Ukuthola nokusabalalisa izakhi zomsoco kuhlanganisa ukudla kwezinambuzane kanye nezici zomlando wempilo.Ukunxephezela ukuntula kwemisoco ethile ezigabeni ezahlukene zokuphila, izinambuzane zingathola lezi zakhi ngokudla okungeziwe, isibonelo, ngokudla uketshezi lwezilwane ezinomgogodla ngenqubo eyaziwa ngokuthi ama-puddles.Umiyane i-Anopheles arabiani ibonakala i-Anopheles arabiani ibonakala ingenamsoco futhi ngenxa yalokho, i-malnoproduction ihlose kokubili. lolu cwaningo bekuwukuhlola ukuthi u-An. Ukuhlukumezeka kwe-arabiensis kumchamo wenkomo ukuze kutholwe izakhi kuthuthukisa izici zomlando wempilo.
Qiniseka ukuthi iphephile.I-arabiensis ikhangwe iphunga lomchamo wenkomo omusha, wamahora angu-24, angu-72, namahora angu-168, kanye nokufuna umninikhaya nokuphakelwa igazi (ukudla kwamahora angu-48 ngemva kokuphuma kwegazi) kwalinganiswa nge-olfactometer ye-Y-tube, futhi abesifazane abakhulelwe bahlolwa futhi kwahlolwa amakhemikhali e-electroological. Izingxube ze-bioactive emchameni wezinkomo kuwo wonke amakilasi eminyaka emine.Izingxube zokwenziwa zezingxube ze-bioactive zahlolwa ku-Y-tube nasezivivinyweni zasensimini.Ukuphenya umchamo wenkomo kanye ne-urea yawo eyinhloko equkethe i-nitrogen njengokudla okungase kube ukudla okwanelisayo kwamagciwane kamalaleveva, imingcele yokudla kanye nezici zomlando wokuphila kwalinganiswa. ukudla, abesifazane bahlolelwa ukuphila, ukundiza okuboshiwe, nokuzala.
Funa igazi kanye nokudla komninikhaya.Ezifundweni zaselabhorethri nezindawo, ama-Arabhu akhangwa iphunga lemvelo nelokwenziwa lomchamo wenkomo omusha nomdala.Abesifazane abakhulelwe babengenandaba nokusabela komchamo wezinkomo ezindaweni zokuzala.Abesifazane abafuna ukusingathwa nabamunca igazi bamunca umchamo wenkomo kanye ne-urea ngenkuthalo futhi bahlukanisele lezi zinsiza njengezindawo zokuhweba zokuphila. ukusinda, noma ukukhiqiza kabusha.
Ukutholwa nokusatshalaliswa komchamo wenkomo i-Anopheles arabinis ukuze kuthuthukiswe izici zomlando wempilo.Ukudla okungeziwe komchamo wenkomo kuthinta umthamo we-vector ngokuqondile ngokwandisa ukuphila kwansuku zonke kanye nokuminyana kwe-vector, futhi ngokungaqondile ngokushintsha umsebenzi wokundiza ngakho-ke kufanele kubhekwe kumamodeli azayo.
Ukuthola nokusabalalisa izakhi zomsoco kuhlanganisa ukuphakelwa kwezinambuzane kanye nezici zomlando wokuphila [1,2,3].Izinambuzane ziyakwazi ukukhetha futhi zithole ukudla futhi zenze ukudla okuyisinxephezelo ngokusekelwe ekutholakaleni kokudla kanye nezidingo zomsoco [1, 3].Ukusatshalaliswa kwezakhi zomzimba kuncike enqubweni yomlando wokuphila futhi kungase kuholele ezidingweni ezahlukene zekhwalithi yokudla kanye nenani lempilo1 compse in life1, compte in life. ukuntuleka kwezakhi ezithile zomsoco, izinambuzane zingathola lezi zakhi ngokudla okungeziwe, njengodaka, indle ehlukahlukene kanye nezimfihlo zezilwane ezinomgogodla, kanye ne-carrion, inqubo eyaziwa ngokuthi ama-puddles [2] .Nakuba izinhlobo ezihlukahlukene zovemvane kanye nebhu zichazwa ngokuyinhloko, izimbobo zokunisela nazo zenzeka kwezinye izilawuli zezinambuzane, futhi ezinye izinhlobo zokudla zingaba nemiphumela ebalulekile yokuphila. izici [2, 4, 5, 6] ,7].Umiyane kamalaleveva u-Anopheles gambiae sensu lato (sl) uvela njengomuntu omdala 'ongondlekile' [8], ngakho ukuchelela kungase kudlale indima ebalulekile ezicini zomlando wokuphila kwawo, kodwa lokhu kuziphatha akunakwa kuze kube manje.
Ukudla okune-nitrogen omiyane abadala besifazane i-Anopheles kulinganiselwe ngenxa yokuqukethwe kwekhalori ephansi okuvela esigabeni sezibungu kanye nokungasetshenziswa kahle kwesidlo segazi [9].I-Female Ann.gambiae sl ngokuvamile inxephezela lokhu ngokufaka ukudla okungeziwe kwegazi [10, 11], lokho kubeka abantu abaningi engozini enkulu yokuthola umiyane Okunye, omiyane bangasebenzisa ukuphakela okwengeziwe kwendle yezilwane ezinomgogodla ukuze bathole izakhi zenitrogen ezithuthukisa ukuzivumelanisa nokundiza, njengoba kuboniswa ezinye izinambuzane [2]. Maqondana nalokhu, ukukhanga okunamandla nokuhlukile kolunye lwezinhlobo eziyizelamani ngaphakathi kwe-An.The Gambian sl species complex, Anow, pheles,12, fresh, 12,12 kuyathakazelisa.I-Anopheles arabinis ingosomathuba ngokuthanda kwayo futhi yaziwa ngokuzihlanganisa nokudla izinkomo.Umchamo wenkomo ungumthombo ocebile ngamakhompiyutha e-nitrogen, urea ibalwa ku-50-95% wenani eliphelele le-nitrogen emchameni omusha [15, 16].Njengoba iminyaka yomchamo wenkomo, i-microorganity inciphisa i-nitrogen emahoreni angu-2 [15] .Ngokwenyuka okusheshayo kwe-ammonia, okuhlotshaniswa nokwehla kwe-nitrogen ephilayo, ama-alkalophilic microorganisms (amaningi awo akhiqiza izinhlanganisela ezinobuthi omiyane) ayachuma [15], okungenzeka ukuthi u-Ann.arabiensis wesifazane ukhangwa ngokukhethekile umchamo oneminyaka engama-24 noma ngaphansi [13, 14].
Kulolu cwaningo, kwabhekwa ama-Ans aphethe kanye nokudla ngegazi.Phakathi nomjikelezo wayo wokuqala we-gonadotropin, i-arabiensis yahlolwa ukuze kutholwe izinhlanganisela ze-nitrogenous, okuhlanganisa i-urea, ngokuxuba umchamo.Okulandelayo, uchungechunge lokuhlola lwenziwa ukuhlola ukuthi omiyane besifazane bawaba kanjani lesi sisetshenziswa esingaba somsoco ukuze kuthuthukiswe ukuzala, ukuzala kanye nokuzala kabusha. Umchamo wenkomo omusha nomdala wahlolwa ukuze kutholakale ukuthi ingabe lezi zinikeza izinkomba ezithembekile zomsingathi kanye no-An ondliwa ngegazi. Ekusesheni kwabo lesi sisetshenziswa esingaba somsoco, i-arabiensis yathola ukuhlobana kwamakhemikhali ngemva kokukhanga okubonwayo. umphumela wephunga lomchamo wezinkomo ezifundeni ezihlukahlukene zomzimba. Ukukhangwa omiyane.Imiphumela etholiwe iqinisekisa ukuthi u-An. i-arabiensis ithola futhi isabalalise izinhlanganisela ze-nitrogenous ezitholakala kumchamo wezilwane ezinomgogodla ukuze zithonye izici zomlando wokuphila.Le miphumela kuxoxwa ngayo kumongo wemiphumela engase ibe khona ye-epidemiological nokuthi ingasetshenziswa kanjani ekugadweni nasekulawuleni i-vector.
Ama-Anopheles arabicans (uhlobo lwe-Dongola) ayenakekelwa ku-25 ± 2 °C, 65 ± 5% RH kanye nomjikelezo wokukhanya okungamahora angu-12:12:ubumnyama. izinkomishi ezingu-30 ml (Nolato Hertila, Åstorp, SE) bese idluliselwa kumakheji e-Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) ukuze kuvunyelwe ukuvela kwabantu abadala.Abadala banikezwe isisombululo se-sucrose esingu-10% sesikhangiso se-ad libitum kuze kube yizinsuku ezi-4 ezinikezwa abasingathi ngemva kokubona ukudla ukuhlola, noma babulawa indlala ngobusuku obubodwa ngamanzi acwebile ngaphambi kokuhlolwa, njengoba kuchazwe ngezansi.Abesifazane abasetshenziselwa ukuhlolwa kweshubhu lendiza babulawa indlala amahora angu-4-6 kuphela nge-ad libitum yamanzi.Ukulungiselela omiyane abamunca igazi ukuze kuhlolwe ama-bioassays alandelayo, abesifazane abangu-4 dpe banikezwe igazi lezimvu le-defibrotic (Håtunalab) usebenzisa i-Workshop system, i-Workshop, i-Workshop. Accrington, UK).Abesifazane ababeminyaniswe ngokugcwele babe sebedluliselwa emakhejini ngamanye futhi banikezwa ukudla ngokuqondile, njengoba kuchazwe ngezansi, noma i-10% ye-sucrose ad libitum yezinsuku ezingu-3 ngaphambi kokuhlolwa okuchazwe ngezansi.Abesifazane bokugcina babesetshenziselwa ama-bioassays e-flight tube futhi badluliselwa elabhorethri, base bene-distilled ad libitum yamanzi ngaphambi kwamahora angu-4-6.
Ukuhlolwa kokuphakelayo kwasetshenziswa ukuze kulinganiswe ukusetshenziswa komchamo nokusetshenziswa kwe-urea kumuntu wesifazane omdala ongum-A.Arab.Abesifazane abafuna ukusingatha futhi abaphakelwa igazi banikezwe ukudla okunomchamo wenkomo ohlanjululiwe ongu-1% omusha nomdala, ukugxiliswa okuhlukahlukene kwe-urea, nezilawuli ezimbili (10% sucrose namanzi) amahora angu-48. Ngaphezu kwalokho, umbala wokudla (1 mgne CASani 2650-17-1; Sigma-Aldrich, Stockholm, SE) yengezwe ekudleni futhi yanikezwa nge-4 × 4 matrix ngamashubhu e-microcentrifuge angu-250 µl (Axygen Scientific, Union City, CA, US; Umfanekiso 1A) Gcwalisa onqenqemeni µ kanye nemiphumela yombala ongase ube khona ukuze ugweme udayi we-moquis (~300). beka omiyane abayi-10 esitsheni esikhulu sePetri (ububanzi obungu-12 cm no-6 cm ukuphakama; i-Semadeni, i-Ostermundigen, i-CH; Umfanekiso 1A) ebumnyameni obuphelele ku-25 ± 2 cm °C no-65 ± 5% umswakama ohlobene.Lezi zilingo ziphindwe izikhathi ezingu-5 kuya kwezingu-10. Ngemva kokuhlaziywa kokudla kwaze kwaba ngu-0 ukuchayeka ku-2 ° C.
Bheka umchamo wezinkomo kanye ne-urea emuncwa isilwane esincelisayo futhi esimunca igazi, u-Anopheles arabianus. Esivivinyweni sokuphakela (A), omiyane besifazane banikezwe ukudla okuhlanganisa umchamo wenkomo omusha nomdala, ukugxila okuhlukahlukene kwe-urea, i-sucrose (10%), namanzi acwecwe (H2O).I-host-seking (B) kanye nezinye i-sucrose ezifunzwa igazi. ukuthi abesifazane abafuna ukusingatha umchamo bamunca umchamo wenkomo wamahora angu-72 ngaphansi komchamo wenkomo wamahora angu-168 (B).Isilinganiso sengqikithi yengqikithi ye-nitrogen (± ukuchezuka okujwayelekile) yomchamo ivezwa ku-inset.Abesifazane abafuna ukusingathwa (D, F) nabamunca igazi (E, G) bathatha i-urea ngendlela encike kumthamo ngendlela ehluke kakhulu kunaleyo nenye i-Dhale. (i-ANOVA yendlela eyodwa isebenzisa ukuhlaziywa kwe-Tukey's post hoc; p <0.05).Amabha amaphutha amelela iphutha elivamile lencazelo (BE).Umugqa wedeshi eqondile umelela umugqa wokuhlehla womugqa welogi (F, G)
Ukuze kukhululwe ukudla okumuncwe, omiyane babekwe ngabodwana kumashubhu e-microcentrifuge angu-1.5 ml aqukethe u-230 µl wamanzi acwecwe futhi isicubu saphazanyiswa kusetshenziswa i-pestle elahlwayo kanye ne-cordless motor (VWR International, Lund, SE), kulandelwa i-centrifugation ngo-10 krpm imizuzu engu-10 . i-microplate (Sigma-Aldrich) kanye ne-absorbence (λ620) kwanqunywa kusetshenziswa isifundi se-microplate esisekelwe ku-spectrophotometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Okunye, omiyane bagaywe ngo-1 ml wamanzi acwecwe, 900 µl okwathi uhlaziyo oluyi-curovethrikhi engu-20 lwadluliswa i-curofted 6 (i-curofted 6 l) nm; UV 1800, Shimadzu, Kista, SE).Ukuze kulinganise ukudla okudlayo, ijika elijwayelekile lalungiswa ngokuhlanjululwa kwe-serial ukuze kukhiqizwe u-0.2 µl kuya ku-2.4 µl ka-1 mg ml-1 xylene cyanide.Kwabe sekuthi, ukuminyana kwe-optical kokudla okwaziwayo kwenani ngalinye le-moquid.
Idatha yevolumu yahlaziywa kusetshenziswa ukuhlaziywa kwendlela eyodwa yokuhluka (i-ANOVA) kwalandelwa ukuqhathanisa kwe-Tukey's post hoc pairwise (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). (I-GraphPad Prism v8.0.0 ye-Mac, i-GraphPad Software, i-San Diego, CA, US).
Cishe u-20 µl wamasampuli omchamo asuka eqenjini ngalinye lobudala ayeboshwe ku-Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) futhi afakwa kumaphilisi kathayela (8 mm × 5 mm). I-Scientific, Waltham, MA, US) ukuze inqume okuqukethwe kwe-nitrogen kumchamo omusha nomdala ngokuya ngephrothokholi yomkhiqizi.Ingqikithi ye-nitrogen (g N l-1) yalinganiswa ngokusekelwe ekugxilweni kwe-urea okwaziwayo okusetshenziswa njengezinga elijwayelekile.
Ukuhlola umphumela wokudla ekusindeni kwabesifazane abafuna ukusingatha nabamunca igazi, omiyane babekwa ngabanye ezitsheni ezinkulu ze-Petri (ububanzi obungu-12 cm no-6 cm ukuphakama; i-Semadeni) enembobo embozwe inezikhala esivalweni (ububanzi obungu-3 cm) kanye Nokungenisa umoya nokudla.Ukudla kwanikezwa ngokuqondile ngemva kokuchama okungu-4 kanye ne-dipe entsha engu-4% kanye ne-dipe entsha ehlanganisiwe i-urea, nezilawuli ezimbili, i-10% ye-sucrose namanzi.Ukudla ngakunye kwafakwa ngepayipi kwi-tampon yamazinyo (DAB Dental AB, Upplands Väsby, SE) efakwe kusirinji engu-5 ml (Thermo Fisher Scientific, Gothenburg, SE), i-plunger ikhishiwe, futhi ibekwe phezu kwe-petri dish yakho ye-petri ngosuku. ngenhla.Omiyane abasaphila babebalwa kabili ngosuku, kuyilapho omiyane abafile balahlwa kwaze kwaba sekufeni umiyane wokugcina (n = 40 ngokwelashwa ngakunye).Ukusinda komiyane abadla ukudla okuhlukahlukene kwahlaziywa ngezibalo kusetshenziswa amajika okusinda e-Kaplan-Meyer kanye nokuhlolwa kwezinga lelogi ukuze kuqhathaniswe ukusinda komiyane be-SPSS. 24.0.0.0).
Igayo elindizayo lomiyane elisekelwe ku-Attisano et al.[17], elenziwe ngamaphaneli e-acrylic acacile angu-5 mm ububanzi (10 cm ububanzi x 10 cm ubude x 10 cm ukuphakama) ngaphandle kwamaphaneli angaphambili nangemuva (Fig. 3: top). inaliti elenga phakathi kwepheya kamagnet we-neodymium ukuhlukana okungamasentimitha angu-9. Ishubhu elivundlile elenziwe ngento efanayo (6.5 cm L) lihlukanise kabili ishubhu eme mpo ukuze lakhe ingalo eboshiwe kanye nengalo ephethe ucezu oluncane lwephepha le-aluminium njengesignali ephazamisa ukukhanya.
Abesifazane abalambile amahora angu-24 banikezwa ukudla okungenhla imizuzu engu-30 ngaphambi kokuvinjelwa. Omiyane besifazane abasuthiswe ngokugcwele babe sebebulawa izinzwa eqhweni imizuzu engu-2-3 futhi banamathisele izikhonkwane zesinambuzane ngengcina yezinyosi (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) bese bexhunywa ezingalweni zamashubhu. I-Mill.Revolutions ngendiza ngayinye yarekhodwa isiqophi sedatha esakhelwe ngokwezifiso, sabe sesigcinwa futhi siboniswa kusetshenziswa isofthiwe ye-PC-Lab 2000™ (v4.01; Velleman, Gavere, BE).Imishini yendiza yafakwa egumbini elilawulwa isimo sezulu (12 h:12 h, ukukhanya: kumnyama, 25 ± 2 °C, 5% RH).
Ukuze ubone ngeso lengqondo iphethini yomsebenzi wendiza, ingqikithi yebanga elindisiwe (m) kanye nenani eliphelele lemisebenzi yendiza elandelanayo ibalwe ngehora esikhathini esingamahora angu-24. Ngaphezu kwalokho, amabanga amaphakathi ahamba ngabesifazane ngabanye aqhathaniswa nokwelashwa futhi ahlaziywa kusetshenziswa indlela eyodwa yokuhlaziya i-ANOVA kanye ne-Tukey's post hoc (JMP Pro, v14.0.0), i-avareji yokwelashwa ibhekwa njenge-Institute ehlukile, i-SAS ibhekwa njengesilinganiso esizimele, i-Inc, i-SAS. factor.Ukwengeza, inani elimaphakathi lemizuliswano libalwa ngokunyuka kwemizuzu eyi-10.
Ukuhlola umphumela wokudla ekusebenzeni kokuzala kwe-An.arabiensis, abesifazane abayisithupha (4 dpe) badluliselwe ngqo kumakheji e-Bugdorm (30 cm × 30 cm × 30 cm) ngemva kokuqoqwa kwegazi base behlinzeka ngokudla kokuhlola amahora angu-48 njengoba kuchazwe ngenhla.Ukudla kwabe sekukhishwa futhi izinkomishi zokuzalanisa (30 ml) zagcwaliswa ngosuku lwesithathu lwe-Nostilled Hertila; Amahora angu-48, ukushintsha njalo emahoreni angu-24. Phinda uhlobo lokudla ngalunye izikhathi ezingu-20-50.Amaqanda abalwa futhi aqoshwa ekhejini ngalinye lokuhlola.Izibonelo zamaqanda zasetshenziselwa ukuhlola ubukhulu besilinganiso kanye nokuhlukahluka kobude bamaqanda ngamanye (n ≥ 200 ngokudla ngakunye) kusetshenziswa i-Dialux-20 microscope (DM1000) ene-Etzlarst, Letzlar equitzlar equitzp; Ikhamera ye-Leica (DFC) 320 R2; I-Leica Microsystems Ltd., DE).Amaqanda asele agcinwe egumbini elilawulwa yisimo sezulu ngaphansi kwezimo ezijwayelekile zokukhulisa amahora angu-24, futhi isampula encane yezibungu zokuqala ezisanda kuvela (n ≥ 200 ngokudla ngakunye) zalinganiswa, njengoba kuchazwe ngenhla.Inani lamaqanda nosayizi wamaqanda nezibungu ziqhathaniswa phakathi kokwelashwa nokusebenzisa i-MP, i-Hockey's analysis v14.0.0, SAS Institute Inc.).
Kwaqoqwa amasampula ezinkomo zakwaZebu, izinhlanga ze-Arsi ukuze kube lula, amasampula omchamo aqoqwa ekuseni kakhulu ngesikhathi izinkomo zisesibayeni. Amasampula omchamo aqoqwa ku-2 ml ngayinye eya ku-010 ml ngayinye yathathwa esbayeni. izikhwama zokubhaka ze-polyamide (Toppits Cofresco, Frischhalteprodukte GmbH and Co., Minden, DE) ku-3 l polyamide enesivalo Emigqomo yepulasitiki ye-vinyl chloride. Ama-volatiles e-headspace avela kusampula ngayinye yomchamo wezinkomo aqoqwe ngokuqondile (fresh) noma ngemva kokuvuthwa endaweni yokushisa engu-164 h 72h, isampula ngayinye engu-72h ibimele iqembu lobudala ngalinye.
Eqoqweni le-headspace volatiles, isistimu ye-loop evaliwe yasetshenziselwa ukuzungeza umfudlana wegesi ehlungiwe nge-carbon eyenziwe yasebenza (100 ml min-1) ngesikhwama se-polyamide ukuya kukholomu ye-adsorption amahora angu-2.5 kusetshenziswa iphampu ye-diaphragm vacuum (KNF Neuberger, Freiburg, DE). yayenziwe ngeshubhu ye-Teflon (5.5 cm x 3 mm id) equkethe u-35 mg we-Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) phakathi kwamapulagi oboya bengilazi.Ngaphambi kokusetshenziswa, ikholomu yayigezwa ngo-1 ml odistilled kabusha we-n-hexane (Merck, Darmstadt 10 ml, GC 9, GC 9, GC Sigma Aldrich).Izimo ezishintshashintshayo ezikhangisiwe zakhishwa ngo-400 μl we-pentane.Ukuqoqwa kwe-Headspace kwahlanganiswa futhi kwagcinwa ku -20°C kuze kusetshenziselwe ukuhlaziya okwengeziwe.
Izimpendulo zokuziphatha zokufuna umsingathi nokudla igazi Izingcaphuno eziguquguqukayo ze-A.Headspace eziqoqwe kusukela kumchamo omusha, 24-h, 72-h, kanye no-168-h-h-ubudala zihlaziywe ukuze kutholwe okukhishwe okuguquguqukayo okuvela omiyane base-Arabidopsis kusetshenziswa ishubhu lengilazi eliqondile i-olfactometer [18]. umsebenzi wokufuna ikhaya.I-Arab [19].I-olfactometer yeshubhu lengilazi (80 cm × 9.5 cm id) yakhanyiswa ngo-3 ± 1 lx wokukhanya okubomvu okuvela phezulu.Ukugeleza komoya okuhlungiwe nokuswakama kwamalahle (25 ± 2 °C, 65 ± 2% umswakama ohlobene) kudlule uchungechunge lwe-bioassay engu-30 cm kudlule i-bioassay yensimbi engu-30 cm. izikrini zemesh, ezidala ukugeleza kwe-laminar kanye nesakhiwo se-plamu efanayo.Isikhiphi se-tampon samazinyo (4 cm × 1 cm; L:D; DAB Dental AB), simiswe ekhoyili engu-5 cm ekupheleni kwe-olfactometer evunguzayo, nezinguquko zesikhuthazi njalo ngemizuzu engu-5. Ukuze kuhlaziywe, 10 μl we-headspace ngayinye esetshenzisiwe i-diluted isamba esingu-1, i-diluted isamba esingu-1 esilinganisiwe ye-pentane yayisetshenziswa njengendlela yokulawula.Izinyosi zomuntu ngamunye ezifuna ukusingatha noma ukumunca igazi zifakwe emakhejini okukhululwa komuntu ngamunye amahora angu-2-3 ngaphambi kokuqala kokuhlolwa.Ikheji yokukhulula yafakwa ohlangothini lwe-downwind ye-olfactometer, futhi omiyane bavunyelwa ukuba bavumelane ne-1 min, bese i-butterfly valve ukuze ikhulule ukwelashwa kwe-caget i-protection ivuliwe. omiyane abahlangane nomthombo phakathi nemizuzu engu-5 yokukhishwa. I-headspace ngayinye ekhishwe futhi yokulawula eguquguqukayo yaphindwa okungenani izikhathi ezingu-30, futhi ukuze kugwenywe imiphumela yanoma yiluphi usuku olulodwa, inani elifanayo lokwelashwa nezilawuli zahlolwa ngosuku ngalunye lokuhlola.Funa izimpendulo ezivela kumsingathi kanye ne-Ans ephakelwa igazi.Isi-Arabhu siqhathanisa nesethi ye-headspace yahlaziywa ngokuhlelwa kabusha kwe-logisticario ye-comp (JMP Pro, v14.0.0, SAS Institute Inc.).
Impendulo yokuzala kwe-An.Okukhishwa kwe-headspace kumchamo wenkomo omusha nomdala kwahlaziywa kumakheji e-Bugdorm (30 cm × 30 cm × 30 cm; Isayensi ye-MegaView).Izinkomishi zepulasitiki (30 mL; Nolato Hertila) ezigcwaliswe ngama-mL angu-20 wamanzi acwebile ahlinzekela indawo yokuzala, ekhoneni elibhekene nengxenye engu-2 yekheji elibhekene ne-4 cm futhi zabekwa endaweni engaphansi kwe-2 yekheji. izinkomishi zalungiswa nge-10 μl ye-headspace ngayinye ekhishwe ku-1: 10 dilution. Inani elilinganayo le-pentane lasetshenziselwa ukulungisa inkomishi yokulawula.Izinkomishi zokwelapha nokulawula zashintshaniswa phakathi kokuhlolwa ngakunye ukuze kulawule imiphumela yesikhundla.Abesifazane abayishumi abaphakelwa igazi bakhululwa emakhejini okuhlola ku-ZT 9-11 futhi amaqanda enkomishini abalwa kamuva. amaqanda abekwe enkomishini yokwelapha – inani lamaqanda abekwe enkomishini yokulawula)/(inani eliphelele lamaqanda abekiwe).Ukwelashwa ngakunye kwaphindwa izikhathi eziyisi-8.
Ukutholwa kwephethini ye-gas chromatographic kanye ne-electron antenna (GC-EAD) ye-An.arabiensis yesifazane kwenziwa njengoba kuchazwe ngaphambilini [20]. Kafushane, izingcaphuno ezintsha ze-headspace eziguqukayo zahlukaniswa kusetshenziswa i-Agilent Technologies 6890 GC (Santa Clara, CA, US) efakwe ikholomu ye-HP-5 ikholomu 5 μd 5 × 30m × 5 mm ukujiya, i-Agilent Technologies). kanye nomchamo ogugayo.I-Hydrogen yasetshenziswa njengesigaba esihambahambayo ngesilinganiso sokugeleza komugqa okuyisilinganiso esingu-45 cm s-1.Isampula ngayinye (2 μl) yajovwa imizuzwana engu-30 ngemodi engahlukaniseki nezinga lokushisa lokungena elingu-225 °C. Izinga lokushisa likahhavini we-GC lahlelwa lisuka ku-35 °C (ukubamba amaminithi angu-3) ukuya ku-GC emizuzwini engu-300 ku-GC -100 °C iminithi. isihlukanisi samanzi angcolile, i-4 psi ye-nitrogen yengezwa futhi yahlukaniswa ngo-1:1 ku-Gerstel 3D/2 ivolumu ephansi yevolumu efile (Gerstel, Mülheim, DE) phakathi komtshina we-ionization yomlilo kanye ne-EAD.I-GC effluent capillary ye-EAD yadluliselwa ngomugqa wokudlulisa we-Gerstel ODP-2), olandelela kuhhavini engu-5 × × C, elandelela kuhhavini engu-5 × × C. lapho wawuxutshwe khona nomoya ocwengiwe nge-carbon, onomswakama (1.5 l min−1).I-antenna yayibekwe u-0.5 cm ukusuka lapho kuphuma khona ithubhu.Umiyane ngamunye ngamunye waba nekhophi eyodwa, kanti komiyane abafuna umninikhaya, kwenziwa okungenani okuphindwe kathathu kumasampula omchamo weminyaka ngayinye.
Ukuhlonzwa kwezinhlanganisela ze-bioactive ekuqoqweni kwe-headspace yomchamo omusha nomdala wezinkomo kusetshenziswa i-GC ehlanganisiwe ne-mass spectrometer (GC-MS; 6890 GC kanye no-5975 MS; Agilent Technologies) ukuze kutholwe izimpendulo zezimpondo ekuhlaziyweni kwe-GC-EAD, esebenza ngemodi ye-ionization yomthelela we-electron ku-70 GC-equipped. ikholomu ye-silica capillary ehlanganisiwe (ububanzi obungaphakathi obungu-60 m × 0.25 mm, ukujiya kwefilimu engu-0.25 μm) kusetshenziswa i-helium njengesigaba esihambahambayo esinesilinganiso sokugeleza somugqa esimaphakathi esingu-35 cm s-1.A 2 μl isampula yajovwa kusetshenziswa izilungiselelo ezifanayo zomjovo kanye nezinga lokushisa likahhavini njengokuhlaziywa kwe-GC-EAD okusekelwe kusisindo sazo kanye nesikhathi sokuhlonishwa kwe-Compounds kutholwe. kumtapo wolwazi wangokwezifiso kanye nomtapo wezincwadi we-NIST14 (Agilent).Izinhlanganisela ezikhonjiwe zaqinisekiswa ngokujova izindinganiso zangempela (Ifayela Lokwengeza 1: Ithebula S2).Ngokwesilinganiso, i-heptyl acetate (10 ng, 99.8% ukuhlanzeka kwamakhemikhali, i-Aldrich) yajovwa njengezinga langaphandle.
Ukuhlola ukusebenza kahle kwengxube yephunga lokwenziwa ehlanganisa izinhlanganisela ze-bioactive ezikhonjwe emchameni omusha nomdala ukuze kuhehe i-Ans.arabiensis efuna umninikhaya futhi emunca igazi, kusetshenziswa i-olfactometer nephrothokholi efanayo njengenhla.Izingxube zokwenziwa zilingise ukwakheka nezilinganiso zamakhompawundi ku-headspace exubile izingcaphuno eziguquguqukayo, amahora angama-24, amahora angama-24, nama-4-ho Umchamo omdala wamahora angu-168 (Umfanekiso 5D-G; Ifayela Lokwengeza 1: Ithebula S2). Ukuze uhlaziye, sebenzisa u-10 μl wengxube engu-1:100 yengxube yokwenziwa ngokuphelele, enezinga lokukhululwa elisuka cishe ku-140-2400 ng h-1, ukuze uhlole ukukhanga komiyane okwenziwa lapho ngemva kokuhlolwa komiyane, lapho ukuhlolwa kwegazi kuqedile. izingxube ezikhiphayo zenhlanganisela eyodwa yengxube ephelele ziyasuswa.Funa izimpendulo ezivela kumsingathi kanye nezingxube ze-Ans eziphakelwa igazi.Izingxube ze-Arab vs zokwenziwa nezikhiphayo zahlaziywa kusetshenziswa ukuhlehla okungokomfanekiso okulandelwa ukuqhathanisa ngakubili kwezilinganiso eziyinqaba (JMP Pro, v14.0.0, SAS Institute Inc.).
Ukuhlola ukuthi umchamo wenkomo ungasebenza yini njengendawo yokuhlala omiyane kamalaleveva, umchamo wenkomo omusha nomdala, oqoqwe njengoba kuchazwe ngenhla, kanye namanzi afakwa emabhakedeni anezikhala ama-3 l (100 ml) futhi afakwa ezicupheni zokuthiya. (inguqulo ye-BG-HDT; i-BioGents, i-Regensburg, i-DE). Izicupho eziyishumi zihlukaniswe ngamamitha angu-50 emadlelweni, amamitha angu-400 ukusuka emphakathini wasemaphandleni (i-Silay, i-Ethiopia, 5°53'24′′N, 37°29'24''E) futhi azikho izinkomo, ezindaweni zokuzalanisa unomphela nasezigodini. ijikeleziswa ubusuku nemini ingqikithi yobusuku obuhlanu.Izinombolo zomiyane ezibanjwe ezingibeni ezibanjwe umchamo weminyaka ehlukene ziqhathaniswe kusetshenziswa ukuhlehla kokuhleleka ngokusatshalaliswa kwe-beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
Esigodini esigcwele umalaleveva eduze kwedolobha laseMaki, esifundeni sase-Oromia, e-Ethiopia (8° 11′ 08″ N, 38° 81′ 70″ E; Umfanekiso 6A). Ucwaningo lwenziwa phakathi no-August namaphakathi no-September ngaphambi kokufuthwa kwensalela yaminyaka yonke yasendlini, kanye nenkathi yemvula ende. zikhethelwe ucwaningo (Fig. 6A) .Imibandela esetshenziselwa ukukhetha izindlu yilezi: azikho izilwane ezivunyelwe endlini, akukho ukuphekela endlini (ukudweba izinkuni noma amalahle) okuvunyelwe (okungenani ngesikhathi sesivivinyo), kanye nezindlu ezihlala abantu ababili, ezilala emithini yokubulala izinambuzane. ngaphansi kwenethi lomiyane elilashiwe.Imvume yezimiso zokuziphatha inikezwe Ibhodi Lokubuyekeza Izimiso Zokucwaninga Kwesikhungo (IRB/022/2016) ye-Faculty of Natural Sciences (CNS-IRB), i-Addis Ababa University, ngokuhambisana nemihlahlandlela esungulwe yi-World Medical Association Declaration of Helsinki.Imvume itholwe yisikhulu sezempilo sasekhaya ngasinye. abaphathi ezingeni lesifunda nesigceme ('kebele').Umklamo wokuhlola ulandele umklamo wesikwele sesiLatini esingu-2 × 2, lapho izingxube zokwenziwa nezilawuli zabelwa ezindlini ezibhanqiwe ngobusuku bokuqala futhi zashintshaniswa phakathi kwezindlu ngobusuku obulandelayo bokuhlola.Le nqubo yaphindwa izikhathi eziyishumi.Ukwengeza, ukulinganisa umsebenzi womiyane ezindlini ezikhethiwe, i-CDC iqhuba izicupho ezinhlanu zokuphela kwensimu ekuqaleni kwenkundla, i-CDC ibeka izicupho ezimaphakathi ebusuku. ngesikhathi esifanayo sosuku.
Ingxube yokwenziwa equkethe ama-bioactive compounds ayisithupha yachithwa ku-heptane (i-97.0% ye-solvent GC grade, i-Sigma Aldrich) futhi yakhululwa ngo-140 ng h-1 isebenzisa i-wick dispenser ye-cotton [20]. Centers for Disease Control and Prevention (CDC) light trap (John W. Hock Company, Gainesville, FL, US; Figure 6A) .Izicupho zazilengiswa ku-0.8 - 1 m ngaphezu komhlabathi, eduze nonyawo lombhede, futhi ivolontiya lalilala ngaphansi kwenethi lomiyane elingalashwa futhi liqhutshwa phakathi kuka-108: 00 no-moquid by 106to30. isimo sokuphila (okungondliwa, okuphakelwe, okukhulelwe, kanye nokukhulelwa [21] kamuva kwahlolwa kusetshenziswa ukuhlaziywa kwe-polymerase chain reaction (PCR) ukuze kuhlonzwe uhlobo lwe-morphologically oluhlonzwe njenge-A. gambiae sl. Amalungu e-complex [23]. control) kwaba umphumela olungisiwe (JMP® 14.0. 0. SAS Institute Inc.) Lapha, sibika u-χ2 kanye namavelu we-p kusuka ekuhlolweni kwesilinganiso sokungenzeka.
Linganisa ukuthi iphephile yini.arabiensis ikwazile ukuthola umchamo, umthombo wayo oyinhloko we-nitrogen, i-urea, ngokudla okuqondile, phakathi kwamahora angu-48 wokuphatha izinsuku ezingu-4 zokuthumela (i-dpe) ukuhlolwa kokufuna ukusingathwa nokuphakelwa igazi kwabesifazane (Fig. 1A) .Kokubili abesifazane abafuna ukusingatha nabamunca igazi bamunca kakhulu i-sucrose kunanoma iyiphi enye, i-F1 = 2, i-2, i-2, i-2 noma i-sucrose yamanzi. I-0.0001 kanye no-F (5,299) = 56.00, p <0.0001, ngokulandelanayo uma kuqhathaniswa nakho konke okunye ukugxila namanzi, kuyilapho kungabonakali kusuka ku-10% sucrose (F(10,813) = 15.72, p <0.0001; Umfanekiso 1D).Lokhu kwakuhluke kakhulu ekuphenduleni kwabesifazane abaphakelwa igazi, abavame ukumunca kakhulu ukudla okuqukethe i-urea kunamanzi, nakuba i-10 (7% ngaphansi kwe-sucrose = i-5% = i-5% engaphansi kakhulu 78.35, p <0.0001; Umfanekiso 1).1E).Ngaphezu kwalokho, lapho kuqhathaniswa phakathi kwezimo ezimbili zomzimba, abesifazane be-phlebotomized bamunca i-urea eningi kunabesifazane abafuna ukusingatha ezindaweni eziphansi kakhulu, futhi laba besifazane bamunca amanani afanayo e-urea ekugxilweni okuphezulu (F(1,980; 80.2)= 1F, G).Ngenkathi ukudla okuqukethe i-urea kubonakala kunamanani afanele (Fig. 1D,E), abesifazane kuzo zombili izifunda zomzimba bakwazile ukulinganisa inani le-urea elimuncwe kulo lonke uhla lokugxiliswa kwe-urea ngendlela yelogi (Fig. 1F,G). ).Ngokufanayo, omiyane babonakala belawula ukumuncwa kwabo kwe-nitrogen ngokulawula inani lomchamo elimuncwa, njengoba inani le-nitrogen emchameni libonakala enanini elimuncwa (Umfanekiso 1B, C no-B).
Ukuhlola imiphumela yomchamo kanye ne-urea ekusindeni komiyane abafuna umninikhaya nabamunca igazi, abesifazane baphakelwa umchamo wayo yonke iminyaka emine (fresh, 24 h, 72 h, and 168 h post-deposition) kanye nohlu lokugxilwa kwe-urea, kanye namanzi acwengekile kanye no-10 % wokuhlaziywa kokudla okusetshenziswe njenge-sucrose yokulawula. umphumela obalulekile ekusindeni okuphelele kwabesifazane abafuna ukusingatha (umchamo: χ2 = 108.5, df = 5, p <0.0001; urea: χ2 = 122.8, df = 5, p <0.0001; Umdwebo 2B, C) kanye nomchamo 2, fd 9 = abesifazane = χd 0. 5, p <0.0001; urea: χ2 = 137.9, df = 5, p <0.0001 Umfanekiso 2D,E) .Kukho konke ukuhlola, abesifazane baphakela ukudla komchamo, urea, namanzi babe namazinga okusinda aphansi kakhulu uma kuqhathaniswa nabesifazane abaphakela i-sucrose Host staeking (Figus-female-B) ukudla okusha. ibonise amazinga okuphila ahlukene, nalabo abaphakelwa umchamo omdala we-72-h (p = 0.016) babe namathuba aphansi okusinda (Fig. 2B). Ngaphezu kwalokho, abesifazane abafuna ukusingatha abaphakelwe i-urea engu-135 mM baphile isikhathi eside kunezilawuli zamanzi (p <0.04) (Uma kuqhathaniswa ne-2C ne-feur water and fresh water). umchamo uphile isikhathi eside (p = 0.001 kanye no-p = 0.012, ngokulandelana; Umfanekiso 2D), kuyilapho abesifazane abondliwe ngomchamo wamahora angu-72 baphila isikhathi eside kunalabo abaphakelwa Umchamo omfushane wesifazane kanye nomchamo wamahora angu-24 ubudala (p <0.0001 kanye no-p = 0.013, uma u-fere 2, u-3, ngokulandelana); abesifazane abondliwe ngegazi baphila isikhathi eside kunakho konke okunye ukugxila kwe-urea namanzi (p <0.013; Umfanekiso 2E).
Ukusinda kwama-Anopheles arabini ezinsikazi ancela igazi ezidla umchamo wenkomo kanye ne-urea.Kucwaningo lwe-bioassay (A), omiyane besifazane banikezwe ukudla okuhlanganisa umchamo wenkomo omusha nomdala, i-urea ehlukahlukene, i-sucrose (10%) kanye namanzi acwecwe (H2O). kuqoshwa njalo emahoreni ayi-12 kuze kube yilapho bonke abesifazane abadla umchamo (B, D) kanye ne-urea (C, E), nezilawuli, i-Sucrose namanzi, sezifile.
Ibanga eliphelele kanye nenani lemijikelezo enqunywe ekuhlolweni komshini wendiza esikhathini esingamahora angu-24 kwahluka phakathi komiyane abafuna umsingathi nabamunca igazi, okubonisa umsebenzi omncane wokundiza jikelele (Fig. 3) .Omiyane abafuna ukusingatha oyo abanikeza umchamo omusha noma osekhulile noma i-sucrose namanzi babonise amaphethini okundiza ahlukile (Fig. 3), kanye nezinsikazi ezi-2 ezisanda kudla ngesikhathi sokuchama okusasha - ukudla okusanhlamvu okusanhlamvu okusanhlamvu okusanhlamvu. Omiyane besifazane ababenikeza i-sucrose noma umchamo wamahora angu-72 babonisa umsebenzi phakathi namahora angu-24, kuyilapho abesifazane ababenikeza amanzi babematasa kakhulu phakathi nenkathi. Umchamo weminyaka engama-72 ube nokwehla okuqhubekayo komsebenzi emahoreni angama-24 (Umfanekiso 3).
Ekuhlolweni komshini wokundiza, omiyane besifazane badla umchamo wenkomo omusha nomdala, i-urea ehlukahlukene, i-sucrose (10%), namanzi acwecwe (H2O) aboshelwe endaweni evundlile, ezulazula ngokuzungeza igazi. (kwesokudla) abesifazane, ingqikithi yebanga nenani lezindiza ngehora lokudla ngakunye esikhathini esingamahora angama-24 kwarekhodwa (okumnyama: okumpunga; ukukhanya: okumhlophe).Ibanga elimaphakathi nenani elimaphakathi lokushaywa kuboniswa kwesokudla segrafu yomsebenzi we-circadian.Amabha wamaphutha amelela iphutha elijwayelekile lencazelo.Ukuhlaziywa kwezibalo bheka umbhalo.
Ngokuvamile, umsebenzi wendiza jikelele wabesifazane abafuna ukusingatha ulandele iphethini efana nebanga lendiza phakathi nesikhathi samahora angu-24. Ibanga lendiza elimaphakathi lalithinteke kakhulu ngokudla okungeniswayo (F(5, 138) = 28.27, p <0.0001), futhi abesifazane abafuna ukusingatha badle amahora angu-72 omchamo wandizela ngokuphawulekayo amanye amabanga amade futhi i-0 <0 kuqhathaniswa nawo wonke amabanga amade0 (p) omiyane abadla i-sucrose bandize isikhathi eside kunokusha (p = 0.022) kanye nomiyane abaphakelwa umchamo oneminyaka engu-24-h (p = 0.022). Ngokuphambene nephethini yomsebenzi wokundiza echazwe ukudla komchamo, abesifazane abafuna i-urea-feed bafuna i-urea babonise umsebenzi wendiza ophikelelayo phakathi nenkathi ye-24-h phakathi nenkathi emnyama. 3).Nakuba amaphethini omsebenzi ayefana, abesifazane abafuna umsingathi baphakele i-urea bakhuphule kakhulu ibanga lendiza elimaphakathi kuye ngokugxilisa okumuncwe (F(5, 138) = 1310.91, p <0.0001).Abesifazane abafuna ukusingatha indawo baphakele noma iyiphi i-urea indize isikhathi eside kunesifazane ephakelwa amanzi noma i-sucrose (p 3). <0.0
Umsebenzi wokundiza usuwonke womiyane abamunca igazi wawuzinzile futhi waqhubeka amahora angu-24 kukho konke ukudla, nokwanda komsebenzi womchamo phakathi nengxenye yesibili yenkathi emnyama kwabesifazane abaphakelwa ngamanzi kanye nakwabesifazane abaphakelwa abasha namahora angu-24 ubudala (isithombe 3). Nakuba ukudla komchamo kuthinte kakhulu ibanga lendiza kwabesifazane abaphakelwa igazi (F.8, 5, 4 = 1). Ukudla kwe-urea akuzange (F(5, 138) = 1.36, p = 0.24) .ngolunye umchamo kanye nokudla okulawulayo (okusha, p = 0.0091; amahora angu-72, p = 0.0022; amahora angu-168, p = 0.001; sucrose, p = 0.H0020, p = 0.H017; d).
Imiphumela yokudla komchamo kanye ne-urea emingceleni yokuzala yahlolwa ekuhlolweni kwe-bioassay yokubeka amaqanda (Umfanekiso 4A) futhi yaphenywa ngokwenani lamaqanda abekwe yinsikazi ngayinye, usayizi weqanda, kanye nezibungu zokuqala ezisanda kuchanyuselwa. Umfanekiso we-4B) .Abesifazane badla umchamo wamahora angu-24, ukudla kwegazi kwabekela amaqanda amaningi kakhulu kunabesifazane abadla okunye ukudla komchamo futhi babefana nalabo abaphakelwa i-sucrose (Fig. 4B) .Ngokufanayo, ubukhulu bamaqanda abekwe abesifazane abadla umchamo bahlukahluka ngokudla (F (5, 209) = 10 , p <4, 209, 12, 209 = 12.8 no-4, p. abesifazane abadla i-sucrose ababeka amaqanda amakhulu kakhulu kunezinsikazi eziphakelwa amanzi, kuyilapho amaqanda ezinsikazi ezondliwe nge-168 h yomchamo ayemancane kakhulu (Fig. 4C) .Ngaphezu kwalokho, ukudla komchamo kwathinta kakhulu usayizi wezibungu (F (5, 187) = 7.86, p <0.0001), namaqanda ama-4 avela ku-laid e-2 ngokuphawulekayo. Ezinsikazi ezincela umchamo amahora angu-72 kunamaqanda azalelwe ezibunguni.Izinsikazi eziphakelwa amanzi namahora angu-168 (Umfanekiso 4D).
Ukusebenza kokuzala kwe-Anopheles arabinis yesifazane idla umchamo wenkomo kanye ne-urea.Omiyane besifazane abaphakelwa igazi baphakelwa ukudla okuhlanganisa umchamo wenkomo omusha nomdala, ukugxila okuhlukahlukene kwe-urea, i-sucrose (10%), namanzi acwecwe (H2O) amahora angu-48 ngaphambi kokufakwa kuma-bioassays kanye nokuthola inombolo yeqanda (Amahora angu-4), iqanda leqanda (Amahora angu-4), iqanda eliphansi usayizi weqanda (C, F) kanye nosayizi wezibungu (D, G) bathikamezeka kakhulu ngokudla okunikeziwe (umchamo wenkomo: BD; urea: EG).Izindlela zepharamitha ngayinye elinganiswa kusetshenziswa amagama ezinhlamvu ezihlukene zazihluke kakhulu kwenye (indlela eyodwa i-ANOVA isebenzisa ukuhlaziywa kwe-Tukey's post hoc; p <0.05).Imigoqo yamaphutha imelela iphutha elijwayelekile lencazelo yencazelo.
Njengengxenye enkulu ye-nitrogenous yomchamo, i-urea, uma inikezwa njengokudla kwabesifazane abaphakelwe igazi, yathinta kakhulu imingcele yokuzala kuzo zonke izifundo.Inani lamaqanda abekwe abesifazane abaphakelwa i-urea, ngemva kokudla kwegazi, kuye ngokugxila kwe-urea (F(11, 360) = 4.69; p <0.0001), ukugxila kwabesifazane phakathi kwe-3Mµ4 m3m urea ephakathi kwe-3Mµ urea. zabekela amaqanda engeziwe (Umfanekiso 4E).Abesifazane abadla ngokugxiliswa kwe-urea engu-134 µM noma ngaphezulu bazalela amaqanda amakhulu kunabesifazane abaphakelwa emanzini (F(10, 4245) = 36.7; p <0.0001; Umfanekiso 4F), nosayizi wezibungu, nakuba kuthinteke ukugxila okufanayo, okungu-3F = 3F; p <0.0001) yayiguquguquka kakhulu (Fig. 4G).
Ukukhanga sekukonke ku-host-seaking bovine umchamo headspace extracts volatile.I-arabiensis ehlolwe ku-glass tube i-olfactometer (Fig. 5A) yathinteka kakhulu iminyaka yomchamo (χ2 = 15.9, df = 4, p = 0.0032; Umdwebo 5B) .Ukuhlaziywa kwe-stale hoc kuqhathaniswa kakhulu namahora we-24 womchamo wabonisa ukuthi izinga eliphezulu le-24 lomchamo liqhathanise kakhulu kuzo zonke ezinye izindlela zokwelapha (amahora angu-72: p = 0.0060, amahora angu-168: p = 0.012, pentane: p = 0.00070), Ngaphandle kwephunga lomchamo omusha (p = 0.13; Umfanekiso 5B) .Nakuba ukukhanga okuphelele komiyane abamunca igazi kwakungeyona into ehlukile, χ8 = iphunga elihlukile lomchamo 8. p = 0.067; Umfanekiso 5C), laba besifazane batholakale bekhanga kakhulu ku-headspace okukhishwa okuguquguqukayo uma kuqhathaniswa nomchamo omdala wamahora angu-72 uma kuqhathaniswa nezilawuli (p = 0.0066; Umfanekiso 5C).
Izimpendulo zokuziphatha kuphunga lomchamo wenkomo wemvelo nowokwenziwa ekufuneni umsingathi kanye negazi lika-Anopheles arabianus.I-Schematic of the glass tube olfactometer (A).Ukuheha ama-headspace ama-volatile extracts avela kumchamo wenkomo omusha nomdala ozobamba (B) nomiyane abamunca igazi (C).Thola i-Lord extract of the tentacle fresh. Ihora elingu-24 (E), ihora elingu-72 (F), kanye nomchamo wenkomo omdala onamahora angu-168 (G) kuyaboniswa.Ukulandelela kokutholwa kwe-antenna ye-electron (EAD) kubonisa izinguquko zamandla kagesi ekuphenduleni izinhlanganisela ze-bioactive ku-headspace ekhishwe ku-chromatograph yegesi futhi itholwe ngomtshina we-ionization we-flame (i-FID ye-Represent Represents) I-FID yesikhathi sokuphendula. (s) isilinganiso somiyane abakhangwa amagama ezinhlamvu ezihlukene sasihluke kakhulu kwenye (i-ANOVA yendlela eyodwa isebenzisa ukuhlaziywa kwe-post hoc ka-Tukey; p <0.05).Amabha wamaphutha amelela iphutha elijwayelekile lesikali
U-Ann.arabiensis wesifazane, i-72 h kanye ne-120 h ngemva kokudla kwegazi, ngesikhathi sokuzala, akukho okuthandwayo okubonisiwe ekukhishweni kwe-headspace okuguquguqukayo okuvela kumchamo wenkomo omusha nomdala uma kuqhathaniswa nezilawuli ze-pentane (χ2 = 3.07, p > 0.05; Ifayela elengeziwe 1: Fig. S1).
Ku-Ann.arabiensis wesifazane, ukuhlaziya kwe-GC-EAD kanye ne-GC-MS kuhlonzwe izinhlanganisela ze-bioactive eziyisishiyagalombili, eziyisithupha, ezintathu nezintathu (Figure 5D-G) .Nakuba umehluko enanini lezinhlanganisela eziveza izimpendulo ze-electrophysiological zabonwa, iningi lalezi zinhlanganisela zazikhona ku-headspace ngayinye ekhishwayo eguquguqukayo eqoqwe kusukela kumchamo omusha kanye nomdala okhiqiza impendulo ngayinye yomchamo wesifazane, ngakho-ke, kuphela, izimpondo ezingaphezulu komkhawulo zifakiwe ekuhlaziyweni okwengeziwe.
Inani eliphelele lokukhululwa kwezinhlanganisela ze-bioactive eqoqweni le-headspace lenyuke lisuka ku-29 µg h-1 kumchamo omusha laya ku-242 µg h-1 kumchamo omdala wamahora angu-168, ikakhulukazi ngenxa yokunyuka kwe-p-cresol ne-m-formaldehyde Phenol kanye ne-phenol. i-decanal, yehla ngokukhula kweminyaka yomchamo, okuhlotshaniswa nokwehla okubonwayo kwe-signal intensity (ubuningi) ku-chromatogram (Fig. 5D) -G iphaneli yesokunxele) kanye nezimpendulo ze-physiological kulezi zinhlanganisela (Fig. 5D-G iphaneli yesokudla).
Sekukonke, ingxube yokwenziwa yayinesilinganiso esifanayo semvelo sezinhlanganisela ze-bioactive ezikhonjwe ekukhishweni okuguquguqukayo kwezikhala zekhanda zomchamo omusha nezindala (I-Fig. 5D-G) futhi ayizange ibonakale iveza isikhalazo esibalulekile ekufuneni umsingathi (χ2 = 8.15, df = 4, p = 0.083; Fig. 4tossucking, 9H) noma i-Fig. df = 4, p = 0.30; Fig. 5I) .Nokho, ukuqhathaniswa kwe-post hoc pairwise phakathi kwemithi yokwelapha kubonise ukuthi omiyane abafuna i-host bekhanga kakhulu kwingxube yokwenziwa yomchamo we-24-h uma kuqhathaniswa nezilawuli ze-pentane (p = 0.0086; Umfanekiso 5H).
Ukuhlola indima yezingxenye ngazinye ezixubeni zokwenziwa zomchamo oneminyaka engu-24-h, izingxube eziyisithupha ezikhiphayo zahlolwa ngokumelene nezingxube eziphelele ku-Y-tube assay, lapho izinhlanganisela ngazinye zakhishwa khona.Ngomiyane abafuna umninikhaya, ukususa izinhlanganisela ngazinye kusukela kunhlanganisela ephelele kwaba nomthelela omkhulu ekuphenduleni kokuziphatha (χ63 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = = I-0.0032; Ifayela elingeziwe le-1: Umfanekiso we-S2A), zonke izingxube ezikhiphayo zazithandeka kakhulu kunezincane kunezixube ngokugcwele.Ngokuphambene, ukususwa kwezinhlanganisela zomuntu ngamunye kusuka engxubeni yokwenziwa ngokugcwele akuzange kuthinte izimpendulo zokuziphatha zomiyane abamunca igazi (χ2 = 11.38, df = 6, p = 0.07 ingxube ephansi) uma kuqhathaniswa nomphumela we-0.07 ophelele Ukukhanga (p = 0.022; Ifayela Elingeziwe 1: Umfanekiso S2B).
Esigodini esigcwele umalaleveva e-Ethiopia, ukusebenza kahle kwengxube yokwenziwa yomchamo wenkomo wamahora angu-24 ekuheheni omiyane ngaphansi kwezimo zensimu kwahlolwa ubusuku obuyishumi (Fig. 6A) .Ingqikithi yomiyane abangu-4,861 babanjwa futhi bahlonzwa, lapho u-45.7% wabo kwakuyi-Anthropus.gambias 18. Ama-35.4% ayeyi-Culex spp.(Ifayela elengeziwe 1: Ithebula S1).I-Anopheles arabinis ukuphela kwelungu le-An.Gambian yezinhlobo zezinhlobo ezihlonzwe ukuhlaziywa kwe-PCR. Ngokwesilinganiso, omiyane abangu-320 babanjwa ngobusuku ngabunye, ngesikhathi lapho izicupho ezinengxubevange yokwenziwa zibamba omiyane abaningi kunengxubevange engu-0 χ9 (312) 170.0, p <0.0001) .Izicupho ezingabanjwanga zabekwa ngobusuku obuhlanu bokulawula ekuqaleni, maphakathi, nasekupheleni kwecala.Izinombolo ezifanayo zomiyane zabanjwa ku-pair of trap, okubonisa ukuthi akukho ukuchema phakathi kwezindlu (χ2 (0, 1665) > 1 nokuncipha kwesibalo sabantu abangu-5 × 0-1 × 0 0 9 × 1. isikhathi sokufunda.Uma kuqhathaniswa nezicupho zokulawula, inani lomiyane ababanjwe ezicupheni eziqukethe inhlanganisela yokwenziwa landa kakhulu: ukufuna i-host (χ2 (0, 2107) = 138.7, p <0.0001), ukondla igazi kwakamuva (χ2 (0, 650) = 32.2, p <0 20) nokukhulelwa (0 = χ20) (0 = 0 20) (0 = 0 20) (0 = χ20) (0). 6.27, p = 0.0123; Ifayela elengeziwe 1: Ithebula S1).Lokhu kubonakala futhi enanini eliphelele lomiyane abathunjiwe: umsingathi ofuna > ukumunca igazi > okhulelwe > okhulelwe kancane > owesilisa.
Ukuhlolwa kwenkundla ukusebenza kahle kwengxube yephunga lomchamo wenkomo yamahora angama-24. Izivivinyo zasensimini zenziwa eningizimu-maphakathi ne-Ethiopia (imephu), eduze nedolobha laseMaki (faka), kusetshenziswa i-Centers for Disease Control (CDC) isicupho esilula (kwesokudla) ezindlini ezibhanqiwe, ezinomklamo wesikwele wesiLatini (isithombe esisemoyeni se-CDC esithwebula isithombe) kanye nesithombe se-CDC esine-photosyps esiheha i-CDC ye-photosyps ehehayo i-Ador-traspheles ye-Anotrasthetic ye-CDC). i-arabesques (B), kodwa hhayi i-Anopheles farroes (C), ngendlela ehlukile, umphumela oncike esimweni sokuphila. Ngaphezu kwalokho, lezi zicupho zibambe amanani enyuke kakhulu omiyane abaphethe i-Culex.(D) Uma kuqhathaniswa nokulawula. kwesokudla simele inkomba yokukhetha emaphakathi ngamapheya ezicupho zokulawula (evuliwe; N = 5). ).Izinkanyezi zibonisa amazinga okubaluleka ezibalo (*p = 0.01 kanye nokuthi ***p <0.0001)
Lezi zinhlobo ezintathu zibanjwe ngokuhlukile ezicupheni eziqukethe izingxube zokwenziwa.Kubheka umsingathi (χ2(1, 1345) = 71.7, p <0.0001), ukuphakelwa kwegazi (χ2(1, 517) = 16.7, p <0.0001) nokukhulelwa (χ2 (1, 1, 6 = 18) = 18. I-.arabiensis yayibanjwe ogibeni olukhulula ingxube yokwenziwa (I-Fig. 6B), kuyilapho inani le-An alizange lihluke.I-Pharoensis ezifundeni ezihlukahlukene ze-physiological yatholakala (Fig. 6C) .Ku-Culex, ukwanda okuphawulekayo kuphela kwenani lomiyane abafuna ababungazi kwatholakala ezicupheni ezibanjwe nge-synthetic χ26 (1) = χ26, ingxube ye-synthetic χ26, i-1 = 1 = χ261, = (p. 0.0004; Umfanekiso we-6D), uma kuqhathaniswa nezicupho zokulawula.
Izicupho ezitholakala ngaphandle kwalabo abangase babambe iqhaza phakathi kwezindawo ezizalela kuzo nemiphakathi yasemaphandleni e-Ethiopia zasetshenziswa ukuze kuhlolwe ukuthi omiyane kamalaleveva basebenzisa iphunga lomchamo wenkomo njengendawo yokuhlala. Uma kungekho zimpawu zokusingatha, ukushisa, kanye nokuba khona noma ngaphandle kwephunga lomchamo wenkomo, abekho omiyane ababanjiwe (Ifayela elengeziwe 1: Kungakhathaliseki ukuthi izinga lokushisa lomchamo wesifazane liphezulu kangakanani, inkomokazi, iphunga le-Sw3). omiyane bakhangwa futhi babanjwa, nakuba ngamanani amancane, azimele kuminyaka yomchamo (χ2 (5, 25) = 2.29, p = 0.13; Ifayela elengeziwe 1: Umfanekiso S3). Ngokuphambene, ukulawulwa kwamanzi akuzange kubambe omiyane be-malaria emazingeni okushisa aphezulu (Ifayela Lokwengeza 1: Umfanekiso S3).
Omiyane bemalariya bathola futhi basabalalise izinhlanganisela eziqukethe i-nitrogen ngokudla okunxeshezelwayo ngomchamo wenkomo (okungukuthi, ama-puddles) ukuze kuthuthukiswe izici zomlando wokuphila, ezifana nezinye izinambuzane [2, 4, 24, 25, 26]. emakhaya asemaphandleni nasezindaweni zokuzalela.Omiyane besifazane bathola lesi sisetshenziswa ngokuhogela futhi bayakwazi ukulawula ukumuncwa kwezakhi ze-nitrogen kumchamo, kuhlanganise ne-urea, ingxenye enkulu ye-nitrogen kumchamo [15, 16]. omiyane, kanye nezici zokusinda nezici zokuzala zabantu abondliwe ngegazi ngesikhathi somjikelezo wokuqala we-gonadotropic.Ngakho-ke, ukuxuba umchamo kudlala indima ebalulekile yokudla okunomsoco kuma-vectors kamalaleveva avaliwe njengabantu abadala abangondlekile [8], njengoba kunikeza omiyane besifazane amandla okuthola izinhlanganisela ezibalulekile ze-nitrogen ngokuhlanganyela ekutholeni okubalulekile kokuphakelayo kwe-epiological yokuphila kwabo okuyingozi kakhulu. isikhathi okulindelekile, umsebenzi kanye nokuphuma kokuzala, konke okuthinta umthamo we-vector.Ngaphezu kwalokho, lokhu kuziphatha kungase kube okuhlosiwe kwezinhlelo zokuphatha i-vector zesikhathi esizayo.