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Inhlanganisela yamakhemikhali abulala izinambuzane avela ezitshalweni ingabonisa ukusebenzisana okusebenzisanayo noma okuphikisayo ngokumelene nezinambuzane. Ngenxa yokusabalala okusheshayo kwezifo ezithwalwa omiyane base-Aedes kanye nokumelana okwandayo kwezinhlobo zomiyane base-Aedes emithini yokubulala izinambuzane yendabuko, inhlanganisela engamashumi amabili nesishiyagalombili yamakhemikhali e-terpene asekelwe kumafutha abalulekile ezitshalo yaklanywa futhi yahlolwa ngokumelene nezigaba zama-larval kanye nezigaba zabantu abadala ze-Aedes aegypti. Amafutha abalulekile ezitshalo amahlanu (ama-EO) aqale ahlolwa ukuze asebenze kahle ekubulaleni ama-larvicidal nasekusetshenzisweni kwabantu abadala, futhi kwatholakala amakhemikhali amabili amakhulu ku-EO ngayinye ngokusekelwe emiphumeleni ye-GC-MS. Kwathengwa amakhemikhali ayinhloko atholakele, okuyi-diallyl disulfide, i-diallyl trisulfide, i-carvone, i-limonene, i-eugenol, i-methyl eugenol, i-eucalyptol, i-eudesmol kanye ne-mosquito alpha-pinene. Inhlanganisela ye-binary yala makhemikhali yabe isilungiswa kusetshenziswa imithamo engaphansi kwebulalayo futhi imiphumela yawo yokubambisana kanye nokuphikisa yahlolwa futhi yanqunywa. Izinhlanganisela ezinhle kakhulu zokubulala ama-larvicidal zitholakala ngokuxuba i-limonene ne-diallyl disulfide, kanti izinhlanganisela ezinhle kakhulu zokubulala ama-larvicidal zitholakala ngokuxuba i-carvone ne-limonene. I-Temphos, i-larvicide yokwenziwa esetshenziswa kwezentengiselwano kanye nomuthi omdala i-Malathion, zihlolwe ngokwehlukana futhi zihlanganiswe ngama-terpenoids. Imiphumela ikhombisile ukuthi inhlanganisela ye-temephos ne-diallyl disulfide kanye ne-malathion kanye ne-eudesmol kwakuyinhlanganisela ephumelela kakhulu. Lezi zinhlanganisela ezinamandla zinamandla okusetshenziswa ngokumelene ne-Aedes aegypti.
Amafutha abalulekile ezitshalo (ama-EO) angama-metabolite esibili aqukethe amakhemikhali ahlukahlukene asebenzayo futhi aya ngokuya ebaluleka njengenye indlela esikhundleni sezibulala-zinambuzane zokwenziwa. Akuwona nje kuphela ukuthi anobungani nemvelo futhi asebenziseka kalula, kodwa futhi ayingxube yamakhemikhali ahlukene asebenzayo, okunciphisa namathuba okuthuthukisa ukumelana nemithi1. Besebenzisa ubuchwepheshe be-GC-MS, abacwaningi bahlola izakhi zamafutha abalulekile ezitshalo ahlukahlukene futhi bathola amakhemikhali angaphezu kuka-3,000 avela ezitshalweni ezingu-17,500 ze-aromatic2, eziningi zazo ezihlolwe izakhiwo zokubulala izinambuzane futhi kubikwa ukuthi zinemiphumela yokubulala izinambuzane3,4. Ezinye izifundo ziqokomisa ukuthi ubuthi bengxenye eyinhloko yenhlanganisela bufana noma bukhulu kunobe-ethylene oxide yayo eluhlaza. Kodwa ukusetshenziswa kwamakhemikhali ngamanye kungaphinde kushiye isikhala sokuthuthukiswa kokumelana, njengoba kunjalo ngama-insecticide amakhemikhali5,6. Ngakho-ke, ukugxila kwamanje kusekulungiseleleni izingxube zamakhemikhali asekelwe ku-ethylene oxide ukuthuthukisa ukusebenza kahle kokubulala izinambuzane nokunciphisa amathuba okumelana nezinambuzane ezihlosiwe. Amakhemikhali asebenzayo ngamanye akhona kuma-EO angabonisa imiphumela yokusebenzisana noma yokuphikisana ngokuhlanganiswa okubonisa umsebenzi ojwayelekile we-EO, iqiniso eligcizelelwe kahle ezifundweni ezenziwe ngabacwaningi bangaphambilini7,8. Uhlelo lokulawula ama-vector luhlanganisa ne-EO kanye nezingxenye zalo. Umsebenzi wokubulala umiyane wamafutha abalulekile ufundwe kabanzi kuma-Culex kanye nama-Anopheles omiyane. Izifundo eziningana ziye zazama ukuthuthukisa izibulala-zinambuzane ezisebenzayo ngokuhlanganisa izitshalo ezahlukahlukene nezibulala-zinambuzane zokwenziwa ezisetshenziswa kwezentengiselwano ukwandisa ubuthi obuphelele nokunciphisa imiphumela emibi9. Kodwa izifundo zamakhemikhali anjalo ngokumelene ne-Aedes aegypti azivamile. Intuthuko kwesayensi yezokwelapha kanye nokuthuthukiswa kwemithi kanye nemijovo yokugoma iye yasiza ekulweni nezinye izifo ezithwalwa yi-vector. Kodwa ukuba khona kwama-serotype ahlukene egciwane, adluliselwa yi-Aedes aegypti omiyane, kuholele ekwehlulekeni kwezinhlelo zokugoma. Ngakho-ke, lapho izifo ezinjalo zenzeka, izinhlelo zokulawula ama-vector ziyindlela kuphela yokuvimbela ukusabalala kwalesi sifo. Kulesi simo samanje, ukulawulwa kwe-Aedes aegypti kubaluleke kakhulu njengoba kuyimbangela eyinhloko yamagciwane ahlukahlukene kanye nama-serotype awo abangela umkhuhlane we-dengue, i-Zika, umkhuhlane we-dengue okopha, umkhuhlane ophuzi, njll. Into ephawuleka kakhulu ukuthi inani lamacala cishe azo zonke izifo ezithwalwa yi-Aedes liyanda minyaka yonke eGibhithe futhi liyanda emhlabeni jikelele. Ngakho-ke, kulo mongo, kunesidingo esiphuthumayo sokuthuthukisa izindlela zokulawula ezinobungani nemvelo nezisebenzayo ze-Aedes aegypti. Abantu abangaba khona kulokhu yi-EOs, izakhi zazo, kanye nokuhlanganiswa kwazo. Ngakho-ke, lolu cwaningo luzame ukuthola inhlanganisela esebenzayo yokusebenzisana yamakhemikhali abalulekile e-EO ezitshalo avela ezitshalweni ezinhlanu ezinezakhiwo zokubulala izinambuzane (okungukuthi, i-mint, i-basil engcwele, i-Eucalyptus spotted, i-Allium sulfur kanye ne-melaleuca) ngokumelene ne-Aedes aegypti.
Zonke i-EO ezikhethiwe zibonise umsebenzi wokubulala izibungu ongaba khona ngokumelene ne-Aedes aegypti nge-LC50 yamahora angu-24 esukela ku-0.42 kuya ku-163.65 ppm. Umsebenzi wokubulala izibungu ophezulu kakhulu uqoshwe ku-peppermint (Mp) EO enenani le-LC50 elingu-0.42 ppm emahoreni angu-24, kulandelwe yigalikhi (As) enenani le-LC50 elingu-16.19 ppm emahoreni angu-24 (Ithebula 1).
Ngaphandle kwe-Ocimum Sainttum, i-Os EO, zonke ezinye i-EO ezine ezihlolwe zibonise imiphumela esobala yokungezwani komzimba, ngamanani e-LC50 asukela ku-23.37 kuya ku-120.16 ppm esikhathini sokuchayeka samahora angama-24. I-Thymophilus striata (Cl) EO iphumelele kakhulu ekubulaleni abantu abadala ngenani le-LC50 elingu-23.37 ppm zingakapheli amahora angama-24 okuchayeka, kulandelwe yi-Eucalyptus maculata (Em) eyayinenani le-LC50 elingu-101.91 ppm (Ithebula 1). Ngakolunye uhlangothi, inani le-LC50 le-Os alikakanqunywa njengoba izinga lokufa eliphakeme kakhulu elingu-53% liqoshwe ngesilinganiso esiphezulu kakhulu (Isithombe Esingeziwe 3).
Ama-compound amabili amakhulu ku-EO ngayinye akhonjwe futhi akhethwa ngokusekelwe emiphumeleni yedathabheyisi yomtapo wolwazi we-NIST, iphesenti lendawo ye-GC chromatogram, kanye nemiphumela ye-MS spectra (Ithebula 2). Ku-EO As, ama-compound ayinhloko akhonjwe kwakuyi-diallyl disulfide kanye ne-diallyl trisulfide; ku-EO Mp ama-compound ayinhloko akhonjwe kwakuyi-carvone kanye ne-limonene, ku-EO Em ama-compound ayinhloko akhonjwe kwakuyi-eudesmol kanye ne-eucalyptol; Ku-EO Os, ama-compound ayinhloko akhonjwe kwakuyi-eugenol kanye ne-methyl eugenol, kanye ne-EO Cl, ama-compound ayinhloko akhonjwe kwakuyi-eugenol kanye ne-α-pinene (Isithombe 1, Izibalo Ezingeziwe 5–8, Ithebula Elingeziwe 1–5).
Imiphumela ye-mass spectrometry yama-terpenoid ayinhloko amafutha abalulekile akhethiwe (i-A-diallyl disulfide; i-B-diallyl trisulfide; i-C-eugenol; i-D-methyl eugenol; i-E-limonene; i-F-aromatic ceperone; i-G-α-pinene; i-H-cineole; i-R-eudamol).
Ama-compound ayisishiyagalolunye (i-diallyl disulfide, i-diallyl trisulfide, i-eugenol, i-methyl eugenol, i-carvone, i-limonene, i-eucalyptol, i-eudesmol, i-α-pinene) ahlonzwe njengama-compound asebenzayo ayizingxenye eziyinhloko ze-EO futhi ahlolwa ngokwawo ngokumelene ne-Aedes aegypti ezigabeni zama-larval. . I-eudesmol ehlanganisiwe yayinomsebenzi ophezulu kakhulu wokubulala ama-larvicidal onenani le-LC50 elingu-2.25 ppm ngemva kwamahora angu-24 okuvezwa. Ama-compound e-diallyl disulfide kanye ne-diallyl trisulfide nawo atholakale enemiphumela engaba khona yokubulala ama-larvicidal, kanye nemithamo engaphansi kwesilinganiso ebangeni eli-10-20 ppm. Umsebenzi ophakathi wokubulala ama-larvicidal uphinde wabonwa kuma-compound eugenol, i-limonene kanye ne-eucalyptol enamanani e-LC50 angu-63.35 ppm, 139.29 ppm. kanye no-181.33 ppm ngemva kwamahora angu-24, ngokulandelana (Ithebula 3). Kodwa-ke, akukho mandla abalulekile okubulala ama-larvicidal e-methyl eugenol kanye ne-carvone atholakale ngisho nasemithini ephezulu kakhulu, ngakho-ke amanani e-LC50 awazange abalwe (Ithebula 3). I-synthetic larvicide i-Temephos yayinezinga eliphakathi lokubulala elingu-0.43 ppm ngokumelene ne-Aedes aegypti emahoreni angu-24 okuvezwa (Ithebula 3, Ithebula Elingeziwe 6).
Ama-compound ayisikhombisa (i-diallyl disulfide, i-diallyl trisulfide, i-eucalyptol, i-α-pinene, i-eudesmol, i-limonene kanye ne-carvone) ahlonzwe njengama-compound ayinhloko e-EO asebenzayo futhi ahlolwa ngawodwana ngokumelene nomiyane abadala base-Egyptian Aedes. Ngokusho kokuhlaziywa kwe-Probit regression, i-Eudesmol itholakale inamandla aphezulu kakhulu ngenani le-LC50 elingu-1.82 ppm, kulandelwe yi-Eucalyptol enenani le-LC50 elingu-17.60 ppm ngesikhathi sokuchayeka emahoreni angu-24. Ama-compound amahlanu asele ahlolwe ayeyingozi ngokulingene kubantu abadala abane-LC50s kusukela ku-140.79 kuya ku-737.01 ppm (Ithebula 3). I-synthetic organophosphorus malathion yayinamandla amancane kune-eudesmol futhi iphakeme kunezinye ama-compound ayisithupha, enenani le-LC50 elingu-5.44 ppm esikhathini sokuchayeka emahoreni angu-24 (Ithebula 3, Ithebula Elingeziwe 6).
Kwakhethwa amakhemikhali ayisikhombisa anamandla e-lead kanye ne-organophosphorus tamephosate ukuze kwakhiwe inhlanganisela ye-binary yemithamo yabo ye-LC50 ngesilinganiso esingu-1:1. Kwalungiswa inhlanganisela ye-binary engu-28 futhi kwahlolwa ukusebenza kwayo kokubulala ama-larvicidal ngokumelene ne-Aedes aegypti. Kwatholakala ukuthi inhlanganisela eyisishiyagalolunye iyasebenza, inhlanganisela engu-14 yayiphikisana, kanti inhlanganisela emihlanu yayingeyona ebulala ama-larvicidal. Phakathi kwenhlanganisela ye-synergistic, inhlanganisela ye-diallyl disulfide kanye ne-temofol yiyona ephumelela kakhulu, kanti ukufa okungu-100% kwabonwa ngemva kwamahora angu-24 (Ithebula 4). Ngokufanayo, ingxube ye-limonene ne-diallyl disulfide kanye ne-eugenol ene-thymetphos ibonise amandla amahle ngokufa kwama-larval okubonwe okungu-98.3% (Ithebula 5). Izinhlanganisela ezine ezisele, okuyi-eudesmol kanye ne-eucalyptol, i-eudesmol kanye ne-limonene, i-eucalyptol kanye ne-alpha-pinene, i-alpha-pinene kanye ne-temephos, nazo zibonise ukusebenza kahle okukhulu kokubulala ama-larvicidal, kanti amazinga okufa abonwe angaphezu kuka-90%. Izinga lokufa elilindelekile liseduze no-60-75%. (Ithebula 4). Kodwa-ke, inhlanganisela ye-limonene ne-α-pinene noma i-eucalyptus ibonise ukusabela okuphikisayo. Ngokufanayo, izingxube ze-Temephos ne-eugenol noma i-eucalyptus noma i-eudesmol noma i-diallyl trisulfide zitholakale zinemiphumela ephikisayo. Ngokufanayo, inhlanganisela ye-diallyl disulfide kanye ne-diallyl trisulfide kanye nenhlanganisela yanoma iyiphi yalezi zinhlanganisela ne-eudesmol noma i-eugenol ziyaphikisana esenzweni sazo sokubulala ama-larvicidal. Ukuphikisana nakho kubikwe ngokuhlanganiswa kwe-eudesmol ne-eugenol noma i-α-pinene.
Kuzo zonke izingxube ezingama-28 ezihlolwe umsebenzi we-asidi yabantu abadala, izingxube ezi-7 zazihambisana, ezi-6 azizange zibe nomphumela, kanti eziyi-15 zaziphikisana. Izingxube ze-eudesmol ne-eucalyptus kanye ne-limonene ne-carvone zitholakale zisebenza kangcono kunezinye izinhlanganisela ezihambisanayo, kanti amazinga okufa angama-76% kanye no-100% emahoreni angama-24, ngokulandelana (Ithebula 5). I-Malathion ibonwe ikhombisa umphumela ohambisanayo nazo zonke izinhlanganisela zamakhemikhali ngaphandle kwe-limonene kanye ne-diallyl trisulfide. Ngakolunye uhlangothi, ukungezwani kutholakale phakathi kwe-diallyl disulfide kanye ne-diallyl trisulfide kanye nokuhlanganiswa kwazo ne-eucalyptus, noma i-eucalyptol, noma i-carvone, noma i-limonene. Ngokufanayo, izinhlanganisela ze-α-pinene ne-eudesmol noma i-limonene, i-eucalyptol ene-carvone noma i-limonene, kanye ne-limonene ene-eudesmol noma i-malathion zibonise imiphumela yokulwa nama-larvicidal. Kulezi zinhlanganisela eziyisithupha ezisele, bekungekho mehluko omkhulu phakathi kokufa okulindelekile nokufa okubonwayo (Ithebula 5).
Ngokusekelwe emiphumeleni yokusebenzisana kanye nemithamo engaphansi kweyokubulala, ubuthi bazo bokubulala ama-larvicidal ngokumelene nenani elikhulu lomiyane be-Aedes aegypti bakhethwa ekugcineni futhi bahlolwa kabanzi. Imiphumela ikhombisile ukuthi ukufa kwama-larval okubonwe kusetshenziswa inhlanganisela ye-binary eugenol-limonene, i-diallyl disulfide-limonene kanye ne-diallyl disulfide-timephos kwaba yi-100%, kanti ukufa okulindelekile kwama-larval kwaba yi-76.48%, 72.16% kanye no-63.4%, ngokulandelana (Ithebula 6). . Inhlanganisela ye-limonene ne-eudesmol yayingasebenzi kahle, kanti ukufa kwama-larval okungu-88% kwabonwa esikhathini sokuchayeka samahora angu-24 (Ithebula 6). Ngamafuphi, inhlanganisela ye-binary ezine ezikhethiwe nazo zibonise imiphumela yokubulala ama-larvicidal ngokumelene ne-Aedes aegypti uma isetshenziswa ngezinga elikhulu (Ithebula 6).
Kukhethwe izinhlanganisela ezintathu ezihambisanayo ze-bioassay yabantu abadala ukulawula inani elikhulu le-Aedes aegypti yabantu abadala. Ukuze sikhethe izinhlanganisela zokuhlola emakoloni amakhulu ezinambuzane, siqale sagxila ezinhlanganisweni ezimbili ezinhle kakhulu ze-terpene ezihambisanayo, okuyi-carvone plus limonene kanye ne-eucalyptol kanye ne-eudesmol. Okwesibili, inhlanganisela engcono kakhulu ehambisanayo ikhethwe ekuhlanganisweni kwe-synthetic organophosphate malathion kanye ne-terpenoids. Sikholelwa ukuthi inhlanganisela ye-malathion kanye ne-eudesmol iyinhlanganisela engcono kakhulu yokuhlola emakoloni amakhulu ezinambuzane ngenxa yokufa okubonwa kakhulu kanye namanani aphansi kakhulu e-LC50 ezithako ezingaba khona. I-Malathion ibonisa ukuhambisana ngokuhlanganiswa ne-α-pinene, i-diallyl disulfide, i-eucalyptus, i-carvone kanye ne-eudesmol. Kodwa uma sibheka amanani e-LC50, i-Eudesmol inenani eliphansi kakhulu (2.25 ppm). Amanani e-LC50 abaliwe e-malathion, i-α-pinene, i-diallyl disulfide, i-eucalyptol kanye ne-carvone ayengu-5.4, 716.55, 166.02, 17.6 kanye no-140.79 ppm. ngokulandelana. Lawa manani abonisa ukuthi inhlanganisela ye-malathion kanye ne-eudesmol iyinhlanganisela efanele ngokwesilinganiso. Imiphumela ikhombisile ukuthi inhlanganisela ye-carvone kanye ne-limonene kanye ne-eudesmol kanye ne-malathion yayinokufa okubonwe okungu-100% uma kuqhathaniswa nokufa okulindelekile okungu-61% kuya ku-65%. Enye inhlanganisela, i-eudesmol kanye ne-eucalyptol, ibonise izinga lokufa elingu-78.66% ngemva kwamahora angu-24 okuvezwa, uma kuqhathaniswa nezinga lokufa elilindelekile elingu-60%. Zonke izinhlanganisela ezintathu ezikhethiwe zibonise imiphumela yokusebenzisana ngisho nalapho zisetshenziswa ngezinga elikhulu ngokumelene ne-Aedes aegypti yabantu abadala (Ithebula 6).
Kulolu cwaningo, izitshalo ezikhethiwe ze-EO ezifana ne-Mp, As, Os, Em kanye ne-Cl zibonise imiphumela ebulalayo ethembisayo ezigabeni zama-larval kanye nezigaba zabantu abadala ze-Aedes aegypti. I-Mp EO yayinomsebenzi ophezulu kakhulu wokubulala ama-larvicidal onenani le-LC50 elingu-0.42 ppm, kulandele i-As, Os kanye ne-Em EOs enenani le-LC50 elingaphansi kwama-50 ppm ngemva kwamahora angama-24. Le miphumela ihambisana nezifundo zangaphambilini zomiyane nezinye izimpukane ezinwabuzelayo10,11,12,13,14. Nakuba amandla okubulala ama-larvicidal e-Cl ephansi kunamanye amafutha abalulekile, enenani le-LC50 elingu-163.65 ppm ngemva kwamahora angama-24, amandla ayo okudala aphezulu kakhulu enenani le-LC50 elingu-23.37 ppm ngemva kwamahora angama-24. Ama-EO e-Mp, As kanye ne-Em nawo abonise amandla amahle okubangela ukungezwani komzimba ngamanani e-LC50 aphakathi kuka-100–120 ppm emahoreni angama-24 okuvezwa, kodwa ayephansi kakhulu kunokusebenza kwawo kokubulala izibungu. Ngakolunye uhlangothi, i-EO Os ibonise umphumela omncane wokubangela ukungezwani komzimba ngisho nangesilinganiso esiphezulu sokwelapha. Ngakho-ke, imiphumela ikhombisa ukuthi ubuthi be-ethylene oxide ezitshalweni bungahluka kuye ngesigaba sokuthuthuka komiyane15. Kuncike futhi esilinganisweni sokungena kwama-EO emzimbeni wesinambuzane, ukusebenzisana kwawo nama-enzyme athile aqondiwe, kanye nekhono lokususa ubuthi komiyane esigabeni ngasinye sokuthuthuka16. Inani elikhulu lezifundo likhombisile ukuthi isakhi esiyinhloko siyisici esibalulekile emsebenzini webhayoloji we-ethylene oxide, njengoba ibala iningi lamakhemikhali aphelele3,12,17,18. Ngakho-ke, sicabangele amakhemikhali amabili ayinhloko ku-EO ngayinye. Ngokusekelwe emiphumeleni ye-GC-MS, i-diallyl disulfide kanye ne-diallyl trisulfide kwatholakala njengezinhlanganisela ezinkulu ze-EO As, okuhambisana nemibiko yangaphambilini19,20,21. Nakuba imibiko yangaphambilini yabonisa ukuthi i-menthol ingenye yezinhlanganisela zayo eziyinhloko, i-carvone kanye ne-limonene kwaphinde kwatholakala njengezinhlanganisela eziyinhloko ze-Mp EO22,23. Iphrofayili yokwakheka kwe-Os EO ibonise ukuthi i-eugenol kanye ne-methyl eugenol yizinhlanganisela eziyinhloko, okufana nokutholakele kwabacwaningi bangaphambilini16,24. I-Eucalyptol kanye ne-eucalyptol kubikwe njengezinhlanganisela eziyinhloko ezikhona ku-Em leaf oil, okuhambisana nokutholakele kwabanye abacwaningi25,26 kodwa ngokuphambene nokutholakele kuka-Olalade et al.27. Ukubusa kwe-cineole kanye ne-α-pinene kwabonwa ku-melaleuca essential oil, okufana nokutholakele kwangaphambilini28,29. Umehluko oqondile ekwakhiweni nasekuqoqweni kwamafutha abalulekile akhishwe ezinhlotsheni ezifanayo zezitshalo ezindaweni ezahlukene uye wabikwa futhi wabonwa kulolu cwaningo, oluthonywa izimo zokukhula kwezitshalo endaweni, isikhathi sokuvuna, isigaba sokukhula, noma ubudala besitshalo. ukubonakala kwezinhlobo ze-chemotypes, njll.22,30,31,32. Amakhemikhali abalulekile atholakele abe esethengwa futhi ahlolwa imiphumela yawo yokubulala ama-larvicidal kanye nemiphumela kuma-Aedes aegypti omiyane abadala. Imiphumela ibonise ukuthi umsebenzi wokubulala ama-larvicidal we-diallyl disulfide wawufana nalowo we-EO As eluhlaza. Kodwa umsebenzi we-diallyl trisulfide uphakeme kune-EO As. Le miphumela ifana naleyo etholwe nguKimbaris et al. 33 kuma-Culex philippines. Kodwa-ke, la makhemikhali amabili awazange abonise umsebenzi omuhle we-autocidal ngokumelene nomiyane abahlosiwe, okuhambisana nemiphumela kaPlata-Rueda et al 34 kuma-Tenebrio molitor. I-Os EO iyasebenza ngokumelene nesigaba sama-larvical se-Aedes aegypti, kodwa hhayi ngokumelene nesigaba sabantu abadala. Kuye kwatholakala ukuthi umsebenzi wokubulala izibungu wamakhemikhali ayinhloko uphansi kunowe-Os EO eluhlaza. Lokhu kusho indima yamanye amakhemikhali kanye nokusebenzisana kwawo ku-ethylene oxide eluhlaza. I-Methyl eugenol yodwa inomsebenzi omncane, kanti i-eugenol yodwa inomsebenzi wokubulala izibungu olinganiselayo. Lesi siphetho siqinisekisa, ngakolunye uhlangothi, 35,36, kanti ngakolunye uhlangothi, siphikisana neziphetho zabacwaningi bangaphambilini 37,38. Umehluko emaqenjini asebenzayo e-eugenol ne-methyleugenol ungabangela ubuthi obuhlukene ezinambuzaneni ezifanayo ezihlosiwe 39. I-Limonene itholakale inomsebenzi wokubulala izibungu olinganiselayo, kuyilapho umphumela we-carvone wawungabalulekile. Ngokufanayo, ubuthi obuphansi be-limonene ezinambuzaneni ezindala kanye nobuthi obuphezulu be-carvone kusekela imiphumela yezifundo ezithile zangaphambilini 40 kodwa kuphikisana nezinye 41. Ukuba khona kwezibopho eziphindwe kabili kuzo zombili izikhundla ze-intracyclic kanye ne-exocyclic kungandisa izinzuzo zalezi zinhlanganisela njenge-larvicides3,41, kuyilapho i-carvone, okuyi-ketone enama-alpha nama-beta carbons angagcwalisiwe, ingabonisa amandla aphezulu obuthi kubantu abadala42. Kodwa-ke, izici zomuntu ngamunye ze-limonene ne-carvone ziphansi kakhulu kune-EO Mp iyonke (Ithebula 1, Ithebula 3). Phakathi kwama-terpenoids ahlolwe, i-eudesmol itholakale inomsebenzi omkhulu kakhulu wokubulala ama-larvicidal kanye nomsebenzi wabantu abadala onenani le-LC50 elingaphansi kuka-2.5 ppm, okwenza kube yikhompawundi ethembisayo yokulawula omiyane be-Aedes. Ukusebenza kwayo kungcono kunokwe-EO Em yonke, yize lokhu kungahambisani nokutholakele kukaCheng et al.40. I-Eudesmol iyi-sesquiterpene enamayunithi amabili e-isoprene angenamandla kakhulu kune-monoterpenes ene-oxygen efana ne-eucalyptus ngakho-ke inamandla amakhulu njengesibulala-zinambuzane. I-Eucalyptol ngokwayo inomsebenzi omkhulu wokubulala ama-larvicidal kubantu abadala kunokubulala ama-larvicidal, futhi imiphumela evela ezifundweni zangaphambilini iyakusekela futhi iyakuphikisa lokhu37,43,44. Umsebenzi wodwa ucishe ufane nowe-EO Cl yonke. Enye i-bicyclic monoterpene, i-α-pinene, inomphumela omncane wokubulala ama-larvicidal kubantu abadala ku-Aedes aegypti kunomphumela wokubulala ama-larvicidal, okuphambene nomphumela we-EO Cl egcwele. Umsebenzi wokubulala izinambuzane ojwayelekile wama-terpenoids uthonywa yi-lipophilicity yawo, ukuguquguquka, ukwakheka kwe-carbon, indawo yokubonakala kwawo, indawo engaphezulu, amaqembu asebenzayo kanye nezikhundla zawo45,46. Lawa ma-compound angasebenza ngokubhubhisa ukuqongelela kwamaseli, ukuvimba umsebenzi wokuphefumula, ukuphazamisa ukudluliselwa kwemizwa yezinzwa, njll.47 I-organophosphate yokwenziwa i-Temephos itholakale inomsebenzi wokubulala ama-larvicidal ophezulu kakhulu onenani le-LC50 elingu-0.43 ppm, okuhambisana nedatha kaLek -Utala48. Umsebenzi wabantu abadala we-synthetic organophosphorus malathion ubikwe ku-5.44 ppm. Nakuba la ma-organophosphate amabili abonise izimpendulo ezinhle ngokumelene nezinhlobo ze-Aedes aegypti zaselabhorethri, ukumelana nomiyane kula makhemikhali kubikwe ezingxenyeni ezahlukene zomhlaba49. Kodwa-ke, akukho mibiko efanayo yokuthuthuka kokumelana nemithi yamakhambi etholakele50. Ngakho-ke, izitshalo zezitshalo zibhekwa njengezindlela ezingaba khona zokusebenzisa izibulala-zinambuzane zamakhemikhali ezinhlelweni zokulawula izinambuzane.
Umphumela wokubulala ama-larvicidal uhlolwe ezinhlanganisweni ezingu-28 ze-binary (1:1) ezilungiselelwe kusuka kuma-terpenoid anamandla nama-terpenoid ane-thymetphos, kanti izinhlanganisela ezingu-9 zitholakale zihambisana, ezingu-14 ziphikisana kanye nezi-5 ziphikisana. Akukho mphumela. Ngakolunye uhlangothi, ekuhlolweni kwamandla omuntu omdala, kutholakale izinhlanganisela ezingu-7 zihambisana, izinhlanganisela ezingu-15 ziphikisana, kanti izinhlanganisela ezingu-6 zibikwe ukuthi azinawo umphumela. Isizathu sokuthi izinhlanganisela ezithile zikhiqize umphumela wokuhambisana kungenzeka ngenxa yokuthi ama-candidate asebenzisana ngesikhathi esisodwa ezindleleni ezibalulekile ezahlukene, noma ekuvinjelweni okulandelanayo kwama-enzyme ayisihluthulelo ahlukene endleleni ethile yezinto eziphilayo51. Ukuhlanganiswa kwe-limonene ne-diallyl disulfide, i-eucalyptus noma i-eugenol kutholakale ukuthi kuyahambisana kokubili ukusetshenziswa okuncane nokukhulu (Ithebula 6), kanti ukuhlanganiswa kwayo ne-eucalyptus noma i-α-pinene kutholakale ukuthi kunemiphumela yokuphikisana kuma-larvae. Ngokwesilinganiso, i-limonene ibonakala iyi-synergist enhle, mhlawumbe ngenxa yokuba khona kwamaqembu e-methyl, ukungena okuhle ku-stratum corneum, kanye nendlela ehlukile yokusebenza52,53. Kuke kwabikwa ngaphambilini ukuthi i-limonene ingabangela imiphumela enobuthi ngokungena ezicutshini zezinambuzane (ubuthi bokuxhumana), okuthinta uhlelo lokugaya ukudla (i-antifeedant), noma okuthinta uhlelo lokuphefumula (umsebenzi we-fumigation),54 kuyilapho ama-phenylpropanoids afana ne-eugenol angathinta ama-enzyme e-metabolic 55. Ngakho-ke, ukuhlanganiswa kwamakhemikhali anezindlela ezahlukene zokusebenza kungandisa umphumela obulalayo wengxube. I-Eucalyptol itholakale isebenzisana ne-diallyl disulfide, i-eucalyptus noma i-α-pinene, kodwa okunye ukuhlanganiswa namanye amakhemikhali kwakungeyona i-larvicidal noma iphikisana. Izifundo zakuqala zibonise ukuthi i-eucalyptol inomsebenzi wokuvimbela ku-acetylcholinesterase (AChE), kanye nama-octamine nama-GABA receptors56. Njengoba ama-monoterpenes ajikelezayo, i-eucalyptol, i-eugenol, njll. angaba nendlela efanayo yokusebenza nomsebenzi wawo onobungozi be-neuro, 57 ngaleyo ndlela anciphise imiphumela yawo ehlangene ngokuvinjelwa okuhlangene. Ngokufanayo, inhlanganisela ye-Temephos ne-diallyl disulfide, i-α-pinene kanye ne-limonene kwatholakala ukuthi kuyasebenzisana, kusekela imibiko yangaphambilini yomphumela wokubambisana phakathi kwemikhiqizo yamakhambi kanye ne-organophosphates yokwenziwa58.
Ukuhlanganiswa kwe-eudesmol ne-eucalyptol kutholakale ukuthi kunomphumela wokubambisana ezigabeni ze-larval neze-adult ze-Aedes aegypti, mhlawumbe ngenxa yezindlela zazo ezahlukene zokusebenza ngenxa yezakhiwo zazo zamakhemikhali ezahlukene. I-Eudesmol (i-sesquiterpene) ingathinta uhlelo lokuphefumula 59 kanti i-eucalyptol (i-monoterpene) ingathinta i-acetylcholinesterase 60. Ukuvezwa kanye kwezithako ezindaweni ezimbili noma ngaphezulu eziqondiwe kungathuthukisa umphumela obulalayo wokuhlanganiswa. Ekuhlolweni kwezinto eziphilayo zabantu abadala, i-malathion itholakale isebenzisana ne-carvone noma i-eucalyptol noma i-eucalyptol noma i-diallyl disulfide noma i-α-pinene, okubonisa ukuthi ihambisana nokwengezwa kwe-limonene ne-di. I-allercide enhle yokubambisana kuyo yonke iphothifoliyo yamakhemikhali e-terpene, ngaphandle kwe-allyl trisulfide. I-Thangam ne-Kathiresan61 nazo zibike imiphumela efanayo yomphumela wokubambisana we-malathion neziqephu zamakhambi. Le mpendulo yokubambisana ingase ibangelwe yimiphumela enobuthi ehlanganisiwe ye-malathion kanye namakhemikhali e-phyto kuma-enzyme okuhlanza izinambuzane. Ama-Organophosphates afana ne-malathion ngokuvamile asebenza ngokuvimbela ama-esterases e-cytochrome P450 kanye nama-monooxygenase62,63,64. Ngakho-ke, ukuhlanganisa i-malathion nalezi zindlela zokusebenza kanye nama-terpenes anezindlela ezahlukene zokusebenza kungathuthukisa umphumela obulalayo ojwayelekile komiyane.
Ngakolunye uhlangothi, ukuphikisana kubonisa ukuthi amakhemikhali akhethiwe awasebenzi kangako ekuhlanganisweni kunekhemikhali ngayinye yodwa. Isizathu sokuphikisana kwezinye izinhlanganisela kungaba ukuthi ikhemikhali eyodwa iguqula ukuziphatha kwenye ngokushintsha izinga lokumuncwa, ukusatshalaliswa, i-metabolism, noma ukuphuma kwayo. Abacwaningi bokuqala babheka lokhu njengembangela yokuphikisana ekuhlanganisweni kwezidakamizwa. Ama-Molecule Indlela engenzeka 65. Ngokufanayo, izimbangela ezingaba khona zokuphikisana zingase zihlobane nezindlela ezifanayo zokusebenza, ukuncintisana kwamakhemikhali ayingxenye ye-receptor efanayo noma indawo eqondiwe. Kwezinye izimo, ukuvinjelwa okungekhona ukuncintisana kweprotheyini eqondiwe nakho kungavela. Kulolu cwaningo, amakhemikhali amabili e-organosulfur, i-diallyl disulfide kanye ne-diallyl trisulfide, abonise imiphumela yokuphikisana, mhlawumbe ngenxa yokuncintisana kwendawo efanayo eqondiwe. Ngokufanayo, lawa makhemikhali amabili e-sulfur abonise imiphumela yokuphikisana futhi awazange abe nomthelela lapho ehlanganiswa ne-eudesmol kanye ne-α-pinene. I-Eudesmol kanye ne-alpha-pinene ngokwemvelo, kanti i-diallyl disulfide kanye ne-diallyl trisulfide ngokwemvelo. Ngokusekelwe esakhiweni samakhemikhali, ukuhlanganiswa kwala ma-compound kufanele kwandise umsebenzi obulalayo jikelele njengoba izindawo zawo eziqondiwe zivame ukuhluka34,47, kodwa ngokuhlola sithole ukungezwani, okungase kube ngenxa yendima yala ma-compound kwezinye izinhlelo zezinto eziphilayo ezingaziwa eziphila nazo ngenxa yokusebenzisana. Ngokufanayo, ukuhlanganiswa kwe-cineole ne-α-pinene kukhiqize izimpendulo eziphikisanayo, yize abacwaningi babike ngaphambilini ukuthi la ma-compound amabili anezinhloso ezahlukene zesenzo47,60. Njengoba womabili la ma-compounds engama-cyclic monoterpenes, kungase kube nezindawo ezithile eziqondiwe ezivamile ezingase zincintisane ngokubopha futhi zithonye ubuthi obuphelele bama-combinatorial pairs afundwe.
Ngokusekelwe kumanani e-LC50 kanye nokufa okubonwe, kukhethwe izinhlanganisela ezimbili ezinhle kakhulu ze-terpene ezisebenzisanayo, okungukuthi amabhangqa e-carvone + limonene kanye ne-eucalyptol + eudesmol, kanye ne-organophosphorus malathion yokwenziwa ene-terpenes. Inhlanganisela engcono kakhulu ye-malathion + Eudesmol compounds ihlolwe ku-bioassay yezinambuzane zabantu abadala. Qondisa amakoloni amakhulu ezinambuzane ukuqinisekisa ukuthi lezi zinhlanganisela ezisebenzayo zingasebenza yini ngokumelene nenani elikhulu labantu ezindaweni ezinkulu zokuchayeka. Zonke lezi zinhlanganisela zibonisa umphumela we-synergistic ngokumelene nezinqwaba ezinkulu zezinambuzane. Imiphumela efanayo itholakale ngenhlanganisela engcono kakhulu ye-synergistic larvicidal ehlolwe ngokumelene nenani elikhulu lezinambuzane ze-Aedes aegypti. Ngakho-ke, kungashiwo ukuthi inhlanganisela ye-synergistic larvicidal kanye ne-adulticidal yamakhemikhali e-EO ezitshalo iyindlela enamandla yokulwa namakhemikhali akhona okwenziwa futhi ingasetshenziswa ukulawula imiphakathi ye-Aedes aegypti. Ngokufanayo, inhlanganisela esebenzayo yama-larvicides okwenziwa noma ama-adulticides ane-terpenes ingasetshenziswa futhi ukunciphisa imithamo ye-thymetphos noma i-malathion enikezwa omiyane. Lezi zinhlanganisela ezinamandla zokubambisana zingase zinikeze izixazululo zezifundo zesikhathi esizayo mayelana nokuvela kokumelana nemithi komiyane base-Aedes.
Amaqanda e-Aedes aegypti aqoqwe e-Regional Medical Research Centre, eDibrugarh, e-Indian Council of Medical Research futhi agcinwa ngaphansi kokushisa okulawulwayo (28 ± 1 °C) kanye nomswakama (85 ± 5%) eMnyangweni weZoology, eGauhati University ngaphansi kwezimo ezilandelayo: I-Arivoli yachazwa nabanye. Ngemva kokuchamisela, izibungu zondliwa ukudla kwezibungu (impuphu yebhisikidi yezinja kanye nemvubelo ngesilinganiso esingu-3:1) kanti abantu abadala bondliwa ngesisombululo se-glucose esingu-10%. Kusukela ngosuku lwesithathu ngemva kokuvela, omiyane besifazane abadala bavunyelwa ukumunca igazi lamagundane angama-albino. Cwilisa iphepha lokuhlunga emanzini engilazini bese ulibeka ekhejini lokubeka amaqanda.
Amasampula ezitshalo akhethiwe okungukuthi amaqabunga e-eucalyptus (Myrtaceae), i-basil engcwele (Lamiaceae), i-mint (Lamiaceae), i-melaleuca (Myrtaceae) kanye nama-allium bulbs (Amaryllidaceae). Aqoqwe eGuwahati futhi ahlonzwa uMnyango Wezitshalo, eGauhati University. Amasampula ezitshalo aqoqwe (500 g) afakwa emanzini acwebezelayo kusetshenziswa umshini we-Clevenger amahora ayi-6. I-EO ekhishwe yaqoqwa emabhodleleni engilazi ahlanzekile futhi yagcinwa ku-4°C ukuze kuqhutshekwe kufundwa.
Ubuthi bokubulala izibungu buhloliwe kusetshenziswa izinqubo ezijwayelekile ze-World Health Organisation 67 ezishintshiwe kancane. Sebenzisa i-DMSO njenge-emulsifier. Ukuhlushwa kwe-EO ngakunye kwahlolwa ekuqaleni ku-100 kanye no-1000 ppm, kwadalulwa izibungu ezingu-20 kokuphindaphindwa ngakunye. Ngokusekelwe emiphumeleni, kwasetshenziswa ububanzi bokuhlushwa futhi ukufa kwaqoshwa kusukela ehoreni eli-1 kuya emahoreni angu-6 (ngezikhathi zehora eli-1), kanye namahora angu-24, amahora angu-48 kanye namahora angu-72 ngemva kokwelashwa. Ukuhlushwa kwe-Sublethal (LC50) kwanqunywa ngemva kwamahora angu-24, 48 kanye nama-72 okuchayeka. Ukuhlushwa ngakunye kwahlolwa kathathu kanye nokulawula okukodwa okungalungile (amanzi kuphela) kanye nokulawula okukodwa okuhle (amanzi aphathwe yi-DMSO). Uma kwenzeka ukuphambana futhi izibungu ezingaphezu kuka-10% zeqembu lokulawula ziyafa, ukuhlolwa kuyaphindwa. Uma izinga lokufa eqenjini lokulawula liphakathi kuka-5-10%, sebenzisa ifomula yokulungisa i-Abbott 68.
Indlela echazwe nguRamar nabanye 69 yasetshenziswa ekuhlolweni kwe-bioassay yabantu abadala ngokumelene ne-Aedes aegypti kusetshenziswa i-acetone njengesinyibilikisi. I-EO ngayinye yahlolwa ekuqaleni ngokumelene nomiyane abadala be-Aedes aegypti ngamanani angu-100 no-1000 ppm. Faka u-2 ml wesisombululo ngasinye esilungisiwe enombolweni ye-Whatman. Iphepha elilodwa lokuhlunga (usayizi 12 x 15 cm2) bese uvumela i-acetone iphele imizuzu eyi-10. Iphepha lokuhlunga eliphathwe nge-acetone engu-2 ml kuphela lasetshenziswa njengokulawula. Ngemva kokuba i-acetone isiphelile, iphepha lokuhlunga eliphathwe kanye nephepha lokulawula lokuhlunga kufakwa epayipini eliyindilinga (elijule ngo-10 cm). Omiyane abayishumi abangadli igazi abanezinsuku ezi-3 kuya kwezi-4 ubudala badluliselwa kuma-triplicate e-concentration ngayinye. Ngokusekelwe emiphumeleni yokuhlolwa kokuqala, kwahlolwa amazinga ahlukahlukene amafutha akhethiwe. Ukufa kwaqoshwa ngehora eli-1, amahora ama-2, amahora ama-3, amahora ama-4, amahora ama-5, amahora ama-6, amahora angama-24, amahora angama-48 kanye namahora angama-72 ngemuva kokukhululwa komiyane. Bala amanani e-LC50 ngezikhathi zokuchayeka zamahora angama-24, amahora angama-48 kanye namahora angama-72. Uma izinga lokufa lendawo yokulawula lidlula u-20%, phinda lonke isivivinyo. Ngokufanayo, uma izinga lokufa eqenjini lokulawula lingaphezu kuka-5%, lungisa imiphumela yamasampula aphathwe usebenzisa ifomula ka-Abbott68.
I-Gas chromatography (Agilent 7890A) kanye ne-mass spectrometry (Accu TOF GCv, Jeol) kwenziwa ukuze kuhlaziywe izakhi zamafutha abalulekile akhethiwe. I-GC yayifakwe i-FID detector kanye nekholomu ye-capillary (HP5-MS). Igesi ethwalayo yayiyi-helium, izinga lokugeleza lalingu-1 ml/min. Uhlelo lwe-GC lubeka i-Allium sativum ku-10:80-1M-8-220-5M-8-270-9M kanye ne-Ocimum Sainttum ku-10:80-3M-8-200-3M-10-275-1M-5 – 280, ye-mint 10:80-1M-8-200-5M-8-275-1M-5-280, ye-eucalyptus 20.60-1M-10-200-3M-30-280, kanye nebomvu. Ngezingqimba eziyinkulungwane, yizo eziyi-10: 60-1M-8-220-5M-8-270-3M.
Ama-compound amakhulu e-EO ngayinye atholakale ngokusekelwe kumaphesenti endawo abalwe kusukela emiphumeleni ye-GC chromatogram kanye ne-mass spectrometry (okubhekiselwe kusizindalwazi sezindinganiso ze-NIST 70).
Ama-compound amabili amakhulu ku-EO ngayinye akhethwe ngokusekelwe emiphumeleni ye-GC-MS futhi athengwa ku-Sigma-Aldrich ngobumsulwa obungu-98-99% ukuze kuhlolwe okwengeziwe nge-bioassays. Ama-compound ahlolwe ukuthi abulala ama-larvicides futhi ayasebenza kahle kubantu abadala ngokumelene ne-Aedes aegypti njengoba kuchaziwe ngenhla. Ama-larvicides okwenziwa asetshenziswa kakhulu i-tamephosate (Sigma Aldrich) kanye ne-malathion yesidakamizwa sabantu abadala (Sigma Aldrich) ahlaziywe ukuze kuqhathaniswe ukusebenza kwawo nama-compounds e-EO akhethiwe, kulandela inqubo efanayo.
Izingxube ezimbili zamakhemikhali e-terpene akhethiwe kanye namakhemikhali e-terpene kanye nama-organophosphate okuhweba (i-tilephos kanye ne-malathion) zalungiswa ngokuxuba umthamo we-LC50 wenhlanganisela ngayinye efanelekile ngesilinganiso esingu-1:1. Izinhlanganisela ezilungiselelwe zahlolwa ezigabeni ze-larval kanye neze-Aedes aegypti zabantu abadala njengoba kuchaziwe ngenhla. Ukuhlolwa ngakunye kwe-bioassay kwenziwa kathathu ngenhlanganisela ngayinye kanye kathathu ngamakhemikhali ngamanye akhona kwinhlanganisela ngayinye. Ukufa kwezinambuzane eziqondiwe kwaqoshwa ngemuva kwamahora angama-24. Bala izinga lokufa elilindelekile lengxube ye-binary usebenzisa ifomula elandelayo.
lapho i-E = izinga lokufa elilindelekile lomiyane be-Aedes aegypti ngenxa yenhlanganisela ye-binary, okungukuthi uxhumano (A + B).
Umphumela wengxube ngayinye enama-binary wabhalwa njengomphumela ohambisanayo, ophikisanayo, noma ongenawo ngokusekelwe enanini le-χ2 elibalwe ngendlela echazwe yi-Pavla52. Bala inani le-χ2 lenhlanganisela ngayinye usebenzisa ifomula elandelayo.
Umphumela wokuhlanganiswa uchazwe njengokuhambisana lapho inani elibaliwe le-χ2 likhulu kunenani lethebula lamadigri enkululeko ahambisanayo (isikhawu sokuzethemba esingu-95%) futhi uma ukufa okubonwayo kutholakale kudlula ukufa okulindelekile. Ngokufanayo, uma inani elibaliwe le-χ2 lanoma iyiphi inhlanganisela lidlula inani lethebula elinamadigri athile enkululeko, kodwa ukufa okubonwayo kuphansi kunokufa okulindelekile, ukwelashwa kubhekwa njengokuphikisayo. Futhi uma kunoma yikuphi ukuhlanganiswa inani elibaliwe le-χ2 lingaphansi kunenani lethebula kumadigri enkululeko ahambisanayo, inhlanganisela ibhekwa njengengenamphumela.
Kukhethwe izinhlanganisela ezintathu kuya kwezine ezingase zivumelane (izibungu eziyi-100 kanye nomsebenzi wezinambuzane ezingama-50 ezibulala izibungu kanye nezinambuzane ezindala) ukuze kuhlolwe inani elikhulu lezinambuzane. Abantu abadala) baqhubeka njengoba kushiwo ngenhla. Kanye nezingxube, izinhlanganisela ngazinye ezikhona ezingxubeni ezikhethiwe nazo zahlolwa enanini elilinganayo lezibungu ze-Aedes aegypti kanye nabantu abadala. Isilinganiso sokuhlanganiswa siyisilinganiso esisodwa se-LC50 senhlanganisela eyodwa efanelekile kanye nesilinganiso se-LC50 senye inhlanganisela eyingxenye. Ekuhlolweni kwe-bioassay yomsebenzi wabantu abadala, izinhlanganisela ezikhethiwe zancibilikiswa ku-acetone ye-solvent futhi zafakwa ephepheni lokuhlunga eligoqwe esitsheni sepulasitiki esingu-1300 cm3. I-acetone yashiswa imizuzu eyi-10 futhi abantu abadala bakhululwa. Ngokufanayo, ekuhlolweni kwe-bioassay ye-larvicidal, izilinganiso zezinhlanganisela ze-LC50 zaqala zancibilikiswa ngamanani alinganayo e-DMSO bese zixutshwa nelitha elilodwa lamanzi eligcinwe ezitsheni zepulasitiki ezingama-1300 cc, futhi izibungu zakhululwa.
Ukuhlaziywa okungenzeka kwedatha yokufa eqoshiwe engu-71 kwenziwa kusetshenziswa i-SPSS (inguqulo 16) kanye nesofthiwe ye-Minitab ukuze kubalwe amanani e-LC50.
Isikhathi sokuthunyelwe: Julayi-01-2024