana autocidal maikaʻi e kūʻē i nā makika i manaʻo ʻia, e kūlike me nā hopena o Plata-Rueda et al 34 ma Tenebrio molitor.Hoʻohana maikaʻi ʻo Os EO i ka pae larval o Aedes aegypti, akā ʻaʻole kūʻē i ka pae makua.Ua hoʻopaʻa ʻia he haʻahaʻa ka hana larvicidal o nā pūhui pākahi nui ma mua o ka Os EO crude.Hōʻike kēia i kahi kuleana no nā pūhui ʻē aʻe a me kā lākou pilina i loko o ka ethylene oxide.ʻO ka methyl eugenol wale nō ka hana liʻiliʻi, akā ʻo ka eugenol wale nō ka hana larvicidal haʻahaʻa.Ke hōʻoia nei kēia hopena, ma kekahiʻaoʻao, 35,36, a ma kekahiʻaoʻao, kū'ē i nā hopena o nā mea noiʻi mua37,38.ʻO nā ʻokoʻa o nā pūʻulu hana o eugenol a me methyleugenol hiki ke hopena i nā mea ʻawaʻawa like ʻole i ka insect target hoʻokahi39.Ua ʻike ʻia ʻo Limonene he hana larvicidal haʻahaʻa, ʻoiai ʻaʻole nui ka hopena o ka carvone.Pēlā nō, ʻo ka limonene haʻahaʻa haʻahaʻa loa i nā inike kanaka makua a me ka ʻawaʻawa kiʻekiʻe o ka carvone e kākoʻo i nā hopena o kekahi mau haʻawina mua40 akā kūʻē i nā mea ʻē aʻe41.ʻO ka loaʻa ʻana o nā paʻa pālua ma nā kūlana intracyclic a me exocyclic hiki ke hoʻonui i ka pono o kēia mau pūhui e like me nā larvicides3,41, ʻoiai ʻo carvone, he ketone me ka alpha unsaturated a me nā kalapona beta, hiki ke hōʻike i kahi kiʻekiʻe kiʻekiʻe o ka ʻona i nā pākeke42.Eia naʻe, ʻoi aku ka haʻahaʻa o nā ʻano o ka limonene a me ke carvone ma mua o ka huina EO Mp (Pakaukau 1, Papa 3).Ma waena o nā terpenoids i hoʻāʻo ʻia, ua ʻike ʻia ʻo eudesmol ka ʻoi loa o ka larvicidal a me ka hana makua me ka waiwai LC50 ma lalo o 2.5 ppm, e lilo ia i mea hoʻohiki no ka mālama ʻana i nā makika Aedes.ʻOi aku ka maikaʻi o kāna hana ma mua o ka EO Em holoʻokoʻa, ʻoiai ʻaʻole kūlike kēia me nā ʻike a Cheng et al.40.He sesquiterpene ʻo Eudesmol me nā ʻāpana isoprene ʻelua i ʻoi aku ka liʻiliʻi ma mua o nā monoterpenes oxygenated e like me ka eucalyptus a no laila ʻoi aku ka nui o ka pesticide.ʻOi aku ka nui o ke kanaka makua ma mua o ka hana larvicidal ʻo Eucalyptol, a ʻo nā hopena mai nā haʻawina mua e kākoʻo a hōʻole i kēia37,43,44.Ua aneane like ka hana me ka EO Cl holoʻokoʻa.ʻO kekahi monoterpene bicyclic, α-pinene, he mea liʻiliʻi ka hopena o ke kanaka makua ma luna o Aedes aegypti ma mua o ka hopena larvicidal, ʻo ia ka mea e kū'ē i ka hopena o ka EO Cl piha.Hoʻopili ʻia ka hana insecticidal holoʻokoʻa o nā terpenoids e ko lākou lipophilicity, volatility, carbon branching, projection area, surface area, functional groups and their positions45,46.Hiki i kēia mau pūhui ke hana ma ka luku ʻana i ka hoʻouluulu cell, ke ālai ʻana i ka hana hanu, ka hoʻopau ʻana i ka hoʻoili ʻana o nā nerve impulses, etc. ʻUtala48.Ua hōʻike ʻia ka hana makua o ka synthetic organophosphorus malathion ma 5.44 ppm.ʻOiai ua hōʻike ʻia kēia mau organophosphate ʻelua i nā pane maikaʻi e kūʻē i nā ʻano mea hoʻokolohua o Aedes aegypti, ua hōʻike ʻia ke kūʻē ʻana o ka makika i kēia mau pūhui ma nā wahi like ʻole o ka honua49.Eia naʻe, ʻaʻole i ʻike ʻia nā hōʻike like o ka hoʻomohala ʻana i ke kūʻē ʻana i nā lāʻau lapaʻau.No laila, manaʻo ʻia nā botanicals he mau mea ʻē aʻe i nā pesticides kemika i nā papahana hoʻokele vector.
Ua hoʻāʻo ʻia ka hopena larvicidal ma 28 hui binary (1: 1) i hoʻomākaukau ʻia mai nā terpenoids potent a me nā terpenoids me thymetphos, a ua ʻike ʻia he 9 hui ʻana he synergistic, 14 antagonistic a me 5 antagonistic.ʻAʻohe hopena.Ma ka ʻaoʻao ʻē aʻe, i ka bioassay potency makua, ua ʻike ʻia nā hui pū ʻana he synergistic, 15 hui pū ʻana he antagonistic, a ua hōʻike ʻia nā hui 6 ʻaʻohe hopena.ʻO ke kumu o ka hana ʻana o kekahi mau hui ʻana i ka hopena synergistic ma muli paha o ka hui pū ʻana o nā moho i ka manawa like ma nā ala koʻikoʻi like ʻole, a i ʻole ke kaohi ʻana o nā enzyme kī like ʻole o kahi ala olaola51.ʻO ka hui pū ʻana o ka limonene me ka diallyl disulfide, eucalyptus a i ʻole eugenol i ʻike ʻia he synergistic i nā noi liʻiliʻi a me ka nui (Papa 6), ʻoiai ʻo kona hui pū ʻana me ka eucalyptus a i ʻole α-pinene i loaʻa i nā hopena antagonistic ma nā larvae.Ma ka awelika, ʻike ʻia ka limonene i kahi synergist maikaʻi, ma muli paha o ke alo o nā hui methyl, ke komo maikaʻi ʻana i ka stratum corneum, a me kahi ʻano hana ʻē aʻe o ka hana52,53.Ua hōʻike mua ʻia e hiki i ka limonene ke hoʻoulu i nā hopena ʻawaʻawa ma ke komo ʻana i nā cuticles insect (contact toxicity), pili i ka ʻōnaehana digestive (antifeedant), a i ʻole pili i ka ʻōnaehana hanu (fumigation activity), 54 aʻo nā phenylpropanoids e like me eugenol hiki ke hoʻopilikia i nā enzyme metabolic 55. No laila, hiki i ka hui pū ʻana me nā ʻano hana like ʻole ke hoʻonui i ka hopena make o ka hui.Ua ʻike ʻia ʻo Eucalyptol he synergistic me ka diallyl disulfide, eucalyptus a i ʻole α-pinene, akā ʻo nā hui ʻē aʻe me nā pūhui ʻē aʻe he non-larvicidal a i ʻole antagonistic.Ua hōʻike ʻia nā haʻawina mua e loaʻa i ka eucalyptol ka hana inhibitory ma ka acetylcholinesterase (AChE), a me ka octaamine a me GABA receptors56.No ka mea, hiki i ka cyclic monoterpenes, eucalyptol, eugenol, etc. ke hana like me ko lakou hana neurotoxic, 57 pela e hoemi ai i ko lakou mau hopena i huiia ma o ke kaohi ana.Pēlā nō, ʻike ʻia ka hui pū ʻana o Temephos me diallyl disulfide, α-pinene a me limonene, e kākoʻo ana i nā hōʻike mua o kahi hopena synergistic ma waena o nā huahana lāʻau a me nā organophosphate synthetic58.
ʻO ka hui pū ʻana o ka eudesmol a me ka eucalyptol ua ʻike ʻia he hopena synergistic i ka larval a me nā pae makua o Aedes aegypti, ma muli paha o kā lākou ʻano hana like ʻole ma muli o ko lākou mau ʻano kemika like ʻole.Hiki i ka Eudesmol (a sesquiterpene) ke hoʻopili i ka ʻōnaehana hanu 59 a me ka eucalyptol (kahi monoterpene) hiki ke hoʻopili i ka acetylcholinesterase 60.Hiki ke hoʻonui i ka hopena make o ka hui pū ʻana o ka hui pū ʻana o nā mea hoʻohui i ʻelua a ʻoi aku paha.I loko o nā bioassays mea makua, ua ʻike ʻia ʻo malathion i ka synergistic me ka carvone a i ʻole eucalyptol a i ʻole eucalyptol a i ʻole diallyl disulfide a i ʻole α-pinene, e hōʻike ana he synergistic me ka hoʻohui ʻana o limonene a me di.ʻO nā moho allercide synergistic maikaʻi no ka waihona holoʻokoʻa o nā pūhui terpene, koe wale nō ka allyl trisulfide.Ua hōʻike pū ʻo Thangam a me Kathiresan61 i nā hopena like o ka hopena synergistic o ka malathion me nā mea kanu lāʻau.ʻO kēia pane synergistic paha ma muli o nā hopena ʻawaʻawa i hui ʻia o ka malathion a me nā phytochemical ma nā enzyme detoxifying insect.ʻO nā organophosphates e like me ka malathion e hana maʻamau ma ke kāohi ʻana i nā esterases cytochrome P450 a me nā monooxygenases62,63,64.No laila, ʻo ka hoʻohui ʻana i ka malathion me kēia mau ʻano hana a me nā terpenes me nā ʻano hana like ʻole e hoʻonui i ka hopena make ma luna o nā makika.
Ma ka ʻaoʻao ʻē aʻe, hōʻike ka antagonism i ka liʻiliʻi o nā pūhui i koho ʻia i ka hui pū ʻana ma mua o kēlā me kēia pūhui wale nō.ʻO ke kumu o ka antagonism i kekahi mau hui pū ʻana, ʻo ia ka hoʻololi ʻana o kekahi pūhui i ke ʻano o ka hui ʻē aʻe ma o ka hoʻololi ʻana i ka wikiwiki o ka absorption, ka hāʻawi ʻana, ka metabolism, a i ʻole ka excretion.Ua manaʻo ka poʻe noiʻi mua ʻo ia ke kumu o ka antagonism i nā hui lāʻau.Molecules Hiki ke hana 65. Pēlā nō, pili paha nā kumu kumu o ka antagonism i nā ʻano hana like, ka hoʻokūkū o nā pūhui constituent no ka loaʻa like ʻana a i ʻole ke kahua i kuhi ʻia.I kekahi mau hihia, hiki i ka hoʻopaʻapaʻa hoʻokūkū ʻole o ka protein target.I loko o kēia noiʻi ʻana, ʻelua pūhui organosulfur, diallyl disulfide a me diallyl trisulfide, i hōʻike i nā hopena kū'ē, ma muli paha o ka hoʻokūkū no ka pae hoʻokahi.Pēlā nō, ua hōʻike kēia mau pūhui sulfur ʻelua i nā hopena antagonistic a ʻaʻohe hopena i ka wā i hui pū ʻia me ka eudesmol a me ka α-pinene.ʻO ka Eudesmol a me ka alpha-pinene he cyclic ma ke ʻano, akā he aliphatic ke ʻano diallyl disulfide a me diallyl trisulfide.Ma muli o ke ʻano kemika, ʻo ka hui pū ʻana o kēia mau pūhui pono e hoʻonui i ka hana lethal holoʻokoʻa no ka mea he ʻokoʻa ko lākou mau wahi i hoʻopaʻa ʻia ma ke ʻano he 34,47, akā ma ka hoʻokolohua ua ʻike mākou i ka antagonism, ʻo ia paha ma muli o ke kuleana o kēia mau pūhui i kekahi mau mea ʻike ʻole i loko o vivo.nā pūnaewele ma muli o ka pilina.Pēlā nō, ʻo ka hui pū ʻana o ka cineole a me ka α-pinene i hoʻopuka i nā pane antagonistic, ʻoiai ua hōʻike mua nā mea noiʻi he ʻokoʻa nā pahuhopu ʻelua o ka action47,60.No ka mea he cyclic monoterpenes nā pūhui ʻelua, aia paha kekahi mau wahi i hoʻopaʻa ʻia e hoʻokūkū ai no ka hoʻopaʻa ʻana a me ka hoʻopili ʻana i ka ʻona o nā hui combinatorial i aʻo ʻia.
Ma muli o nā waiwai LC50 a me ka nānā ʻana i ka make, ua koho ʻia nā hui ʻelua synergistic terpene maikaʻi loa, ʻo ia hoʻi nā hui o carvone + limonene a me eucalyptol + eudesmol, a me ka synthetic organophosphorus malathion me terpenes.Ua hoʻāʻo ʻia ka hui synergistic maikaʻi loa o nā pūhui malathion + Eudesmol i kahi bioassay insecticide makua.E hoʻopaʻa i nā kolo pepeke nui e hōʻoia inā hiki i kēia mau hui pū ʻana ke hana kūʻē i ka nui o nā kānaka ma luna o nā wahi ākea ākea.Hōʻike kēia mau hui ʻana a pau i ka hopena synergistic e kūʻē i nā ʻōpala nui.Ua loaʻa nā hopena like no ka hui pū ʻana o nā larvicidal synergistic maikaʻi loa i hoʻāʻo ʻia e kūʻē i nā heluna nui o nā larvae Aedes aegypti.No laila, hiki ke ʻōlelo ʻia ʻo ka hui ʻana o ka larvicidal synergistic maikaʻi a me ka hui ʻana o nā mea kanu EO pūhui he moho ikaika e kūʻē i nā kemika synthetic i loaʻa a hiki ke hoʻohana hou ʻia e hoʻomalu i ka heluna Aedes aegypti.Pēlā nō, hiki ke hoʻohana ʻia ka hui pū ʻana o nā larvicides synthetic a i ʻole adulticides me terpenes e hōʻemi i ka nui o thymetphos a i ʻole malathion i hāʻawi ʻia i nā makika.Hiki i kēia mau hui hoʻohui ikaika ke hāʻawi i nā hoʻonā no nā noiʻi e hiki mai ana i ka ulu ʻana o ke kūʻē ʻana i ka lāʻau i nā makika Aedes.
Ua hōʻiliʻili ʻia nā hua manu o Aedes aegypti mai ka Regional Medical Research Center, Dibrugarh, Indian Council of Medical Research a mālama ʻia ma lalo o ka mālama ʻana i ka mahana (28 ± 1 °C) a me ka haʻahaʻa (85 ± 5%) ma ke Keʻena ʻo Zoology, Gauhati University ma lalo o ka ma hope o nā kūlana: Ua wehewehe ʻia ʻo Arivoli et al.Ma hope o ka paʻi ʻana, hānai ʻia nā larvae i ka meaʻai larva (ʻo ka pauka biscuit ʻīlio a me ka hū ma ka ratio 3:1) a ua hānai ʻia nā pākeke i ka hopena glucose 10%.E hoʻomaka ana i ka lā 3 ma hope o ka puka ʻana, ua ʻae ʻia nā makika wahine makua e omo i ke koko o nā ʻiole albino.E hoʻopaʻa i ka pepa kānana i ka wai i loko o ke aniani a waiho i loko o ka hale hoʻomoe hua.
ʻO nā laʻana mea kanu i koho ʻia ʻo ia nā lau eucalyptus (Myrtaceae), basil hemolele (Lamiaceae), mint (Lamiaceae), melaleuca (Myrtaceae) a me nā ʻōpala allium (Amaryllidaceae).Ua hōʻiliʻili ʻia mai Guwahati a ʻike ʻia e ke Keʻena ʻo Botany, Ke Kulanui ʻo Gauhati.Hoʻopili ʻia nā mea kanu i hōʻiliʻili ʻia (500 g) i ka hydrodistillation me ka hoʻohana ʻana i kahi mea Clevenger no 6 mau hola.Ua hōʻiliʻili ʻia ka EO i unuhi ʻia i loko o nā hue aniani maʻemaʻe a mālama ʻia ma 4 ° C no ke aʻo hou ʻana.
Ua aʻo ʻia ka lāʻau ʻawaʻawa me ka hoʻohana ʻana i nā kaʻina hana maʻamau o ka World Health Organization 67.E hoʻohana i ka DMSO ma ke ʻano he emulsifier.Ua ho'āʻo mua ʻia kēlā me kēia manaʻo EO ma 100 a me 1000 ppm, e hōʻike ana i 20 larvae i kēlā me kēia hana.Ma muli o nā hopena, ua hoʻopili ʻia kahi pae ʻike a ua hoʻopaʻa ʻia ka make mai ka hola 1 a hiki i 6 mau hola (ma nā hola 1 hola), a ma nā hola 24, 48 a me 72 mau hola ma hope o ka mālama ʻana.Ua hoʻoholo ʻia nā manaʻo sublethal (LC50) ma hope o 24, 48 a me 72 mau hola o ka ʻike.Ua hoʻāʻo ʻia kēlā me kēia manaʻo i ka triplicate me hoʻokahi mana maikaʻi ʻole (wai wale nō) a me hoʻokahi mana maikaʻi (wai mālama ʻia DMSO).Inā loaʻa ka pupation a ʻoi aku ma mua o 10% o nā larvae o ka pūʻulu hoʻomalu e make, e hana hou ʻia ka hoʻokolohua.Inā ma waena o 5-10% ka nui o ka make ma ka hui hoʻomalu, e hoʻohana i ke kumu hoʻoponopono Abbott 68.
ʻO ke ʻano i wehewehe ʻia e Ramar et al.Ua hoʻohana ʻia ʻo 69 no kahi bioassay makua e kūʻē iā Aedes aegypti me ka hoʻohana ʻana i ka acetone ma ke ʻano he mea hoʻoheheʻe.Ua hoʻāʻo mua ʻia kēlā me kēia EO e kūʻē i nā makika Aedes aegypti makua ma ka ʻike o 100 a me 1000 ppm.E hoʻopili i 2 ml o kēlā me kēia hopena i hoʻomākaukau ʻia i ka helu Whatman.1 ʻāpana pepa kānana (ka nui 12 x 15 cm2) a hoʻokuʻu i ka acetone no 10 mau minuke.ʻO ka pepa kānana i mālama ʻia me 2 ml wale nō o ka acetone i hoʻohana ʻia ma ke ʻano he mana.Ma hope o ka pau ʻana o ka acetone, waiho ʻia ka pepa kānana i mālama ʻia a me ka pepa kānana mana i loko o kahi paipu cylindrical (10 knm hohonu).Ua hoʻoili ʻia nā makika hānai koko ʻole he ʻumi 3 a 4 mau lā i ʻekolu ʻāpana o kēlā me kēia ʻano.Ma muli o nā hopena o nā hoʻokolohua mua, ua hoʻāʻo ʻia nā ʻano ʻokoʻa o nā aila i koho ʻia.Hoʻopaʻa ʻia ka make ma 1 hola, 2 hola, 3 hola, 4 hola, 5 hola, 6 hola, 24 hola, 48 hola a me 72 mau hola ma hope o ka hoʻokuʻu ʻia ʻana o ka makika.E helu i nā waiwai LC50 no nā manawa hoʻolaha o 24 mau hola, 48 hola a me 72 mau hola.Inā ʻoi aku ka nui o ka make o ka ʻāpana hoʻomalu ma mua o 20%, e hana hou i ka hoʻāʻo holoʻokoʻa.Pēlā nō, inā ʻoi aku ka nui o ka make ma ka hui hoʻomalu ma mua o 5%, e hoʻoponopono i nā hopena no nā laʻana i mālama ʻia me ka hoʻohana ʻana i ka formula68 a Abbott.
Ua hana ʻia ka chromatography kinoea (Agilent 7890A) a me ka spectrometry nui (Accu TOF GCv, Jeol) no ka nānā ʻana i nā pūhui constituent o nā aila pono i koho ʻia.Ua hoʻolako ʻia ka GC me kahi mea ʻike FID a me kahi kolamu capillary (HP5-MS).ʻO ke kinoea halihali he helium, ʻo ka kahe kahe ʻana he 1 ml / min.Hoʻonohonoho ka polokalamu GC iā Allium sativum i 10:80-1M-8-220-5M-8-270-9M a me Ocimum Sainttum i 10:80-3M-8-200-3M-10-275-1M-5 - 280, no ka mint 10:80-1M-8-200-5M-8-275-1M-5-280, no ka eucalyptus 20.60-1M-10-200-3M-30-280, a no ka ʻulaʻula No nā papa hoʻokahi tausani lākou. 10: 60-1M-8-220-5M-8-270-3M.
Ua ʻike ʻia nā pūhui nui o kēlā me kēia EO ma muli o ka pākēneka ʻāpana i helu ʻia mai ka GC chromatogram a me nā hualoaʻa spectrometry mass (i kuhikuhi ʻia i ka NIST 70 standard database).
Ua koho ʻia nā pūhui nui ʻelua i kēlā me kēia EO ma muli o nā hopena GC-MS a kūʻai ʻia mai Sigma-Aldrich ma 98-99% maʻemaʻe no nā bioassays hou aʻe.Ua hoʻāʻo ʻia nā pūhui no ka larvicidal a me ka pono o nā pākeke e kūʻē iā Aedes aegypti e like me ka mea i wehewehe ʻia ma luna.ʻO ka mea maʻamau i hoʻohana ʻia i nā larvicides synthetic tamephosate (Sigma Aldrich) a me ka malathion lāʻau makua (Sigma Aldrich) i kālai ʻia e hoʻohālikelike i ko lākou pono me nā pūhui EO i koho ʻia, ma muli o ke kaʻina hana like.
Ua hoʻomākaukau ʻia nā hui binary o nā pūhui terpene i koho ʻia a me nā pūhui terpene a me nā organophosphates pāʻoihana (tilephos a me malathion) ma ka hoʻohui ʻana i ka LC50 o kēlā me kēia hui moho ma kahi ratio 1:1.Ua hoʻāʻo ʻia nā hui i hoʻomākaukau ʻia ma nā pae larval a me nā pae makua o Aedes aegypti e like me ka mea i wehewehe ʻia ma luna.Ua hana ʻia kēlā me kēia bioassay i ʻekolu no kēlā me kēia hui ʻana a i ʻekolu no nā pūhui pākahi i loaʻa i kēlā me kēia hui.Ua hoʻopaʻa ʻia ka make ʻana o nā iniseti ma hope o 24 mau hola.E helu i ka helu make i manaʻo ʻia no kahi hui binary me ka hoʻohana ʻana i ke ʻano ma lalo.
kahi E = ka nui o ka make i manaʻo ʻia o nā makika Aedes aegypti ma ka pane ʻana i kahi hui binary, ʻo ia hoʻi ka pilina (A + B).
Ua kapa ʻia ka hopena o kēlā me kēia hui binary ma ke ʻano he synergistic, antagonistic, a ʻaʻohe hopena e pili ana i ka waiwai χ2 i helu ʻia e ke ʻano i wehewehe ʻia e Pavla52.E helu i ka waiwai χ2 no kēlā me kēia hui ʻana me ka hoʻohana ʻana i ke kumu hoʻohālike.
Ua wehewehe ʻia ka hopena o ka hui ʻana ma ke ʻano he synergistic i ka wā i ʻoi aku ka nui o ka helu χ2 ma mua o ke kumukūʻai papa no nā degere like o ke kūʻokoʻa (95% confidence interval) a inā ʻike ʻia ka make i ʻike ʻia ma mua o ka make i manaʻo ʻia.Pēlā nō, inā ʻoi aʻe ka waiwai χ2 i helu ʻia no kēlā me kēia hui ʻana i ka waiwai papa me kekahi mau degere o ke kūʻokoʻa, akā ʻoi aku ka haʻahaʻa o ka make i ʻike ʻia ma mua o ka make i manaʻo ʻia, ua manaʻo ʻia ka lāʻau antagonistic.A inā ʻoi aku ka liʻiliʻi o ka waiwai i helu ʻia o χ2 i loko o kekahi hui ma mua o ka waiwai papa ma nā degere like o ke kūʻokoʻa, ua manaʻo ʻia ʻaʻohe hopena o ka hui ʻana.
Ua koho ʻia ʻekolu a ʻehā mau hui like ʻole (100 larvae a me 50 larvicidal a me ka hana ʻino kanaka makua) no ka hoʻāʻo ʻana i ka nui o nā pepeke.ʻO nā mākua) e hoʻomau e like me luna.Me ka hui pū ʻana, ua hoʻāʻo ʻia nā pūhui pākahi i loko o nā hui i koho ʻia ma nā helu like o nā ʻōhua Aedes aegypti a me nā pākeke.ʻO ka lākiō hoʻohui ʻia he ʻāpana LC50 o ka hui moho hoʻokahi a me kahi ʻāpana LC50 o ka pūhui mea ʻē aʻe.I loko o ka bioassay hana kanaka makua, ua hoʻoheheʻe ʻia nā pūhui i koho ʻia i loko o ka solvent acetone a hoʻohana ʻia i ka pepa kānana i ʻōwili ʻia i loko o kahi pahu plastik cylindrical 1300 cm3.Hoʻopau ʻia ka acetone no 10 mau minuke a hoʻokuʻu ʻia nā pākeke.Pēlā nō, i loko o ka larvicidal bioassay, ua hoʻoheheʻe mua ʻia nā doses o nā pūhui moho LC50 i nā helu like o DMSO a laila hui pū ʻia me 1 lita o ka wai i mālama ʻia i loko o 1300 cc ipu plastik, a ua hoʻokuʻu ʻia nā larvae.
Ua hana ʻia ka loiloi probabilistic o 71 i hoʻopaʻa ʻia i ka ʻikepili make me ka hoʻohana ʻana i ka SPSS (version 16) a me Minitab lako polokalamu e helu ai i nā waiwai LC50.
Ka manawa hoʻouna: Jul-01-2024