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Udibaniso lwemixube ephuma kwisityalo esibulala izinambuzane lusenokubonisa intsebenziswano okanye intsebenziswano echasene nezinambuzane.Ngenxa yokusasazeka ngokukhawuleza kwezifo ezithwalwa ziingcongconi ze-Aedes kunye nokuxhathisa okwandayo kweengcongconi ze-Aedes kwizitshabalalisi zemveli, imidibaniso engamashumi amabini anesibhozo yeekhompawundi ze-terpene esekwe kwioyile ebalulekileyo yezityalo zayilwa kwaye zavavanywa ngokuchasene nenqanaba lombungu kunye nabantu abadala be-Aedes aegypti.Izityalo ezintlanu ze-oyile ezibalulekileyo (ii-EOs) zaqala ukuvandlakanywa kwi-larvicidal kunye nokusetyenziswa kwabantu abadala, kwaye iikhompawundi ezimbini ezinkulu zachongwa kwi-EO nganye ngokusekelwe kwiziphumo ze-GC-MS.Iikhompawundi eziphambili ezichongiweyo zathengwa, ezizezi diallyl disulfide, diallyl trisulfide, carvone, limonene, eugenol, methyl eugenol, eucalyptol, eudesmol kunye nemosquito alpha-pinene.Iindibaniselwano zeBinary zezi khompawundi zaye zalungiswa kusetyenziswa iidosi ezingaphantsi kwe-sublethal kunye neziphumo zabo ze-synergistic kunye ne-antagonistic zavavanywa kwaye zamiselwa.Iingqungquthela ezingcono kakhulu ze-larvicidal zifunyenwe ngokuxuba i-limonene kunye ne-diallyl disulfide, kunye neengqungquthela ezingcono kakhulu ze-addicidal zifunyenwe ngokuxuba i-carvone kunye ne-limonene.I-larvicide esetyenzisiweyo kurhwebo i-Temphos kunye nechiza labantu abadala iMalathion zavavanywa ngokwahlukeneyo kwaye kwindibaniselwano yokubini kunye ne-terpenoids.Iziphumo zibonise ukuba ukudibanisa i-temephos kunye ne-diallyl disulfide kunye ne-malathion kunye ne-eudesmol yayiyeyona nto isebenzayo.Ezi ndibaniselwano zinamandla zibambe amandla okusetyenziswa ngokuchasene ne-Aedes aegypti.
Izityalo ze-oyile eziyimfuneko (EOs) ziimetabolites zesibini eziqulethe iikhompawundi ezahlukeneyo ze-bioactive kwaye ziya zisibaluleka ngakumbi njengenye indlela kwizibulali-zinambuzane zokwenziwa.Ayipheleli nje ekukhuseleni kokusingqongileyo kwaye isebenziseka lula, kodwa ikwangumxube weekhompawundi ezahlukeneyo ze-bioactive, nto leyo ekwanciphisa ukuba nokwenzeka kokuphuhlisa ukunganyangeki kumachiza1.Ukusebenzisa iteknoloji ye-GC-MS, abaphandi bavavanya amacandelo ezityalo ezahlukeneyo ze-oyile ezibalulekileyo kwaye bachonga ngaphezu kwe-3,000 iikhompawundi ezivela kwi-17,500 izityalo ezivumba 2, ezininzi zazo zavavanywa kwiipropati ze-insecticidal kwaye zichazwe ukuba zineempembelelo ze-insecticidal3,4.Olunye uphononongo lubonisa ukuba ubutyhefu beyona nxalenye iphambili yekhompawundi buyafana okanye bungaphezulu kwaleyo ye-ethylene oxide yayo ekrwada.Kodwa ukusetyenziswa kweekhompawundi zomntu ngamnye kunokuphinda kushiye indawo yokuphuhliswa kokuchasana, njengoko kunjalo nge-insecticide yamachiza5,6.Ngoko ke, ugxininiso lwangoku lulungiselela imixube ye-ethylene oxide-based compounds ukuphucula ukusebenza kwe-insecticidal kunye nokunciphisa amathuba okuchasana kwiindawo ezijoliswe kuzo.Iikhompawundi ezisebenzayo ezizimeleyo ezikhoyo kwii-EOs zinokubonisa iziphumo ze-synergistic okanye ezichasayo kwiindibaniso ezibonisa umsebenzi wonke we-EO, inyaniso egxininiswe kakuhle kwizifundo eziqhutywe ngabaphandi bangaphambili7,8.Inkqubo yokulawula i-vector iquka i-EO kunye namacandelo ayo.Umsebenzi we-mosquitocidal weeoli eziyimfuneko sele ifundwe ngokubanzi kwiingcongconi zeCulex kunye ne-Anopheles.Uphononongo oluninzi luzamile ukuphuhlisa izibulali-zinambuzane ezisebenzayo ngokudibanisa izityalo ezahlukeneyo kunye nezibulali-zinambuzane ezisetyenziselwa urhwebo ukunyusa ubutyhefu kunye nokunciphisa iziphumo ebezingalindelekanga9.Kodwa izifundo zeekhompawundi ezinjalo ngokuchasene ne-Aedes aegypti zihlala zinqabile.Inkqubela phambili kwinzululwazi yezonyango kunye nophuhliso lwamachiza kunye nezitofu zokugonya ziye zanceda ukulwa nezifo ezithwalwa zizilwanyana.Kodwa ubukho bee-serotypes ezahlukeneyo zentsholongwane, ezithunyelwa yi-Aedes aegypti mosquito, kuye kwabangela ukungaphumeleli kweenkqubo zokugonywa.Ngoko ke, xa izifo ezinjalo zisenzeka, iinkqubo zokulawula i-vector kuphela kwendlela yokuthintela ukusasazeka kwesi sifo.Kwimeko yangoku, ulawulo lwe-Aedes aegypti lubaluleke kakhulu njengoko luyi-vector engundoqo yeentsholongwane ezahlukeneyo kunye ne-serotypes yazo ebangela i-dengue fever, i-Zika, i-dengue fever hemorrhagic fever, i-yellow fever, njl. phantse zonke izifo ezithwalwa zii-Aedes-ezithwala izifo ziyanda minyaka le e-Egypt kwaye ziyanda kwihlabathi jikelele.Ngoko ke, kulo mxholo, kukho imfuneko engxamisekileyo yokuphuhlisa imilinganiselo yokulawula indalo kunye nokulawula okusebenzayo kubantu base-Aedes aegypti.Abantu abanokuthi bagqale kulo mba zii-EOs, iikhompawundi eziyinxalenye yazo, kunye nendibaniselwano yazo.Ngoko ke, olu phononongo luzame ukuchonga imidibaniso esebenzayo ye-synergistic yezityalo eziphambili ze-EO compounds ezivela kwizityalo ezihlanu ezineempawu ze-insecticidal (okt, i-mint, i-basil engcwele, i-Eucalyptus ebonakalayo, i-Allium sulphur kunye ne-melaleuca) ngokuchasene ne-Aedes aegypti.
Zonke ii-EO ezikhethiweyo zibonise umsebenzi onokuthi ube ne-larvicidal ngokuchasene ne-Aedes aegypti ene-24-h LC50 ukusuka kwi-0.42 ukuya kwi-163.65 ppm.Umsebenzi ophezulu we-larvicidal wabhalwa kwi-peppermint (Mp) EO enexabiso le-LC50 le-0.42 ppm kwi-24 h, ilandelwa yigalikhi (As) ngexabiso le-LC50 le-16.19 ppm kwi-24 h (Itheyibhile 1).
Ngaphandle kwe-Ocimum Sainttum, i-Os EO, zonke ezinye ii-EO ezine ezihlolwe zibonise iziphumo ezicacileyo ze-allercidal, kunye nexabiso le-LC50 ukusuka kwi-23.37 ukuya kwi-120.16 ppm kwixesha le-24-exposure yeeyure.I-Thymophilus striata (Cl) EO yayisebenza kakhulu ekubulaleni abantu abadala abanexabiso le-LC50 le-23.37 ppm kwiiyure ezingama-24 zokuvezwa, ilandelwa yi-Eucalyptus maculata (Em) eyayinexabiso le-LC50 le-101.91 ppm (Itheyibhile 1).Ngakolunye uhlangothi, ixabiso le-LC50 le-Os alikacaciswa njengelona nani liphezulu lokufa kwe-53% lirekhodwe kwi-dose ephezulu (i-Supplementary Figure 3).
Iikhompawundi ezimbini eziphambili ze-EO nganye zichongiwe zaza zakhethwa ngokusekelwe kwiziphumo zedatha yethala leencwadi le-NIST, ipesenti yendawo yechromatogram yeGC, kunye neziphumo ze-MS spectra (Itheyibhile 2).Kwi-EO Njengoko, iikhompawundi eziphambili ezichongiweyo ziyi-diallyl disulfide kunye ne-diallyl trisulfide;kwi-EO Mp iikhompawundi eziphambili ezichongiweyo yayiyi-carvone kunye ne-limonene, ye-EO Em iikhompawundi eziphambili ezichongiweyo yayiyi-eudesmol kunye ne-eucalyptol;Kwi-EO Os, ii-compounds eziphambili ezichongiweyo ziyi-eugenol kunye ne-methyl eugenol, kunye ne-EO Cl, iikhompawundi eziphambili ezichongiweyo ziyi-eugenol kunye ne-α-pinene (Umfanekiso 1, iFigures Supplementary Figures 5-8, i-Supplementary Table 1-5).
Iziphumo ze-mass spectrometry yee-terpenoids eziphambili zeeoli ezikhethiweyo ezikhethiweyo (A-diallyl disulfide; B-diallyl trisulfide; C-eugenol; D-methyl eugenol; E-limonene; F-aromatic ceperone; G-α-pinene; H-cineole R-eudamol).
Iikhompawundi ezisithoba (i-diallyl disulfide, i-diallyl trisulfide, i-eugenol, i-methyl eugenol, i-carvone, i-limonene, i-eucalyptol, i-eudesmol, i-α-pinene) zichongiwe njengeekhompawundi ezisebenzayo ezingawona malungu aphambili e-EO kwaye zaye zavavanywa ngokuzimeleyo ngokuchasene ne-Aedes aegypti kwi-law izigaba..I-eudesmol edibeneyo inomsebenzi ophezulu we-larvicidal kunye nexabiso le-LC50 le-2.25 ppm emva kweeyure ezingama-24 zokuvezwa.Iikhompawundi ze-diallyl disulfide kunye ne-diallyl trisulfide nazo zifunyenwe zineempembelelo ezinokuthi zibe ne-larvicidal, kunye needosi eziphantsi kwe-sublethal kuluhlu lwe-10-20 ppm.Umsebenzi ophakathi we-larvicidal waphinda wabonwa kwi-eugenol, i-limonene kunye ne-eucalyptol enexabiso le-LC50 ye-63.35 ppm, i-139.29 ppm.kunye ne-181.33 ppm emva kweeyure ze-24, ngokulandelanayo (iThebhile 3).Nangona kunjalo, akukho mandla abalulekileyo e-larvicidal ye-methyl eugenol kunye ne-carvone efunyenwe nakwidosi ephezulu, ngoko ke amaxabiso e-LC50 awazange abalwe (Itheyibhile 3).I-synthetic larvicide iTemephos yayinoxinzelelo olubi lwe-0.43 ppm ngokuchasene ne-Aedes aegypti ngaphezulu kweeyure ezingama-24 zokuvezwa (Itheyibhile 3, iTheyibhile eyoNgezelelweyo 6).
Iikhompawundi ezisixhenxe (i-diallyl disulfide, i-diallyl trisulfide, i-eucalyptol, i-α-pinene, i-eudesmol, i-limonene kunye ne-carvone) zichongiwe njengezona zixhobo eziphambili ze-EO ezisebenzayo kwaye zavavanywa ngabanye kwiimiyane zase-Aedes zaseYiputa ezindala.Ngokutsho kohlalutyo lwe-Probit regression, i-Eudesmol ifunyenwe inamandla aphezulu kunye nexabiso le-LC50 le-1.82 ppm, ilandelwa yi-Eucalyptol enexabiso le-LC50 le-17.60 ppm kwixesha le-24-exposure time.Iikhompawundi eziseleyo ezintlanu ezivavanyiweyo ziyingozi ngokulinganayo kubantu abadala abane-LC50 ukusuka kwi-140.79 ukuya kwi-737.01 ppm (Itheyibhile 3).I-synthetic organophosphorus malathion yayingaphantsi kwe-eudesmol kwaye iphezulu kunezinye iikhompawundi ezintandathu, kunye nexabiso le-LC50 le-5.44 ppm kwixesha leeyure ezingama-24 zokuvezwa (iThebhile 3, iTheyibhile eyoNgezelelweyo yesi-6).
Iikhompawundi ezisixhenxe ezikhokelayo ezinamandla kunye ne-organophosphorus tamephosate zakhethwa ukuba zenze imidibaniso yokubini yeedosi zabo ze-LC50 kwi-1: umlinganiselo we-1.Iyonke i-28 yendibaniselwano yokubini yalungiswa kwaye yavavanywa ukusebenza kwayo kokubulala imibungu ngokuchasene ne-Aedes aegypti.Iindibaniselwano ezilithoba zafunyaniswa zine-synergistic, iindibaniselwano ezili-14 bezichasene, kwaye indibaniselwano emihlanu yayingeyombumbulu.Phakathi kweenhlanganisela ze-synergistic, indibaniselwano ye-diallyl disulfide kunye ne-temofol yayiyeyona isebenzayo, kunye ne-100% yokufa okubonwe emva kweeyure ze-24 (Itheyibhile 4).Ngokufanayo, imixube ye-limonene kunye ne-diallyl disulfide kunye ne-eugenol ene-thymetphos ibonise amandla amahle kunye nokufa okubonwayo kwe-larval ye-98.3% (Itheyibhile 5).Iindibaniso ezi-4 eziseleyo, ezizezi, i-eudesmol kunye ne-eucalyptol, i-eudesmol kunye ne-limonene, i-eucalyptol kunye ne-alpha-pinene, i-alpha-pinene kunye ne-temephos, nazo zibonise ukusebenza ngempumelelo kwe-larvicidal, kunye namazinga okusweleka aqatshelweyo angaphezu kwama-90%.Izinga lokufa elilindelekileyo lisondele kwi-60-75%.(Uluhlu 4).Nangona kunjalo, indibaniselwano ye-limonene kunye ne-α-pinene okanye i-eucalyptus ibonise ukusabela okuchasayo.Ngokunjalo, imixube yeTemephos ene-eugenol okanye i-eucalyptus okanye i-eudesmol okanye i-diallyl trisulfide ifunyenwe ineziphumo ezichasayo.Ngokunjalo, indibaniselwano ye-diallyl disulfide kunye ne-diallyl trisulfide kunye nokudityaniswa nokuba yeyiphi kwezi khompawundi kunye ne-eudesmol okanye i-eugenol ziyachasa kwisenzo sazo sokubulala.Ukuchasana kuye kwabikwa kunye nokudibanisa i-eudesmol kunye ne-eugenol okanye i-α-pinene.
Kuyo yonke imixube engama-28 yokubini evavanyelwe ukusebenza kweasidi yabantu abadala, isi-7 sendibaniselwano yayiyi-synergistic, i-6 ayinasiphumo, kwaye i-15 yayichasene.Imixube ye-eudesmol kunye ne-eucalyptus kunye ne-limonene ene-carvone ifunyenwe isebenza ngakumbi kunezinye iindibaniselwano ze-synergistic, kunye namazinga okufa kwiiyure ze-24 ze-76% kunye ne-100%, ngokulandelanayo (iThebhile 5).I-Malathion iye yabonwa ukuba ibonise umphumo we-synergistic kunye nayo yonke indibaniselwano yeekhompawundi ngaphandle kwe-limonene kunye ne-diallyl trisulfide.Kwelinye icala, ukuchasana kufunyenwe phakathi kwe-diallyl disulfide kunye ne-diallyl trisulfide kunye nokudityaniswa nokuba yeyiphi enye ne-eucalyptus, okanye i-eucalyptol, okanye i-carvone, okanye i-limonene.Ngokufanayo, ukudibanisa kwe-α-pinene kunye ne-eudesmol okanye i-limonene, i-eucalyptol ene-carvone okanye i-limonene, kunye ne-limonene ene-eudesmol okanye i-malathion ibonise imiphumo echasene ne-larvicidal.Kwimidibaniso emithandathu eseleyo, akukho mahluko ubalulekileyo phakathi kokusweleka okulindelekileyo kunye nokuqwalaselwayo (Itheyibhile 5).
Ngokusekwe kwiziphumo ze-synergistic kunye ne-sublethal doses, i-larvicidal toxicity ngokuchasene nenani elikhulu le-Aedes aegypti mosquitoes ekugqibeleni yakhethwa kwaye yavavanywa ngakumbi.Iziphumo zibonise ukuba ukufa kombungu okubonwayo usebenzisa i-binary indibaniselwano eugenol-limonene, diallyl disulfide-limonene kunye ne-diallyl disulfide-timephos yayingu-100%, ngelixa ukufa okulindelweyo kwakungu-76.48%, 72.16% kunye ne-63.4%, ngokulandelanayo (Itheyibhile 6)..Indibaniselwano ye-limonene kunye ne-eudesmol ayizange isebenze ngokufanelekileyo, ngama-88% okufa kombungu okuqatshelwe kwisithuba seeyure ezingama-24 sesichengeni (Itheyibhile 6).Isishwankathelo, imidibaniso emine ekhethiweyo yokubini nayo ibonise iziphumo ze-synergistic larvicidal ngokuchasene ne-Aedes aegypti xa isetyenziswe kwisikali esikhulu (iThebhile 6).
Iindibaniselwano ezintathu ze-synergistic zakhethwa kuvavanyo lwebhayoloji yabantu abadala ukulawula inani elikhulu labantu abadala i-Aedes aegypti.Ukukhetha indibaniselwano yokuvavanya kwiindawo ezinkulu zezinambuzane, siqale sajolisa kwezona ndibaniselwano zimbini ze-synergistic terpene, ezizezi, i-carvone kunye ne-limonene kunye ne-eucalyptol kunye ne-eudesmol.Okwesibini, eyona ndibaniselwano ye-synergistic yakhethwa kwindibaniselwano ye-synthetic organophosphate malathion kunye ne-terpenoids.Sikholelwa ukuba indibaniselwano ye-malathion kunye ne-eudesmol yeyona ndibaniselwano yovavanyo kwiikholoni ezinkulu zezinambuzane ngenxa yokubona okuphezulu kokusweleka kunye nexabiso eliphantsi kakhulu le-LC50 lezithako zabaviwa.I-Malathion ibonisa i-synergism ngokudibanisa ne-α-pinene, i-diallyl disulfide, i-eucalyptus, i-carvone kunye ne-eudesmol.Kodwa ukuba sijonga amaxabiso eLC50, i-Eudesmol inexabiso eliphantsi (2.25 ppm).I-LC50 ixabiso elibaliweyo le-malathion, i-α-pinene, i-diallyl disulfide, i-eucalyptol kunye ne-carvone yayiyi-5.4, 716.55, 166.02, 17.6 kunye ne-140.79 ppm.ngokulandelelanayo.La maxabiso abonisa ukuba indibaniselwano ye-malathion kunye ne-eudesmol yeyona ndibaniselwano ifanelekileyo ngokwedosi.Iziphumo zibonise ukuba indibaniselwano ye-carvone plus limonene kunye ne-eudesmol plus malathion ibe ne-100% yokufa okuqwalaselweyo xa kuthelekiswa nokufa okulindelweyo okungama-61% ukuya kuma-65%.Enye indibaniselwano, i-eudesmol kunye ne-eucalyptol, ibonise izinga lokufa kwe-78.66% emva kweeyure ze-24 zokuvezwa, xa kuthelekiswa nezinga lokufa elilindelekileyo lama-60%.Zontathu iindibaniselwano ezikhethiweyo zibonise iziphumo ze-synergistic naxa zisetyenziswe kwinqanaba elikhulu ngokuchasene ne-Aedes aegypti yabantu abadala (Itheyibhile 6).
Kule sifundo, ii-EO zezityalo ezikhethiweyo ezifana ne-Mp, As, Os, Em kunye ne-Cl zibonise iziphumo ezithembisayo ezibulalayo kwi-larval kunye nezigaba zabantu abadala be-Aedes aegypti.I-Mp EO yayinomsebenzi ophezulu we-larvicidal kunye nexabiso le-LC50 le-0.42 ppm, ilandelwa yi-As, Os kunye ne-Em EOs enexabiso le-LC50 elingaphantsi kwe-50 ppm emva kwe-24 h.Ezi ziphumo zihambelana nezifundo zangaphambili zeengcongconi kunye nezinye iimpukane ze-dipterous10,11,12,13,14.Nangona i-larvicidal potency ye-Cl iphantsi kunamanye amafutha abalulekileyo, kunye nexabiso le-LC50 le-163.65 ppm emva kweeyure ze-24, amandla ayo omntu omdala awona aphezulu kunye nexabiso le-LC50 le-23.37 ppm emva kweeyure ze-24.I-Mp, As kunye ne-Em EOs nazo zibonise amandla alungileyo e-allercidal kunye namaxabiso e-LC50 kuluhlu lwe-100-120 ppm kwi-24 h yokuvezwa, kodwa yayingaphantsi kunokusebenza kwayo kokubunjwa kwe-larvicidal.Kwelinye icala, i-EO Os ibonise isiphumo esinganyangekiyo se-allercidal nakwidosi ephezulu yonyango.Ngaloo ndlela, iziphumo zibonisa ukuba i-toxicity ye-ethylene oxide kwizityalo inokuhluka ngokuxhomekeka kwinqanaba lokuphuhlisa i-mosquitoes15.Kukwaxhomekeke kwisantya sokungena kwee-EOs kumzimba wesinambuzane, ukusebenzisana kwazo kunye nee-enzyme ezijoliswe kuzo, kunye nomthamo wokukhupha ingcongconi kwinqanaba ngalinye lophuhliso16.Inani elikhulu lezifundo libonise ukuba i-compound yecandelo eliphambili yinto ebalulekileyo kumsebenzi we-biological we-ethylene oxide, kuba ibala uninzi lwee-compounds ezipheleleyo3,12,17,18.Ngoko ke, siqwalasele iikhompawundi ezimbini eziphambili kwi-EO nganye.Ngokusekelwe kwiziphumo ze-GC-MS, i-diallyl disulfide kunye ne-diallyl trisulfide zichongiwe njengezona zixhobo eziphambili ze-EO As, ezihambelana neengxelo zangaphambili19,20,21.Nangona iingxelo zangaphambili zibonise ukuba i-menthol yayiyenye yeengqungquthela zayo eziphambili, i-carvone kunye ne-limonene ziphinde zachongwa njengezona zixhobo eziphambili ze-Mp EO22,23.Iprofayili yokubunjwa kwe-Os EO ibonise ukuba i-eugenol kunye ne-methyl eugenol zizona zixhobo eziphambili, ezifana neziphumo zabaphandi bangaphambili16,24.I-Eucalyptol kunye ne-eucalyptol ziye zaxelwa njengezona zixhobo eziphambili ezikhoyo kwi-Em leaf oil, ehambelana neziphumo zabaphandi abathile25,26 kodwa ngokuchasene neziphumo ze-Olalade et al.27.Ulawulo lwe-cineole kunye ne-α-pinene yabonwa kwi-melaleuca yeoli ebalulekileyo, efana nezifundo zangaphambili28,29.Umahluko we-Intraspecific ekuqulunqweni nasekuxutyweni kwee-oyile ezibalulekileyo ezitsalwa kwiintlobo zezityalo ezifanayo kwiindawo ezahlukeneyo ziye zaxelwa kwaye zabonwa kolu phononongo, oluphenjelelwa ziimeko zokukhula kwezityalo, ixesha lokuvuna, inqanaba lokukhula, okanye ubudala bezityalo.ukubonakala kweekhemotypes, njl.22,30,31,32.Iikhompawundi eziphambili ezichongiweyo zaye zathengwa kwaye zavavanyelwa iziphumo zazo ze-larvicidal kunye neziphumo kwiingcongconi ze-Aedes aegypti zabantu abadala.Iziphumo zabonisa ukuba umsebenzi we-larvicidal we-diallyl disulfide wawuthelekiseka naleyo ye-EO ekrwada.Kodwa umsebenzi we-diallyl trisulfide uphezulu kune-EO As.Ezi ziphumo zifana nezo zifunyenwe nguKimbaris et al.33 kwiCulex philippines.Nangona kunjalo, ezi zimbini azizange zibonise umsebenzi omhle we-autocidal ngokuchasene neemiyane ezijoliswe kuyo, ezihambelana neziphumo zePlata-Rueda et al 34 kwi-Tenebrio molitor.I-Os EO iyasebenza ngokuchasene nesigaba sombungu se-Aedes aegypti, kodwa hayi ngokuchasene nenqanaba labantu abadala.Kuye kwafunyaniswa ukuba umsebenzi we-larvicidal we-compounds eziphambili zomntu ungaphantsi kune-Os EO ekrwada.Oku kuthetha indima yezinye iikhompawundi kunye nokusebenzisana kwazo kwi-ethylene oxide ekrwada.I-Methyl eugenol iyodwa inomsebenzi ongathandabuzekiyo, ngelixa i-eugenol iyodwa inomsebenzi ophakathi we-larvicidal.Esi sigqibo siqinisekisa, kwelinye icala, i-35,36, kwaye kwelinye icala, iyaphikisana nezigqibo zabaphandi bangaphambili37,38.Ukwahluka kumaqela asebenzayo e-eugenol kunye ne-methyleugenol kunokubangela ubuthi obuhlukeneyo kwinambuzane enye ekujoliswe kuyo39.I-Limonene yafunyanwa inomsebenzi ophakathi we-larvicidal, ngelixa umphumo we-carvone wawungabalulekanga.Ngokufanayo, ubuthi obuphantsi be-limonene kwizinambuzane zabantu abadala kunye netyhefu ephezulu ye-carvone ixhasa iziphumo zezifundo zangaphambili40 kodwa ziphikisana nezinye41.Ubukho bemibhobho ephindwe kabini kwiindawo zombini ze-intracyclic kunye ne-exocyclic kunokunyusa inzuzo yala ma-compounds njenge-larvicides3,41, ngelixa i-carvone, i-ketone ene-alpha kunye ne-beta carbons engasetyenziswanga, ingabonisa amandla aphezulu okuba netyhefu kubantu abadala42.Nangona kunjalo, iimpawu zomntu ngamnye we-limonene kunye ne-carvone ziphantsi kakhulu kune-EO Mp epheleleyo (Itheyibhile 1, iThebhile 3).Phakathi kwe-terpenoids evavanyiweyo, i-eudesmol yafunyanwa inomsebenzi omkhulu we-larvicidal kunye nomntu omdala kunye nexabiso le-LC50 elingaphantsi kwe-2.5 ppm, okwenza kube yinto ethembisayo yokulawula i-Aedes mosquitoes.Ukusebenza kwayo kubhetele kune-EO Em yonke, nangona oku kungahambelani neziphumo ze-Cheng et al.40.I-Eudesmol yi-sesquiterpene eneeyunithi ezimbini ze-isoprene ezingaguquguquki kangako kune-oxygen monoterpenes ezifana ne-eucalyptus kwaye ngoko ke inamandla amakhulu njenge-pesticide.I-Eucalyptol ngokwayo inomntu omdala omkhulu kunomsebenzi we-larvicidal, kwaye iziphumo ezivela kwizifundo zangaphambili zombini zixhasa kwaye ziyayiphikisa le37,43,44.Umsebenzi uwodwa uphantse uthelekiseke naleyo ye-EO Cl iyonke.Enye i-bicyclic monoterpene, i-α-pinene, inempembelelo engaphantsi komntu omdala kwi-Aedes aegypti kunempembelelo ye-larvicidal, echasene nefuthe le-EO Cl epheleleyo.Umsebenzi we-insecticidal jikelele we-terpenoids uthonywa yi-lipophilicity, i-volatility, i-carbon branching, indawo yokubonisa, indawo ephezulu, amaqela asebenzayo kunye nezikhundla zabo45,46.Ezi zixhobo zinokuthi zisebenze ngokutshabalalisa ukuqokelela kweeseli, ukuthintela umsebenzi wokuphefumula, ukuphazamisa ukuhanjiswa kweempembelelo ze-nerve, njl njl. Utala48.Umsebenzi wabantu abadala we-synthetic organophosphorus malathion wabikwa kwi-5.44 ppm.Nangona ezi zimbini ze-organophosphates zibonise iimpendulo ezithandekayo ngokuchasene nohlobo lwelabhoratri ye-Aedes aegypti, ukuxhathisa kweengcongconi kwezi khompawundi kuye kwaxelwa kwiindawo ezahlukeneyo zehlabathi49.Nangona kunjalo, akukho ngxelo zifanayo zophuhliso lokunganyangeki kumayeza esintu ezifunyenweyo50.Ke ngoko, i-botanicals ithathwa njengenye indlela enokubakho kwimichiza yokubulala izinambuzane kwiinkqubo zolawulo lwe-vector.
Impembelelo ye-larvicidal yavavanywa kwi-28 indibaniselwano yokubini (1: 1) elungiswe ukusuka kwi-terpenoids enamandla kunye ne-terpenoids ene-thymetphos, kunye ne-9 indibaniselwano yafunyanwa i-synergistic, i-14 echasayo kunye ne-5 echasayo.Akukho siphumo.Ngakolunye uhlangothi, kwi-bioassay ye-potency yabantu abadala, i-7 indibaniselwano ifunyenwe i-synergistic, i-15 indibaniselwano yayichasene, kwaye i-6 indibaniselwano ichazwe ukuba ayinayo umphumo.Isizathu sokuba kutheni imidibaniso ethile ivelisa umphumo we-synergistic ingaba ngenxa ye-compounds yabaviwa esebenzisana ngexesha elifanayo kwiindlela ezibalulekileyo ezahlukeneyo, okanye ukuvinjelwa okulandelelanayo kwee-enzymes eziphambili zendlela ethile yezinto eziphilayo51.Ukudibanisa i-limonene kunye ne-diallyl disulfide, i-eucalyptus okanye i-eugenol ifunyenwe i-synergistic kwizicelo ezincinci kunye nezinkulu (Itheyibhile 6), ngelixa ukudibanisa kwayo ne-eucalyptus okanye i-α-pinene ifunyenwe ibe nemiphumo echasayo kwi-larvae.Ngokomyinge, i-limonene ibonakala iyi-synergist efanelekileyo, mhlawumbi ngenxa yobukho bamaqela e-methyl, ukungena kakuhle kwi-stratum corneum, kunye nendlela eyahlukileyo yokusebenza52,53.Kuye kwaxelwa kwangaphambili ukuba i-limonene inokubangela imiphumo enobuthi ngokungena kwi-cuticles yezinambuzane (i-toxity yoqhagamshelwano), echaphazela inkqubo yokutya (i-antifeedant), okanye ichaphazela inkqubo yokuphefumula (umsebenzi wokuphefumula), i-54 ngelixa i-phenylpropanoids efana ne-eugenol inokuchaphazela i-enzyme ye-metabolic 55. Ke ngoko, ukudityaniswa kweekhompawundi ezineendlela ezahlukeneyo zesenzo kunokunyusa isiphumo esibulalayo somxube.I-Eucalyptol yafunyanwa i-synergistic kunye ne-diallyl disulfide, i-eucalyptus okanye i-α-pinene, kodwa ezinye iindibaniselwano kunye nezinye iikhompawundi zazingekho i-larvicidal okanye i-antagonistic.Izifundo zangaphambili zibonise ukuba i-eucalyptol inomsebenzi wokuthintela kwi-acetylcholinesterase (AChE), kunye ne-octaamine kunye ne-GABA receptors56.Ekubeni i-cyclic monoterpenes, i-eucalyptol, i-eugenol, njl.Ngokufanayo, ukudibanisa kweTemephos kunye ne-diallyl disulfide, i-α-pinene kunye ne-limonene ifunyenwe i-synergistic, ixhasa iingxelo zangaphambili zempembelelo ye-synergistic phakathi kweemveliso ze-herbal kunye ne-synthetic organophosphates58.
Ukudityaniswa kwe-eudesmol kunye ne-eucalyptol kwafunyaniswa ukuba nefuthe le-synergistic kwizigaba ze-larval kunye nabantu abadala be-Aedes aegypti, mhlawumbi ngenxa yeendlela zabo ezahlukeneyo zokwenza ngenxa yezakhiwo zabo zeekhemikhali ezahlukeneyo.I-Eudesmol (i-sesquiterpene) inokuchaphazela inkqubo yokuphefumula i-59 kunye ne-eucalyptol (i-monoterpene) inokuchaphazela i-acetylcholinesterase 60.Ukuvezwa ngokubambisana kwezithako kwiindawo ekujoliswe kuzo ezibini okanye ngaphezulu kunokonyusa isiphumo esibulalayo sisonke sendibaniselwano.Kwii-bioassays zezinto zabantu abadala, i-malathion ifunyenwe i-synergistic kunye ne-carvone okanye i-eucalyptol okanye i-eucalyptol okanye i-diallyl disulfide okanye i-α-pinene, ebonisa ukuba i-synergistic kunye nokongezwa kwe-limonene kunye ne-di.Abaviwa abalungileyo be-synergistic allercide kuyo yonke ipotfoliyo yeekhompawundi zeterpene, ngaphandle kwe allyl trisulfide.UThangam kunye noKathiresan61 baphinde baxela iziphumo ezifanayo zempembelelo ye-synergistic ye-malathion kunye ne-herbal extracts.Le mpendulo ye-synergistic ingaba ngenxa yeempembelelo ezinobuthi ezidibeneyo ze-malathion kunye ne-phytochemicals kwi-insect detoxifying enzymes.Ii-Organophosphates ezifana ne-malathion ngokuqhelekileyo zisebenza ngokunqanda i-cytochrome P450 esterases kunye ne-monooxygenases62,63,64.Ke ngoko, ukudibanisa i-malathion kunye nezi ndlela zokwenza kunye ne-terpenes ngeendlela ezahlukeneyo zesenzo kunokuphucula isiphumo esibulalayo kwiingcongconi.
Kwelinye icala, ukuchasana kubonisa ukuba imixube ekhethiweyo ayisebenzi kangako xa idibene kunekhompawundi nganye yodwa.Isizathu sokuchasana kwezinye iindibaniselwano inokuba kukuba enye ikhompawundi iguqula indlela yokuziphatha yenye ikhompawundi ngokutshintsha izinga lokufunxa, ukusasazwa, imetabolism, okanye ukukhutshelwa ngaphandle.Abaphandi bangaphambili bathathela ingqalelo oku njengonobangela wokuchasana kwindibaniselwano yamachiza.Iimolekyuli Indlela enokwenzeka 65. Ngokufanayo, izinto ezinokubangela ukuchasana zinokuthi zihambelane neendlela ezifanayo zokwenza, ukhuphiswano lweekhompawundi ezidibeneyo kwi-receptor efanayo okanye indawo ekujoliswe kuyo.Kwezinye iimeko, ukuvinjelwa okungenakukhuphisana kweprotheni ekujoliswe kuyo kunokuthi kwenzeke.Kulo cwaningo, iikhompawundi ezimbini ze-organosulfur, i-diallyl disulfide kunye ne-diallyl trisulfide, zibonise iziphumo ezichasayo, mhlawumbi ngenxa yokhuphiswano lwendawo ekujoliswe kuyo.Ngokufanayo, ezi zimbini zesulfure zibonise iziphumo ezichasayo kwaye azizange zibe nefuthe xa zidibene ne-eudesmol kunye ne-α-pinene.I-Eudesmol kunye ne-alpha-pinene ziyi-cyclic kwindalo, ngelixa i-diallyl disulfide kunye ne-diallyl trisulfide ziyindalo ye-aliphatic.Ngokusekelwe kulwakhiwo lweekhemikhali, ukudityaniswa kwezi zixhobo kufuneka kwandise umsebenzi obulalayo ngokubanzi kuba iindawo ekujoliswe kuzo zihlala zihluke34,47, kodwa ngokuvavanya sifumene inkcaso, enokuthi ibe ngenxa yendima yezi zixhobo kwezinye izinto ezingaziwayo kwi-vivo.iinkqubo ngenxa yentsebenziswano.Ngokufanayo, udibaniso lwe-cineole kunye ne-α-pinene luvelise iimpendulo ezichasayo, nangona abaphandi bachaza ngaphambili ukuba iikhompawundi ezimbini zineethagethi ezahlukeneyo zesenzo47,60.Kuba zombini iikhompawundi ziyi-cyclic monoterpenes, kunokubakho iindawo ekujoliswe kuzo eziqhelekileyo ezinokukhuphisana ngokubophelela kunye nokuphembelela ityhefu iyonke yezibini ezidityanisiweyo ezifundiweyo.
Ngokusekwe kumaxabiso e-LC50 kunye nokubonwa kokusweleka, ezona ndibaniselwano zimbini ze-synergistic terpene zakhethwa, ezizezi, izibini ze-carvone + limonene kunye ne-eucalyptol + eudesmol, kunye ne-synthetic organophosphorus malathion ene-terpenes.Udibaniso olufanelekileyo lwe-synergistic ye-malathion + i-Eudesmol ikhompawundi yavavanywa kwi-bioassay ye-insecticide yabantu abadala.Kujongwe kwiikholoni ezinkulu zezinambuzane ukuqinisekisa ukuba ezi ndibaniselwano zisebenzayo zingasebenza na kumanani amakhulu abantu kwiindawo ezinkulu zokuchanabeka.Zonke ezi ndibaniselwano zibonisa impembelelo ye-synergistic ngokuchasene namabubu amakhulu ezinambuzane.Iziphumo ezifanayo zifunyenwe kwindibaniselwano ye-synergistic larvicidal evavanyiweyo ngokuchasene nombungu omkhulu we-Aedes aegypti.Ngaloo ndlela, kunokutshiwo ukuba i-synergistic larvicidal esebenzayo kunye ne-adulticidal indibaniselwano yezityalo ze-EO compounds ngumgqatswa oqinileyo ngokuchasene neekhemikhali zokwenziwa ezikhoyo kwaye zinokusetyenziswa ngakumbi ukulawula abantu base-Aedes aegypti.Ngokukwanjalo, indibaniselwano esebenzayo ye-synthetic larvicides okanye i-addicides ene-terpenes inokusetyenziselwa ukunciphisa idosi ye-thymetphos okanye i-malathion elawulwa kwiingcongconi.Ezi ndibaniselwano zinamandla ze-synergistic zinokubonelela ngezisombululo zezifundo zexesha elizayo malunga nokuvela kokumelana neziyobisi kwiingcongconi ze-Aedes.
Amaqanda e-Aedes aegypti aqokelelwa kwiZiko loPhando lwezoNyango lweNgingqi, eDibrugarh, iBhunga laseIndiya loPhando lwezoNyango kwaye agcinwe phantsi kobushushu obulawulwayo (28 ± 1 ° C) kunye nomswakama (85 ± 5%) kwiSebe leZoology, iYunivesithi yaseGauhati phantsi iimeko ezilandelayo: Arivoli zachazwa et al.Emva kokuqandusela, imibungu yayityiswa ukutya kwe-larval (i-dog biscuit powder kunye ne-yeast kwi-3: 1 ratio) kwaye abantu abadala batyiswa isisombululo se-10% se-glucose.Ukuqala ngomhla we-3 emva kokuvela, iingcongconi zabasetyhini abadala bavunyelwe ukufunxa igazi leempuku ze-albino.Gxuma iphepha lokucoca emanzini eglasini kwaye ulibeke kwikheji yokubeka iqanda.
Iisampulu zezityalo ezikhethiweyo ezingamagqabi e-eucalyptus (Myrtaceae), i-basil engcwele (Lamiaceae), i-mint (Lamiaceae), i-melaleuca (Myrtaceae) kunye ne-allium bulbs (Amaryllidaceae).Iqokelelwe eGuwahati kwaye ichongiwe liSebe leBotany, iYunivesithi yaseGauhati.Iisampulu zezityalo eziqokelelweyo (500 g) ziphantsi kwe-hydrodistillation usebenzisa i-Clevenger apparatus kwiiyure ze-6.I-EO ekhutshiweyo iqokelelwe kwiiglasi ezicocekileyo zeglasi kwaye zigcinwe kwi-4 ° C ukuze kufundwe ngokuqhubekayo.
Ityhefu yeLarvicidal yaphononongwa kusetyenziswa iinkqubo ze-67 zeWorld Health Organization ezilungelelanisiweyo.Sebenzisa i-DMSO njenge-emulsifier.I-concentration nganye ye-EO yaqala ukuvavanywa kwi-100 kunye ne-1000 ppm, iveza izibungu ezingama-20 kwi-replicate nganye.Ngokusekwe kwiziphumo, uluhlu loxinaniso luye lwasetyenziswa kwaye ukufa kwarekhodwa ukusuka kwiyure e-1 ukuya kwiiyure ze-6 (ngexesha leyure eli-1), kwaye kwiiyure ezingama-24, iiyure ezingama-48 kunye neeyure ezingama-72 emva konyango.I-sublethal concentrations (LC50) yagqitywa emva kwe-24, i-48 kunye ne-72 iiyure zokuvezwa.Ugxininiso ngalunye luye lwavavanywa ngokuphindwe kathathu kunye nolawulo olubi (amanzi kuphela) kunye nolawulo olulodwa oluhle (amanzi aphethwe yi-DMSO).Ukuba i-pupation iyenzeka kwaye ngaphezu kwe-10% ye-larvae yeqela lolawulo iyafa, uvavanyo luyaphindwa.Ukuba izinga lokufa kwiqela lolawulo liphakathi kwe-5-10%, sebenzisa ifomula yokulungiswa kwe-Abbott 68.
Indlela echazwe nguRamar et al.I-69 yayisetyenziselwa i-bioassay yabantu abadala ngokuchasene ne-Aedes aegypti isebenzisa i-acetone njengesinyibilikisi.I-EO nganye yaqala ukuvavanywa ngokuchasene neengcongconi ze-Aedes aegypti ezigxininiswe kwi-100 kunye ne-1000 ppm.Faka i-2 ml yesisombululo ngasinye esilungisiweyo kwinombolo ye-Whatman.Iphepha eli-1 lokucoca (ubukhulu be-12 x 15 cm2) kwaye uvumele i-acetone iphuphe imizuzu eyi-10.Iphepha lokucoca eliphathwe nge-2 ml ye-acetone kuphela lisetyenziswe njengolawulo.Emva kokuba i-acetone iqhume, iphepha lokucoca eliphathwayo kunye nephepha lokucoca lifakwe kwityhubhu ye-cylindrical (10 cm ubunzulu).Iingcongconi ezilishumi elineentsuku ezi-3 ukuya kwezi-4 ubudala ezingatyiswanga gazi zaye zatshintshelwa kwiindidi ezintathu zoxinaniso ngalunye.Ngokusekelwe kwiziphumo zovavanyo lwangaphambili, ukugxilwa kweeoli ezikhethiweyo zavavanywa.Ukufa kwarekhodwa ngeyure enye, iiyure ezi-2, iiyure ezi-3, iiyure ezi-4, iiyure ezi-5, iiyure ezi-6, iiyure ezingama-24, iiyure ezingama-48 kunye neeyure ezingama-72 emva kokukhululwa kweengcongconi.Bala amaxabiso eLC50 ngamaxesha okuvezwa kweeyure ezingama-24, iiyure ezingama-48 kunye neeyure ezingama-72.Ukuba izinga lokufa kweqashiso lolawulo lidlula i-20%, phinda uvavanyo lonke.Ngokufanayo, ukuba izinga lokufa kwiqela lolawulo likhulu kune-5%, lungisa iziphumo zeesampuli eziphathwayo usebenzisa i-formula68 ka-Abbott.
I-chromatography yegesi (i-Agilent 7890A) kunye ne-mass spectrometry (i-Accu TOF GCv, i-Jeol) yenziwa ukuhlalutya iikhompawundi ze-oyile ezibalulekileyo ezikhethiweyo.I-GC yayixhotyiswe nge-FID detector kunye nekholomu ye-capillary (HP5-MS).Irhasi yokuthwala yayiyi-helium, izinga lokuhamba laliyi-1 ml / min.Inkqubo yeGC ibeka i-Allium sativum kwi-10: 80-1M-8-220-5M-8-270-9M kunye ne-Ocimum Sainttum ukuya kwi-10: 80-3M-8-200-3M-10-275-1M-5 - 280, i-mint 10:80-1M-8-200-5M-8-275-1M-5-280, i-eucalyptus 20.60-1M-10-200-3M-30-280, kunye nobomvu Kwiwaka leeleya ngabo 10: 60-1M-8-220-5M-8-270-3M.
Iikhompawundi ezinkulu ze-EO nganye zichongiwe ngokusekelwe kwipesenti yendawo ebalwe kwi-GC chromatogram kunye neziphumo ze-mass spectrometry (kubhekiselwa kwisiseko sedatha ye-NIST 70).
Iikhompawundi ezimbini ezinkulu kwi-EO nganye zikhethwe ngokusekelwe kwiziphumo ze-GC-MS kwaye zithengwe kwi-Sigma-Aldrich kwi-98-99% yococeko lokuqhubela phambili i-bioassays.Iikhompawundi zavavanywa kwi-larvicidal kunye nokusebenza kwabantu abadala ngokuchasene ne-Aedes aegypti njengoko kuchazwe ngasentla.I-larvicides esetyenziswa ngokuqhelekileyo i-synthetic tamephosate (i-Sigma Aldrich) kunye ne-malathion yeziyobisi zabantu abadala (i-Sigma Aldrich) zahlaziywa ukuthelekisa ukusebenza kwazo kunye neengqungquthela ze-EO ezikhethiweyo, ngokulandela inkqubo efanayo.
Imixube yeBinary yeekhompawundi ezikhethiweyo zeterpene kunye neekhompawundi zeterpene kunye ne-organophosphates yorhwebo (i-tilephos ne-malathion) zalungiswa ngokuxuba idosi ye-LC50 yekhompawundi yomviwa ngamnye kumlinganiselo we-1:1.Iindibaniselwano ezilungiselelwe zavavanywa kwizigaba zombungu kunye nabadala be-Aedes aegypti njengoko kuchaziwe ngasentla.Uvavanyo lwebhayoloji ngalunye lwenziwa ngokuphindwe kathathu kwindibaniselwano nganye kwaye ngokuphindwe kathathu kwiikhompawundi ezikhoyo kwindibaniselwano nganye.Ukufa kwezinambuzane ekujoliswe kuzo kwabhalwa emva kweeyure ezingama-24.Bala umlinganiselo wokusweleka olindelekileyo kumxube wokubini usebenzisa le fomula ilandelayo.
apho u-E = izinga lokufa elilindelekileyo leengcongconi ze-Aedes aegypti ekuphenduleni indibaniselwano yokubini, okt uxhumano (A + B).
Isiphumo somxube ngamnye wokubini ubhalwe njenge-synergistic, inchasi, okanye akukho siphumo esisekelwe kwixabiso le-χ2 elibalwe ngendlela echazwe yi-Pavla52.Bala ixabiso le-χ2 kwindibaniselwano nganye usebenzisa le fomula ilandelayo.
Isiphumo sendibaniselwano sichazwe njenge-synergistic xa ixabiso elibaliweyo χ2 lalilikhulu kunexabiso letheyibhile leedigri zenkululeko ezihambelanayo (isithuba sokuzithemba esingama-95%) kwaye ukuba ukufa okuqatshelweyo kwafunyaniswa kudlula inani lokufa elilindelweyo.Ngokufanayo, ukuba ixabiso elibaliweyo elingu-χ2 layo nayiphi na indibaniselwano lingaphezulu kwexabiso letheyibhile kunye nezidanga ezithile zenkululeko, kodwa ukufa okuqatshelweyo kusezantsi kunoko kulindelweyo lokufa, unyango luthathwa njengoluchasayo.Kwaye ukuba kuyo nayiphi na indibaniselwano ixabiso elibaliweyo le-χ2 lingaphantsi kwexabiso letafile kwiidigri ezihambelanayo zenkululeko, ukudibanisa kuthathwa njengokungenasiphumo.
Imidibaniso emithathu ukuya kwezine enokubanakho ukusebenzisana (imibungu eli-100 kunye nama-50 okubulala imibungu kunye nemisebenzi yezinambuzane ezindala) zikhethelwe ukuvavanywa kwinani elikhulu lezinambuzane.Abantu abadala) qhubela phambili njengoko kungasentla.Kanye kunye nemixube, iikhompawundi zomntu ngamnye ezikhoyo kwimixube ekhethiweyo nazo zavavanywa ngamanani alinganayo e-Aedes aegypti larvae kunye nabantu abadala.Umlinganiselo odityanisiweyo yinxalenye enye ye-LC50 idosi yekhompawundi yomgqatswa omnye kunye nenxalenye ye-LC50 yedosi yenye ikhompawundi ekhoyo.Kwi-bioassay yomsebenzi wabantu abadala, iikhompawundi ezikhethiweyo zachithwa kwi-acetone ye-solvent kwaye isetyenziswe kwiphepha lokucoca elifakwe kwi-1300 cm3 yeplastiki ye-cylindrical.I-acetone yatshitshiswa imizuzu eyi-10 kwaye abantu abadala bakhululwa.Ngokufanayo, kwi-larvicidal bioassay, iidosi ze-LC50 zekhompawundi zabaviwa zaqala zanyibilika kwimiqulu elinganayo ye-DMSO kwaye emva koko yaxutywa ne-1 ilitha yamanzi agcinwe kwi-1300 cc izitya zeplastiki, kwaye imibungu yakhululwa.
Uhlalutyo olunokwenzeka lwedatha yokufa erekhodiweyo engama-71 lwenziwa kusetyenziswa i-SPSS (uguqulelo 16) kunye nesoftware yeMinitab ukubala amanani eLC50.
Ixesha lokuposa: Jul-01-2024