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 iyingculazi, ngakho-ke idinga ukukhiqizwa kabusha kwe-malnouri. hlola 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 ubudala, kanye nokufuna umninikhaya nokuphakelwa igazi (ukudla kwamahora angu-48 ngemva kokuphuma kwegazi) abesifazane balinganiswa nge-olfactometer ye-Y-tube, futhi abesifazane abakhulelwe bahlola amakhemikhali asebenzayo futhi kwahlolwa i-electrophysical test. w umchamo kuwo wonke amakilasi eminyaka emine.Izingxube zokwenziwa zezingxube ze-bioactive zahlolwa ku-Y-tube nasezivivinyweni zasensimini.Ukuphenya umchamo wenkomo kanye ne-urea eyinhlanganisela eyinhloko ene-nitrogen njengokudla okungase kube ukudla okuhambisana nezifo zikamalaleveva, imingcele yokudla kanye nezici zomlando wokuphila kwalinganiswa.Ingxenye yomiyane besifazane kanye nenani lomchamo wesifazane owamunca umchamo wenkomokazi owasetshenziselwe umchamo. i-vival, indiza eboshiwe, nokukhiqizwa kabusha.
Funa igazi kanye nokondleka komninikhaya.Ezifundweni zaselabhorethri nezasensimini, ama-Arabhu akhangwa iphunga lemvelo nelokwenziwa lomchamo wenkomo omusha nomdala.Abesifazane abakhulelwe babengenandaba nokusabela komchamo wezinkomo ezindaweni zokuzala.Abesifazane abafuna ukusingatha nabamunca igazi bamunca umchamo wenkomo kanye ne-urea ngenkuthalo futhi bahlukanisele lezi zinsiza njengezindawo zokudayisa ukuze baphile 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 ukudla 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 kwemisoco kuncike enqubweni yomlando wokuphila futhi kungase kuholele ezidingweni ezihlukene zekhwalithi yokudla kanye nobuningi bezinga lokudla okuhlukene1, ukuntula ngokwehlukana kwezigaba zokuphila. izakhi ezithile, 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 zevemvane nebhu zichazwa ngokuyinhloko, izimbobo zokunisela nazo zenzeka kwezinye izilokazane zezinambuzane, futhi ukukhangwa kwezilwane ezifuywayo kanye nezilwane ezifuywayo, lezi zinhlobo ze-5 zezempilo zingaba nemiphumela ephawulekayo yokuphila. , 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 komiyane besifazane asebekhulile i-Anopheles kulinganiselwe ngenxa yokuqukethwe kwekhalori ephansi okuvela esigabeni sezibungu kanye nokusetshenziswa ngendlela engafanele kwesidlo segazi [9].I-Female Ann.gambiae sl ngokuvamile inxephezela lokhu ngokufaka ukudla okungeziwe kwegazi [10, 11], ngaleyo ndlela kubeka abantu abaningi engcupheni yokungenwa yizifo ezithathelwanayo. I-es ingasebenzisa ukuphakelwa okungeziwe kwendle yezilwane ezinomgogodla ukuze zithole izakhi ze-nitrogenous ezithuthukisa ukuzivumelanisa nokuhamba kwendiza, njengoba kuboniswa ezinye izinambuzane [2].Kulokhu, ukukhanga okunamandla nokuhlukile kolunye lwezinhlobo eziyizelamani ngaphakathi kwe-An.I-Gambian sl species complex, i-Anopheles arabinis, inkomo esanda kuzalwa nendala iyathakazelisa3, i-opheles, i-opheles,12 umchamo wezinkomo uyisisetshenziswa esicebile nge-nitrogenous compounds, ne-urea ehlanganisa u-50-95% wengqikithi ye-nitrogen emchamweni omusha [15, 16].Njengoba iminyaka yomchamo wenkomo, ama-microorganisms asebenzisa lezi zinsiza ukunciphisa inkimbinkimbi ye-nitrogenous compounds ekhuphuka ngokushesha phakathi kwamahora we-nitrogen, anda i-nitrogen ngokushesha ngaphakathi kwe-24 am. ama-alkalophilic microorganisms (amaningi awo akhiqiza izinhlanganisela ezinobuthi omiyane) ayachuma [15], okungenzeka ukuthi u-Ann.arabiensis wesifazane ukhangwa kakhulu umchamo oneminyaka engama-24 noma ngaphansi [13, 14].
Kulolu cwaningo, kwabhekwa ama-Ans asokhaya kanye nama-Ans adla igazi. Phakathi nomjikelezo wayo wokuqala we-gonadotropin, i-arabiensis yahlolelwa ukutholwa kwezinhlanganisela ze-nitrogenous, okuhlanganisa i-urea, ngokuxuba umchamo. Okulandelayo, uchungechunge lokuhlola lwenziwa ukuze kuhlolwe ukuthi omiyane besifazane bawaba kanjani lesi sisetshenziswa esingaba somsoco ukuze kuthuthukiswe ukuzala kabusha nokuzala kabusha kwe-cooval. Umchamo wahlolwa ukuze kutholakale ukuthi ingabe lezi zinikeza izinkomba ezinokwethenjelwa zomsingathi futhi ondliwa ngegazi u-An. Ekusesheni kwabo lesi sisetshenziswa esingaba somsoco, i-arabiensis yathola ukuhlobana kwamakhemikhali ngemva kokukhanga okubonwayo. ezimweni ezihlukahlukene ze-physiological.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) ayegcinwe ku-25 ± 2 °C, 65 ± 5% RH kanye nomjikelezo wokukhanya okungamahora angu-12:12:ubumnyama. izinkomishi (Nolato Hertila, Åstorp, SE) bese zidluliselwa kumakheji e-Bugdorm (30 cm × 30 cm × 30 cm; I-MegaView Science, Taichung, Taiwan) ukuze kuvunyelwe ukuvela kwabantu abadala.Abadala banikezwe i-10% sucrose solution ad libitum kuze kube yizinsuku ezi-4 lapho i-host-emergence inikezwa khona ngaphambi kokuhlolwa kosuku, noma ukuhlolwa kwabesifazane ngaphambi kwesikhathi. amanzi athulisiwe ngaphambi kokuhlolwa, njengoba kuchazwe ngezansi.Abesifazane abasetshenziselwa ukuhlolwa kwe-flight tube babulawa indlala amahora angu-4-6 kuphela nge-ad libitum yamanzi.Ukulungiselela omiyane abamunca igazi ukuze kuhlolwe i-bioassays elandelayo, abesifazane abangu-4 be-dpe banikezwe igazi lezimvu ze-defibrotic (Håtunalab, Bro, SE) kusetshenziswa i-membrane feeding system, Discovered female feeding system (Hescormonge) I-Workshop ye-Workshop i-Accrymonge i-Workshop feeding system (Heyington cod. obomvu kumakheji ngamanye futhi unikeze ukudla ngokuqondile, njengoba kuchazwe ngezansi, noma i-10% ye-sucrose ad libitum yezinsuku ezingu-3 ngaphambi kokuhlolwa okuchazwe ngezansi.Abesifazane bokugcina basetshenziselwa ama-bioassays e-flight tube futhi badluliselwa elabhorethri, bese beba ne-distilled water ad libitum amahora angu-4-6 ngaphambi kokuhlolwa.
Ukuhlolwa kokuphakelayo kwasetshenziswa ukuze kulinganiswe ukusetshenziswa komchamo ne-urea kumuntu wesifazane omdala ongum-A.Arab.Abesifazane abafuna ukusingatha futhi abaphakelwa igazi banikezwe ukudla okuqukethe umchamo wenkomo ohlanjululiwe ongu-1% omusha nomdala, ukugxila okuhlukahlukene kwe-urea, kanye nezilawuli ezimbili (10% sucrose namanzi) amahora angu-48. Ngaphezu kwalokho, umbala wokudla (1 mgne61-CASani 1; I-gma-Aldrich, Stockholm, SE) yengezwe ekudleni futhi yanikezwa nge-4 × 4 matrix kumashubhu e-microcentrifuge angu-250 µl (Axygen Scientific, Union City, CA, US; Umfanekiso 1A) Gcwalisa ekugcineni (~300 µl). Ukuze ugweme ukuncintisana phakathi kwezilwane ezifuywayo ezinomiyane ku-0 umbala womiyane (isilwane esifuywayo) 12 cm ububanzi no-6 cm ukuphakama, i-Semadeni, i-Ostermundigen, i-CH; Umfanekiso 1A) ebumnyameni obuphelele ku-25 ± 2 cm °C kanye nomswakama ohlobene no-65 ± 5%.
Bheka umchamo wezinkomo kanye ne-urea emuncwa isilwane esincelisayo esincela igazi futhi esimunca igazi u-Anopheles arabianus. Esivivinyweni sokuphakela (A), omiyane besifazane banikezwe ukudla okuhlanganisa umchamo wenkomo omusha nomdala, i-urea ehlukahlukene, i-sucrose (10%), namanzi acwecwe (H2O). abesifazane bamunca umchamo wenkomo wamahora angu-72 ngaphansi komchamo wenkomo wamahora angu-168 (B).Inani eliphelele lokuqukethwe kwe-nitrogen (± ukuchezuka okujwayelekile) lomchamo limelelwa ku-inset.Abesifazane abafuna ukusingathwa (D, F) nabamunca igazi (E, G) bathatha i-urea ngendlela encike kumthamo.Inani eliphelele lokuqukethwe kwe-nitrogen (± ukuchezuka okujwayelekile) lomchamo limelelwa ku-inset.Abesifazane abafuna ukusingathwa (D, F) nabamunca igazi (E, G) bathatha i-urea ngendlela encike kumthamo.IMean inhaled ehlukene nge-postkey amagama ahlukene kakhulu ne-postkey' ngayinye ukuhlaziywa kwe-hoc; p <0.05).Amabha wephutha amelela iphutha elijwayelekile lencazelo (BE).Umugqa wedeshi eqondile umelela umugqa wokuhlehla womugqa welogi (F, G)
Ukuze kukhululwe ukudla okumuncwe, omiyane babekwe ngabanye 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 . ma-Aldrich) kanye nokumunca (λ620) kwanqunywa kusetshenziswa isifundi se-microplate esisekelwe ku-spectrophotometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm).Okunye, omiyane bagaywe ngo-1 ml wamanzi acwebile, u-900 µl wawo adluliselwe ku-spectrometer ye-UV60m1 ye-UVrometric, i-dzurometric engu-0pho8 ye-UV602; , Kista, SE).Ukuze kulinganiswe 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. Bese, ukuminyana kokubona kokugxiliswa kukadayi okwaziwayo kwasetshenziswa ukuze kunqunywe inani lokudla okumunyisiwe yi-mosquito ngayinye.
Idatha yevolumu yahlaziywa kusetshenziswa ukuhlaziya kwendlela eyodwa yokuhluka (ANOVA) kwalandelwa ukuqhathanisa kokubili kuka-Tukey okuthunyelwe kokubili (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). I-v8.0.0 ye-Mac, i-GraphPad Software, i-San Diego, i-CA, i-US).
Cishe u-20 µl wamasampuli omchamo asuka eqenjini ngalinye lobudala ayeboshelwe ku-Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) futhi afakwa kumakhepsuli kathayela (8 mm × 5 mm). Amakhepsule afakwe egunjini elivuthayo le-analsh000000000000Fish, i-analsh, i-Fish, i-Fish, i-Fish, i-ChNS ltham, MA, US) ukuze kunqunywe okuqukethwe kwe-nitrogen emchameni 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 kwe-4 1% yomchamo omusha kanye nama-dipe amabili amasha kanye nama-dipe amabili ahlanganisiwe. izilawuli, u-10% we-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 kwesitsha se-petri esihlanjululwe ngenhla (isibalo 1) Chaza ukudla kwakho okuchazwe ngenhla. ama-es ayebalwa kabili ngosuku, kuyilapho omiyane abafile balahlwa kuze kube yilapho umiyane wokugcina ufa (n = 40 ngokwelashwa) .Ukusinda komiyane abadla ukudla okuhlukahlukene kwahlaziywa ngezibalo kusetshenziswa ama-curve okusinda e-Kaplan-Meyer kanye nokuhlolwa kwe-log-rank ukuze kuqhathaniswe ukuqhathaniswa kokusabalalisa kokusinda phakathi kwe-IBM20SS Statistics.
Isigayo esindizayo sikamiyane esisekelwe ku-Attisano et al.[17], esakhiwe ngamaphaneli e-acrylic acacile angu-5 mm (ububanzi obungu-10 cm x 10 cm ubude x 10 cm ukuphakama) ngaphandle kwamaphaneli angaphambili nangemuva (Fig. 3: top). d 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 abaphakelwe ngokugcwele babe sebebulawa izinzwa eqhweni ngamaminithi angu-2-3 futhi banamathisele izikhonkwane zezinambuzane ngengcina yezinyosi (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) base berekhodwa izingalo ze-Mill Revolution. ihlelwe isiloga sedatha esakhelwe ngokwezifiso, bese sigcinwa futhi siboniswe kusetshenziswa isofthiwe ye-PC-Lab 2000™ (v4.01; Velleman, Gavere, BE).Imishini yokugaya indiza yafakwa egumbini elilawulwa isimo sezulu (12 h:12 h, ukukhanya: kumnyama, 25 ± 2 °C, 65 ± 5% RH).
Ukuze ubone ngeso lengqondo iphethini yomsebenzi wendiza, isamba sebanga elindisiwe (m) kanye nenani eliphelele lemisebenzi yendiza elandelanayo ibalwe ngehora esikhathini esingamahora angu-24. Ngaphezu kwalokho, amabanga amaphakathi ahanjiswa abesifazane ngabanye aqhathaniswe kuzo zonke izindlela zokwelapha futhi ahlaziywa kusetshenziswa indlela eyodwa yokuhlaziya i-ANOVA kanye ne-Tukey's post hoc (JMP Pro, v14.0.0), i-avareji ye-Addient Institute ibhekwa njenge-advantage Inc. , inani elimaphakathi lemizuliswano libalwa ngokunyuka kwamaminithi angu-10.
Ukuhlola umthelela wokudla ekusebenzeni kokuzala kwe-An.arabiensis, abesifazane abayisithupha (4 dpe) badluliselwa 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 amahora angu-4 e-Dilato Hertila nge-8 ngosuku, i-Nostilled ye-0 yesithathu yanikezwa amahora angu-2 shintsha njalo emahoreni angu-24.Phinda uhlobo lokudla ngalunye izikhathi ezingu-20-50.Amaqanda abalwa futhi aqoshwa ekhejini ngalinye lokuhlola.Amasampula amaqanda asetshenziswa ukuze kuhlolwe usayizi osho nokuhlukahluka kobude bamaqanda ngamanye (n ≥ 200 ngokudla ngakunye) kusetshenziswa isibonakhulu se-Dialux-20 (DM1000; Wetzlarst aquitzD) I-Letzlarst (i-Ernstlar aitzDFC) LequitzD FC Lequitz Leitzlar 320 R2;I-Leica Microsystems Ltd., DE).Amaqanda asele agcinwe ekamelweni elilawulwa yisimo sezulu ngaphansi kwezimo ezijwayelekile zokukhulisa amahora angu-24, futhi isampula encane yezibungu ezisanda kuvela ezisanda kuvela (n ≥ 200 ngokudla ngakunye) zalinganiswa, njengoba kuchazwe ngenhla.Inani lamaqanda nobukhulu bamaqanda nezibungu zaqhathaniswa phakathi kokwelashwa nokusebenzisa i-PANOVA eyodwa, i-v. SAS Institute Inc.).
Kwaqoqwa amasampula ezinkomo ze-Zebu, izinhlanga ze-Arsi ukuze kube lula, amasampula omchamo aqoqwa ekuseni kakhulu ngesikhathi izinkomo zisesibayeni. Amasampula omchamo aqoqwe kumasampula ezinkomo zeZebu, izinhlanga ze-Arsi ukuze kube lula, amasampula omchamo aqoqwa ekuseni ngesikhathi izinkomo zisesibayeni. izikhwama (Toppits Cofresco, Frischhalteprodukte GmbH and Co., Minden, DE) in 3 l polyamide with lid In vinyl chloride plastic drums.Ama-headspace volatiles avela kusampula ngayinye yomchamo wezinkomo aqoqwe ngokuqondile (fresh) noma ngemva kokuvuthwa ekamelweni lokushisa amahora angu-24 h, isampula yomchamo eyi-72 ngayinye yayiyi-72 yeqembu 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). n ishubhu (5.5 cm x 3 mm id) equkethe u-35 mg we-Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) phakathi kwamapulagi ovolo wengilazi.Ngaphambi kokusetshenziswa, ikholomu yayihlanjululwe ngo-1 ml redistilled n-hexane (Merck, Darmstadt, DE) kanye ne-1.9 lgd GC puree 1.9 mld GC ukuntengantenga kombhede kwakhishwa 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, we-24-h, 72-h, kanye no-168-h-h-ubudala kwahlaziywa ukuze kutholwe okukhishwe okuguquguqukayo omiyane base-Arabidopsis kusetshenziswa ishubhu lengilazi eliqondile i-olfactometer [18]. [19].I-olfactometer yeshubhu lengilazi (i-id engu-80 cm × 9.5 cm) yakhanyiswa nge-3 ± 1 lx yokukhanya okubomvu okuvela phezulu.Ukugeleza komoya okuhlungiwe namalahle okumswakama (25 ± 2 °C, 65 ± 2% umswakama ohlobene) kudlule i-bioassay ku-30 ​​cm i-lamina yensimbi futhi idlula uchungechunge lwe-steel ye-30 cm. isakhiwo se-plume esilinganayo.Isikhiphi se-tampon yamazinyo (4 cm × 1 cm; L:D; DAB Dental AB), simisiwe kukhoyili engu-5 cm ekugcineni kwe-olfactometer, esishintshashintsha isikhuthazi njalo ngemizuzu emi-5. Ukuze kuhlaziywe, u-10 μl wokukhishwa kwe-headspace ngayinye, uhlanjululwe 1:10, i-stimulus control of the host. noma omiyane abamunca igazi bafakwa emakhejini okukhulula ngawodwana amahora angu-2-3 ngaphambi kokuqala kokuhlolwa.Ikheji yokukhulula yabekwa ohlangothini lomoya ophansi we-olfactometer, futhi omiyane bavunyelwa ukuba bajwayelane neminithi elingu-1, kwase kuvulelwa ivalve uvemvane wekheji ukuze kukhululwe.Ukukhangwa ekwelashweni noma ekulawuleni kwahlaziywa lapho ukukhishwa kwe-moquiesation kwemizuzu5 kwahlaziywa ukukhishwa kwe-moquies. ukukhishwa nokulawula okuguquguqukayo kwe-headspace kwaphindaphindwa okungenani izikhathi ezingu-30, futhi ukuze kugwenywe imiphumela yanoma yiluphi usuku olulodwa, inani elifanayo lokwelashwa nezilawuli zahlolwa ngosuku ngalunye lokuhlola.Izimpendulo zokufuna kumsingathi kanye namasethi e-Ans.Arabic ngokumelene ne-headspace anikezwe igazi ahlaziywa kusetshenziswa ukuhlehla kwezinto ezilandelwayo okulandelwa ukuqhathaniswa okubili kwe-odd.JMP0 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 acwecwe ahlinzekela i-substrate yokuzalanisa, i-substrate engaphansi engu-4 ebekwe eceleni kwenkomishi engu-4 cm futhi yafakwa enye inkomishi yekheji. I-10 μl yokukhishwa kwe-headspace ngayinye nge-dilution ye-1: 10. Inani elilinganayo le-pentane lasetshenziselwa ukulungisa indebe yokulawula.Izinkomishi zokwelashwa nokulawula zashintshaniswa phakathi kokuhlolwa ngakunye ukuze kulawuleke imiphumela yesimo.Izinsikazi eziyishumi ezondliwa ngegazi zikhishwe emakhejini okuhlola ku-ZT 9-11 futhi amaqanda ezinkomishini abalwa emahoreni angu-24 we-spawn inkomba - inombolo ye-spawn inkomba kamuva. amaqanda abekwe enkomishini yokulawula)/(inani eliphelele lamaqanda abekiwe).Ukwelashwa ngakunye kwaphindwa izikhathi eziyisi-8.
Ukuhlaziywa kwephethini ye-gas chromatographic ne-electron antenna (GC-EAD) kwe-An.arabiensis yesifazane kwenziwa njengoba kuchazwe ngaphambilini [20]. Kafushane, izingcaphuno ezintsha ze-headspace eziguquguqukayo zahlukaniswa kusetshenziswa i-Agilent Technologies 6890 GC (Santa Clara, CA, US) efakwe ikholomu ye-HP-5, ifilimu ye-id 5 × 30m × 30m Tech. ama-chnologies).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-3000 min. , i-4 psi ye-nitrogen yengezwe futhi yahlukaniswa ngo-1:1 ku-Gerstel 3D/2 isiphambano sevolumu ephansi efile (Gerstel, Mülheim, DE) phakathi komtshina we-ionization yomlilo kanye ne-EAD.I-GC effluent capillary ye-EAD yadluliswa ngomugqa wokudlulisa we-Gerstel ODP-2, olandelela i-oven ye-GC, i-oven ye-GC, izinga lokushisa elixutshwe nengilazi engu-8 × 8 mm, ishubhu le-carbon elixutshwe ngu-8 × 8 mm umoya omanzi (1.5 l min−1).I-antenna yayibekwe u-0.5 cm ukusuka lapho kuphuma khona ishubhu.Umiyane ngamunye ubalwa ngokuphindaphinda okukodwa, kanti komiyane abafuna umninikhaya, kwenziwa okungenani iziphindaphindo ezintathu 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-electron impact ionization ku-70 GC-casipped ye-Silica-epipped ye-HP. Ikholomu ye-llary (60 m × 0.25 mm ububanzi bangaphakathi, 0.25 μm ukujiya kwefilimu) kusetshenziswa i-helium njengesigaba esihambahambayo esinesilinganiso sokugeleza somugqa esimaphakathi esingu-35 cm s-1.A 2 μl isampula yajovwa kusetshenziswa izilungiselelo ezifanayo zomjovo nezinga lokushisa likahhavini njengokuhlaziywa kwe-GC-EAD.Izinhlanganisela zikhonjwe ngokususelwe kumtapo wezincwadi we-NISTvten futhi waqhathaniswa nesikhathi selabhulali ye-NISTvten futhi yaqhathaniswa nenkomba ye-NISTvten yangokwezifiso (Agilent).Izinhlanganisela ezikhonjiwe ziqinisekiswe ngokujova izindinganiso eziyiqiniso (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 kuhehwe i-Ans.arabiensis efuna umninikhaya futhi emunca igazi, kusetshenziswa i-olfactometer nephrothokholi efanayo engenhla.Izingxube zokwenziwa zilingise ukwakheka nezilinganiso zamakhompiyutha kusikhala esixubile esikhishwayo esishisayo, esingamahora angu-6, 24hourho, amahora angu-24 no-24-hourho umchamo omdala (Umdwebo 5D-G; Ifayela Lokwengeza 1: Ithebula S2). Ukuze uhlaziye, sebenzisa u-10 μl we-dilution engu-1:100 yengxube eyenziwe ngokuphelele, enesilinganiso sokukhulula esisuka cishe ku-140-2400 ng h-1, ukuze uhlole ukukhanga kokusingatha kanye nokumunca igazi, izingxube zomiyane eziphelele, okwenziwa ngemva kokuhlolwa komiyane okuncane. ingxube iyasuswa.Izimpendulo zokufuna ezisuka kumsingathi kanye nezingxube ze-Ans eziphakelwa ngegazi.Izingxube ze-Arab vs zokwenziwa nezisusayo zahlaziywa kusetshenziswa ukuhlehla kwezinto 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, DE).Izicupho eziyishumi zihlukaniswe ngamamitha angu-50 emadlelweni, amamitha angu-400 ukusuka emphakathini wendawo (Silay, Ethiopia, 5°53'24′′N, 37°29′24′′E) futhi azikho izinkomo, ezindaweni zokuzalanisa unomphela nasezigodini. ubusuku obuhlanu buphelele.Izinombolo zomiyane ezibanjwe ezingibeni ezibanjwe umchamo wobudala obuhlukene ziqhathaniswe kusetshenziswa ukuhlehla kwezinto ezinokusabalalisa 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. Izingxenye ezinhlanu zocwaningo zazikhethelwe amapheya amahlanu esigodini (amapheya angama-20) atholakala endaweni yocwaningo. (Fig. 6A).Imibandela eyayisetshenziswa ekukhetheni izindlu kwakuyilezi: azikho izilwane ezivunyelwe endlini, akukho ukuphekela endlini (ukudweba izinkuni noma amalahle) okuvunyelwe (okungenani ngesikhathi sokuvivinywa), kanye nezindlu ezihlala abantu ababili, ezilala emithini yokubulala izinambuzane.ngaphansi kwenethi lomiyane elilashiwe.Imvume yezimiso zokuziphatha inikezwe Ibhodi Lokubuyekeza Izimiso Zokucwaninga Ngesikhungo (IRB/022/2016) ye-Faculty of Natural Sciences (CNS-IRB), i-Addis Ababa University, ngokuhambisana neziqondiso ezisungulwe yi-World Medical Association Declaration of Helsinki.Imvume itholwe kubaphathi bendawo yonke inhloko yezempilo yendawo ngayinye. kanye nezinga lesigceme ('kebele').Umklamo wokuhlola wawulandela umklamo wesikwele wesiLatini we-2 × 2, lapho izingxube zokwenziwa kanye nezilawuli zabelwa ezindlini ezibhanqiwe ngobusuku bokuqala futhi zashintshaniswa phakathi kwezindlu ngobusuku obulandelayo bokuhlola.Le nqubo yaphindwa izikhathi eziyishumi.Ukwengeza, ukulinganisa umsebenzi womiyane ezindlini ezikhethiwe, izicupho ze-CDC zazibekwe ukuze ziphele phakathi nosuku oluyisihlanu olulandelanayo ekuqaleni kosuku olufanayo lwesivivinyo.
Ingxube yokwenziwa equkethe ama-bioactive compounds ayisithupha yachithwa ku-heptane (i-97.0% ye-solvent GC grade, i-Sigma Aldrich) futhi yakhululwa ku-140 ng h-1 isebenzisa i-wick dispenser ye-cotton [20]. I-Prevention (CDC) light trap (John W. Hock Company, Gainesville, FL, US; Figure 6A). Izicupho zazilengiswe ku-0.8 - 1 m ngaphezu komhlaba, eduze nonyawo lombhede, futhi ivolontiya lalilala ngaphansi kwenethi lomiyane elalingelashiwe futhi lahlinzwa phakathi kuka-18:00 no-06:00 kanye no-06:00. abakhulelwe, nabakhulelwe [21] kamuva bahlolwa kusetshenziswa ukuhlaziywa kwe-polymerase chain reaction (PCR) ukuze kuhlonzwe uhlobo lwe-morphologically oluhlonzwe njenge-A. gambiae sl. Amalungu e-complex [23]. Ocwaningweni lwasensimini, ukucupha ugibe lwezindlu ezibhanqiwe kwahlaziywa kusetshenziswa imodeli ye-logistic fit ye-nominal, lapho ukukhanga kwakuwumphumela wokulawula okuguquguqukayo (J0 nokulawulwa okuguquguqukayo kwe-J0 MP).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 (dpe) ukuhlolwa kokufuna ukusingathwa nokuphakelwa igazi kwabesifazane (Fig. 1A).Kokubili abesifazane abafuna ukusingatha nabamunca igazi bamunca ngokuphawulekayo i-sucrose eningi kunanoma iyiphi enye i-sucrose (F10 = 20, 40 = 20, 2, 2, 2, 2, 2, 2, 2, 2, 20) noma i-sucrose. I-01 kanye ne-F (5,299) = 56.00, p <0.0001, ngokulandelana; Umdwebo 1B, C) .Ngaphezu kwalokho, abesifazane abafuna ukusingatha badla kancane emchameni emahoreni angu-72 uma kuqhathaniswa nomchamo emahoreni angu-168 (Fig. 1B) .Lapho kunikezwa ukudla okuqukethe i-urea, i-host-search iqhathaniswa ne-2 ye-concentration ye-2, i-host-seance i-concentration ye-6 iqhathaniswa ne-mrea ye-2 ngokuphawulekayo. okungabonakali ku-10% sucrose (F(10,813) = 15.72, p <0.0001; Umfanekiso 1D).Lokhu kwakungafani nempendulo yabesifazane abaphakelwa igazi, abavame ukumunca kakhulu ukudla okuqukethe i-urea kunamanzi, nakuba kungaphansi kakhulu kuka-10% we-sucrose, 70 = 510; 10% 0, 5 51; 51; 10% 0,51; i-gure 1).1E).Ngaphezu kwalokho, lapho kuqhathaniswa phakathi kwezimo ezimbili ze-physiological, abesifazane be-phlebotomized bamunca i-urea eningi kunabesifazane abafuna ukusingatha ezindaweni eziphansi kakhulu, futhi laba besifazane badonsa amanani afanayo e-urea ekugxilweni okuphezulu (F(1,953)= 78.82, p <0.0001;Fig. 1F, G).Ngenkathi ukudla okuqukethe i-urea kubonakala kunamanani alungile (Fig. 1D,E), abesifazane kuzo zombili izifunda zomzimba bakwazile ukulinganisa inani le-urea elimuncwe kulo lonke uhla lokugxila 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 lokugxiliswa kwe-urea, kanye namanzi acwengekile kanye no-10 % wokuhlaziywa kokudla okunomphumela obalulekile wokulawula i-2 (i-Aval 2 ibonise umphumela obalulekile wokulawula). ukusinda kwabesifazane abafuna ukusingatha (umchamo: χ2 = 108.5, df = 5, p <0.0001; urea: χ2 = 122.8, df = 5, p <0.0001; Fig. 2B, C) kanye nabesifazane abondliwe ngegazi (2,0 = f 9 = 0,0 = 1, 0 = 0. ; urea: χ2 = 137.9, df = 5, p <0.0001; Umfanekiso 2D,E).Kukho konke ukuhlola, abesifazane bondle ukudla komchamo, urea, namanzi babe namazinga okusinda aphansi kakhulu uma kuqhathaniswa nabesifazane abaphakela ukudla okune-sucrose (Umfanekiso 2B-E). -h umchamo omdala (p = 0.016) onamathuba aphansi okusinda (Fig. 2B) .Ngaphezu kwalokho, abesifazane abafuna ukusingatha abaphakela i-urea engu-135 mM baphile isikhathi eside kunezilawuli zamanzi (p <0.04) (Fig.2C) .Uma kuqhathaniswa namanzi, abesifazane abondliwe ngomchamo omusha kanye nomchamo wamahora angu-24 baphila isikhathi eside (p = 0.001 kanye p = 0.012, ngokulandelana; Umfanekiso 2D), kuyilapho abesifazane abondliwe ngomchamo wamahora angu-72 baphila isikhathi eside kunalabo abaphakelwa Umchamo omfushane wesifazane kanye ne-24-ihora le-0 <0 = 0 p. gure 2D).Lapho kudliswa i-urea engu-135 mM, abesifazane abondliwe ngegazi baphila isikhathi eside kunawo wonke amanye amaqoqo e-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). amahora kuze kube yilapho bonke abesifazane abadla umchamo (B, D) kanye ne-urea (C, E), nezilawuli, i-Sucrose namanzi, zifile.
Ibanga eliphelele kanye nenani lemijikelezo enqunywe ekuhlolweni komshini wendiza esikhathini samahora angu-24 kwahluka phakathi komiyane abafuna umninikhaya nabamunca igazi, okubonisa umsebenzi omncane wokundiza jikelele (Fig. 3) .Omiyane abafuna ukusingathwa abahlinzeka ngomchamo omusha noma omdala noma i-sucrose namanzi babonise amaphethini endiza ahlukene (Fig. 3), kanye ne-dint 6 esebenzayo ngesikhathi sokuchama kwe-2 , kanye ne-2 dawn fesquid lapho i-24 idla umchamo omusha, kanye ne-24 yabesifazane abasha abadlayo. Omiyane besifazane ababenikeza i-sucrose noma umchamo wamahora angu-72 babonisa umsebenzi phakathi namahora angu-24, kuyilapho abesifazane ababenikeza amanzi babematasa kakhulu phakathi nesikhathi esimaphakathi. ngibe nokwehla okuqhubekayo komsebenzi emahoreni angama-24 (Umfanekiso 3).
Ekuhlolweni komshini wokundiza, omiyane besifazane badla umchamo wenkomo omusha nosebekhulile, i-urea ehlukahlukene, i-sucrose (10%), namanzi acwengiwe (H2O) aboshelwe endaweni evundlile, ezulazula kwesokudla (ukuzulazula kwesokudla) kwabesifazane (ukuzulazula kwesokudla). ingqikithi yebanga nenani lezindiza ngehora lokudla ngakunye esikhathini esingamahora angama-24 zarekhodwa (okumnyama: okumpunga; ukukhanya: okumhlophe).Ibanga elimaphakathi nesilinganiso senani lokuhamba kuboniswa kwesokudla segrafu yomsebenzi we-circadian.Imigoqo yamaphutha imelela iphutha elivamile lencazelo.Ukuhlaziywa kwezibalo bheka umbhalo
Ngokuvamile, umsebenzi wokundiza uwonke wabesifazane abafuna ukusingatha ulandele iphethini efana nebanga lendiza phakathi nenkathi yamahora 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-moquid ye-0000 yonke i-moquid ye-0 kuqhathaniswa nawo wonke amabanga amade. I-es indize isikhathi eside kunokusha (p = 0.022) kanye nomchamo we-24-h-aged (p = 0.022) -omiyane abondliwe. Ngokuphambene nephethini yomsebenzi wendiza echazwe ngokudla komchamo, abesifazane abafuna i-urea-fed bafuna i-ureaugh babonise umsebenzi wendiza ophikelelayo phakathi nenkathi ye-24-h, ukukhuphuka kwamaphethini wesigaba sesibili se-fig, i-fig ye-fig ye-female ye-3 yesigaba esifanayo. i-rea ikhulise kakhulu ibanga lendiza elimaphakathi kuye ngokugxilisa okumuncwe (F(5, 138) = 1310.91, p <0.0001).Abesifazane abafuna ukusingatha le midlalo baphakele noma yikuphi ukuhlangana kwe-urea endizayo isikhathi eside kunesifazane ephakelwa amanzi noma i-sucrose (p <0.03).
Umsebenzi wokundiza uwonke womiyane abamunca igazi wawuzinzile futhi ugcinwe ngaphezu kwamahora angu-24 kuzo zonke izidlo, nokwanda komsebenzi womchamo phakathi nengxenye yesibili yenkathi emnyama kwabesifazane abaphakelwa emanzini kanye nakwabesifazane abaphakelwa abasha namahora angu-24 ubudala (isithombe 3) .Ngenkathi ukudla komchamo kuthinteke kakhulu ibanga lendiza kubantu besifazane abaphakelwa igazi (F.8, 0 = 4, 0 = 30 = 40 = I-3 urea = 3, 0 = 4. (F(5, 138) = 1.36, p = 0.24) .nomunye umchamo kanye nokudla okulawulayo (okusha, p = 0.0091; amahora angu-72, p = 0.0022; amahora angu-168, p = 0.001; i-sucrose, p = 0.0017; dH6 = 0.3, p.
Imiphumela yokudla komchamo kanye ne-urea emingceleni yokuzala yahlolwa ekuhlolweni kwe-bioassays yokubeka amaqanda (Umfanekiso 4A) futhi yaphenywa ngokwenani lamaqanda abekwe owesifazane ngamunye, usayizi weqanda, kanye nezibungu zokuqala ezisanda kuchanyuselwa. B) .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 bahluka ngokudla (F (5, 209) = 12.85, p0-2010 0-209, p0-10, p0-10, p0-209, p0-209, p0-10. amaqanda amakhulu kakhulu kunezinsikazi eziphakelwa amanzi , kuyilapho amaqanda ezinsikazi adla nge-168 h yomchamo ayemancane kakhulu (Fig. 4C) .Ngaphezu kwalokho, ukudla komchamo kwathinta kakhulu usayizi wezibungu (F (5, 187) = 7.86, p <0.0001), enezibungu ezinkulu kakhulu eziphuma kumaqanda ama-24 abesifazane abaphuma ku-urine-feed laid kusuka kumaqanda ama-24 namaqanda e-urine laid. kusukela emaqandeni izibungu.Izinsikazi eziphakelwa amanzi kanye namahora angu-168 ezondleka ngomchamo (Umfanekiso 4D).
Ukusebenza kokuzala kwe-Anopheles arabinis yesifazane idla umchamo wenkomo kanye ne-urea.Omiyane besifazane abadla igazi baphakelwa ukudla okuhlanganisa umchamo wenkomo omusha nomdala, ukugxila okuhlukahlukene kwe-urea, i-sucrose (10%), namanzi acwengekile (H2O) amahora angama-48 ngaphambi kokufakwa kuma-bioassays amaqanda kanye nokuthola inombolo yeqanda (Amahora angu-4), iqanda leqanda (Amahora angu-4). , F) kanye nosayizi wezibungu (D, G) bathinteke kakhulu ngokudla okunikeziwe (umchamo wenkomo: BD; urea: EG).Izindlela zepharamitha ngayinye elinganiswa kusetshenziswa amagama ezinhlamvu ezihlukene zazihluke kakhulu komunye nomunye (i-ANOVA yendlela eyodwa isebenzisa ukuhlaziywa kwe-Tukey's post hoc; p <0.05).Amabha amaphutha amelela iphutha elijwayelekile lencazelo yencazelo.
Njengengxenye enkulu ye-nitrogenous yomchamo, i-urea, uma inikezwa njengokudla kwabesifazane abaphakelwa igazi, ithinte kakhulu imingcele yokuzala kuzo zonke izifundo.Inani lamaqanda abekwe abesifazane abaphakelwa i-urea, ngemva kokudla kwegazi, kuye ngokuthi i-urea igxiliwe (F(11, 360) = 4.69; p <0.0001), amaqanda okugxilwa kwe-fed 4M 3M ngaphezulu kwe-3M3 urea. Umfanekiso 4E).Abesifazane abasuthiswa nge-urea yokugxila engu-134 µM noma ngaphezulu bazalela amaqanda amakhudlwana kunabesifazane abaphakelwa emanzini (F(10, 4245) = 36.7; p <0.0001; Umfanekiso 4F), nosayizi wezibungu, nakuba kuthinteke ukugxiliswa okufanayo kwe-urea komama, 0 3(0) = 3 (F.19) = 3(19) kwaba variable more (Fig. 4G).
Ukukhangwa okuphelele ekufuneni i-hospine umchamo wezinkomo izingcaphuno eziguquguqukayo.I-arabiensis ehlolwe ku-glass tube olfactometer (Fig. 5A) yathinteka kakhulu iminyaka yomchamo (χ2 = 15.9, df = 4, p = 0.0032; Umfanekiso (Amahora angu-72: p = 0.0060, amahora angu-168: p = 0.012, i-pentane: p = 0.00070), Ngaphandle kwephunga lomchamo omusha (p = 0.13; Umfanekiso 5B).Nakuba ukukhanga okuphelele komiyane abamunca igazi ephunga lomchamo kwakungeyona into ehluke kakhulu (7 = 8, p. g. 5C), laba besifazane batholakale bekhanga kakhulu ku-headspace okukhishwe 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 kumchamo wenkomo omusha nomdala ozobamba (B) nokumunca igazi (C) omiyane.Thola4Isikhala se-Lord esikhipha i-Lord 2014 i-isoncleur entsha. (E), amahora angu-72 (F), kanye nomchamo wenkomo omdala wamahora angu-168 (G) kuyaboniswa. Ukulandelelwa 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 yomlilo (FID).Ibha yesikali imelela izici zokukhishwa kwe-remVg (ukukhishwa kwe-mVg). U-h-1) wezinhlanganisela zezinto eziphilayo uyaboniswa.Inkanyezi eyodwa (*) ibonisa impendulo ye-amplitude ephansi engaguquki.Izinkanyezi ezimbili (**) zibonisa izimpendulo ezingenakuphindaphindeka.Thola umsingathi (H) kanye nokumunca igazi (I) I-Arabiensis inokukhanga okuhlukile emixubeni yokwenziwa yezinhlamvu ezihlukene zomchamo wezinhlamvu ezigugile kakhulu. ihluke komunye nomunye (i-ANOVA yendlela eyodwa isebenzisa ukuhlaziywa kwe-Tukey's post hoc;p <0.05).Amabha wephutha 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 (Umfanekiso we-5D-G) .Nakuba umehluko enanini lamakhompiyutha aveza izimpendulo ze-electrophysiological wabonwa, iningi lalawa makhompiyutha lalikhona ku-headspace ngayinye ekhishwayo eguquguqukayo eqoqwe kusuka kumchamo omusha nomdala okhiqiza impendulo yomchamo wesifazane kuphela, ngakho-ke, i-anten ngayinye ikhiqiza impendulo yomchamo wesifazane. umkhawulo wafakwa ekuhlaziyweni okwengeziwe.
Inani eliphelele lokukhululwa kwezinhlanganisela ze-bioactive ekuqoqweni kwe-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. ubudala bomchamo, obuhlotshaniswa nokuncipha okuphawuliwe kokuqina kwesignali (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 (Fig. 5D-G) futhi ayizange ibonakale iveza isikhalazo esibalulekile ekusesheni umsingathi (χ2 = 8.15, df = 4, p = 0.083; Fig. 4tosd = 2 H) noma i-fig. 4, p = 0.30; Fig. 5I).Nokho, ukuqhathaniswa kwe-post hoc pairwise phakathi kokwelashwa kubonise ukuthi omiyane abafuna ukusingatha babethandeka kakhulu engxubeni yokwenziwa yomchamo omdala we-24-h uma kuqhathaniswa nezilawuli ze-pentane (p = 0.0086; Umfanekiso 5H).
Ukuhlola indima yezingxenye zomuntu ngamunye ezixubeni zokwenziwa zomchamo we-24-h-aguge, 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 (χ3 p 60 = ifayela le-1 = 1 = 60 = 1 = 1 = 1 = 1 = 60 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 1 = 0 = 1 = 0 = 1 = 1 = 1 = 6 ; I-1: Umfanekiso we-S2A), zonke izingxube ezikhiphayo zazikhanga kakhulu kuneziNcane kunokuxutshwa ngokugcwele.Ngokuphambene, ukususwa kwezinhlanganisela zomuntu ngamunye kusukela engxubeni yokwenziwa ngokuphelele akuzange kuthinte izimpendulo zokuziphatha komiyane abamunca igazi (χ2 = 11.38, df = 6, p = 0.077), ngaphandle kwengxube ehlanganisiwe ye-2 = Ifayela le-2 eliphansi, lapho kuqhathaniswa ne-2 Ifayela eliphansi. : Umfanekiso S2B).
Esigodini esigcwele umalaleveva e-Ethiopia, ukusebenza kwenhlanganisela 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 3% i-Anthropus. I-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 ezinengxube yokwenziwa zibambe omiyane abaningi kunezicupho ezibhanqiwe ngaphandle kwengxube, 0120 = 0120 = 0120. .Izicupho ezingezona izicupho zabekwa ngobusuku obuhlanu bokulawula obuyisihlanu ekuqaleni, phakathi, nasekupheleni kwecala.Izinombolo ezifanayo zomiyane zabanjwa kubhangqa ngalinye lezicupho, okubonisa ukuthi akukho ukwenzelela phakathi kwezindlu (χ2 (0, 1665) = 9 × 10-13, p > 0.0Compared to the period of the traps in the study of traps. izicupho eziqukethe ingxube yokwenziwa zanda kakhulu: ukufuna umsingathi (χ2(0, 2107) = 138.7, p <0.0001), ukuphakelwa kwegazi kwakamuva (χ2(0, 650) = 32.2, p <0.0001) nokukhulelwa (χ2(0, 227) = 2.0 = 2.2;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 zenkundla zenziwa eningizimu-maphakathi ne-Ethiopia (imephu), eduze nedolobha laseMaki (faka), kusetshenziswa i-Centers for Disease Control (CDC) i-trap light (kwesokudla) ezindlini ezibhanqiwe, ezinomklamo wesikwele wesiLatini (isithombe esisemoyeni se-CDC) (isithombe se-Aerial photo) se-Ador-Bpheleips ye-Anodorbes eheha i-CDC ye-Anordish ekhanga i-CDC yesithombe se-Anordish ekhangayo ye-Anordis-Baphelei . kodwa hhayi u-Anopheles farroes (C), ngendlela ehlukile, umphumela wokuziphilisa oncike esimweni sokuphila. Ngaphezu kwalokho, lezi zicupho zithwebule amanani anda kakhulu omiyane abasingethe i-Culex.(D) Uma kuqhathaniswa nokulawula.Amabha angakwesokunxele amelela inkomba yokukhetha emaphakathi yomiyane ababanjwe ngamapheya esiyelo esinuka kamnandi (green) kanye nokulawula (i-avareji yokukhetha i-bars = i-bars) imele (i-avareji yokukhetha) lawula izicupho (ezivulekile; N = 5).).Izinkanyezi zibonisa amazinga okubaluleka ezibalo (*p = 0.01 kanye nokuthi ***p <0.0001)
Lezi zinhlobo ezintathu zibanjwe ngokuhlukile ezicupheni eziqukethe izingxube zokwenziwa.Ifuna umsingathi (χ2(1, 1345) = 71.7, p <0.0001), ukuphakelwa kwegazi (χ2(1, 517) = 16.7, p <0.0001) nokukhulelwa (χ2(1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1, 1. sis wayevaleleke ogibeni ekhulula ingxube yokwenziwa (Fig. 6B), kuyilapho inani An alizange lihluke.I-Pharoensis ezifundeni ezihlukahlukene ze-physiological yatholakala (Fig. 6C) .Ku-Culex, ukwanda okuphawulekayo kuphela kwenani lomiyane abafuna abanikazi batholakala ezicupheni eziboshwe ngengxube yokwenziwa, i-130 = 130 = 130; 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 yezilwane ezifuywayo. ihlelwe futhi ibanjwe, nakuba ngezinombolo ezincane, ngaphandle kweminyaka yobudala 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]. Omiyane besifazane bathola lesi sisetshenziswa ngokuhogela futhi bayakwazi ukulawula ukumuncwa kwezakhi ze-nitrogen kumchamo, okuhlanganisa i-urea, ingxenye enkulu yenitrogen emchamweni [15, 16]. Ngokuya ngesimo somzimba womiyane wesifazane, imisoco emchameni inikezwa ukuze ikwazi ukuphila kahle njengomiyane. nezici zokuzala zabantu abondliwe ngegazi phakathi nomjikelezo 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 ama-nitrogenous compounds abalulekile ngokuzibandakanya ekondleni okuyingozi ephansi.Lokhu kutholakala kunomphumela obalulekile we-epidemiological, ukwandisa umsebenzi wabo we-epidemiological, ukwandisa umsebenzi wabo we-epidemiological, ukwandisa umsebenzi wabo we-female olindelekile kanye nemiphumela yempilo yabo yokukhiqiza. ngaphezulu, lokhu kuziphatha kungase kube okuhlosiwe kwezinhlelo zesikhathi esizayo zokuphatha ama-vector.