Ukufumana isondlo kunye nokuhanjiswa kudibanisa ukufunyanwa kwezinambuzane kunye neempawu zembali yobomi.Ukuhlawula iintsilelo 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 kunye ne-metabolism yophando. jonga ukuba u-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 abafazi abakhulelweyo baye bavavanywa kwi-biophysical test kunye nohlalutyo lwe-electrophysical oluye lwavavanywa. Umchamo womchamo kuzo zone iiklasi zobudala.Imixube yokwenziwa kweekhompawundi ze-bioactive iye yavavanywa kwi-Y-tube kunye novavanyo lwasendle.Ukuphanda ngomchamo wenkomo kunye neyona nto iphambili ye-nitrogen equlathe urea yekhompawundi njengokutya okunokongeza kwizifo zesifo seengcongconi, imilinganiselo yokutyisa kunye neempawu zembali yobomi zaye zalinganiswa.Umlinganiselo weengcongconi ezibhinqileyo kunye nomthamo osetyenzisiweyo wokucoca umchamo kwimazi kunye nomthamo womchamo osetyenzisiweyo. i-vival, i-tethered flight, kunye nokuveliswa kwakhona.
Funa igazi lomkhosi kunye nesondlo.Kwizifundo zaselabhoratri kunye nentsimi, ama-Arabhu atsalwa kwivumba lendalo kunye nelokwenziwa komchamo wenkomo.Abafazi abakhulelweyo babengenamdla kwiimpendulo zomchamo wenkomo kwiindawo zokuzala. okanye ukuvelisa.
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 zesondlo [1, 3]. izondlo ezithile, izinambuzane ziyakwazi ukufumana ezi zondlo ngokutya okongeziweyo, njengodaka, inkunkuma eyahlukeneyo kunye ne-secretions ye-vertebrates, kunye ne-carrion, inkqubo eyaziwa ngokuba yi-puddles [2] .Nangona iintlobo zeentlobo zebhabhathane kunye ne-nundu zichazwe ngokuyinhloko, imingxuma yokunkcenkceshela iyenzeka nakwezinye ii-oda zezinambuzane, kunye nokukhangayo kunye nemithombo yezempilo kunye ne-5 yezondlo zobomi, i-2 imiphumo ebalulekileyo yempilo, i-2. , 6] ,7].Ingcongconi yemalariya u-Anopheles gambiae sensu lato (sl) uvela njengomntu omdala 'ongondlekanga' [8], ngoko ke ukunkcenkceshela kunokudlala indima ebalulekileyo kwiimpawu zembali yobomi bayo, kodwa le ndlela yokuziphatha iye yatyeshelwa ngoku.
Ukuthathwa kwe-nitrojeni kwiingcongconi ze-Anopheles ezibhinqileyo kulinganiselwe ngenxa yeekhalori ezisezantsi 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 se-moquid kunye nomngcipheko omkhulu wokufumana isifo. I-es ingasebenzisa ukondla okongeziweyo kwilindle le-vertebrate ukufumana iikhompawundi ezinenitrogen eziphucula ukuziqhelanisa nokuhamba kwenqwelomoya, njengoko kubonisiwe zezinye izinambuzane [2]. Kulo mba, umtsalane owomeleleyo nowahlukileyo wolunye lweentlobo ezizalanayo ngaphakathi kwe-An.I-Gambian sl species complex, i-Anopheles arabinis, inkomo entsha nekhulileyo, i-urine e-unis12 inika umdla,14Abinis unip12. inginginya ezikhethwayo kwaye yaziwa ukudibanisa kunye nokutya kwinkomo.Umchamo wenkomo ngumthombo otyebileyo kwiikhompawundi zenitrogen, kunye ne-urea ethatha i-50-95% ye-nitrogen iyonke kumchamo omtsha [15, 16].Njengoko umchamo wenkomo uneminyaka yobudala, ii-microorganisms zisebenzisa ezi zixhobo ukunciphisa ubunzima be-nitrogenous compounds ukuhla kweeyure ze-nitrogenous ezihambelanayo kunye nokwanda kwe-nitrogenous ngaphakathi kwe-24 am. i-alkalophilic microorganisms (ezininzi zazo ezivelisa iikhompawundi eziyityhefu kwiingcongconi) ziyachuma [15], enokuba ngabasetyhini u-Ann.arabiensis utsalwa ngokukhethekileyo kumchamo oneminyaka engama-24 ubudala okanye ngaphantsi [13, 14].
Kolu phononongo, i-host kunye nee-Ans ezondliwa ngegazi zaye zajongwa. Ngexesha lomjikelo wokuqala we-gonadotropin, i-arabiensis yavavanywa ukufumana i-nitrogenous compounds, kuquka i-urea, ngokuxuba umchamo. Umchamo wavavanywa ukufumanisa ukuba ngaba ezi zibonelela ngemikhondo ethembekileyo yokusingatha kunye nokutyiswa ngegazi An. Kukhangelo lwabo lwesi sixhobo sesondlo esinokubakho, i-arabiensis yafumanisa ulungelelwaniso lweekhemikhali emva kokubonwa okunomtsalane. kwiimeko 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) yagcinwa ku-25 ± 2 °C, 65 ± 5% RH kunye nokukhanya kwe-12:12 h: umjikelo omnyama. Imibungu yayikhuliswe kwiitreyi zeplastiki (20 cm × 18 cm × 7 cm) izaliswe ngamanzi adiyiweyo kwaye ifakwe kwi-Tetramin yokutya, i-Ml 3 iqokelela intlanzi kwi-3 ml. iikomityi (Nolato Hertila, Åstorp, SE) kwaye emva koko idluliselwe Bugdorm amakheji (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) ukuvumela ukuvela abadala.Abadala babonelelwa nge-10% sucrose isisombululo ad libitum kude kube 4 iintsuku post-ekuveleni apho kwanikwa i-host-emergence kwi-finary pre-emergence kwi-experiment ye-dietnight okanye i-dietnight ngokukhawuleza. amanzi amisiweyo ngaphambi kovavanyo, njengoko kuchazwe 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 zabonelelwa ngegazi lezimvu ze-defibrotic (Håtunalab, Bro, SE) usebenzisa i-membrane ye-female feeding, i-Discovered ye-Workshop ye-Workshop, i-Workshop ye-Workshop ye-discovered, i-Workshop yokudlulisa i-Workshop ye-Workshop. obomvu kwiikheji zomntu ngamnye kunye nokubonelela ngokutya ngokuthe ngqo, njengoko kuchazwe ngezantsi, okanye i-10% ye-sucrose ad libitum yeentsuku ze-3 ngaphambi kovavanyo oluchazwe ngezantsi.Abafazi bokugqibela basetyenziselwa i-bioassays ye-flight tube kwaye idluliselwe kwi-laboratory, kwaye emva koko ine-distilled yamanzi ad libitum kwiiyure ze-4-6 ngaphambi kovavanyo.
Uvavanyo lokutyisa lusetyenziselwa ukulinganisa umchamo kunye nokusetyenziswa kwe-urea kumntu omdala we-An.Arab female.Abafazi abafuna umkhosi kunye negazi banikwa ukutya okuqukethe i-1% ehlanjululweyo yomchamo wenkomo entsha kunye nekhulile, imilinganiselo eyahlukeneyo ye-urea, kunye nokulawula okubini (10% sucrose kunye namanzi) kwi-48 h.Ukongezelela, umbala wokutya (1 mgne-61-CASanide 6; gma-Aldrich, Stockholm, SE) yongezwa ekutyeni kwaye yanikezelwa kwi-4 × 4 matrix kwi-250 µl microcentrifuge tubes (Axygen Scientific, Union City, CA, US; Figure 1A) Gcwalisa emphethweni (~300 µl). Ukunqanda ukhuphiswano phakathi kweengcongconi ezinombala kunye nefuthe leengcongconi ezinokuba khona1 I-12 cm ububanzi kunye ne-6 cm ubude, i-Semadeni, i-Ostermundigen, i-CH; Umzobo 1A) ebumnyameni obupheleleyo kwi-25 ± 2 cm ° C kunye ne-65 ± 5% yomswakama ohambelanayo.Olu vavanyo lwaphindwa ngamaxesha angama-5 ukuya kwe-10. Emva kokuvezwa kokutya, iingcongconi zabekwa kwi-20 ° C kude kube ngaphezulu -20 ° C.
Khangela umchamo wenkomo kunye ne-urea efunxelwe ngumnini kunye nemazi efunxa igazi u-Anopheles arabianus.Kulingo lokutyisa (A), iingcongconi ezingamamazi zanikwa ukutya okuquka umchamo wenkomo osemtsha nomdala, umlinganiselo owahlukeneyo we-urea, i-sucrose (10%), kunye namanzi adityanisiweyo (H2O) ). iimazi zifunxa umchamo wenkomo iiyure ezingama-72 ngaphantsi kwe-168 yeyure yomchamo wenkomo (B) Uhlalutyo lwe-hoc p <0.05)
Ukukhulula ukutya okufunxayo, iingcongconi zafakwa ngabanye kwiityhubhu ze-1.5 ml ze-microcentrifuge eziqukethe i-230 µl yamanzi adibeneyo kwaye izicubu zaphazamiseka ngokusebenzisa i-pestle elahlayo kunye ne-cordless motor (VWR International, Lund, SE), ilandelwa yi-centrifugation kwi-10 krpm ye-10 min . i-ma-Aldrich) kunye ne-absorbeance (λ620) yamiselwa kusetyenziswa i-spectrophotometer-based microplate reader (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Ngenye indlela, iingcongconi zaziphantsi kwe-1 ml yamanzi adibeneyo, i-900 µl leyo idluliselwe kwi-spectrometer ye-UV600 ye-UV608; , Kista, SE) .Ukulinganisa ukutya okudliwayo, i-curve esemgangathweni yalungiswa nge-serial dilution ukuze ivelise i-0.2 µl ukuya kwi-2.4 µl ye-1 mg ml-1 i-xylene cyanide. Emva koko, ubuninzi be-optical optical concentrations yedayi eyaziwayo yayisetyenziselwa ukugqiba ubungakanani bokutya okufakwe kwi-mosquito nganye.
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). v8.0.0 for Mac, GraphPad Software, San Diego, CA, 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) . ltham, 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 kwi-lid (3 cm ububanzi) kunye Ukungena komoya kunye nokubonelela ngokutya. Ukutya kwanikezelwa ngokuthe ngqo emva kwe-4 kunye ne-concentration ye-4% kunye ne-cograted entsha kunye ne-dislu ukulawula, i-10% ye-sucrose kunye namanzi.Ukutya ngakunye kufakwe ngombhobho kwi-tampon yamazinyo (DAB Dental AB, Upplands Väsby, SE) ifakwe kwisirinji ye-5 ml (i-Thermo Fisher Scientific, Gothenburg, SE), i-plunger isusiwe, kwaye ibekwe phezu kwesitya se-petri . I-es yayibalwe kabini ngosuku, ngelixa i-mosquito efile yalahlwa kwaze kwaba yilapho i-mosquito yokugqibela yafa (n = 40 ngonyango) Ukusinda kweengcongconi ezondliwe kwizidlo ezahlukahlukeneyo kwahlalutywa ngokwezibalo kusetyenziswa i-Kaplan-Meyer i-curves yokusinda kunye neemvavanyo ze-log-rank ukuthelekisa ukulinganisa ukusabalalisa kokusinda. I-IBM20SS I-Diettistics.
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 iglu idinga L. d 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 ebotshiweyo kunye nengalo ephethe iqhekeza elincinci lefoyile yealuminiyam njengophawu lokuphazamisa ukukhanya.
Iimazi zeeyure ezingama-24 ezilambileyo zanikwa ukutya okungentla kwemizuzu ye-30 ngaphambi kokuthintela. Iingcongconi ezibhinqileyo ezondliwe ngokupheleleyo zaza zafakwa i-anesthetized emkhenkceni nge-2-3 min kwaye zifakwe kwi-insect pins kunye ne-bewax (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) kunye neengalo ezirekhodiweyo ze-Frizol kwi-recording ye-Frizol. ihlelwe yi-logger yedatha eyakhelwe ngokwezifiso, emva koko igcinwe kwaye iboniswe usebenzisa i-PC-Lab 2000 ™ software (v4.01; Velleman, Gavere, BE) .I-flight mill yafakwa kwigumbi elilawulwa yimozulu (12 h: 12 h, ukukhanya: kumnyama, 25 ± 2 ° C, 65 ± 5% RH).
Ukujonga umfanekiso wendlela yokusebenza kwendiza, umgama opheleleyo ohambayo (m) kunye nenani elipheleleyo lemisebenzi yendiza elandelelanayo ibalwa ngeyure kwisithuba seeyure ezingama-24. Ukongezelela, umyinge ophakathi oqhutywe ngamabhinqa ngamanye afaniswa kuwo wonke unyango kwaye uhlalutywe kusetyenziswa indlela enye ye-ANOVA kunye ne-Tukey's post hoc analysis (JMP Pro, v14.0.0), i-avareji ye-Avareji ye-Advant, i-Avareji ye-Add, i-Avareji ye-Add, i-Avareji ye-Add, i-Advance Inc. , inani eliqhelekileyo lemijikelo 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 igazi kwaye emva koko babonelele ngokutya kovavanyo lwe-48 h njengoko kuchazwe ngasentla.Izidlo zaye zasuswa kwaye iikomityi zokuzala (30 ml) zazaliswa ngeeyure ze-4 ze-distilled ze-20 ze-Nolato, i-20 ml yamanzi e-distilled ngosuku lwe-3 ye-Hertila. ukuguqula zonke iiyure ze-24. Phinda irejimeni yokutya nganye ngamaxesha angama-20-50. Amaqanda abalwe kwaye abhalwa kwikheji nganye yovavanyo.Iisampula zamaqanda zazisetyenziselwa ukuvavanya ubungakanani bentsingiselo kunye nobude benguqu yeqanda ngalinye (n ≥ 200 ngokutya ngokutya) usebenzisa i-microscope ye-Dialux-20 (DM1000; Wetzlarst aquitzD) I-Letzlarst, i-Ernstlar ye-equitzDFC, i-Letzlarst, i-Ernstlar ye-equitzDFC, i-Letzlar 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 apha ngasentla.Inani lamaqanda kunye nobukhulu bamaqanda kunye nezibungu zifaniswa phakathi kwonyango kunye ne-PANOVA ye-4, uhlalutyo lwe-4, i-v0, i-v0, i-MP, i-v0, i-v. SAS Institute Inc.).
I-Headspace volatiles evela kwi-fresh (iyure eyi-1 emva kokuthatha isampuli), i-24 hr, i-72 yeyure kunye ne-168 yomchamo omdala yaqokelelwa kwiisampuli eziqokelelwe kwiinkomo ze-Zebu, iintlanga ze-Arsi. Ukuze kube lula, iisampulu zomchamo zaqokelelwa kusasa ngelixa iinkomo zisesedlelweni. iingxowa (Toppits Cofresco, Frischhalteprodukte GmbH kunye ne-Co., Minden, DE) kwi-3 l polyamide enesiciko Kwi-vinyl chloride imigqomo yeplastiki. I-volatiles ye-headspace evela kwisampulu yomchamo wenkomo nganye yaqokelelwa ngokuthe ngqo (entsha) okanye emva kokuvuthwa kwiqondo lokushisa kwegumbi le-24 h, i-isampuli ye-72 ye-urine nganye kunye ne-168 yeqela ngalinye.
Ukuqokelela i-headspace volatiles, inkqubo ye-loop evaliweyo yayisetyenziselwa 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-polyamide ye-polyamide yenziwe nge-polyamide engenanto. I-tubing (5.5 cm x 3 mm id) equkethe i-35 mg ye-Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) phakathi kweeplagi zoboya beglasi.Ngaphambi kokusetyenziswa, ikholamu yayihlanjululwe nge-1 ml i-redistilled n-hexane (Merck, Darmstadt, DEGRE), kunye ne-9 mld GC i-puresol, i-9 mld, i-puree, i-9 mld GC, i-puresol, i-1. Ukuguquguquka kweebhedi kuye kwakhutshwa nge-400 μl ye-pentane.Iingqokelela zezithuba zentloko ziye zadityaniswa zaza zagcinwa ku -20°C de zisetyenziswe kuhlalutyo olongezelelweyo.
Iimpendulo zokuziphatha zokufuna umkhosi kunye nokutya igazi I-An.Headspace isicatshulwa esiguquguqukayo esiqokelelwe kwi-fresh, i-24-h, i-72-h, kunye ne-168-h-agugile umchamo yahlalutywa kwiicatshulwa eziguquguqukayo ezivela kwiimiyane ze-Arabidopsis usebenzisa i-tube yeglasi eqondileyo i-olfactometer [18]. [19] .I-tube yeglasi ye-olfactometer (80 cm × 9.5 cm id) yakhanyiswa nge-3 ± 1 lx yokukhanya okubomvu ukusuka phezulu. I-Charcoal ehluziweyo kunye ne-humidified air flow (25 ± 2 ° C, i-65 ± 2% i-humidity ehambelanayo) idlulise i-bioassay kwi-30 cm ye-steel ye-stainless, idlulisa i-serial ye-stainless kwi-30 cm. isakhiwo se-plume esifanayo.Isixhobo sokuhambisa i-tampon yamazinyo (4 cm × 1 cm; L: D; DAB Dental AB), inqunyanyiswe kwi-coil ye-5 cm ekupheleni kwe-olfactometer yomoya, kunye ne-stimulator utshintsho rhoqo ngemizuzu emi-5. Uhlalutyo, i-10 μl yesicatshulwa se-headspace nganye, ihlanjululwe 1:10, isetyenziswe ukulawulwa kwe-hostee elinganayo njenge-pensee. okanye iingcongconi ezifunxa igazi zifakwe kwiikheji zokukhululwa komntu ngamnye kwiiyure ze-2-3 ngaphambi kokuqala kovavanyo.I-cage yokukhulula ibekwe kwicala le-downwind ye-olfactometer, kunye neengcongconi zavunyelwa ukuba zilungelelanise i-1 min, kwaye emva koko i-valve butterfly ye-cage yavulwa ukuba ikhulule. I-headspace i-extract volatile extract kunye nokulawulwa kwaphinda kwahlaziywa ubuncinane ngamaxesha e-30, kwaye ukuphepha imiphumo nayiphi na imini enye, inani elifanayo lonyango kunye nokulawulwa kwavavanywa ngosuku ngalunye lovavanyo.Fumana iimpendulo ezivela kwi-host kunye ne-Ans.I-Arab kunye neeseti ze-headspace zahlaziywa ngokusebenzisa i-nominal logistic regression elandelwa yi-pairwise comparisons for odd Institute, v14.
Impendulo yokuzala kwe-An. Izicatshulwa ze-Headspace ezivela kumchamo wenkomo omtsha kunye nomdala zahlalutywa kwiikheji ze-Bugdorm (30 cm × 30 cm × 30 cm; i-MegaView Science) .Iikomityi zeplastiki (30 mL; Nolato Hertila) zizaliswe nge-20 mL yamanzi adiyiweyo zinike indawo yokuzala, i-substrate edibeneyo ye-2 yekomityi yabekwa kwikona ye-2 ye-sparked kwikona kunye ne-2 ye-cuatment ye-cup kunye ne-2 ye-substrate ye-cuat kunye ne-2 ye-squat ye-cup. I-10 μl yesicatshulwa ngasinye se-headspace kwi-1: 10 dilution.Umlinganiselo olinganayo wepentane wasetyenziselwa ukulungelelanisa indebe yokulawula.Unyango kunye neekomityi zokulawula zatshintshiswa phakathi kovavanyo ngalunye ukulawula imiphumo yesimo.Abafazi abalishumi begazi bakhululwe kwiikheji zokulinga kwi-ZT 9-11 kunye namaqanda kwiikomityi abalwa kwiikomityi ze-24 emva koko. amaqanda abekwe kwikomityi yokulawula)/(inani lilonke lamaqanda abekiweyo).Unyango ngalunye lwaphindwa izihlandlo ezisi-8.
Uhlalutyo lwegesi yechromatographic kunye ne-electron ye-elektroni yokukhangela ipateni (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, ifilimu ye-5 × 5 × 5 × ifilimu ye-5 × 30m. iikhnoloji).kunye nomchamo ogugayo.I-Hydrogen yasetyenziswa njengesigaba esihambahambayo kunye nesantya sokuhamba kwe-avareji yomgca we-45 cm s-1.Isampuli nganye (2 μl) yatofwa imizuzwana engama-30 kwimowudi engacalulwanga kunye neqondo lokushisa lokungena lika-225 °C. Ubushushu be-oven ye-GC yacwangciswa ukusuka kwi-35 °C (imizuzu emi-3 ibambe) ukuya kwi-3000 ngomzuzu we-GC kwi-1000 °C ibamba i-1000 °C ngomzuzu. , I-4 psi ye-nitrogen yongezwa kwaye yahlula i-1: i-1 kwi-Gerstel 3D / 2 i-low-dead volume cross (Gerstel, Mülheim, DE) phakathi kwe-flame ionization detector kunye ne-EAD.I-GC effluent capillary ye-EAD idluliselwe kumgca wokudlulisa iGerstel ODP-2, elandelela i-oven ye-GC, i-oven ye-GC, i-oven ye-GC, i-oven ye-GC, i-oven edibeneyo ye-0, i-carbon edibeneyo kunye ne-5 × i-intshi ye-5 × , i-tube edibeneyo yeglasi kunye ne-5 × . i-humidified air (1.5 l min-1) .I-antenna ifakwe kwi-0.5 cm ukusuka kwi-outlet ye-tube.Ingcongconi nganye iphendule enye i-replicate, 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-UGCIMS ye-silili ye-70 e-equipped e-U. ikholamu yellari (i-60 m × 0.25 mm ububanzi bangaphakathi, 0.25 μm ubukhulu befilimu) kusetyenziswa i-helium njengenqanaba elihambayo elinomlinganiselo wokuhamba komndilili ophakathi kwama-35 cm s-1.A 2 μl isampulu yatofwa kusetyenziswa useto lwe-injector efanayo kunye nobushushu be-oveni njengohlahlelo lweGC-EAD. Iicompounds zaye zachongwa ngokusekwe kwilayibrari ye-NISTvten kunye nexesha eliqhelekileyo lokujonga ilayibrari ye-NISTvten (Agilent) .Iikhompawundi ezichongiweyo ziqinisekiswa ngokujova imigangatho eyiyo (iFayile eyoNgezelelweyo 1: Itheyibhile S2) .Ngobungakanani, i-heptyl acetate (10 ng, 99.8% yokuhlanjululwa kweekhemikhali, i-Aldrich) yajongwa njengomgangatho wangaphandle.
Ukuvavanya ukusebenza komxube wokwenziwa kwevumba elibandakanya iikhompawundi ze-bioactive ezichongiweyo kumchamo omtsha kunye nomdala ukutsala umkhosi-ukufuna kunye nokufunxa igazi i-Ans.arabiensis, usebenzisa i-olfactometer efanayo kunye neprotocol njengangentla.Imixube ye-Synthetic yalinganisa ukubunjwa kunye nobungakanani beekhompawundi kwi-headspace edibeneyo i-extracts eguqukayo ye-freshur, 6-24ho-24ho-24ho-24ho-ho-24ho-24ho umchamo omdala (Umfanekiso 5D-G; Ifayile eyongezelelweyo 1: Itheyibhile S2). Uhlalutyo, sebenzisa i-10 μl ye-1: 100 dilution yomxube owenziwe ngokupheleleyo, kunye nomlinganiselo wokukhulula ngokubanzi osuka malunga ne-140-2400 ng h-1, ukuvavanya umtsalane wokubamba kunye nokufunxa igazi, imixube yeengcongconi ezigqityiweyo emva komxube owenziweyo ogqityiweyo. umxube uyasuswa.Fumana iimpendulo kwinginginya kunye nezondliwa ngegazi i-Ans.Arab vs synthetic and subtractive mixes zahlalutywa kusetyenziswa uhlengahlengiso lwegama elilandelwa luthelekiso lwe-pairwise 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, DE).Izibatha ezilishumi zabeka i-50 m phakathi kwamadlelo, i-400 m ukusuka ekuhlaleni kwidolophana (i-Silay, i-Ethiopia, 5°53´24′′N, 37°29′24′′E) kwaye akukho nkomo, kwiindawo zokuzalela ezisisigxina kunye nakwiilali. ly ubusuku obuhlanu buphelele.Amanani engcongconi abanjwe kwimigibe ebanjwe ngumchamo weminyaka eyahlukeneyo ubudala athelekiswe kusetyenziswa ukuhlehliswa kwezinto kunye nokusasazwa 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 naphakathi kuSeptemba phambi kokutshizwa kwentsalela yangaphakathi ngonyaka, kunye nexesha elide lemvula. (Umfanekiso 6A) .Iikhrayitheriya ezisetyenziselwa ukukhetha izindlu zaziyi: akukho zilwanyana ezivunyelwe endlwini, akukho ukupheka kwangaphakathi (ukuzoba iinkuni okanye amalahle) kuvunyelwe (ubuncinci ngexesha lovavanyo), kunye nezindlu ezinabemi ababini abaninzi, balala kwi-uninsecticides.phantsi komnatha ophathwayo weengcongconi.Imvume yokuziphatha inikwe yiBhodi yeZiko loPhando lweeNqobo 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 lwezempilo yesithili yonke ifunyenwe kwintloko yabasebenzi bendawo yonke. kunye newadi ('kebele') inqanaba.Uyilo lokuhlola lulandele i-2 × 2 yesiLatini uyilo lwesikwere, apho imixube yokwenziwa kunye nolawulo lwabelwa izindlu ezidibeneyo ngobusuku bokuqala kwaye zatshintshwa phakathi kwezindlu ngobusuku obulandelayo bokulinga.Le nkqubo iphindwe kalishumi.Ukongezelela, ukuqikelela umsebenzi wengcongconi kwizindlu ezikhethiweyo, izibambo ze-CDC zabekwa ukuba ziqhube phakathi kweentsuku ezintlanu zobusuku obulandelelanayo ngexesha elifanayo lexesha lokulingwa.
Umxube owenziweyo oqukethe i-bioactive compounds ezintandathu zachithwa kwi-heptane (i-97.0% ye-solvent grade GC, i-Sigma Aldrich) kwaye yakhululwa kwi-140 ng h-1 isebenzisa i-cotton wick dispenser [20]. Ukuthintela (CDC) umgibe wokukhanya (uJohn W. Hock Inkampani, iGainesville, FL, US; Umzobo 6A) .Imigibe yaxhonywa kwi-0.8 - 1 m ngaphezu komhlaba, kufuphi nonyawo lwebhedi, kwaye ivolontiya yalala phantsi kwenethi engaphendulwanga yomiyane kwaye iqhutywe phakathi kwe-18: 00 kunye ne-06: i-modfesid kunye ne-06: i-mosquid kunye ne-06: i-mi-pregnant, kunye nokukhulelwa [21] emva koko bahlolwe ngokusebenzisa i-polymerase chain reaction (PCR) uhlalutyo ukuchonga iintlobo ze-morphologically ezichongiweyo njenge-A. gambiae sl.0. SAS Institute Inc.).Apha, sixela i-χ2 kunye ne-p-amaxabiso ukusuka kuvavanyo lomlinganiselo wokunokwenzeka.
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 iimazi ezifuna umkhosi kunye negazi zifunxa kakhulu i-sucrose ngaphezu kwayo nayiphi na enye i-sucrose (F60 = 20, i-20, i-20, i-5, i-5, i-2, i-p. I-01 kunye ne-F (5,299) = 56.00, p <0.0001, ngokulandelanayo; Umzobo 1B, C) .Ngaphezu koko, abafazi abafuna umkhosi badla ukutya okuncinci kumchamo kwiiyure ze-72 xa kuthelekiswa nomchamo kwiiyure ze-168 (umzobo 1B) .Xa unikezwa ukutya okuqukethe i-urea, i-host-ifuna i-6 i-concentration xa ithelekisa i-mx ye-concentration ye-6 ithelekisa yonke i-m2 ye-concentration kunye ne-mx . Ukungabonakali kwi-10% ye-sucrose (F (10,813) = 15.72, p <0.0001; Umfanekiso we-1D) .Oku kwakuchasene nempendulo yabasetyhini abatya igazi, abadla ngokuthatha ngokubonakalayo ukutya okuqukethe i-urea kunamanzi, nangona kuncinci ngaphantsi kwe-10% = 0,510; 510; 10% 0,51 (70,51; 51; 51; i-gure 1) .1E) .Ngaphezu koko, xa kuthelekiswa phakathi kwamazwe amabini e-physiological, amabhinqa e-phlebotomized athatha i-urea engaphezulu kuneentombi ezifuna umkhosi kwiindawo eziphantsi kakhulu, kwaye ezi zibhinqa zithatha inani elifanayo le-urea kwiindawo eziphezulu (F (1,953) = 78.82, p <0.0001;Umzobo we-1F, G) Ngelixa ukuthathwa kokutya okune-urea kubonakala kunemilinganiselo efanelekileyo (Umfanekiso we-1D, E), amabhinqa kuwo omabini amazwe e-physiological akwazi ukumodareyitha isixa se-urea efunxwe kulo lonke uluhlu lokugxilwa kwe-urea kwi-log-linear fashion (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 efunxa igazi, amabhinqa asondliwa ngomchamo wabo bonke ubudala obune (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 olubonisa ukuba i-figure yokulawula i-2 ibonise i-sucrose yokulawula i-2. ukusinda kubafazi abafuna umkhosi (umchamo: χ2 = 108.5, df = 5, p <0.0001; urea: χ2 = 122.8, df = 5, p <0.0001; Umzobo 2B, C) kunye nabasetyhini abanegazi (2,0 = 3,0 = f χ.0,0 = 1,0 = 9 = f. Urea: χ2 = 137.9, df = 5, p <0.0001; Umfanekiso 2D, E).Kuzo zonke iimvavanyo, amabhinqa atyisa ukutya komchamo, urea, kunye namanzi ayenamazinga okuphila aphantsi kakhulu xa kuthelekiswa nabasetyhini abondla ukutya kwe-sucrose (Umfanekiso we-2B-E). -h umchamo we-stale (p = 0.016) onethuba eliphantsi lokusinda (Umfanekiso we-2B) .Ngaphezu koko, abafazi abafuna umkhosi bondla i-135 mM urea baphila ixesha elide kunolawulo lwamanzi (p <0.04) (Fig.I-2C) .Xa kuthelekiswa namanzi, abafazi abondliwe ngomchamo omtsha kunye nomchamo weeyure ze-24 baphila ixesha elide (p = 0.001 kunye p = 0.012, ngokulandelanayo; Umfanekiso we-2D), ngelixa abafazi bondliwa ngomchamo weeyure ze-72 baphila ixesha elide kunezo zondliwe Umchamo omfutshane kunye ne-24-iyure kunye ne-24-iyure kunye ne-24-iyure kunye ne-0. i-gure 2D) .Xa isondliwa i-135 mM urea, amabhinqa anegazi ahlala ixesha elide kunawo onke amanye amanqanaba e-urea kunye namanzi (p <0.013; Umfanekiso 2E).
Ukusinda komkhosi kunye nemazi efunxa igazi i-Anopheles arabinis etyisa kumchamo wenkomo kunye ne-urea.Kwi-bioassay (A), iingcongconi ezimazi zibonelelwe ngokutya okuquka umchamo omtsha nomdala wenkomo, imilinganiselo eyahlukeneyo ye-urea, i-sucrose (10%) kunye namanzi adibeneyo (H2O). iiyure de zonke iimazi ezondliwa ngomchamo (B, D) kunye ne-urea (C, E), kunye nolawulo, i-Sucrose kunye namanzi, zifile.
Umgama opheleleyo kunye nenani leemijikelo ezinqunywe kuvavanyo lokusila ngenqwelomoya kwisithuba seeyure ezingama-24 zahluka phakathi kweengcongconi ezifuna umkhosi kunye negazi, ezibonisa umsebenzi omncinci wokundiza ngokubanzi (umzobo 3) .Iingcongconi ezifuna umkhosi ezibonelela ngomchamo omtsha kunye nomdala okanye i-sucrose kunye namanzi abonisa iipatheni zokubhabha ezahlukileyo (umzobo 3), kunye ne-female ye-18 esebenzayo kunye ne-18 entsha, kunye ne-feines 2 esebenzayo, kunye ne-18 ye-female esebenzayo, kunye ne-fig. Iingcongconi eziziimazi ezinikezela nge-sucrose okanye umchamo weeyure ezingama-72 zabonisa umsebenzi kulo lonke ixesha leeyure ezingama-24, ngelixa iimazi ezinikezela ngamanzi zazisebenza ngakumbi ngexesha eliphakathi. andifumananga ukwehla okuthe chu kumsebenzi kwiiyure ezingama-24 (Umfanekiso 3).
Kuvavanyo lwe-flight mill, iingcongconi ezimazi zityiswe ngomchamo omtsha nomdala wenkomo, i-urea, i-sucrose (10%), kunye namanzi adiyiweyo (H2O) abotshelelwe kwindawo ethe tyaba, ejikeleza ngokukhululekileyo, ejikelezisa iingalo zemazi. umgama opheleleyo kunye nenani leenqwelomoya ngeyure kwisidlo ngasinye kwisithuba seeyure ezingama-24 zarekhodwa (kumnyama: ngwevu; ukukhanya: okumhlophe). Umyinge womgama kunye nenani eliqhelekileyo lokuhamba liboniswa ngasekunene kwegrafu yomsebenzi we-circadian.Iimpazamo zemivalo zibonisa impazamo eqhelekileyo yentsingiselo.Uhlalutyo lwamanani jonga umbhalo.
Ngokuqhelekileyo, umsebenzi wendiza jikelele wabasetyhini abafuna umkhosi ulandele iphethini efana neyomgama wendiza ngexesha leeyure ze-24. Umgama wokuhamba ngenqwelomoya wawuchaphazeleka kakhulu ngokutya okutyayo (F (5, 138) = 28.27, p <0.0001), kunye nabasetyhini abafuna umkhosi bangene kwiiyure ze-72 zomchamo zahamba ngokuphawulekayo ezinye iimda ezide kakhulu kunye ne-moquid 0000 xa kuthelekiswa nayo yonke imigama emide ye-moquid. I-es yahamba ixesha elide kune-fresh (p = 0.022) kunye ne-24-h-umchamo oneminyaka engama-24 (p = 0.022) -imiyane ezondliwe. Ngokwahlukileyo kwindlela yokusebenza yendiza echazwe kukutya komchamo, abafazi abafuna i-urea-fed bafuna i-urea babonise umsebenzi oqhubekayo wokuhamba ngendiza kwixesha le-24-h, i-peaking ngexesha le-24-h, i-peaking ngexesha lesigaba sesibini se-female, i-fig ye-fig-fig-fig-figment, i-fig ye-fig-fig. irea yanda kakhulu umgama wokubhabha ngokuxhomekeke kugxininiso olufunxayo (F (5, 138) = 1310.91, p <0.0001) .Abafazi abafuna umkhosi bondla nayiphi na inxinaniso ye-urea ibhabha ixesha elide kunokuba abafazi besondla amanzi okanye i-sucrose (p <0.03).
Umsebenzi wendiza jikelele weemiyane ezifunxa igazi zazinzile kwaye zigcinwe kwiiyure ze-24 kuzo zonke izidlo, kunye nokwanda komsebenzi womchamo ngexesha lesibini lexesha elimnyama labasetyhini abondliwe emanzini kunye nabasetyhini abondliwe amatsha kunye neeyure ze-24 ubudala (umfanekiso we-3) .Ngelixa ukutya komchamo kuchaphazeleke kakhulu umgama wokubhabha kwi-female ezondliwa ngegazi (F.8, 0 = 4, 0 = 3 Urea = 3 Urea = 3, 0 = 4, 4) (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 = 0.0017; dH6 = 0.30, p = 0.3, p.
Imiphumo yokutya komchamo kunye ne-urea kwiiparameters zokuzala zavavanywa kwii-bioassays zokubeka iqanda (Umfanekiso 4A) kwaye zaphandwa ngokwenani lamaqanda abekwe ngowesifazane ngamnye, ubungakanani beqanda, kunye nezibungu ezisandul 'ukuqanduselwa.Inani lamaqanda abekwe. B) .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) .Ngokufanayo, ubungakanani bamaqanda abekwe ngamabhinqa atyalwe ngumchamo ahluka ngokutya (F (5, 209) = 12.85, p0-2000, p0-10, p0-10, p0-209, p0-urine-urine-urine-hour, i-urine-uringyi-0. amaqanda amakhulu kakhulu kunabafazi abondliwe ngamanzi, ngelixa amaqanda amabhinqa asondliwa nge-168 h yomchamo ayencinci kakhulu (umzobo 4C) .Ukongezelela, ukutya komchamo kuchaphazele kakhulu ubungakanani be-larval (F (5, 187) = 7.86, p <0.0001), kunye ne-larvae enkulu kakhulu ephuma kwi-24 yamaqanda e-urine-laid ephuma kwi-24 yamaqanda e-urine-feed-laid ephuma kwi-24 yamaqanda e-urine. ukusuka kwimibungu yamaqanda.Zondliwe ngamanzi kunye neemazi eziyi-168-h ezondliwa ngumchamo (Umfanekiso 4D).
Ukusebenza kokuzala kwemazi ye-Anopheles arabinis etyisa umchamo wenkomo kunye ne-urea.Iingcongconi zemazi ezondliwa ngegazi zatyiswa ukutya okubandakanya umchamo wenkomo omtsha kunye nomdala, ubuninzi be-urea, i-sucrose (10%), kunye namanzi adibeneyo (H2O) kwiiyure ezingama-48 ngaphambi kokubeka kwii-bioassays kunye neeyure ze-egg (i-egg-egg kunye nokufumana inombolo ye-Egg, i-Egg-iyure) , F) kunye nobukhulu beembungu (D, G) zichaphazeleke kakhulu ngokutya okubonelelweyo (umchamo wenkomo: BD; urea: EG) . Iindlela zeparameter nganye ezilinganiswe ngokusebenzisa amagama ahlukeneyo oonobumba zahluke kakhulu komnye nomnye (indlela enye i-ANOVA isebenzisa uhlalutyo lwe-post hoc ye-Tukey; p <0.05) .
Njengenxalenye enkulu ye-nitrogen yomchamo, i-urea, xa inikezelwa njengokutya kwabasetyhini abondliwa ngegazi, ichaphazele kakhulu iiparameters zokuzala 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-female i-3M i-3M idla phakathi kwe-3M amaqanda angaphezulu kwe-3M 30001, i-concentrations ye-3M ¥ i-4M i-1M idla i-1M urea ngaphezulu. Umzobo 4E) .Abafazi abondliwa kumlinganiselo we-urea we-134 µM okanye ngaphezulu beka amaqanda amakhulu kuneemazi ezondliwe emanzini (F (10, 4245) = 36.7; p <0.0001; Umfanekiso 4F), kunye nobukhulu bombungu, nangona buchatshazelwa yimilinganiselo efanayo ye-urea koomama, i-0 3 (0) = 3 (F.19) = 3 (F.19) = 0 (0) yayiguquguquka ngakumbi (Fig. 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 ye-24 yohlalutyo oluphezulu lubonise ukuba uhlalutyo oluphezulu lwe-24 lubonisa ukuba uhlalutyo oluphezulu lwe-24 lwe-urine lubonisa ukuba i-stale ye-hocdor yonyango ifanise kakhulu. (iiyure ezingama-72: p = 0.0060, iiyure ze-168: p = 0.012, pentane: p = 0.00070), Ngaphandle kwephunga lomchamo omtsha (p = 0.13; Umfanekiso 5B) .Nangona umtsalane jikelele weemiyane ezifunxa igazi kwi-urine ivumba lalingekho , 6 = 7 = 0, p8 = 0, p8 = 0, p. g. 5C), aba bafazi bafunyaniswa ukuba banomtsalane ngakumbi kwi-headspace isicatshulwa esiguquguqukayo xa kuthelekiswa nomchamo we-72 weyure xa kuthelekiswa nolawulo (p = 0.0066; Umfanekiso 5C).
Iimpendulo zokuziphatha kwivumba lomchamo wenkomo yendalo kunye neyokwenziwa kukhangelo lomkhosi kunye negazi le-Anopheles arabianus.Schematic of the glass tube olfactometer (A) .Umtsalane we-headspace isicatshulwa esiguquguqukayo esivela kumchamo wenkomo omtsha kunye nomdala ukuya kubamba (B) kunye negazi lokufunxa igazi (C) kwiingcongconi.Fumana i-4 ye-Actres ye-Anducle ye-Lord (E), i-72-iyure (F), kunye ne-168-iyure (G) i-urine yenkomo endala ibonisiwe.Ukukhangela kwe-elektroni ye-eriyali (EAD) kubonisa utshintsho lombane ekuphenduleni i-bioactive compounds kwi-headpace ekhutshwe kwi-chromatograph yegesi kwaye ifunyenwe ngumtshini we-ionization yomlilo (FID) . I-h-1) yeekhompawundi ze-biologically ezisebenzayo zibonisiwe.I-asterisk enye (*) ibonisa impendulo ehambelanayo ye-amplitude ephantsi.Iiinkwenkwezi ezimbini (**) zibonisa iimpendulo ezingenakulinganiswa.Fumana inginginya (H) kunye ne-blood-sucking (I) I-Arabiensis inezibheno ezahlukeneyo kwimixube yokwenziwa komchamo ococekileyo kunye namagama akhulileyo. ezahlukeneyo omnye komnye (indlela enye ANOVA usebenzisa Tukey uhlalutyo post hoc;p <0.05) .Iimpazamo zemivalo zibonisa imposiso eqhelekileyo yesikali
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 zafumanisa iimpendulo ze-electrophysiological, ininzi yale michiza yayikhona kwi-headspace nganye isicatshulwa esiguquguqukayo esiqokelelwe kwi-fresh and aged compounds eqokelelwe kwi-extract fresh and aggiant , i-extracts ye-extracts ye-female kunye ne-extracts engaphezulu kwi-extract yomchamo we-female, i-extracts ye-extracts ephuma kwi-extracts ye-female. umqobo wabandakanywa kuhlalutyo olongezelelweyo.
Ixabiso elipheleleyo lokukhululwa kwe-bioactive compounds kwiqoqo le-headspace linyuke 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. ubudala bomchamo, obuhambelana nokuhla okubonwayo kwi-signal intensity (ubuninzi) kwi-chromatogram (Umfanekiso we-5D) -G ipaneli yasekhohlo) kunye neempendulo ze-physiological kule ngqungquthela (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-1, i-moquisf = i-2H = i-5H) 4, p = 0.30; Umfanekiso we-5I) .Nangona kunjalo, uthelekiso lwe-post hoc pairwise phakathi kwezonyango lubonise 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 eyongezelelweyo kwi-Y-tube assay, apho iikhompawundi zomntu ngamnye zaye zasuswa. Kuba iingcongconi ezifuna umkhosi, ukukhupha iikhompawundi zomntu ngamnye kumxube opheleleyo kunempembelelo ebalulekileyo kwiimpendulo zokuziphatha (χ3 p 60 = ifayile ye-1 = 1 = 1 = 0 ; I-1: Umzobo we-S2A), yonke imixube ekhuphayo yayithandeka ngakumbi kuneNcinane kunokuxutywa ngokupheleleyo.Ngokwahlukileyo, ukususwa kweekhompawundi zomntu ngamnye kumxube wokwenziwa ngokupheleleyo akuzange kuthintele iimpendulo zokuziphatha kweemiyane ezifunxa igazi (χ2 = 11.38, df = 6, p = 0.077), ngaphandle kwe-0. : Umfanekiso we-S2B).
Kwilali ye-malaria e-Ethiopia, ukusebenza komxube wokwenziwa komchamo wenkomo yeeyure ezingama-24 ekutsaleni iingcongconi phantsi kweemeko zentsimi kwavavanywa ubusuku obulishumi (umzobo 6A) .Itotali ye-4,861 yemiyane yabanjwa kwaye ichongiwe, apho i-45.7% yayiyi-Anthropus%, i-Anoroen 18% kunye ne-Anthropus. Culex spp.(Ifayile eyongezelelweyo 1: Itheyibhile S1).Anopheles arabinis kuphela kwelungu An.Gambian iintlobo complex ezichongwe ngohlalutyo PCR.Ngomndilili, 320 iingcongconi zabanjwa ngobusuku ngabunye, ngelo xesha imigibe kunye izithiyelo zokwenziwa umxube babambisa iingcongconi ngaphezu idityaniswe imigibe ngaphandle komxube, χ0120 = 0110 = 0120. .Imigibe engabonakaliyo ibekwe kubusuku obuhlanu bokulawula ekuqaleni, phakathi, nasekupheleni kwetyala.Amanani afanayo omiyane abanjwe kwibini ngalinye lemigibe, ebonisa ukuba akukho cala phakathi kwezindlu (χ2 (0, 1665) = 9 × 10-13, p> 0.05 yehla, i-0.05 iyancipha kwi-traps kunye nenani le-traps ebanjwe kwi-traps kunye nenani labantu. izibatha eziqulethe umxube wokwenziwa zanda kakhulu: ukufuna inginginya (χ2 (0, 2107) = 138.7, p <0.0001), ukutya okutsha nje kwegazi (χ2 (0, 650) = 32.2, p <0.0001) kunye nokukhulelwa (χ2 (0, 227) = 2.0 = 2.0 = 2.0 = 6.Ifayile eyongezelelweyo 1: Itheyibhile S1) .Oku kubonakaliswe kwinani elipheleleyo leengcongconi ezithinjiweyo: i-host looking> bloodsucking > ikhulelwe > 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 ojikelezayo weCDC) kunye ne-Ador-Bphelei arabeleation ye-CDC (umfanekiso we-CDC) obhinqileyo, i-Ador-Baphelei i-Anocotral kodwa hayi Anopheles farroes (C), ngendlela eyahlukileyo, i physiological state-ezixhomekeke isiphumo.Ukongezelela, le migibe babanjwa ngokuphawulekayo anda amanani omkhosi Culex iingcongconi.(D) Xa kuthelekiswa control.I imivalo ngasekhohlo imele i-avareji yokukhetha index iingcongconi abanjwe ngababini of odorant bait (eluhlaza) kunye nolawulo (i-avareji yokukhetha i-i-avareji ye-Nu) kunye nolawulo (i-avareji yokukhetha i-open = i-open) ukulawula imigibe (evulekileyo; N = 5).).Iinkwenkwezi zibonisa amanqanaba okubaluleka kweenkcukacha-manani (*p = 0.01 kunye ne-***p <0.0001)
Ezi ntlobo zintathu zibanjwe ngokwahlukileyo kwimigibe equlethe imixube eyenziweyo. 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, 6, p. sis wayevaleleke emgibeni ekhulula umxube wokwenziwa (umzobo 6B), ngelixa inani le-An alizange lihluke.I-Pharoensis kwiimeko ezahlukeneyo zomzimba zafunyanwa (umzobo 6C) . Kuba Culex, kuphela ukwanda okubalulekileyo kwinani leengcongconi ezifuna imikhosi yafunyanwa kwimigibe exutywe kunye nomxube wokwenziwa, i-p120 = i-p10 = i-p121 = i-p121 = i-p120 = i-p10 = i-p10; 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 zemalariya zisebenzisa ivumba lomchamo wenkomo njengendawo yokuhlala yokuhlala. i-ed kwaye ibanjwe, nangona amanani amancinci, azimeleyo 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]. spawning sites.Iingcongconi Female indawo lo vimba ngevumba kwaye bayakwazi ukulawula ukufunyaniswa iikhompawundi nitrogen kumchamo, kuquka urea, icandelo nitrogenous enkulu kumchamo [15, 16].Kuxhomekeka ingcongconi physiological imo, izondlo emchameni zabelwa umsebenzi wokuphila njengeengcongconi nokuphucula ukuphila, ingcongconi inika umsebenzi kunye nokuphucula i-mosquito. kunye neempawu zokuzala zabantu abondliwa ngegazi ngexesha lomjikelezo wokuqala we-gonadotropic.Ngoko ke, ukuxuba umchamo kudlala indima ebalulekileyo yesondlo kwii-malaria ze-malaria ezivaliweyo njengabantu abadala abangondlekanga [8], njengoko ibonelela ngeengcongconi zabasetyhini ngokukwazi ukufumana iikhompawundi ezibalulekileyo ze-nitrogen ngokubandakanyeka kwisondlo esisezantsi. ngakumbi, le ndlela yokuziphatha inokuba kujoliswe kuyo kwiinkqubo zolawulo lwe-vector yexesha elizayo.