Ukufumana isondlo kunye nokuhanjiswa kwesondlo kudibanisa ukutya kwezinambuzane kunye neempawu zembali yobomi.Ukubuyisela ukusilela kwizondlo ezithile kwizigaba ezahlukeneyo zobomi, izinambuzane zinokufumana ezi zondlo ngokutya okongeziweyo, umzekelo, ngokutya kwiimfihlo ze-vertebrate kwinkqubo eyaziwa ngokuba yi-puddles.Ingcongconi ye-Anopheles arabianished ibonakala ngathi yi-malnoproduction, ngoko ke, i-metabolism ifuna i-malnoproduction. olu phononongo yayikukuvavanya ukuba An. Ukuphazamiseka kwe-arabiensis kumchamo wenkomo ukuze kufumaneke izondlo kuphucula iimpawu zembali yobomi.
Qinisekisa ukuba ikhuselekile.arabiensis yatsalwa livumba elitsha, iiyure ezingama-24, iiyure ezingama-72, kunye neyure ye-168 yenkomo endala yomchamo, kunye nokufuna umkhosi kunye nokutyiswa kwegazi (iyure ezingama-48 emva kwesidlo segazi) abafazi balinganiswa kwi-olfactometer ye-Y-tube, kwaye abasetyhini abakhulelweyo baye bavavanywa kunye nohlalutyo lwe-electrophysiological. Iikhompawundi ze-bioactive kumchamo wenkomo kuzo zonke iiklasi ezine ubudala.Imixube yokwenziwa kweekhompawundi ze-bioactive yavavanywa kwi-Y-tube kunye nezilingo zentsimi.Ukuphanda umchamo wenkomo kunye ne-urea edibeneyo equlethe i-nitrogen njengokutya okunokuncedisa kwi-malaria, imilinganiselo yokutyisa kunye neempawu zembali yobomi zaye zalinganiswa. ukondla, amabhinqa avavanyelwa ukuphila, ukubhabha okubotshelelwayo, kunye nokuzala.
Funa igazi lomkhosi kunye nesondlo.Kwizifundo zebhubhoratri nakwintsimi, ama-Arabhu atsalwa kwivumba lendalo kunye nelokwenziwa komchamo wenkomo omtsha kunye nomdala.Abafazi abakhulelweyo babengenamdla kwiimpendulo zomchamo wenkomo kwiindawo zokuzala.Iimazi ezifunayo kunye negazi-ezifunxa ngenkuthalo umchamo wenkomo kunye ne-urea kwaye zabela ezi zixhobo zokusebenza njenge-flight-off ye-flighting ye-flight-off-off ukusinda, okanye ukuzala.
Ukufumana nokusasazwa komchamo wenkomo i-Anopheles arabinis ukuphucula iimpawu zembali yobomi.Ukondla okongeziweyo komchamo wenkomo kuchaphazela umthamo wezilwanyana ngokuthe ngqo ngokwandisa ubomi bemihla ngemihla kunye nokuxinana kwezilwanyana, kwaye ngokungangqalanga ngokuguqula umsebenzi wokubhabha kwaye ke kufuneka kuqwalaselwe kwimizekelo yexesha elizayo.
Ukufumana isondlo kunye nokusabalalisa isondlo kudibanisa izinambuzane kunye neempawu zembali yobomi [1,2,3].Izinambuzane ziyakwazi ukukhetha nokufumana ukutya kunye nokwenza ukutya okuhlawulelwayo ngokusekelwe kubukho bokutya kunye neemfuno zezondlo [1, 3]. ukusilela kwizondlo ezithile, izinambuzane ziyakwazi ukufumana ezi zondlo ngokutya okongeziweyo, njengodaka, inkunkuma eyahlukeneyo kunye nemfihlo yezilwanyana ezinomqolo, kunye ne-carrion, inkqubo eyaziwa ngokuba yi-puddles [2] .Nangona iintlobo zebhabhathane kunye ne-nundu zichazwe ngokuyinhloko, imingxuma yokunkcenkceshela iyenzeka nakwezinye ii-oda zezinambuzane kunye nezixhobo zokutya ezinokuthi zibe nefuthe elibalulekileyo kwimpilo yezilwanyana kunye nezilwanyana ezinokuthi zibe nemiphumo ebalulekileyo yokuphila. iimpawu [2, 4, 5, 6] ,7] .Ingcongconi yemalariya u-Anopheles gambiae sensu lato (sl) uvela njengomntu omdala 'ongondlekanga' [8], ngoko ke ukunkcenkceshela kunokudlala indima ebalulekileyo kwiimpawu zembali yobomi, kodwa le ndlela yokuziphatha iye yatyeshelwa ngoku.
Ukuthathwa kwe-nitrojeni kwimazi yeengcongconi ezidala i-Anopheles kulinganiselwe ngenxa yogcino lweekhalori eziphantsi ezithathwe kwinqanaba le-larval kunye nokungasebenzi kakuhle kwesidlo segazi [9].I-Female Ann.gambiae sl ihlawulela oku ngokuncedisa kwizidlo zegazi ezongezelelweyo [10, 11], ngaloo ndlela ibeka abantu abaninzi emngciphekweni wokufumana isifo kunye ne-moquito. Kungenjalo, iingcongconi zinokusebenzisa ukongezelela kwilindle le-vertebrate ukufumana iikhompawundi ezinenitrogen eziphucula ukuziqhelanisa nokubhabha, njengoko kubonisiwe zezinye izinambuzane [2]. Kulo mba, umtsalane onamandla nowahlukileyo wolunye lweentlobo ezizalanayo ngaphakathi kwe-An.The Gambian sl species complex, Anow, pheles, 12, 14, 12 inika umdla.I-Anopheles arabinis inethuba ngokukhetha kwayo kwaye iyaziwa ngokunxulumana kunye nokutya iinkomo. Umchamo wenkomo ngumthombo otyebileyo kwiikhompawundi zenitrogen, kunye ne-urea ibalwa kwi-50-95% yenitrogen epheleleyo kumchamo omtsha [15, 16]. Njengoko iminyaka yomchamo wenkomo, i-microorganisms ye-nitrogen iyancipha kwiiyure ezi-2 ze-microorganisms ze-nitrogen [15] .Ngokunyuka ngokukhawuleza kwe-ammonia, ehambelana nokuhla kwe-nitrogen ephilayo, i-alkalophilic microorganisms (ezininzi ezivelisa iikhompawundi ezinetyhefu kwiingcongconi) zikhula [15], ezinokuthi zibe ngu-Ann.arabiensis obhinqileyo unomdla kakhulu kumchamo oneminyaka eyi-24 iiyure okanye ngaphantsi [13, 14].
Kulo cwaningo, i-host host kunye nee-Ans ezondliwa ngegazi zaye zajongwa.Ngethuba lokuqala lomjikelo we-gonadotropin, i-arabiensis yavavanywa ukufumana i-nitrogenous compounds, kuquka i-urea, ngokuxuba umchamo.Okulandelayo, uchungechunge lweemvavanyo lwenziwe ukuhlola indlela iimiyane ezibhinqileyo ezinikezela ngayo le mithombo yezondlo ezinokuthi zibekho, ukuphuculwa kwe-osurvival kunye nokuphucula i-osurvival. Umchamo wenkomo omtsha nomdala wavavanywa ukufumanisa ukuba ngaba ezi zibonelela ngemikhondo ethembekileyo yomkhosi kunye ne-An etyiswe ngegazi. Ekufuneni kwabo lo mthombo wesondlo onokubakho, i-arabiensis yafumanisa ulungelelwaniso lweekhemikhali emva kokubonwa okunomtsalane. impembelelo yevumba lomchamo wenkomo kwiindawo ezahlukeneyo zomzimba. Umtsalane weengcongconi.Iziphumo ezifunyenweyo ziqinisekisa ukuba u-An. i-arabiensis ifumana kwaye isasaze iikhompawundi ze-nitrogenous ezifumaneka kumchamo we-vertebrate ukuchaphazela iimpawu zembali yobomi.Ezi ziphumo zixutyushwa kumxholo weziphumo ze-epidemiological ezinokuthi zingasetyenziselwa ukucupha kunye nokulawula i-vector.
I-Anopheles arabicans (i-Dongola strain) yayigcinwe ku-25 ± 2 °C, 65 ± 5% RH kunye ne-12: 12 h ukukhanya: umjikelo omnyama. Imibungu yayikhuliswe kwiitreyi zeplastiki (20 cm × 18 cm × 7 cm) izaliswe ngamanzi adiyiweyo kwaye ifakwe ukutya, i-Tetramin, i-DE iqokelelwa ukutya, i-Tetramin iqokelelwa. kwiikomityi ze-30 ml (Nolato Hertila, Åstorp, SE) kwaye emva koko idluliselwe kwiikheji ze-Bugdorm (30 cm × 30 cm × 30 cm; i-MegaView Science, Taichung, Taiwan) ukuvumela ukuvela kwabantu abadala.Abadala babonelelwa nge-10% yesisombululo se-sucrose ad libitum kude kube yi-4 yeentsuku ze-post-seemerge, apho abantu ababhinqileyo banikezela ngeentsuku ze-4 emva kokubonana. umfuniselo, okanye walamba ngobusuku ngamanzi adityanisiweyo ngaphambi kovavanyo, njengoko kuchaziwe ngezantsi.Abafazi abasetyenziselwa iimvavanyo zetyhubhu yendiza babulawa yindlala kuphela iiyure ze-4-6 ngamanzi ad libitum.Ukulungiselela iingcongconi ezifunxa igazi kwii-bioassays ezilandelayo, iimazi ze-4 dpe zanikwa igazi lezimvu ze-defibrotic (Håtunalab usebenzisa inkqubo ye-Discovery, i-Workshopy feed, i-SEO) I-Accrington, UK) .Abafazi abagcwele ngokupheleleyo bathunyelwa kwiikheji zomntu ngamnye kwaye banikezela ngokutya ngokuthe ngqo, njengoko kuchazwe ngezantsi, okanye i-10% ye-sucrose ad libitum kwiintsuku ze-3 ngaphambi kovavanyo oluchazwe ngezantsi.Amabhinqa amva asetyenziselwa i-bioassays ye-flight tube kwaye adluliselwe kwi-laboratory, kwaye emva koko abe ne-distilled ad libitum yamanzi ngaphambi kweeyure ze-4-6.
Uvavanyo lokutyisa lusetyenziselwa ukulinganisa umchamo kunye nokusetyenziswa kwe-urea kumntu omdala we-A.Arab female.Abafazi abafuna umkhosi kunye negazi banikwa ukutya okuqukethe i-1% ehlanjululweyo yomchamo wenkomo entsha kunye nekhulile, imilinganiselo eyahlukeneyo ye-urea, kunye nolawulo olubini (10% sucrose kunye namanzi) kwi-48 h.Ukongezelela, umbala wokutya (1 mgne-CASani ; I-2650-17-1; i-Sigma-Aldrich, i-Stockholm, i-SE) yongezwa kwi-4 × 4 matrix kwi-250 µl microcentrifuge tubes (i-Axygen Scientific, i-Union City, i-US; Umzobo 1A) Gcwalisa kwi-edge µes kunye nemiphumo ye-moquies ukuze ugweme ukukhuphisana (~300). beka iingcongconi ezili-10 kwisitya esikhulu sePetri (i-12 cm ububanzi kunye no-6 cm ubude; Semadeni, Ostermundigen, CH; Figure 1A) ebumnyameni obupheleleyo kwi-25 ± 2 cm °C kunye ne-65 ± 5% ukufuma okuhlobene.Le mifuniselo yaphindwa 5 ukuya ku-10 amaxesha.Emva kokuhlalutya okungaphezulu kwe-2.
Khangela umchamo wenkomo kunye ne-urea efunxelwe ngumnini kunye nemazi efunxa igazi u-Anopheles arabianus.Kulingo lokutyisa (A), iingcongconi ezizimazi zanikwa ukutya okuquka umchamo wenkomo osemtsha nomdala, ubuninzi be-urea, i-sucrose (10%), kunye namanzi adibeneyo (H2O) ). ukuba iimazi ezifuna umkhosi zifunxa umchamo wenkomo weeyure ezingama-72 ngaphantsi kwe-168 yeyure yomchamo wenkomo (B) .Intsingiselo ye-nitrogen epheleleyo (± ukuphambuka okusemgangathweni) yomchamo imelwe kwi-inset.I-Host-seking (D, F) kunye nokufunxa igazi (E, G) amabhinqa athatha i-urea kwi-dose-exhomekeke kwi-dose-dependent volume. (indlela enye i-ANOVA isebenzisa uhlalutyo lwesithuba se-Tukey; p <0.05). Iibha zeemposiso zibonisa imposiso eqhelekileyo yentsingiselo (BE).Umgca odayiweyo othe tye umele umgca wobuyiselo welog (F, G)
Ukukhulula ukutya okufunxayo, iingcongconi zafakwa ngabanye kwiityhubhu ze-1.5 ml ze-microcentrifuge eziqukethe i-230 µl yamanzi adibeneyo kwaye i-tissue yaphazamiseka ngokusebenzisa i-pestle elahlayo kunye ne-cordless motor (VWR International, Lund, SE), ilandelwa yi-centrifugation kwi-10 krpm ye-10 min . i-microplate (Sigma-Aldrich) kunye ne-absorbence (λ620) yamiselwa kusetyenziswa i-spectrophotometer-based microplate reader (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Ngenye indlela, iingcongconi zagatywa kwi-1 ml yamanzi adiyitiweyo, i-900 µl apho uhlalutyo lwe-curovetrometric6 ludluliselwe kwi-curovethi ye-2 i-curovethi nm; UV 1800, Shimadzu, Kista, SE).Ukulinganisa ukutya okudliwayo, ijika eliqhelekileyo lalungiswa nge-serial dilution ukuvelisa i-0.2 µl ukuya kwi-2.4 µl ye-1 mg ml-1 i-xylene cyanide.
Idatha yomthamo yahlalutywa kusetyenziswa uhlalutyo lwendlela enye yokwahluka (ANOVA) ilandelwa luthelekiso lwe-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, i-CA, i-US).
Ngokumalunga ne-20 µl yeesampuli zomchamo ukusuka kwiqela ngalinye leminyaka yayiboshwe kwi-Chromosorb® W / AW (10 mg 80/100 mesh, Sigma Aldrich) kwaye ifakwe kwii-capsules ze-tin (8 mm × 5 mm) . Inzululwazi, iWaltham, MA, US) ukumisela isiqulatho senitrogen kumchamo omtsha nomdala ngokweprothokholi yomenzi.Itotali yenitrogen (g N l-1) yabalwa ngokusekelwe kugxininiso lwe-urea olwaziwayo olusetyenziswa njengomgangatho.
Ukuvavanya isiphumo sokutya ekufuneni umkhosi kunye nokusinda kwabasetyhini abafunxa igazi, iingcongconi zafakwa ngokukodwa kwisitya esikhulu sePetri (i-12 cm ububanzi kunye ne-6 cm ubude; i-Semadeni) kunye nomngxuma ogqunywe ngumnatha kwisiciko (i-3 cm ububanzi) kunye Nokungena komoya kunye nokutya. urea, kunye nolawulo olubini, i-10% ye-sucrose kunye namanzi.Ukutya ngakunye kwakufakwe umbhobho kwi-tampon yamazinyo (DAB Dental AB, Upplands Väsby, SE) efakwe kwi-5 ml yesirinji (Thermo Fisher Scientific, Gothenburg, SE), i-plunger isusiwe, kwaye ibekwe phezu kwe-petri dish yakho ye-dish1b echazwe yonke imihla. ngasentla.Iingcongconi ezisindileyo zazibalwa kabini ngosuku, ngelixa iingcongconi ezifileyo zalahlwa kwaze kwaba yilapho ingcongconi yokugqibela yafa (n = 40 ngonyango) Ukusinda kweengcongconi ezondliwe kwizidlo ezahlukahlukeneyo kwahlalutywa ngokwezibalo kusetyenziswa i-Kaplan-Meyer curves kunye ne-log-rank tests ukuthelekisa ukuhanjiswa kwe-Statistics ye-SPSS. 24.0.0.0).
A isiko ingcongconi flying lokusila esekelwe Attisano et al. [17], eyenziwe 5 mm ubukhulu ecacileyo panels acrylic (10 cm ububanzi x 10 cm ubude x 10 cm ukuphakama) ngaphandle panels ngaphambili nangasemva (Fig. 3: phezulu) .Indibano pivot kunye ityhubhu nkqo eyenziwe ngegesi chromatography ikholamd (0.25 mm intlama intlama L. inaliti exhonywe phakathi kweperi yemagnethi ye-neodymium ukuqelelana ngeesentimitha ezili-9. Ityhubhu ethe tyaba eyenziwe ngento efanayo (6.5 cm L) yacanda kabini ityhubhu ethe nkqo ukuze yenze ingalo ebotshelelweyo kunye nengalo ephethe iqhekeza elincinci lefoyile yealuminiyam njengophawu lokuphazamisa ukukhanya.
Iimazi ezilambileyo zeeyure ezingama-24 zanikwa ukutya okungasentla kwimizuzu engama-30 ngaphambi kokunqanda. Iingcongconi ezibhinqileyo ezondliwe ngokupheleleyo zaza zafakwa i-anesthetized nganye emkhenkceni kangangemizuzu eyi-2-3 zaza zancanyathiselwa kwizikhonkwane zezinambuzane nge-bewax (uJoel Svenssons Vaxfabrik AB, Munka Ljungby, SE) kwaye emva koko wabopha iityhubhu ze-horzontal. I-Mill.Revolutions ngendiza nganye irekhodwe yi-logger yedatha eyakhelwe ngokwezifiso, emva koko igcinwe kwaye iboniswe usebenzisa i-software ye-PC-Lab 2000™ (v4.01; Velleman, Gavere, BE) .I-flight mill yafakwa kwigumbi elilawulwa yimozulu (12 h: 12 h, ukukhanya: kumnyama, 25 ± 2 ° C, 5% R ± 65).
Ukujonga umfanekiso wendlela yokusebenza kwendiza, umgama opheleleyo ohambayo (m) kunye nenani elipheleleyo lemisebenzi yendiza elandelelanayo ibalwa ngeyure kwisithuba seeyure ezingama-24. Ukongezelela, umgama oqhelekileyo ohamba ngeenqwelo-moya ngabasetyhini ngabanye uthelekiswe kunyango kwaye uhlalutywe kusetyenziswa indlela enye ye-ANOVA kunye ne-Tukey's post hoc analysis (JMP Pro, v14.0.0), i-avareji ye-Institute yonyango yayiqwalaselwa, i-avareji ye-SAS, i-Inc, i-Avareji yonyango ixhomekeke kwi-Institute. factor.Ukongezelela, inani eliphakathi kwemijikelo libalwa kwi-10-minute increments.
Ukuvavanya umphumo wokutya ekusebenzeni kokuzala kwe-An.arabiensis, abafazi abathandathu (i-4 dpe) badluliselwe ngokuthe ngqo kwi-Bugdorm cages (30 cm × 30 cm × 30 cm) emva kokuqokelela kwegazi kwaye emva koko babonelele ngokutya kovavanyo lwe-48 h njengoko kuchaziwe ngasentla.Izidlo zaye zasuswa kwaye iikomityi zokuzala (30 ml) zazaliswa ngosuku lwesithathu lwe-Hertila; Iiyure ze-48, ukuguqula zonke iiyure ze-24. Phinda kwakhona irejimeni yokutya nganye ngamaxesha angama-20-50. Amaqanda abalwa kwaye abhalwa kwi-cage nganye yovavanyo.Iisampula zamaqanda zisetyenziselwa ukuvavanya ubungakanani bentsingiselo kunye nobude benguqu yamaqanda ngamanye (n ≥ 200 ngokutya) usebenzisa i-Dialux-20 microscope (DM1000; Ikhamera yeLeica (DFC) 320 R2; I-Leica Microsystems Ltd., DE) .Amaqanda asele agcinwe kwigumbi elilawulwa yimozulu phantsi kweemeko eziqhelekileyo zokukhulisa i-24 h, kunye ne-subsample ye-1st instar larvae esandul 'ukuvela (n ≥ 200 ngokutya) ilinganiswe, njengoko kuchazwe ngasentla.Inani lamaqanda kunye nobukhulu bamaqanda kunye nezibungu zifaniswa phakathi kwonyango kunye nokusebenzisa i-PANOVA ye-hockey enye (i-hoc-way) v14.0.0, SAS Institute Inc.).
Ukuvuthuluka kwe-Headspace ukusuka kwi-fresh (iyure eyi-1 emva kokuthathwa kwesampuli), i-24 hr, i-72 yeyure kunye ne-168 yomchamo omdala yaqokelelwa kwiisampuli eziqokelelwe kwiinkomo ze-Zebu, i-Arsi races. Ukuze kube lula, iisampulu zomchamo zaqokelelwa kusasa ngelixa iinkomo zisesedlelweni. iingxowa zokubhaka ze-polyamide (i-Toppits Cofresco, i-Frischhalteprodukte GmbH kunye ne-Co., i-Minden, i-DE) kwi-3 l polyamide enesiciko Kwi-vinyl chloride imigqomo yeplastiki. I-Volatiles ye-Headspace evela kwisampuli nganye yomchamo wenkomo yaqokelelwa ngokuthe ngqo (entsha) okanye emva kokuvuthwa kwi-172h isampuli yokushisa kunye ne-172h isampuli ye-urineh, i-172h isampuli ye-urineh kunye ne-172h lalimele iqela ngalinye lobudala.
Ukuqokelela i-headspace volatiles, inkqubo evaliweyo isetyenziselwa ukujikeleza i-carbon-filtered gas stream (100 ml min-1) ngokusebenzisa isikhwama se-polyamide ukuya kwikholamu ye-adsorption ye-2.5 h ngokusebenzisa i-diaphragm vacuum pump (KNF Neuberger, Freiburg, DE) .Njengoko iqoqo le-polyad ye-polyaption yayiyi-bag engenanto yokulawula, i-polyad iqoqo le-polyap. yenziwe nge-tubing ye-Teflon (5.5 cm x 3 mm id) equkethe i-35 mg ye-Porapak Q (i-50/80 mesh; i-Waters Associates, i-Milford, MA, i-US) phakathi kweeplagi zoboya beglasi.Ngaphambi kokusetyenziswa, ikholamu ihlanjululwe nge-1 ml i-redistilled n-hexane (i-Merck, i-Darmstadt 9 ml, i-peni ye-9 ml, i-pure, i-DE, i-DE, i-pure, i-DE, i-puree 9 ml, i-DE) Sigma Aldrich).I-adsorbed volatiles yahluthwa nge-400 μl ye-pentane.Iingqokelela ze-headspace zadityaniswa zaza zagcinwa ku--20°C de zisetyenziselwe uhlalutyo olongezelelweyo.
Iimpendulo zokuziphatha zokufuna umkhosi kunye nokutya igazi I-A.Headspace isicatshulwa esiguquguqukayo esiqokelelwe kwi-fresh, i-24-h, i-72-h, kunye ne-168-h-agugile umchamo yahlalutywa kwiicatshulwa eziguquguqukayo ezivela kwiingcongconi ze-Arabidopsis zisebenzisa i-tube yeglasi eqondileyo i-olfactometer [18]. umsebenzi wokufuna ikhaya.I-Arab [19].Ityhubhu yeglasi ye-olfactometer (80 cm × 9.5 cm id) yakhanyiswa nge-3 ± 1 lx yokukhanya okubomvu ukusuka phezulu.I-Charcoal ehluziweyo kunye nokuhamba komoya okufumileyo (25 ± 2 °C, 65 ± 2% umswakama ohambelanayo) udlulise i-30cm ye-bioassay yentsimbi. izikrini zemesh, ukudala ukuhamba kwe-laminar kunye nesakhiwo esifanayo se-plume. I-pentane yayisetyenziswe njengolawulo.Iimiyane ezifuna umkhosi okanye i-mosquito ezifunxa igazi zifakwe kwiindawo zokukhulula umntu ngamnye kwiiyure ze-2-3 ngaphambi kokuqala kovavanyo.I-cage yokukhulula ifakwe kwicala le-downwind ye-olfactometer, kwaye iimiyane zavunyelwa ukuba zivumelane ne-1 min, kwaye emva koko i-butterfly valve yokukhulula i-valve yokulawula i-caget ivuliwe okanye i-valve ye-butterfly ivule i-protection. yeengcongconi eziye zadibana nomthombo phakathi kwemizuzu ye-5 yokukhululwa.I-headspace nganye isicatshulwa esiguquguqukayo kunye nolawulo lwenziwa ngokuphindaphindiweyo ngamaxesha angama-30, kwaye ukuphepha imiphumo nayiphi na imini enye, inani elifanayo lonyango kunye nolawulo lwavavanywa ngosuku ngalunye lovavanyo.Fumana iimpendulo ezivela kwi-host kunye ne-Ans. (JMP Pro, v14.0.0, SAS Institute Inc.).
Impendulo yokuzala kwe-An.Izicatshulwa ze-headspace ezivela kumchamo wenkomo omtsha kunye nomdala zahlalutywa kwiikheji ze-Bugdorm (30 cm × 30 cm × 30 cm; iSayensi yeMegaView).Iikomityi zeplastiki (30 mL; Nolato Hertila) ezizaliswe yi-20 mL yamanzi adiyiweyo zanikezela i-sparking substrate, i-substrate ye-2 echasene ne-cm ye-caatment ejongene ne-2 cm kunye ne-20 cm ye-sparking. iikomityi zahlengahlengiswa kunye ne-10 μl yesicatshulwa ngasinye se-headspace kwi-1: 10 dilution.Ixabiso elilinganayo le-pentane lisetyenziselwe ukulungelelanisa indebe yokulawula.Unyango kunye neekomityi zokulawula zatshintshwa phakathi kovavanyo ngalunye lokulawula imiphumo yesimo.Abafazi abalishumi begazi bakhululwa kwiikheji zokulinga kwi-ZT 9-11 kunye namaqanda kwiikomityi zibalwe emva kweeyure ze-24. amaqanda abekwe kwikomityi yonyango – inani lamaqanda abekwe kwikomityi yokulawula)/(inani lilonke lamaqanda abekiweyo).Unyango ngalunye lwaphindwa izihlandlo ezisi-8.
Uhlalutyo lwerhasi yechromatographic kunye ne-elektroni ye-antenna (GC-EAD) uhlalutyo lwe-an.arabiensis yowesifazane lwenziwa njengoko kuchaziwe ngaphambili [20] .Ngokufutshane, i-headspace entsha isicatshulwa esiguquguqukayo sahlulwa kusetyenziswa i-Agilent Technologies 6890 GC (Santa Clara, CA, US) ixhotyiswe ngekholamu ye-HP-5 μd 5 μd 5 μd 5 mm × 2. ubukhulu, Agilent Technologies). kunye nomchamo ogugayo.I-Hydrogen isetyenziswe njengesigaba esihambayo kunye nesantya sokuhamba kwe-avareji yomgca we-45 cm s-1.Isampuli nganye (2 μl) yatofwa imizuzwana ye-30 kwimowudi engacalulwanga kunye nobushushu bokungena kwe-225 °C. Ubushushu be-oven ye-GC yacwangciswa ukusuka kwi-35 °C (imizuzu eyi-3 ibambe) ukuya kwi-3000 imizuzu ye-GC kwi-100 °C ngomzuzu. i-spluent splitter, i-4 psi ye-nitrogen yongezwa kwaye yahlula i-1: 1 kwi-Gerstel 3D / 2 i-low volume cross cross (Gerstel, Mülheim, DE) phakathi kwe-flame ionization detector kunye ne-EAD.I-GC effluent capillary ye-EAD idluliselwe kumgca wokudlulisa weGerstel ODP-2, olandelela kwiglasi ye-5 × C, i-oven ye-5 mm, i-oven ye-5 mm apho yayixutywe ne-carbon-filtered, air humidified (1.5 l min-1) .I-antenna yafakwa kwi-0.5 cm ukusuka kwi-outlet ye-tube.I-mosquito nganye yayibalelwa kwi-replicate enye, kunye neengcongconi ezifuna umkhosi, ubuncinane ii-replicates ezintathu zenziwa kwiisampuli zomchamo zeminyaka yobudala.
Ukuchongwa kweekhompawundi ze-bioactive kwingqokelela ye-headspace yomchamo omtsha kunye nomdala wenkomo usebenzisa i-GC edibeneyo kunye ne-mass spectrometer (GC-MS; 6890 GC kunye ne-5975 MS; i-Agilent Technologies) ukufumana iimpendulo ze-antennal kuhlalutyo lwe-GC-EAD, esebenza kwimowudi ye-electron yempembelelo ye-ionization kunye ne-UGCS ye-70 equipped. ikholamu ye-silica capillary edityanisiweyo (i-60 m × 0.25 mm ubukhulu bangaphakathi, 0.25 μm ubukhulu befilimu) kusetyenziswa i-helium njengenqanaba elihambayo elinomyinge wokuhamba komndilili we-35 cm s-1.A 2 μl isampulu yatofwa kusetyenziswa useto lwe-injector efanayo kunye nobushushu be-oveni njenge-GC-EAD yobunzima bohlalutyo kunye nohlalutyo lobunzima oluchongiwe ngokusekwe kuhlahlelo lwe-Compounds ye-Compounds ethelekisiweyo. kwilayibrari yesiko kunye nelayibrari ye-NIST14 (i-Agilent) .Iikhompawundi ezichongiweyo ziqinisekiswe ngokufaka imigangatho eyiyo (iFayile eyoNgezelelweyo 1: Itheyibhile S2) .Ngobungakanani, i-heptyl acetate (i-10 ng, i-99.8% yokuhlanjululwa kweekhemikhali, i-Aldrich) yajongwa njengomgangatho wangaphandle.
Ukuvavanya ukusebenza komxube wokwenziwa kwevumba elibandakanya iikhompawundi ze-bioactive ezichongiweyo kumchamo omtsha kunye nomdala ukutsala i-Ans.arabiensis efuna umkhosi kunye nokufunxa igazi, usebenzisa i-olfactometer efanayo kunye neprotocol njengangentla.Imixube ye-Synthetic yalinganisa ukubunjwa kunye nomlinganiselo weekhompawundi kwi-headspace edibeneyo isicatshulwa esiguquguqukayo se-freshur, i-24-ho-ho-24ho-ho-24ho Umchamo omdala weyure eziyi-168 (Umfanekiso 5D-G; Ifayile eyongezelelweyo 1: Itheyibhile S2). Uhlalutyo, sebenzisa i-10 μl ye-1: 100 yokuxutywa komxube owenziwe ngokupheleleyo, kunye nezinga lokukhulula ngokubanzi eliqala malunga ne-140-2400 ng h-1, yokuvavanya umtsalane wokubamba i-moquito emva kovavanyo lwegazi kunye novavanyo lwegazi. imixube ethotywayo yeekhompawundi enye yomxube opheleleyo iyasuswa.Fumana iimpendulo kwinginginya kunye negazi le-Ans.Arab vs imixube yokwenziwa kunye neyokukhuphayo yahlalutywa kusetyenziswa uhlengahlengiso oluqhelekileyo olulandelwa luthelekiso lwesibini lwemilinganiselo engaqhelekanga (JMP Pro, v14.0.0, SAS Institute Inc.).
Ukuvavanya ukuba ingaba umchamo wenkomo ungasebenza na njengendawo egcina ingcongconi yemalariya, umchamo omtsha kunye nomdala wenkomo, oqokelelwe njengoko kuchaziwe ngasentla, kunye namanzi afakwe kwii-emele ezi-3 zelitha (100 ml) ezinemingxuma kwaye zibekwe kwizirhintyelo zokuthiyela. (inguqulo ye-BG-HDT; i-BioGents, i-Regensburg, i-DE).Izibatha ezilishumi zabeka i-50 m phakathi kwedlelo, i-400 m ukusuka kuluntu lwelali (i-Silay, i-Ethiopia, 5°53´24′′N, 37°29′24′′E) kwaye akukho nkomo, kwiindawo zokuzalela ezisisigxina kunye nakwiilali. ijikeleziswa ebusuku ubusuku obuhlanu bubonke.Amanani engcongconi abanjwe kwimigibe ebanjwe ngumchamo weminyaka eyahlukeneyo ubudala athelekiswa kusetyenziswa uhlengahlengiso kunye nokuhanjiswa kwe-beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
Kwilali egquba isifo seengcongconi kufutshane nedolophu yaseMaki, kwingingqi yeOromia, Ethiopia (8° 11′ 08″ N, 38° 81′ 70″ E; Figure 6A). Uphononongo lwaqhutywa phakathi ku-Agasti nophakathi kuSeptemba phambi kokutshizwa kwentsalela yangaphakathi ngonyaka, kunye nexesha elide lemvula. zikhethwe kwisifundo (umzobo 6A) .Iikhrayitheriya ezisetyenziselwa ukukhetha izindlu zaziyi: akukho zilwanyana ezivunyelwe endlwini, akukho kupheka kwangaphakathi (ukuzoba iinkuni okanye amalahle) kuvunyelwe (ubuncinci ngexesha lovavanyo), kunye nezindlu ezinabemi ababini abaninzi, balala kwi-uninsecticides. phantsi komnatha wengcongconi onyangwayo.Imvume yokuziphatha inikwe yiBhodi yeZiko loPhando lweeNdlela zokuziphatha (IRB/022/2016) yeFaculty of Natural Sciences (CNS-IRB), iYunivesithi yaseAddis Ababa, ngokuhambelana nezikhokelo ezisekwe yi-World Medical Association Declaration of Helsinki.Imvumelwano ifunyenwe yinkqubo yolwandiso lwendlu nganye ifunyenwe kwintloko yabasebenzi bendawo yonke. ulawulo kwisithili kunye newadi ('kebele') nqanaba.Uyilo lokulinga lulandele i-2 × 2 yoyilo lwesikwere sesiLatini, apho imixube yokwenziwa kunye nokulawulwa kwabelwa izindlu ezidibeneyo ngobusuku bokuqala kwaye zitshintshe phakathi kwezindlu ngobusuku obulandelayo bokulinga.Le nkqubo yaphindwa izihlandlo ezilishumi.Ukongezelela, ukuqikelela umsebenzi wengcongconi kwizindlu ezikhethiweyo, i-CDC iqhuba i-traps ye-CDC ekupheleni kobusuku obuphakathi, i-CDC ibeka i-traps ye-trial trips kwi-traps ye-wiringa ngexesha elifanayo losuku.
Umxube owenziweyo oqulethe i-bioactive compounds ezintandathu zachithwa kwi-heptane (i-97.0% ye-solvent grade ye-GC, i-Sigma Aldrich) kwaye ikhutshwe kwi-140 ng h-1 isebenzisa i-cotton wick dispenser [20]. Amaziko oLawulo lweSifo kunye noThintelo (CDC) umgibe wokukhanya (i-John W. Hock Inkampani, iGainesville, FL, US; Umzobo 6A) .Imigibe yayixhonywe kwi-0.8 - 1 m ngaphezu komhlaba, kufuphi neenyawo zombhede, kwaye ivolontiya yalala phantsi kwe-mosquito net engakhange iphathwe kwaye iqhutywe phakathi kwe-106to30 kunye ne-sex. isimo somzimba (okungondliwayo, ukondliwa, ukukhulelwa okuncinci, kunye nokukhulelwa [21] emva koko zahlolwa kusetyenziswa i-polymerase chain reaction (PCR) uhlalutyo ukuchonga iintlobo ze-morphologically ezichongiweyo njenge-A. gambiae sl. Amalungu e-complex [23]. ulawulo) yaba sisiphumo esisigxina (JMP® 14.0. 0. SAS Institute Inc.).
Vavanya ukuba ngaba ikhuselekile.arabiensis yakwazi ukufumana umchamo, umthombo wayo oyintloko we-nitrogen, i-urea, ngokutya ngokuthe ngqo, ngaphakathi kwe-48 h yolawulo lweentsuku ze-4 post (dpe) izilingo zokufuna umkhosi kunye negazi lokutya kwabasetyhini (umzobo 1A) .Zombini iintombi ezifuna umkhosi kunye negazi zifunxa kakhulu i-sucrose ngaphezu kwayo nayiphi na enye i-sucrose (F6 = 5) i-2, i-p5, i-p2, i-2, i-p. I-0.0001 kunye ne-F (5,299) = 56.00, p <0.0001, ngokulandelanayo, i-Fig. 1B, C) .Ngaphezu koko, abafazi abafuna umkhosi badla ngaphantsi komchamo kwiiyure ze-72 xa kuthelekiswa nomchamo kwiiyure ze-168 (Umfanekiso 1B) . xa kuthelekiswa nazo zonke ezinye izigxina kunye namanzi, ngelixa lingenakulinganiswa kwi-10% sucrose (F (10,813) = 15.72, p <0.0001; Umfanekiso we-1D) .Oku kwakuchasene nempendulo yabasetyhini abatya igazi, abadla ngokugqithisa kakhulu ukutya okuqukethe i-urea kunamanzi, nangona kunjalo i-5% i-sucrose ingaphantsi kwe-5% (i-5% i-sucrose, kodwa i-5% 0, i-sucrose, i-10%, i-10, i-10, i-10, i-1) 78.35, p <0.0001; Umzobo 1) .Ngaphezu koko, xa kuthelekiswa phakathi kwamazwe amabini e-physiological, abafazi be-phlebotomized bathatha i-urea ngaphezu kwabasetyhini abafuna umkhosi kwiindawo eziphantsi kakhulu, kwaye aba basetyhini bathatha inani elifanayo le-urea kwiindawo eziphezulu (F (1,950; 1F, G).Ngelixa ukuthathwa kokutya okune-urea kubonakala kunamaxabiso afanelekileyo (Fig. 1D,E), amabhinqa kuwo omabini la mazwe akwifiziyoloji akwazile ukumodareyitha isixa se-urea efunxwe kulo lonke uluhlu lokugxilwa kwe-urea ngendlela yelog-linear (Fig. 1F, G). .Ngokufanayo, iingcongconi zibonakala zilawula ukufunyanwa kwazo kwe-nitrogen ngokulawula umlinganiselo womchamo ofunyenweyo, njengoko ubuninzi be-nitrogen kumchamo bubonakaliswe kwixabiso elifunyenweyo (Umfanekiso 1B, C kunye no-B).
Ukuvavanya imiphumo yomchamo kunye ne-urea ekufuneni i-host-sucking mosquito kunye ne-mosquito yokufunxa igazi, abafazi bondliwa umchamo wabo bonke abane ubudala (olutsha, i-24 h, i-72 h, kunye ne-168 h post-deposition) kunye noluhlu lwe-urea concentrations, kunye namanzi adibeneyo kunye ne-10 % uhlalutyo oluye lwabonisa ukuba i-Aval surcrose yayinolawulo lwe-2. impembelelo ebalulekileyo ekusindeni ngokubanzi kwabasetyhini abafuna umkhosi (umchamo: χ2 = 108.5, df = 5, p <0.0001; urea: χ2 = 122.8, df = 5, p <0.0001; Umzobo 2B, C) kunye negazi-2 = χd 9 abafazi = i-fd = f. 5, p <0.0001; urea: χ2 = 137.9, df = 5, p <0.0001 Umzobo 2D, E) .Kuzo zonke iimvavanyo, amabhinqa atya ukutya komchamo, urea, kunye namanzi ayenamazinga aphantsi kakhulu okuphila xa kuthelekiswa nabasetyhini bondla i-sucrose-E-female ye-female-E-female-E-female. ibonise amazinga okuphila ahlukeneyo, kunye nalawo ondliwe i-72-h umchamo omdala (p = 0.016) eneyona nto iphantsi yokusinda (umzobo 2B) .Ngaphezu koko, abafazi abafuna umkhosi wondla i-135 mM urea baphila ixesha elide kunolawulo lwamanzi (p <0.04) (Xa kuthelekiswa ne-2C kunye namanzi ahlambulukileyo kunye namanzi ahlambulukileyo). umchamo wasinda ixesha elide (p = 0.001 kunye p = 0.012, ngokulandelelanayo; Umfanekiso 2D), ngelixa abafazi bondliwa ngomchamo weeyure ezingama-72 baphila ixesha elide kunabo bondliwa Umchamo omfutshane Osetyhini kunye nomchamo weeyure ezingama-24 ubudala (p <0.0001 kunye p = 0.013, xa fere 1M 1D, ngokulandelanayo, i-1M ye-1D). amabhinqa anegazi ahlala ixesha elide kunawo onke amanye ama-urea kunye namanzi (p <0.013; Umfanekiso 2E).
Ukusinda komkhosi kunye nemazi efunxa igazi i-Anopheles arabinis etyisa umchamo wenkomo kunye ne-urea.Kwi-bioassay (A), iingcongconi ezimazi zibonelelwe ngokutya okuquka umchamo omtsha kunye nomdala wenkomo, imilinganiselo eyahlukeneyo ye-urea, i-sucrose (10%) kunye namanzi adibeneyo (H2O). irekhodwa rhoqo kwiiyure ze-12 de bonke ababhinqileyo abondliwa ngomchamo (B, D) kunye ne-urea (C, E), kunye nolawulo, i-Sucrose kunye namanzi, bafile.
Umgama opheleleyo kunye nenani lemijikelo echongiweyo kuvavanyo lokusila ngenqwelomoya kwisithuba seyure ye-24 yahluka phakathi kokufuna umkhosi kunye neengcongconi ezifunxa igazi, ezibonisa umsebenzi omncinci wokubhabha jikelele (Umfanekiso 3) .Iingcongconi ezifuna umkhosi ezibonelela ngomchamo omtsha kunye nomdala okanye i-sucrose kunye namanzi abonisa iipatheni zokubhabha ezahlukileyo (umzobo 3), kunye ne-feines esebenzayo kwi-2 yasekuseni, kunye ne-feines esebenzayo kwi-2 yamandulo, kunye ne-feine esebenzayo kwi-2 yasekhaya. kunye ne-168-iyure ubudala IiMosquitos ezazisondla ngomchamo zibonise iipatheni zokubhabha ezahlukeneyo kwaye ngokuyintloko zaziyi-diurnal.Iingcongconi ezizibhinqa ezibonelela nge-sucrose okanye umchamo weeyure ezingama-72 zibonise umsebenzi kulo lonke ixesha leeyure ezingama-24, ngelixa iimazi ezinikezela ngamanzi zazisebenza ngakumbi ngexesha eliphakathi. Umchamo weeyure ezingama-72 ufumene ukwehla okuthe gqolo kumsebenzi kwiiyure ezingama-24 (Umfanekiso 3).
Kuvavanyo lwe-flight mill, iingcongconi ezizimazi zityiswa ngomchamo omtsha nomdala wenkomo, i-urea, i-sucrose (10%), kunye namanzi adiyiweyo (H2O) abotshelelwe kwindawo ethe tyaba, ejikeleza ngokukhululekileyo (ngokujikeleza igazi). (ekunene) abafazi, umgama opheleleyo kunye nenani leenqwelomoya ngeyure kwisidlo ngasinye kwisithuba seeyure ezingama-24 zarekhodwa (kumnyama: ngwevu; ukukhanya: okumhlophe). Umyinge womgama kunye nenani eliqhelekileyo lokubhobhoza liboniswa ngasekunene kwegrafu yomsebenzi we-circadian.Iimpazamo zemivalo zibonisa imposiso eqhelekileyo yentsingiselo.Uhlalutyo lwamanani bona isicatshulwa.
Ngokuqhelekileyo, umsebenzi wenqwelomoya jikelele wabasetyhini abafuna umkhosi ulandele iphethini efana neyomgama wendiza ngexesha leeyure ze-24. Umgama wokuhamba ngenqwelomoya wawuchaphazeleka kakhulu ngokutya okufakwe kwi-F (5, 138) = 28.27, p <0.0001), kunye nabasetyhini abafuna umkhosi bathatha iiyure ze-72 zomchamo zahamba ngokuphawulekayo ezinye iimgama ezide kakhulu kunye ne-0 (0) xa kuthelekiswa nayo yonke imigama emide ye-0 (p 0). I-sucrose-fed mosquitoes indiza ixesha elide kune-fresh (p = 0.022) kunye ne-24-h-aged umchamo (p = 0.022) -imiyane ezondliwe. Ngokuchasene nephethini yomsebenzi wokubhabha ochazwe kukutya komchamo, abafazi abafuna i-urea-feed urea babonisa umsebenzi wendiza oqhubekayo phezu kwexesha le-24 yesigaba sesibini se-peg. 3) .Nangona iipatheni zemisebenzi zifana, abafazi abafuna umkhosi bondla i-urea ngokuphawulekayo banda kakhulu umgama wokubhabha ngokuxhomekeke kwi-concentration efunyenweyo (F (5, 138) = 1310.91, p <0.0001)
Umsebenzi wokubhabha opheleleyo weengcongconi ezifunxa igazi uzinzile kwaye uzinzile kwiiyure ze-24 kuzo zonke izidlo, kunye nokunyuka komsebenzi womchamo ngexesha lesibini lexesha elimnyama labasetyhini abondliwe emanzini kunye nabasetyhini abondliwe amatsha kunye neeyure ze-24 ubudala (umfanekiso 3) .Ngelixa ukutya komchamo kuchaphazeleke kakhulu umgama wokubhabha kwi-female ezondliwa ngegazi (F.8, 0 = 4), 38 = 0 = 1.8 = 0 = 1.0 = 1.8 = 0 = 1.0 = 4.8 = 0 = 1.8 = 0 = 1.0 = 1.0 = 1.8 = 0 = 4. ukutya kwe-urea akuzange (F (5, 138) = 1.36, p = 0.24) .kunye nomchamo kunye nokulawula ukutya (okutsha, p = 0.0091; iiyure ze-72, p = 0.0022; iiyure ze-168, p = 0.001; i-sucrose, p = 0.H0027, p = 0.H020.
Imiphumo yokutya komchamo kunye ne-urea kwiiparameters zokuzala zavavanywa kwii-bioassays ze-egg-laying (Umfanekiso 4A) kwaye zaphandwa ngokwenani lamaqanda abekwe yimazi nganye, ubukhulu beqanda, kunye ne-larvae yokuqala esanda kuqanduselwa.Inani lamaqanda abekwe. Umzobo we-4B) .Abafazi bondla umchamo weeyure ze-24, ukutya kwegazi kwabeka amaqanda amaninzi ngakumbi kunokuba amabhinqa atya ukutya okunye umchamo kwaye ayefana nalawo ondla i-sucrose (umzobo 4B) .Ngokunjalo, ubungakanani bamaqanda abekwe ngamabhinqa aphethwe ngumchamo ahluka ngokutya (F (5, 209) = 12.0 , 209 = 12.0, 209 = 12.0 no-urine, 209 = 12.0, 209, 12.0, 12, 209, = 12.0, no-4, p. I-sucrose-feed females ibeka amaqanda amakhulu kakhulu kunamanzi athotyiweyo, ngelixa amaqanda amabhinqa asondla nge-168 h yomchamo ayencinci kakhulu (umzobo 4C) .Ukongezelela, ukutya komchamo kwachaphazela kakhulu ubungakanani be-larval (F (5, 187) = 7.86, p <0.0001), kunye namaqanda e-4 avela kwi-lar-2 enkulu kakhulu. Iimazi ezineeyure ezingama-72 ezityiswa ngomchamo kunamaqanda abekwe emaqandeni imibungu.Ondliwa ngamanzi kunye neemazi eziyi-168 ezondliwa ngumchamo (Umfanekiso 4D).
Ukusebenza kokuzala kwemazi u-Anopheles arabinis etyisa umchamo wenkomo kunye ne-urea.Iingcongconi ezizithokazi ezondliwe ngegazi zatyiswa ukutya okubandakanya umchamo wenkomo omtsha kunye nomdala, umlinganiselo ohlukeneyo we-urea, i-sucrose (10%), kunye namanzi adibeneyo (H2O) kwiiyure ezingama-48 ngaphambi kokubeka kwii-bioassays kunye neeyure ze-Egg (i-Bst, i-Egg-iyure) Ubungakanani beqanda (C, F) kunye nobukhulu bombungu (D, G) bachaphazeleka kakhulu ngokutya okubonelelweyo (umchamo wenkomo: BD; urea: EG) . Iindlela zeparameter nganye ezilinganiswe ngokusebenzisa amagama ahlukeneyo anobumba zahluke kakhulu omnye komnye (indlela enye i-ANOVA isebenzisa uhlalutyo lwe-Tukey ye-post hoc; p <0.05) .
Njengenxalenye enkulu ye-nitrogen yomchamo, i-urea, xa inikezelwa njengokutya kwabasetyhini abondliwa ngegazi, ichaphazele kakhulu imilinganiselo yokuzala kuzo zonke izifundo.Inani lamaqanda abekwe ngabasetyhini abondliwe i-urea, emva kokutya kwegazi, kuxhomekeke kwi-concentration ye-urea (F (11, 360) = 4.69; p <0.0001), i-concentrations ye-3M 4M phakathi kwe-3M µ urea 4M3M urea i-4. zabeka amaqanda amaninzi (Figure 4E) .Abafazi bondliwa kwi-urea concentrations ye-134 µM okanye ngaphezulu bazala amaqanda amakhulu kunabafazi abondliwe emanzini (F (10, 4245) = 36.7; p <0.0001; Umfanekiso 4F), kunye nobukhulu bombungu, nangona bechatshazelwa ngumama ofanayo, i-30 (70.19); p <0.0001) yayitshintshile ngakumbi (Umfanekiso 4G).
Umtsalane ngokubanzi kwi-host-funa i-headspace yomchamo wenkomo i-extracts eguquguqukayo.I-arabiensis ehlolwe kwi-glass tube i-olfactometer (i-Fig. 5A) yayichatshazelwa kakhulu ngumdala womchamo (χ2 = 15.9, df = 4, p = 0.0032; Umzobo. 5B) .I-Post 2 uhlalutyo olubonisa ukuba uhlalutyo oluphezulu lwe-urine lubonisa ukuba i-stale hocdor ithelekisa kakhulu kwiiyure ze-2 ezikhangayo. kuzo zonke ezinye iindlela zonyango (iiyure ze-72: p = 0.0060, iiyure ze-168: p = 0.012, pentane: p = 0.00070), Ngaphandle kwevumba lomchamo omtsha (p = 0.13; Umfanekiso 5B) .Nangona umtsalane jikelele weengcongconi ezifunxa igazi , i-8 yayingekho, i-4 ivumba, i-4 = ivumba elibi, i-4 ivumba = i-8, i-4 ivumba. p = 0.067; Umzobo we-5C), ezi zibhinqa zifunyenwe zikhangeleke kakhulu kwi-headspace ezicatshulwayo eziguquguqukayo xa kuthelekiswa nomchamo we-72 weyure xa kuthelekiswa nokulawula (p = 0.0066; Umfanekiso 5C).
Iimpendulo zokuziphatha kwivumba lomchamo wenkomo yendalo kunye neyokwenziwa kukhangelo lomkhosi kunye negazi le-Anopheles arabianus.I-Schematic ye-glass tube olfactometer (A) .Umtsalane we-headspace isicatshulwa esiguquguqukayo esivela kumchamo wenkomo osandul 'ukuzalwa ukuya kubamba (B) kunye negazi lokufunxa igazi (C) kwiingcongconi.Fumana isicatshulwa se-Tentacle entsha yeNkosi. I-24-iyure (E), i-72-iyure (F), kunye ne-168-iyure (G) i-urine yenkomo endala ibonakalisiwe.Ukukhangela i-antenna ye-Electron (EAD) ibonisa utshintsho lwe-voltage ekuphenduleni i-bioactive compounds kwi-headpace ekhutshwe kwi-chromatograph yegesi kwaye ifunyenwe yi-flame ionization detector (i-FID) i-replition ye-scale ye-FID (impendulo ye-FID) ye-FID. (s) Imilinganiselo yeengcongconi ezitsalwa kumagama ahlukeneyo anoonobumba zahluke kakhulu kwenye (indlela enye i-ANOVA isebenzisa uhlalutyo lwe-Tukey ye-post hoc; p <0.05).
I-Female Ann.arabiensis, i-72 h kunye ne-120 h emva kokutya kwegazi, ngexesha lokuzala, akukho kukhethwa kuboniswe kwi-headspace isicatshulwa esiguquguqukayo esivela kumchamo wenkomo omtsha kunye nomdala xa kuthelekiswa nolawulo lwe-pentane (χ2 = 3.07, p> 0.05; Ifayile eyongezelelweyo 1: Umzobo S1).
Kwi-Ann.arabiensis yabasetyhini, i-GC-EAD kunye ne-GC-MS ihlalutya ezichongiweyo ezisibhozo, ezintandathu, ezintathu kunye nezintathu ze-bioactive compounds (Umfanekiso we-5D-G) .Nangona ukungafani kwenani leembumba eziye zafumana iimpendulo ze-electrophysiological zabonwa, ininzi yale michiza yayikhona kwi-headspace nganye isicatshulwa esiguquguqukayo esiqokelelwe kwi-fresh and aged compounds eqokelelwa kwi-extracts ye-extracts evela kwi-extracts ye-female kunye ne-extracts ekhulile kwi-extracts yomchamo ngamnye owenziwe ngumchamo. i-eriyali ngaphezu komda zaye zabandakanywa kuhlalutyo olongezelelweyo.
Ixabiso elipheleleyo lokukhululwa kwe-bioactive compounds kwi-headspace collection yanda ukusuka kwi-29 µg h-1 kumchamo omtsha ukuya kwi-242 µg h-1 kwi-168-iyure ye-urine ubudala, ngokukodwa ngenxa ye-p-cresol kunye ne-m-formaldehyde Phenol inyuka kunye ne-phenol. i-decanal, yehla ngokunyuka kweminyaka yomchamo, ehambelana nokuhla okubonwayo kwi-signal intensity (ubuninzi) kwi-chromatogram (umzobo we-5D) -G ipaneli yasekhohlo) kunye neempendulo ze-physiological kule mixube (Umfanekiso we-5D-G ipaneli yasekunene).
Ngokubanzi, umxube we-synthetic wawunomlinganiselo ofanayo wendalo we-bioactive compounds ezichongiweyo kwiincindi eziguquguqukayo ze-headspaces zomchamo omtsha kunye nomdala (Umfanekiso we-5D-G) kwaye awuzange ubonakale wenze isibheno esibalulekileyo ekufuneni umkhosi (χ2 = 8.15, df = 4, p = 0.083; Umzobo-4, i-2, i-moquis, i-2H) df = 4, p = 0.30; Umfanekiso 5I) .Nangona kunjalo, ukuthelekiswa kwe-post hoc pairwise phakathi kwezonyango kubonise ukuba iingcongconi ezifuna umkhosi zazinomtsalane kakhulu kumxube wokwenziwa komchamo we-24-h xa kuthelekiswa nolawulo lwe-pentane (p = 0.0086; Umfanekiso 5H).
Ukuvavanya indima yamacandelo ngamnye kwimixube yokwenziwa komchamo we-24-h-ubudala, imixube emithandathu ethotywayo yavavanywa ngokuchasene nemixube epheleleyo kwi-Y-tube assay, apho iikhompawundi zomntu ngamnye zaye zasuswa. Kuba iingcongconi ezifuna umkhosi, ukukhupha iikhompawundi zomntu ngamnye kumxube opheleleyo kunempembelelo ebalulekileyo kwiimpendulo zokuziphatha (χ63 = 1, p. I-0.0032; Ifayile eyongezelelweyo ye-1: Umfanekiso we-S2A), yonke imixube ekhuphayo yayithandeka ngakumbi kuneNcinane kunokuxutywa ngokupheleleyo. Ngokwahlukileyo, ukususwa kweekhompawundi zomntu ngamnye kumxube owenziwe ngokupheleleyo akuzange kuthintele iimpendulo zokuziphatha kweemiyane ezifunxa igazi (χ2 = 11.38, df = 6, p = kunye ne-0.07 ephantsi komxube we-0.07) Umtsalane (p = 0.022; Ifayile eyongezelelweyo ye-1: Umfanekiso we-S2B).
Kwidolophana yase-malaria e-Ethiopia, ukusebenza komxube wokwenziwa komchamo wenkomo yeeyure ezingama-24 ekutsaleni iingcongconi phantsi kweemeko zentsimi zavandlakanywa ubusuku obulishumi (umzobo 6A) .Itotali ye-4,861 yomiyane yabanjwa kwaye ichongiwe, apho i-45.7% yayiyi-Anthropus%, i-Anotroenpheles kunye ne-Anoroenphelee. I-35.4% yayiyiCulex spp. (Ifayile eyongezelelweyo 1: Itheyibhile S1) .Anopheles arabinis kuphela kwelungu leentlobo ze-An.Gambian ezichongiweyo ngokuhlalutya kwe-PCR.Ngomndilili, iingcongconi ezingama-320 zabanjwa ngobusuku ngabunye, ngexesha apho izibatha ezineengcongconi ezenziweyo zabanjwa ngaphezulu kweengcongconi kune-312 yomxube (i-312) 170.0, p <0.0001) .Imigibe engabonakaliyo ibekwe kubusuku obuhlanu bokulawula ekuqaleni, phakathi, nasekupheleni kwetyala.Amanani afanayo omiyane abanjwe kwimigibe nganye, ebonisa ukuba akukho cala phakathi kwezindlu (χ2 (0, 1665) > 0 0 - 1665 > 0 0 - 1 × 1665 > 0 0 - 1 × 1665 = 0 0 × 0 1 × 1 p. ixesha lokufunda.Xa kuthelekiswa nemigibe yokulawula, inani leemiyane ezibanjwe kwimigibe equkethe umxube wokwenziwa kwandiswe kakhulu: ukufuna i-host (χ2 (0, 2107) = 138.7, p <0.0001), ukutya okutsha nje kwegazi (χ2 (0, 650) = 32.2, p <0 20) nokukhulelwa (χ20) (0) (0 = χ20) (0) (0 χ20.0). 6.27, p = 0.0123; Ifayile eyongezelelweyo 1: Itheyibhile S1) .Oku kukwabonakaliswa kwinani lilonke leengcongconi ezithinjiweyo: inginginya efuna > ukufunxa igazi > ukhulelwe > i-semi-pregnant > indoda.
Uvavanyo lwasendle lwentsebenzo yomxube wevumba lomchamo wenkomo weeyure ezingama-24. Ulingo lwasendle lwenziwa kumzantsi-mbindi we-Ethiopia (imephu), kufutshane nedolophu yaseMaki (faka), kusetyenziswa amaZiko oLawulo lweSifo (CDC) umgibe wokukhanya (ekunene) kwizindlu ezidityanisiweyo, kunye noyilo lwesikwere lesiLatini (umfanekiso ophaphazelayo weCDC kunye ne-Ador-baphelei ye-CDC ithatha i-Ador-baphelei itsalela i-CDC). arabesques (B), kodwa hayi i-Anopheles farroes (C), ngendlela eyahlukileyo, i-physiological state-dependent effect.Ukongezelela, le migibe ibambe ngokuphawulekayo amanani anda kakhulu eengcongconi ze-Culex.(D) Xa kuthelekiswa nolawulo.Imivalo esekhohlo imele i-avareji yokukhetha isalathiso seengcongconi ezibanjwe ngababini be-odorrant bait (i-NBA) kunye ne-bait yokulawula i-bait (i-N 0 eluhlaza) kunye ne-traps yokulawula i-bait (i-0) ilungelo limele isalathisi sokukhetha esiphakathi kwizibini zemigibe yokulawula (evulekileyo; N = 5). ).Iinkwenkwezi zibonisa amanqanaba okubaluleka kweenkcukacha-manani (*p = 0.01 kunye ne-***p <0.0001)
Ezi ntlobo zintathu zibanjwe ngokungafaniyo kwimigibe equkethe imixube yokwenziwa. Ukukhangela i-host (χ2 (1, 1345) = 71.7, p <0.0001), ukutya kwegazi (χ2 (1, 517) = 16.7, p <0.0001) kunye 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, p, 1, 1, 1, p, 1, 1, 1, 1, 1, p. I-arabiensis yayibanjwe kumgibe wokukhulula umxube we-synthetic (umzobo 6B), ngelixa inani le-An alizange lihluke.I-Pharoensis kwiindawo ezahlukeneyo ze-physiological zafunyanwa (umzobo 6C) .Ku-Culex, kuphela ukwanda okubalulekileyo kwinani leengcongconi ezifuna imikhosi efunyenwe kwimigibe ebanjwe kunye ne-synthetic χ26 (p16) = χ26 umxube we-synthetic (p16 = 1) = 1. 0.0004; Umfanekiso we-6D), xa kuthelekiswa nemigibe yokulawula.
I-host bait trap ibekwe ngaphandle kwemikhosi enokubakho phakathi kweendawo zokuzalela kunye noluntu lwasemaphandleni e-Ethiopia yayisetyenziselwa ukuvavanya ukuba iingcongconi ze-malaria zisebenzisa ivumba lomchamo wenkomo njengendawo yokuhlala yokuhlala. iimiyane zakhangwa kwaye zabanjwa, nangona ngamanani amancinci, azimele kumchamo ubudala (χ2 (5, 25) = 2.29, p = 0.13; Ifayile eyongezelelweyo 1: Umfanekiso S3) Ngokwahlukileyo, ulawulo lwamanzi aluzange lubambe iimiyane ze-malaria kwiqondo lokushisa eliphezulu (Ifayile eyongezelelweyo 1: Umfanekiso S3).
Iingcongconi zeMalariya zifumana kwaye zisasaze iikhompawundi ezinenitrogen ngokutya okuhlawulelwayo kumchamo wenkomo (okt, amadibi) ukuphucula iimpawu zembali yobomi, ezifana nezinye izinambuzane [2, 4, 24, 25, 26]. kumakhaya asemaphandleni kunye neendawo zokuzala.Iingcongconi eziziimazi zifumane lo vimba ngevumba kwaye ziyakwazi ukulawula ukufunyaniswa kweekhompawundi zenitrogen kumchamo, kubandakanya urea, eyona nxalenye inkulu yenitrogen kumchamo [15, 16] .Kuxhomekeke kwimeko yempilo yengcongconi yemazi, izondlo kumchamo ophuculweyo wokuphucula umchamo kunye nomsebenzi we-allo-umchamo. iingcongconi, kunye nokusinda kunye neempawu zokuzala zabantu abondliwa ngegazi ngexesha lomjikelo wokuqala we-gonadotropic.Ngoko ke, ukuxuba umchamo kudlala indima ebalulekileyo yesondlo kwiintsholongwane ze-malaria ezivaliweyo njengabantu abadala abangondlekanga [8], njengoko kunika iimiyane ezibhinqileyo ngokukwazi ukufumana i-nitrogenous compounds ebalulekileyo ngokubandakanyeka kwi-low-risk yokwandisa i-female i-low-risk kwandisa ubomi babo be-female. ukulindela, umsebenzi kunye nemveliso yokuzala, yonke into echaphazela umthamo we-vector.Ngaphezu koko, le ndlela yokuziphatha ingaba yinjongo yeenkqubo zokulawula i-vector ezizayo.