Kev tau txais cov khoom noj khoom haus thiab kev faib khoom sib xyaw ua ke cov kab foraging thiab lub neej keeb kwm zoo.Yuav kom them nyiaj rau qhov tsis txaus ntawm cov as-ham ntawm lub neej sib txawv, kab tuaj yeem tau txais cov as-ham los ntawm kev pub mis ntxiv, piv txwv li, los ntawm kev pub mis rau vertebrate secretions nyob rau hauv cov txheej txheem hu ua puddles.Cov yoov tshaj cum Anopheles arabiani zoo nkaus li yog li ntawd, ob qho tib si kev tshawb fawb thiab kev noj qab haus huv. sess seb An.arabiensis agitation ntawm nyuj tso zis rau cov khoom noj khoom haus txhim kho lub neej keeb kwm yam ntxwv.
Xyuas kom tseeb tias nws muaj kev nyab xeeb.arabiensis tau ntxim rau cov ntxhiab tsw tshiab, 24-teev, 72-teev, thiab 168-teev laus nyuj tso zis, thiab tus tswv tsev nrhiav thiab noj ntshav (48-teev tom qab ntshav qab zib) cov poj niam tau ntsuas hauv Y-tube olfactometer, thiab cov poj niam cev xeeb tub tau siv tshuaj ntsuam xyuas biochemical. Cov active compounds nyob rau hauv nyuj tso zis nyob rau hauv tag nrho plaub lub hnub nyoog chav kawm.Synthetic mixs ntawm bioactive compounds raug soj ntsuam nyob rau hauv Y-tube thiab teb trials.To soj ntsuam nyuj cov zis thiab nws lub ntsiab muaj nitrogen-muaj compound urea raws li tej zaum yuav supplemental diets rau malaria vectors, pub tsis tau thiab lub neej keeb kwm yam ntxwv tau ntsuas. Cov poj niam raug soj ntsuam rau kev ciaj sia, tethered davhlau, thiab kev yug me nyuam.
Nrhiav rau tus tswv tsev cov ntshav thiab khoom noj khoom haus.Nyob rau hauv lub chaw soj nstuam thiab cov kev tshawb fawb, Arabs tau kos rau lub ntuj thiab hluavtaws tsw ntawm cov nyuj tshiab thiab cov laus tso zis. Cov poj niam cev xeeb tub tsis xav txog cov lus teb rau nyuj tso zis ntawm qhov chaw spawning.Host-nrhiav thiab ntshav nqus cov poj niam no nquag nqus nyuj tso zis ntawm lub neej thiab kev ua haujlwm as-physiosa. ological xeev rau ya davhlau, ciaj sia taus, los yog yug me nyuam.
Anopheles arabinis tau txais thiab kev faib tawm ntawm nyuj cov zis rau kev txhim kho lub neej keeb kwm yam ntxwv.Kev pub mis ntxiv ntawm cov zis nyuj cuam tshuam rau vector muaj peev xwm ncaj qha los ntawm kev ua kom muaj sia nyob txhua hnub thiab vector ntom, thiab tsis ncaj los ntawm kev hloov pauv kev ua si thiab yuav tsum yog li ntawd yuav tsum tau txiav txim siab hauv cov qauv yav tom ntej.
Kev tau txais cov khoom noj thiab kev faib khoom sib xyaw ua ke cov kab foraging thiab lub neej keeb kwm yam ntxwv [1,2,3].Cov kab muaj peev xwm xaiv thiab tau txais cov zaub mov thiab ua cov khoom noj khoom haus raws li cov zaub mov muaj thiab cov khoom noj khoom haus [1, 3].Kev faib cov as-ham nyob ntawm cov txheej txheem ntawm kev ua neej nyob thiab tej zaum yuav ua rau muaj kev sib txawv ntawm kev noj haus zoo thiab ntau theem ntawm lub neej. Hauv cov khoom noj tshwj xeeb, kab tuaj yeem tau txais cov khoom noj los ntawm kev pub mis ntxiv, xws li hauv av nkos, ntau yam excrement thiab secretions ntawm vertebrates, thiab carrion, txheej txheem hu ua puddles [2].Txawm hais tias ntau hom kab npauj npaim thiab npauj tau piav qhia, watering qhov kuj tshwm sim nyob rau hauv lwm yam kab txiav txim, thiab attraction ntawm lub neej thiab kev noj qab haus huv hom. 4, 5, 6] ,7].Tus malaria yoov tshaj cum Anopheles gambiae sensu lato (sl) tshwm sim raws li ib tug neeg laus 'malnourished' [8], yog li watering yuav ua tau ib tug tseem ceeb luag hauj lwm nyob rau hauv nws lub neej keeb kwm yam ntxwv, tab sis tam sim no tus cwj pwm no twb tsis quav ntsej .Kev siv agitation raws li ib tug txhais tau tias ntawm kev noj zaub mov kom tsawg ntawm cov khoom tseem ceeb hauv cov khoom tseem ceeb no.
Kev noj tshuaj nitrogen hauv cov poj niam laus Anopheles yoov tshaj cum yog txwv vim tsis muaj calories ntau nqa los ntawm lub cev xeeb tub thiab kev siv cov ntshav tsis zoo [9].Cov poj niam Ann.gambiae sl feem ntau them nyiaj rau qhov no los ntawm kev ntxiv cov zaub mov ntxiv rau ntshav [10, 11], yog li muab ntau tus neeg muaj kev pheej hmoo ntawm kab mob yoov tshaj cum. siv cov khoom noj ntxiv ntawm vertebrate excreta kom tau txais nitrogenous compounds uas txhim khu kev yoog thiab ya davhlau maneuverability, raws li pom los ntawm lwm yam kab [2].Nyob rau hauv no hais txog, lub zog thiab txawv attraction ntawm ib tug ntawm cov kwv tij hom nyob rau hauv An.The Gambian sl hom complex, Anopheles arabinis, tshiab thiab aged nyuj 13porta, nthuav tawm [13] . Nyob rau hauv nws cov tswv nyiam thiab paub txog kev koom tes nrog thiab pub rau nyuj.Noj cov zis yog ib qho chaw nplua nuj nyob rau hauv nitrogenous tebchaw, nrog urea accounting rau 50-95% ntawm tag nrho cov nitrogen nyob rau hauv cov zis tshiab [15, 16].Raws li nyuj tso zis muaj hnub nyoog, microorganisms siv cov kev pab cuam los txo cov complexity nyob rau hauv ib tug 2 hli ntawm nitrogen. poob ntawm cov organic nitrogen, alkalophilic microorganisms (ntau yam uas tsim cov tshuaj lom rau yoov tshaj cum) vam meej [15], uas tej zaum yuav poj niam Ann.arabiensis nyiam nyiam cov zis hnub nyoog 24 teev lossis tsawg dua [13, 14].
Hauv txoj kev tshawb no, tus tswv tsev thiab cov ntshav pub Ans tau saib rau.Thaum thawj lub voj voog gonadotropin, arabiensis raug soj ntsuam rau kev nrhiav tau cov nitrogenous compounds, suav nrog urea, los ntawm cov zis sib tov.Nom qab no, cov kev sim tau ua los soj ntsuam seb cov poj niam yoov tshaj cum faib cov peev txheej ntawm cov khoom noj muaj txiaj ntsig zoo thiab muaj txiaj ntsig ntxiv rau kev txhim kho kev noj qab haus huv. Cov neeg laus nyuj cov zis tau raug soj ntsuam los txiav txim seb cov no puas muab cov ntsiab lus txhim khu kev qha rau cov tswv tsev thiab cov ntshav pub An.Nyob rau hauv lawv cov kev tshawb fawb txog cov khoom noj muaj peev xwm no, arabiensis tau tshawb pom cov tshuaj sib raug zoo tom qab pom qhov sib txawv ntawm qhov zoo nkauj.Synthetic odor mixs ntawm volatile organic compounds (VOCs) tau txheeb xyuas nyob rau hauv 24-teev cov txiaj ntsig tau txais txiaj ntsig ntxiv nyob rau hauv 24-teev cov txiaj ntsig ntawm evaporation. cov nyhuv ntawm bovine tso zis tsw ntawm txawv physiological xeev.Mosquito attraction.Cov txiaj ntsig tau lees paub tias An.arabiensis tau txais thiab faib cov tshuaj nitrogenous uas muaj nyob hauv cov zis vertebrate los cuam tshuam cov yam ntxwv ntawm lub neej.Cov txiaj ntsig no tau tham txog hauv cov ntsiab lus ntawm qhov muaj feem cuam tshuam txog kev kis kab mob thiab yuav ua li cas lawv tuaj yeem siv rau vector soj ntsuam thiab tswj.
Anopheles arabicans (Dongola hom) tau khaws cia ntawm 25 ± 2 ° C, 65 ± 5% RH thiab lub teeb 12:12 h: tsaus nti.Larvae tau muab tso rau hauv cov tais yas (20 cm × 18 cm × 7 cm) ntim nrog dej distilled thiab pub Meletramin® 0 ml ntawm cov khoom noj. khob (Nolato Hertila, Åstorp, SE) thiab tom qab ntawd hloov mus rau Bugdorm tawb (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) kom tso cai rau cov neeg laus tshwm sim. Cov neeg laus tau muab 10% sucrose tov ad libitum mus txog rau 4 hnub tom qab tshwm sim (dpe), thaum lub hnub qub tau muab cov poj niam ua ntej los yog kev sim ua ntej. dej ua ntej qhov kev sim, raws li tau piav qhia hauv qab no.Cov poj niam siv rau kev sim davhlau ya davhlau sim tau tshaib plab tsuas yog 4-6 teev nrog dej ad libitum.Rau npaj cov ntshav nqus yoov tshaj cum rau cov poj niam tom qab bioassays, 4 dpe poj niam tau muab cov ntshav yaj yaj defibrotic (Håtunalab, Bro, SE) siv cov txheej txheem kev noj qab haus huv, kev ua haujlwm ntawm UK. tom qab ntawd hloov mus rau ib tus neeg lub tawb thiab muab cov khoom noj ncaj qha, raws li tau piav qhia hauv qab no, lossis 10% sucrose ad libitum rau 3 hnub ua ntej qhov kev sim tau piav qhia hauv qab no.Cov poj niam tom kawg tau siv rau lub dav hlau raj bioassays thiab xa mus rau hauv chav kuaj, thiab tom qab ntawd tau distilled dej ad libitum rau 4-6 teev ua ntej kev sim.
Kev ntsuam xyuas pub mis yog siv los ntsuas cov zis thiab urea noj hauv cov neeg laus An.Arab poj niam.Host-nrhiav thiab cov poj niam noj ntshav tau muab cov zaub mov uas muaj 1% diluted tshiab thiab laus nyuj tso zis, ntau qhov ntau ntawm urea, thiab ob qho kev tswj hwm (10% sucrose thiab dej) rau 48 h.Ntxiv rau, zaub mov-200 mg-200000000000 nbsp; 1; Sigma-Aldrich, Stockholm, SE) tau muab tso rau hauv cov khoom noj thiab muab tso rau hauv 4 × 4 matrix hauv 250 µl microcentrifuge hlab (Axygen Scientific, Union City, CA, US; Daim duab 1A) Sau rau ntawm ntug (~ 300 µl). Kom tsis txhob muaj kev sib tw ntawm cov kab mob loj ntawm cov yoov tshaj cum 1 (100 μl). cm nyob rau hauv txoj kab uas hla thiab 6 cm nyob rau hauv qhov siab; Semadeni, Ostermundigen, CH; Daim duab 1A) nyob rau hauv tag nrho tsaus ntuj ntawm 25 ± 2 cm ° C thiab 65 ± 5% txheeb ze humidity.Cov kev sim no tau rov ua dua 5 mus rau 10 zaug.Tom qab raug kev noj zaub mov, yoov tshaj cum tau muab tso rau ntawm -20 ° C kom txog rau thaum kev soj ntsuam ntxiv.
Saib bovine cov zis thiab urea absorbed los ntawm tus tswv tsev thiab cov ntshav nqus cov poj niam Anopheles arabianus.Nyob rau hauv kev sim pub mis (A), poj niam yoov tshaj cum tau muab cov zaub mov uas muaj cov zis tshiab thiab cov laus laus, ntau qhov ntau ntawm urea, sucrose (10%), thiab dej distilled (H2O) ntau dua (H2O). .Nco ntsoov tias cov poj niam nrhiav tus tswv tsev nqus 72-teev nyuj tso zis tsawg dua 168-teev nyuj tso zis (B).Qhov nruab nrab cov ntsiab lus nitrogen tag nrho (± tus qauv sib txawv) ntawm cov zis yog sawv cev rau hauv inset.Host-nrhiav (D, F) thiab nqus ntshav (E, G) cov poj niam noj li -Mean cov ntsiab lus tseem ceeb nyob rau hauv ib qho kev sib txawv ntawm E. ly txawv ntawm ib leeg (ib-txoj kev ANOVA siv Tukey's post hoc tsom xam; p < 0.05).Kev ua yuam kev bar sawv cev tus qauv kev ua yuam kev ntawm qhov nruab nrab (BE).Txoj kab ncaj nraim sawv cev rau kab-linear regression (F, G)
Txhawm rau tso cov khoom noj uas nqus tau, yoov tshaj cum tau muab tso rau hauv 1.5 ml microcentrifuge hlab uas muaj 230 µl ntawm cov dej distilled thiab cov ntaub so ntswg raug cuam tshuam los ntawm kev siv cov pestle pov tseg thiab cordless lub cev muaj zog (VWR International, Lund, SE), tom qab ntawd centrifugation ntawm 10 krpm rau 10 min (20 μl) superwell hloov mus rau 9 micron. ma-Aldrich) thiab absorbance (λ620) tau txiav txim siab siv lub spectrophotometer-based microplate nyeem ntawv (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Tsis tas li ntawd, yoov tshaj cum tau hauv av hauv 1 ml ntawm dej distilled, 900 µl ntawm cov uas tau pauv mus rau ib qho kev tshuaj ntsuam xyuas Shitomad 2 nm; ta.
Cov ntaub ntawv ntim tau raug tshuaj xyuas siv ib txoj kev tsom xam ntawm qhov sib txawv (ANOVA) ua raws li Tukey's post hoc pairwise kev sib piv (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007).Linear regression analyses piav txog concentration-dependent urea hosting cov ntshav thiab cov lus teb (piv txwv. v8.0.0 rau Mac, GraphPad Software, San Diego, CA, US).
Kwv yees li ntawm 20 µl ntawm cov zis kuaj los ntawm txhua pawg hnub nyoog raug khi rau ntawm Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) thiab muab ntim rau hauv tin capsules (8 mm × 5 mm).Capsules tau muab tso rau hauv lub combustion chamber ntawm a 00rfic, CHNS / O. am, MA, US) los txiav txim siab cov ntsiab lus nitrogen hauv cov zis tshiab thiab muaj hnub nyoog raws li cov chaw tsim khoom raws tu qauv.Tag nrho cov nitrogen (g N l-1) tau ntsuas raws li paub cov urea concentrations siv raws li tus qauv.
Txhawm rau soj ntsuam cov txiaj ntsig ntawm kev noj zaub mov ntawm tus tswv tsev nrhiav thiab ntshav nqus cov poj niam ciaj sia, yoov tshaj cum tau muab tso rau hauv cov tais loj Petri (12 cm inch thiab 6 cm nyob rau hauv qhov siab; Semadeni) nrog ib lub mesh-pob qhov nyob rau hauv lub hau (3 cm inch) nrog rau qhov cua thiab khoom noj khoom haus. Cov zaub mov tau muab ncaj qha tom qab 4 aged 1% diluted thiab diluted tshiab concentration. thiab ob qho kev tswj hwm, 10% sucrose thiab dej.Txhua cov khoom noj tau muab tso rau hauv cov hniav tampon (DAB Dental AB, Upplands Väsby, SE) tso rau hauv 5 ml syringe (Thermo Fisher Scientific, Gothenburg, SE), plunger tshem tawm, thiab muab tso rau saum ib lub petriang. txhua hnub. Cov yoov tshaj cum uas muaj sia nyob raug suav ob zaug hauv ib hnub, thaum yoov tshaj cum tuag raug muab pov tseg kom txog thaum yoov tshaj cum tuag (n = 40 ib txoj kev kho mob).Kev ciaj sia ntawm yoov tshaj cum noj ntawm ntau yam zaub mov tau txheeb xyuas siv Kaplan-Meyer survival curves thiab cov ntawv xeem ntsuas los sib piv cov ciaj sia taus kev sib piv ntawm cov khoom noj. SP0SS.
Ib qho kev cai yoov tshaj cum ya raws li Attisano li al.[17], ua los ntawm 5 hli tuab ntshiab acrylic panels (10 cm dav x 10 cm ntev x 10 cm siab) tsis muaj pem hauv ntej thiab nram qab panels (Fig. 3: saum).A pivot los ua ke nrog ib tug ntsug raj ua los ntawm ib tug roj chromatography kem (0.25 mm kua nplaum yuav tsum tau nyob rau hauv ib tug khub L. Neodymium hlau nplaum 9 cm sib nrug.Ib kab rov tav raj ua los ntawm tib yam khoom (6.5 cm L) bisected lub ntsug raj los tsim ib tug tethered caj npab thiab ib sab caj npab uas nqa ib tug me me daim ntawm txhuas ntawv ci raws li ib tug teeb liab cuam tshuam.
Cov poj niam tshaib plab 24-teev tau noj cov zaub mov saum toj no rau 30 feeb ua ntej kev txwv.Fully noj yoov tshaj cum poj niam ces muab tshuaj loog ib leeg rau ntawm cov dej khov rau 2-3 min thiab txuas rau kab pins nrog beeswax (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE.R. ions ib lub davhlau tau sau tseg los ntawm kev tsim cov ntaub ntawv logger, tom qab ntawd khaws cia thiab tso tawm siv PC-Lab 2000™ software (v4.01; Velleman, Gavere, BE).Lub davhlau zeb tau muab tso rau hauv chav tswj kev nyab xeeb (12 h: 12 h, lub teeb: tsaus, 25 ± 2 ° C, 65% R ± 5% ).
Txhawm rau pom cov qauv ntawm kev ua si hauv lub davhlau, tag nrho qhov kev ncua deb ya (m) thiab tag nrho cov kev sib law liag sib law liag tau suav ib teev nyob rau lub sijhawm 24-teev. Tsis tas li ntawd, qhov kev ncua deb ntawm ib tus poj niam tau muab piv rau kev kho mob thiab tshuaj xyuas siv ib txoj kev ANOVA thiab Tukey's post hoc tsom xam (JMP Pro, v14.0.0), qhov nruab nrab ntawm kev kho mob SAS vari Inc. itionally, qhov nruab nrab ntawm cov rounds yog xam nyob rau hauv 10-feeb increments.
Txhawm rau ntsuam xyuas cov txiaj ntsig ntawm kev noj zaub mov ntawm kev ua me nyuam ntawm An.arabiensis, rau tus poj niam (4 dpe) raug xa ncaj qha mus rau Bugdorm tawb (30 cm × 30 cm × 30 cm) tom qab sau cov ntshav thiab muab cov kev sim noj rau 48 h raws li tau piav qhia saum toj no. Cov zaub mov tau muab tshem tawm thiab muab cov khob cij (30 ml 4 hnub thib peb) ntawm Nolato distil 4 hnub. Cov xuab moos, hloov txhua 24 teev. Rov ua dua txhua txoj kev noj zaub mov 20-50 zaug.Cov qe raug suav thiab sau tseg rau txhua qhov kev sim tawb.Cov qe qe tau siv los ntsuas qhov ntsuas qhov loj thiab qhov ntev ntawm cov qe sib txawv (n ≥ 200 rau ib qho kev noj haus) siv Dialux-20 microscope (DM1000; Wetz, FC, Leitz) 3 lub koob yees duab, Ernlarst Leitz. R2;Leica Microsystems Ltd., DE).Cov qe seem tau muab khaws cia rau hauv chav tswj kev nyab xeeb raws li tus qauv txhawb nqa rau 24 h, thiab ib qho piv txwv ntawm tsis ntev los no tau tshwm sim 1st instar larvae (n ≥ 200 ib qho khoom noj) tau ntsuas, raws li tau piav qhia saum toj no.Cov qe thiab qhov loj ntawm cov qe thiab cov larvae tau muab piv rau ib qho kev soj ntsuam ntawm kev kho mob thiab cov larvae (NOMP). 0.0, SAS Institute Inc.).
Headspace volatiles los ntawm cov tshiab (1 hr post-sampling), 24 hr, 72 hr thiab 168 hr laus zis tau sau los ntawm cov qauv sau los ntawm Zebu nyuj, Arsi haiv neeg.Rau qhov yooj yim, cov zis kuaj tau sau thaum sawv ntxov thaum cov nyuj tseem nyob rau hauv lub barn.Urine kuaj tau sau los ntawm ib tug neeg ntawm 2 ml ntawm polyamide mus rau 10 10 baum. hnab (Toppits Cofresco, Frischhalteprodukte GmbH thiab Co., Minden, DE) nyob rau hauv 3 l polyamide nrog lub hau Nyob rau hauv vinyl chloride yas drums.Headspace volatiles los ntawm txhua bovine zis coj mus kuaj los yog tom qab maturation ntawm chav tsev kub rau 24 h, 7 2 h.
Rau headspace volatiles sau, kaw-voj kaw lus tau siv los ncig lub tshuab ua pa roj carbon lim dej (100 ml min-1) los ntawm lub hnab polyamide mus rau sab adsorption rau 2.5 h los ntawm kev siv lub tshuab nqus tsev diaphragm (KNF Neuberger, Freiburg, DE). (5.5 cm x 3 mm id) uas muaj 35 mg ntawm Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) nruab nrab ntawm iav ntaub plaub plugs.Ua ntej siv, lub kem yog flushed nrog 1 ml redistilled n-hexane (Merck, Darmstadt, C 0% tanegrade, 1 ml) thiab 1 ml. Cov adsorbed volatiles tau eluted nrog 400 μl ntawm pentane.Headspace collections tau pooled thiab ces khaws cia ntawm -20 ° C kom txog thaum siv rau kev soj ntsuam ntxiv.
Cov lus teb coj tus cwj pwm ntawm tus tswv tsev nrhiav thiab noj ntshav An.Headspace volatile extracts sau los ntawm cov zis tshiab, 24-h, 72-h, thiab 168-h-hnub nyoog zis tau soj ntsuam rau cov tsis muaj tshuaj tiv thaiv los ntawm Arabidopsis yoov tshaj cum siv lub iav raj ncaj qha olfactometer [18].Lub sijhawm ua haujlwm ntawm Z.15-lub sijhawm ua haujlwm. Arab [19].A iav raj olfactometer (80 cm × 9.5 cm id) tau illuminated nrog 3 ± 1 lx ntawm lub teeb liab los ntawm saum toj no.Charcoal lim thiab humidified cua ntws (25 ± 2 ° C, 65 ± 2% txheeb ze av noo) dhau lub bioassay ntawm 30 cms ntawm cov hlau tsis sib xws, thiab ib qho kev sib txawv ntawm cov hlau tsis sib xws los ntawm lam. plume structure.Dental tampon dispenser (4 cm × 1 cm; L: D; DAB Dental AB), raug tshem tawm los ntawm 5 cm coil ntawm lub windward kawg ntawm lub olfactometer, nrog stimulator hloov txhua 5 feeb.Rau kev tshuaj ntsuam, 10 μl ntawm txhua lub taub hau extract, diluted 1:10, yog siv raws li ib tug stimulation ntawm cov ntshav los yog ib tug sib npaug zos hostess. Cov yoov tshaj cum tau muab tso rau hauv ib tus neeg tso lub tawb 2-3 teev ua ntej pib qhov kev sim.Qhov kev tso tawm lub tawb tau muab tso rau ntawm lub downwind side ntawm lub olfactometer, thiab yoov tshaj cum raug tso cai rau acclimate rau 1 min, thiab ces tus npauj npaim valve ntawm lub tawb tau qhib rau kev tso tawm.Txoj kev kho mob los yog tswj tau soj ntsuam raws li qhov kev faib ua feem ntawm kev sib cuag nrog cov tshuaj tua kab mob thiab cov tshuaj tua kab mob. plicated tsawg kawg yog 30 zaug, thiab kom tsis txhob muaj kev cuam tshuam ntawm ib hnub twg, tib tus naj npawb ntawm kev kho mob thiab kev tswj hwm tau sim nyob rau txhua hnub kev sim.Nrhiav cov lus teb los ntawm tus tswv tsev thiab cov ntshav pub Ans.Arabic piv rau headspace tau txheeb xyuas siv nominal logistic regression ua raws li kev sib piv sib piv rau qhov sib piv tsis zoo (JMP Pro, Inc., 0, AS).
Ib qho lus teb spawning.Headspace extracts los ntawm cov nyuj tshiab thiab laus laus tau soj ntsuam nyob rau hauv Bugdorm cages (30 cm × 30 cm × 30 cm; MegaView Science).Plastic khob (30 mL; Nolato Hertila) ntim nrog 20 mL ntawm distilled dej muab lub spawning substrate thiab muab tso rau nyob rau hauv ib tug sib sib zog nqus khob, 4 cm ntawm cov ces kaum. μl ntawm txhua lub taub hau extract ntawm 1:10 dilution.Ib qho sib npaug ntawm pentane tau siv los kho lub khob tswj.Kev kho thiab tswj lub khob tau sib pauv ntawm txhua qhov kev sim los tswj rau txoj hauj lwm cuam tshuam.Kaum cov poj niam noj cov ntshav tau tso rau hauv kev sim tawb ntawm ZT 9-11 thiab cov qe hauv khob tau suav 24 teev tom qab.Cov qe yog cov mis nyuj rau kev kho mob ntawm cov qe. id nyob rau hauv lub khob tswj) / (tag nrho cov qe nteg).Txhua qhov kev kho mob tau rov ua dua 8 zaug.
Gas chromatographic thiab electron antenna pattern detection (GC-EAD) kev soj ntsuam poj niam An.arabiensis tau ua raws li tau piav qhia yav dhau los [20] .Liab, tshiab headspace volatile extracts tau sib cais siv Agilent Technologies 6890 GC (Santa Clara, CA, US) nruab nrog ib kab HP-5 mm, thickness 0.5 m2 (3 μm. ilent Technologies).thiab cov zis aging.Hydrogen tau siv los ua lub sijhawm txawb nrog qhov nruab nrab linear flow rate ntawm 45 cm s-1.Txhua tus qauv (2 μl) tau txhaj rau 30 vib nas this hauv splitless hom nrog inlet kub ntawm 225 ° C.The GC qhov cub kub yog programmed los ntawm 35 ° C (3 feeb tuav) mus rau 300 ° C) ntawm 1 min. splitter, 4 psi ntawm nitrogen tau ntxiv thiab faib 1: 1 nyob rau hauv ib tug Gerstel 3D / 2 tsawg tuag ntim ntoo khaub lig (Gerstel, Mülheim, DE) nruab nrab ntawm cov nplaim ionization ntes thiab EAD.Lub GC effluent capillary rau EAD tau dhau los ntawm Gerstel ODP-2 hloov kab, uas taug qab GC qhov cub kub, 1 hli × 8 tere. ed, humidified huab cua (1.5 l min-1).Lub kav hlau txais xov tau muab tso rau 0.5 cm los ntawm lub qhov hluav taws xob ntawm lub raj.Txhua tus yoov tshaj cum suav nrog ib tus neeg rov ua dua, thiab rau cov yoov tshaj cum nrhiav tswv tsev, tsawg kawg yog peb qhov rov ua dua ntawm cov zis kuaj txhua hnub.
Kev txheeb xyuas cov bioactive compounds nyob rau hauv headspace collections ntawm cov tshiab thiab cov laus bovine tso zis siv ib tug ua ke GC thiab loj spectrometer (GC-MS; 6890 GC thiab 5975 MS; Agilent Technologies) mus elicit antennal teb nyob rau hauv GC-EAD tsom xam, kev khiav hauj lwm nyob rau hauv electron cuam tshuam ionization hom 5 MS . ica capillary kem (60 m × 0.25 mm puab txoj kab uas hla, 0.25 μm zaj duab xis thickness) siv helium raws li lub sij hawm mobile nrog ib tug nruab nrab linear ntws tus nqi ntawm 35 cm s-1.A 2 μl qauv raug txhaj tshuaj siv tib lub injector nqis thiab qhov cub kub raws li rau GC-EAD tsom xam.Compounds tau txais kev cai retention ntawm lub sij hawm thiab identified. ST14 lub tsev qiv ntawv (Agilent). Cov tshuaj tiv thaiv tau lees paub los ntawm kev txhaj tshuaj cov qauv tseeb (Ntxiv Cov Ntaub Ntawv 1: Table S2).Rau quantification, heptyl acetate (10 ng, 99.8% tshuaj purity, Aldrich) tau txhaj tshuaj raws li tus qauv sab nraud.
Kev ntsuas qhov ua tau zoo ntawm cov tshuaj tsw qab hluavtaws uas muaj cov tshuaj bioactive uas pom hauv cov zis tshiab thiab muaj hnub nyoog kom nyiam tus tswv-nrhiav thiab ntshav nqus Ans.arabiensis, siv tib lub olfactometer thiab raws tu qauv raws li saum toj no.Synthetic mixtures mimicked cov muaj pes tsawg leeg thiab proportions ntawm cov tebchaw nyob rau hauv mixed headspace volaourh, 4 tshiab-4h extracts, thiab 168-teev cov zis uas muaj hnub nyoog (Daim duab 5D-G; Cov ntaub ntawv ntxiv 1: Table S2).Rau kev soj ntsuam, siv 10 μl ntawm 1:100 dilution ntawm cov khoom sib xyaw ua ke, nrog rau tag nrho cov nqi tso tawm txij li kwv yees li 140-2400 ng h-1, rau kev soj ntsuam cov ntshav thiab ua kom tiav-kev ua kom zoo nkauj. Cov khoom sib xyaw ua ke, nyob rau hauv uas cov sib tov sib xyaw ntawm ib qho sib xyaw ntawm qhov sib xyaw ua tiav raug tshem tawm.Nrhiav cov lus teb los ntawm tus tswv tsev thiab cov ntshav noj Ans.Arab vs cov khoom sib xyaw thiab cov khoom sib cais tau raug tshuaj xyuas siv nominal logistic regression ua raws li kev sib piv piv rau qhov sib txawv ntawm qhov sib txawv (JMP Pro, v14.0.0, SAS Institute Inc.).
Txhawm rau ntsuam xyuas seb nyuj tso zis puas tuaj yeem ua tus tswv tsev cue rau cov yoov tshaj cum malaria, cov zis nyuj tshiab thiab muaj hnub nyoog, sau raws li tau piav qhia saum toj no, thiab dej tau muab tso rau hauv meshed 3 l thoob (100 ml) thiab muab tso rau hauv cov cuab yeej cuab tam.(BG-HDT version; BioGents, Regensburg, DE).Kaum lub ntxiab tso 50 m sib nrug hauv pasture, 400 m ntawm lub zos zej zog (Silay, Ethiopia, 5° 53´24´´N, 37° 29´24´´E) thiab tsis muaj nyuj, ntawm qhov chaw yug me nyuam mus tas li thiab lub zos raug cua sov.Fives muaj cua sov. ach qhov chaw kho mob yog tig ib hmos rau tag nrho tsib hmos.Mosquito nab npawb ntes nyob rau hauv cov ntxiab baited nrog zis ntawm lub hnub nyoog sib txawv tau muab piv nrog logistic regression nrog ib tug beta binomial faib (JMP Pro, v14.0.0, SAS Institute Inc.).
Nyob rau hauv ib lub zos malaria-cov kab mob ze ntawm lub nroog Maki, Oromia cheeb tsam, Ethiopia (8° 11′ 08″ N, 38° 81′ 70″ E; Daim duab 6A). Txoj kev tshawb no tau ua nyob rau nruab nrab lub yim hli ntuj thiab nruab nrab lub Cuaj Hli ua ntej lub xyoo nyob rau hauv tsev nyob rau hauv cov tshuaj tsuag, nrog rau ib tug ntev lub sij hawm los nag. s ntawm lub zos tau raug xaiv rau txoj kev kawm (Fig. 6A).Cov qauv siv los xaiv cov tsev yog: tsis pub tsiaj nyob hauv tsev, tsis muaj kev ua noj sab hauv tsev (kos taws los yog charcoal) raug tso cai (tsawg kawg thaum lub sij hawm sim), thiab cov tsev uas muaj ntau tshaj ob tus neeg nyob, pw hauv cov tshuaj tua kab mob.raws li kev kho yoov tshaj cum.Ethical pom zoo tau tso cai los ntawm Pawg Saib Xyuas Kev Tshawb Fawb Txog Kev Ncaj Ncees (IRB/022/2016) ntawm Kws Qhia Ntawv ntawm Natural Sciences (CNS-IRB), Addis Ababa University, raws li cov lus qhia tsim los ntawm World Medical Association tshaj tawm ntawm Helsinki.Cov kev tso cai los ntawm txhua lub taub hau ntawm cov neeg ua hauj lwm hauv cheeb tsam tau txais kev pab ntxiv. 'kebele') theem.Qhov kev sim tsim ua raws li 2 × 2 Latin square tsim, uas cov khoom sib tov sib xyaw thiab kev tswj tau raug xa mus rau cov tsev ua ke nyob rau thawj hmo thiab sib pauv ntawm cov tsev nyob rau yav tom ntej kev sim hmo ntuj.Cov txheej txheem no tau rov ua dua kaum zaug. Tsis tas li ntawd, txhawm rau kwv yees cov yoov tshaj cum hauv cov tsev xaiv, CDC cov cuab yeej raug teeb tsa mus sib tw tsib hnub ntawm qhov kawg ntawm lub sijhawm nruab nrab thiab nruab nrab.
Ib qho hluavtaws sib xyaw uas muaj rau cov khoom siv bioactive tau yaj hauv heptane (97.0% hnyav GC qib, Sigma Aldrich) thiab tso tawm ntawm 140 ng h-1 siv cov paj rwb wick dispenser [20].Lub wick dispenser tso cai rau tag nrho cov tebchaw tso tawm nyob rau hauv tas li proportions thoob plaws hauv lub 12 teev kev sim. Tswj thiab Tiv Thaiv (CDC) lub teeb cuab (John W. Hock Company, Gainesville, FL, US; Daim duab 6A). Cov ntxiab tau dai 0.8 - 1 m saum toj no hauv av, ze ntawm ko taw ntawm lub txaj, thiab ib tus neeg tuaj yeem pab dawb tau pw hauv qab cov yoov tshaj cum tsis kho thiab ua haujlwm nruab nrab ntawm 18:00 thiab 06:30.Mos physiotherapy, kev sib deev thiab kev sib deev. , thiab cev xeeb tub [21] tau soj ntsuam tom qab siv cov tshuaj polymerase saw cov tshuaj tiv thaiv (PCR) txhawm rau txheeb xyuas hom kab mob morphologically txheeb xyuas raws li A. gambiae sl. Cov tswv cuab ntawm cov complex [23]. Hauv kev tshawb fawb, cuab ntxiab ntawm cov tsev ua ke tau txheeb xyuas siv tus qauv logistic haum, qhov kev nyiam yog qhov sib txawv ntawm J0 thiab kev kho mob (tshwj xeeb 1 MP.0. SAS Institute Inc.).Ntawm no, peb tshaj tawm qhov χ2 thiab p-tus nqi los ntawm qhov kev xeem qhov piv txwv.
Ntsuam xyuas seb nws puas muaj kev nyab xeeb.arabiensis tau tso zis, nws qhov chaw nitrogen tseem ceeb, urea, los ntawm kev pub mis ncaj qha, hauv 48 h ntawm kev tswj hwm rau 4 hnub tom qab (dpe) tus tswv tsev nrhiav thiab ntshav pub poj niam noj kev sim (Daim duab 1A).Ob tus tswv tsev-nrhiav thiab cov poj niam nqus ntshav. p < 0.0001 thiab F(5,299) = 56.00, p < 0.0001, ntsig txog; Fig. 1B, C). Tsis tas li ntawd, cov poj niam nrhiav tus tswv tsev noj cov zis tsawg dua ntawm 72 teev piv rau cov zis ntawm 168 teev (Fig. 1B).Thaum noj zaub mov muaj txiaj ntsig zoo, cov poj niam noj zaub mov muaj txiaj ntsig zoo. ntawm 2.69 mM piv rau tag nrho lwm cov concentrations thiab dej, thaum indistinguishable los ntawm 10% sucrose (F(10,813) = 15.72, p < 0.0001; Daim duab 1D).Qhov no yog qhov sib piv rau cov lus teb ntawm cov poj niam noj ntshav, uas feem ntau absorbed roses ntau tshaj li 1% sucrose (feem ntau yog urea 10% dej) 10,557) = 78.35, p < 0.0001; Daim duab 1.1E). Tsis tas li ntawd, thaum muab piv ntawm ob lub cev lub cev, cov poj niam phlebotomized absorbed urea ntau dua li cov poj niam nrhiav tus tswv tsev ntawm qhov qis tshaj plaws, thiab cov poj niam no absorbed ntau npaum li cas ntawm urea (p 7 8.0), siab dua concentration ntawm urea 5 . 1;Fig. 1F, G).Thaum noj los ntawm cov khoom noj uas muaj urea zoo nkaus li muaj txiaj ntsig zoo (Daim duab 1D, E), cov poj niam hauv ob lub cev muaj zog tuaj yeem hloov kho cov nyiaj urea absorbed tshaj tag nrho cov ntau yam ntawm urea concentrations nyob rau hauv lub log-linear zam (Fig. 1F, G).) Ib yam li ntawd, yoov tshaj cum zoo li tswj lawv cov nitrogen uptake los ntawm kev tswj cov zis nqus, vim tias cov nitrogen hauv cov zis tau cuam tshuam txog qhov nqus tau (Daim duab 1B, C thiab B insets).
Txhawm rau ntsuas qhov tshwm sim ntawm cov zis thiab urea ntawm tus tswv tsev nrhiav thiab nqus cov yoov tshaj cum muaj sia nyob, cov poj niam tau tso zis ntawm tag nrho plaub lub hnub nyoog (tshiab, 24 h, 72 h, thiab 168 h tom qab tso tawm) thiab ntau yam ntawm urea concentrations, nrog rau cov dej distilled thiab 10% kev noj zaub mov muaj txiaj ntsig zoo. ntawm tag nrho cov ciaj sia nyob rau hauv cov poj niam nrhiav tswv tsev (cov zis: χ2 = 108.5, df = 5, p < 0.0001; urea: χ2 = 122.8, df = 5, p < 0.0001; Fig. 2B, C) thiab cov poj niam uas muaj ntshav qab zib 2 = 0, 9 pf 3. ; urea: χ2 = 137.9, df = 5, p < 0.0001; Daim duab 2D, E).Nyob rau hauv txhua qhov kev sim, cov poj niam tau noj zaub mov ntawm cov zis, urea, thiab dej tau qis qis qis dua li cov poj niam noj sucrose noj zaub mov (Daim duab 2B-Essee, cov poj niam survival survival sib txawv). nrog cov uas pub 72-h stale tso zis (p = 0.016) muaj qhov yuav muaj sia nyob qis tshaj (Fig. 2B). Tsis tas li ntawd, cov poj niam nrhiav tswv tsev pub 135 mM urea muaj sia nyob ntev dua li kev tswj dej (p <0.04) (Fig.2C).Piv nrog cov dej, cov poj niam noj cov zis tshiab thiab cov zis 24-teev muaj sia nyob ntev dua (p = 0.001 thiab p = 0.012, raws li; Daim duab 2D), thaum cov poj niam pub nrog cov zis 72-teev muaj sia nyob ntev dua li cov poj niam noj cov zis tshiab thiab < 20.3 teev = 24-hour. Raws li; Daim duab 2D).Thaum pub 135 mM urea, cov poj niam noj cov ntshav muaj sia nyob ntev dua li lwm cov concentration ntawm urea thiab dej (p <0.013; Daim duab 2E).
Kev ciaj sia ntawm tus tswv tsev thiab cov ntshav nqus cov poj niam Anopheles arabinis pub rau nyuj tso zis thiab urea.Nyob rau hauv bioassay (A), poj niam yoov tshaj cum tau muab cov zaub mov uas muaj cov zis tshiab thiab muaj hnub nyoog nyuj, ntau qhov ntau ntawm urea, sucrose (10%) thiab cov dej distilled (H2O).Qhov ciaj sia ntawm cov neeg tua tsiaj, cov ntshav (Cesquies) txhua tus ciaj sia. 12 teev kom txog thaum tag nrho cov poj niam noj cov zis (B, D) thiab urea (C, E), thiab tswj, sucrose thiab dej, tuag.
Tag nrho cov kev ncua deb thiab tus naj npawb ntawm cov rounds txiav txim nyob rau hauv lub davhlau mills nyob rau hauv lub sij hawm 24-teev sib txawv ntawm tus tswv tsev nrhiav thiab nqus cov yoov tshaj cum, uas qhia tau hais tias tsis tshua muaj kev ya davhlau tag nrho (Fig. 3). Cov yoov tshaj cum-nrhiav uas muab cov zis tshiab thiab muaj hnub nyoog los yog sucrose thiab dej pom cov qauv sib txawv ntawm cov poj niam (daim duab 2, 3). Cov yoov tshaj cum uas muaj hnub nyoog 4- thiab 168-teev uas tau tso zis tso zis pom cov qauv sib txawv thiab feem ntau yog diurnal.Cov poj niam yoov tshaj cum uas muab sucrose lossis 72-teev tso zis tau pom kev ua si thoob plaws hauv 24-teev, thaum cov poj niam uas muab dej muaj zog ntau dua thaum hmo ntuj sucrose nyob rau theem siab tshaj plaws ntawm kev ua haujlwm ntawm Mos. thaum sawv ntxov, thaum cov neeg uas noj cov zis 72-teev-teev muaj kev poob qis hauv kev ua haujlwm dhau 24 teev (Daim duab 3).
Kev ua tau zoo ntawm cov neeg yos hav zoov nrhiav ntshav nqus cov poj niam Anopheles arabinis pub rau nyuj tso zis thiab urea.Nyob rau hauv lub davhlau zeb kuaj, cov poj niam yoov tshaj cum pub rau cov nyuj tshiab thiab cov laus tso zis, ntau qhov ntau ntawm urea, sucrose (10%), thiab cov dej distilled (H2O) tau tethering, pub dawb (horizatleov). ) thiab cov ntshav nqus dej (txoj cai) cov poj niam, tag nrho cov kev ncua deb thiab tus naj npawb ntawm kev ya mus rau ib teev rau txhua qhov kev noj zaub mov hauv 24-teev tau sau tseg (tsaus: grey; lub teeb: dawb). Qhov nruab nrab qhov deb thiab qhov nruab nrab ntawm qhov sib tw yog qhia rau sab xis ntawm lub circadian kev ua ub no graph.Error bars sawv cev tus qauv kev ua yuam kev ntawm lub ntsiab lus.Statistical tsom xam saib cov ntawv nyeem.
Feem ntau, tag nrho cov kev ua ub no ntawm tus tswv tsev nrhiav poj niam ua raws li tus qauv zoo ib yam li kev ya davhlau nyob rau lub sijhawm 24-teev.Mean ya davhlau deb tau cuam tshuam loj heev los ntawm kev noj zaub mov noj (F(5, 138) = 28.27, p < 0.0001), thiab cov poj niam nrhiav tus tswv tsev tau noj 72 teev ntawm kev noj zaub mov ntev dua 0sflew. yoov tshaj crose yoov tshaj qhov tshiab (p = 0.022) thiab 24-h-hnub tso zis (p = 0.022)-fed yoov tshaj cum. Txawm hais tias cov qauv kev ua ub no zoo sib xws, cov poj niam nrhiav tswv yim pub urea nce qhov dav dav dav nyob ntawm qhov nqus tau (F(5, 138) = 1310.91, p < 0.0001). Cov poj niam nrhiav tus tswv tsev pub cov concentration ntawm urea ya ntev dua cov poj niam noj dej los yog sucrose (p <0.
Zuag qhia tag nrho kev ua si ntawm yoov tshaj cum ntshav tau ruaj khov thiab txhawb nqa ntau dua 24 teev thoob plaws txhua qhov kev noj haus, nrog rau kev tso zis ntau ntxiv thaum lub sijhawm thib ob ntawm lub sijhawm tsaus ntuj rau cov poj niam noj dej nrog rau cov poj niam noj tshiab thiab 24 teev (daim duab 3).Thaum tso zis muaj feem cuam tshuam rau lub davhlau nrug ntawm cov poj niam noj ntshav (F 1 4 0.30, 5, 1.380, 5. Kev noj zaub mov tsis zoo (F(5, 138) = 1.36, p = 0.24).nrog rau lwm cov zis thiab tswj kev noj zaub mov (tshiab, p = 0.0091; 72 teev, p = 0.0022; 168 teev, p = 0.001; sucrose, p = 0.0017, 3.
Cov kev cuam tshuam ntawm cov zis thiab urea pub rau kev ua me nyuam tsis tau raug soj ntsuam hauv cov qe nteg bioassays (Daim duab 4A) thiab tau tshawb xyuas raws li tus naj npawb ntawm cov qe nteg los ntawm txhua tus poj niam, qe loj, thiab tshiab hatched thawj instar larvae.Tus naj npawb ntawm cov qe nteg.Urine-fed Arab poj niam varied los ntawm kev noj zaub mov 5,20 F = 0.20 F. ig. 4B).Cov poj niam tau tso zis 24-teev, cov ntshav pluas noj tau tso cov qe ntau dua li cov poj niam pub lwm cov zis tso zis thiab zoo ib yam li cov qe uas tau noj sucrose (Fig. 4B). Ib yam li ntawd, qhov loj ntawm cov qe tso los ntawm cov poj niam noj cov zis muaj ntau qhov sib txawv ntawm kev noj zaub mov (F(5, 208), ur 208, 204, p. Cov poj niam rose-fed nteg qe loj dua li cov poj niam noj dej, thaum cov qe ntawm cov poj niam pub nrog 168 h cov zis tau me me (Fig. 4C). Tsis tas li ntawd, cov zis tso zis cuam tshuam loj heev rau cov me nyuam mos (F(5, 187) = 7.86, p < 0.0001h - peb cov qe loj los ntawm 0.0001h), nrog rau cov qe loj loj los ntawm peb lub lard. Cov poj niam laus tso zis ntau dua li ntawm cov qe nteg qe los ntawm cov qe larvae. Dej-piv thiab 168-h tso zis poj niam (Daim duab 4D).
Kev yug me nyuam ntawm poj niam Anopheles arabinis pub rau nyuj tso zis thiab urea.Cov poj niam yoov tshaj cum tau noj cov zaub mov uas muaj cov zis tshiab thiab hnub nyoog nyuj, ntau cov ntsiab lus ntawm urea, sucrose (10%), thiab dej distilled (H2O) rau 48 teev ua ntej muab tso rau hauv cov qe qe, 8 teev (8 teev) Eggestays. Lub qe loj (C, F) thiab larvae loj (D, G) tau cuam tshuam loj heev los ntawm kev noj zaub mov (nqaij zis: BD; urea: EG).Txhais rau txhua qhov ntsuas ntsuas uas siv cov npe sib txawv yog qhov sib txawv ntawm ib leeg (ib-txoj kev ANOVA siv Tukey's post hoc tsom xam; p < 0.05). Cov kab lus yuam kev sawv cev rau tus qauv kev ua yuam kev.
Raws li cov ntsiab lus nitrogenous tseem ceeb ntawm cov zis, urea, thaum muab cov khoom noj rau cov poj niam uas muaj ntshav qab zib, cuam tshuam rau cov poj niam cev xeeb tub hauv txhua qhov kev tshawb fawb.Tus naj npawb ntawm cov qe tso los ntawm cov poj niam noj urea, tom qab noj ntshav, nyob ntawm urea concentration (F(11, 360) = 4.69; p < 0.0001 m3 a µs 4 m3 μs 3 m 3 μs 4 μs 1. id ntau cov qe (Daim duab 4E). Cov poj niam pub rau ntawm urea concentrations ntawm 134 µM los yog saum toj no nteg qe loj dua cov poj niam pub rau hauv dej (F(10, 4245) = 36.7; p < 0.0001; Daim duab 4F), thiab larval loj, txawm hais tias cuam tshuam los ntawm cov concentrations zoo sib xws ntawm 30 p. 30 (30 . 0001) muaj ntau qhov sib txawv (Fig. 4G).
Zuag qhia tag nrho attractiveness rau host-nrhiav bovine urine headspace volatile extracts.The arabiensis soj ntsuam nyob rau hauv lub iav raj olfactometer (Fig. 5A) tau ho cuam tshuam los ntawm cov zis muaj hnub nyoog (χ2 = 15.9, df = 4, p = 0.0032; Fig. 5A) muaj kev cuam tshuam loj heev ntawm cov zis muaj hnub nyoog (χ2 = 15.9, df = 4, p = 0.0032; Fig. 5B) qhov kev ntsuam xyuas tseem ceeb ntawm P. theem ntawm attractiveness piv rau tag nrho lwm yam kev kho mob (72 teev: p = 0.0060, 168 teev: p = 0.012, pentane: p = 0.00070), Tsuas yog cov tsis hnov ​​tsw ntawm cov zis tshiab (p = 0.13; Daim duab 5B).Txawm hais tias tag nrho cov attraction ntawm cov ntshav-nqus tsw 8.Txawm hais tias tag nrho cov attraction ntawm ntshav-nqus tsw. = 4, p = 0.067; Fig. 5C), cov poj niam no tau pom tias muaj kev ntxim nyiam ntau dua rau lub taub hau tsis muaj zog ntxiv piv nrog 72-teev hnub nyoog tso zis piv rau kev tswj hwm (p = 0.0066; Daim duab 5C).
Cwj pwm teb rau tej yam ntuj tso thiab hluavtaws nyuj tso zis tsw nyob rau hauv kev tshawb nrhiav rau tus tswv tsev thiab cov ntshav pub Anopheles arabianus.Schematic ntawm lub iav raj olfactometer (A).Tshem tawm ntawm headspace volatile rho tawm los ntawm cov tshiab thiab laus nyuj tso zis rau host (B) thiab ntshav-sucking (C) cov tshuaj tiv thaiv ntawm tus Tswv (C) cov tshuaj tiv thaiv kab mob. 24-teev (E), 72-teev (F), thiab 168-teev (G) lub hnub nyoog nyuj tso zis tau qhia.Electron kav hlau txais xov (EAD) kab qhia pom cov kev hloov pauv ntawm cov lus teb rau bioactive compounds nyob rau hauv lub headspace eluted los ntawm cov roj chromatograph thiab kuaj pom los ntawm ib tug nplaim ionization detector (FID) lub sij hawm (FID) reacts. tso tawm tus nqi (µg h-1) ntawm cov tshuaj lom neeg lom neeg tau tshwm sim.Ib lub hnub qub (*) qhia tau hais tias muaj cov lus teb qis-amplitude.Double asterisks (**) qhia tias cov lus teb tsis zoo.Nrhiav tus tswv tsev (H) thiab cov ntshav nqus dej (I) An.arabiensis muaj qhov sib txawv ntawm cov mos mos thiab cov odors ntawm cov khoom sib xyaw tshiab. Cov npe ntawv sib txawv tau sib txawv ntawm ib leeg (ib-txoj kev ANOVA siv Tukey's post hoc tsom xam;p < 0.05). Cov kab yuam kev sawv cev tus qauv yuam kev ntawm qhov ntsuas
Poj niam Ann.arabiensis, 72 h thiab 120 h tom qab ntshav noj, thaum lub sij hawm spawning, tsis muaj kev nyiam rau headspace volatile extracts los ntawm tshiab thiab laus nyuj zis piv nrog pentane tswj (χ2 = 3.07, p > 0.05; Ntxiv cov ntaub ntawv 1: Fig. S1).
Rau cov poj niam Ann.arabiensis, GC-EAD thiab GC-MS kev tshuaj ntsuam xyuas pom yim, rau, peb thiab peb bioactive compounds (Daim duab 5D-G).Txawm hais tias qhov sib txawv ntawm cov tebchaw uas elicited electrophysiological teb tau pom, feem ntau ntawm cov tebchaw no muaj nyob rau hauv txhua lub taub hau volatile rho tawm los ntawm cov tshiab thiab cov laus, refores physiological teb. antennae saum toj kawg nkaus tau suav nrog hauv kev tshuaj xyuas ntxiv.
Tag nrho cov volatile tso tawm tus nqi ntawm bioactive compounds nyob rau hauv lub headspace sau tau nce los ntawm 29 µg h-1 nyob rau hauv cov zis tshiab mus rau 242 µg h-1 nyob rau hauv 168-teev cov zis muaj hnub nyoog, feem ntau yog vim p-cresol thiab m-formaldehyde Phenol nce raws li phenol.In contrast, lub tso tawm tus nqi xws li clone-2-can-txo lwm yam. lub hnub nyoog tso zis ntau ntxiv, uas cuam tshuam nrog qhov pom qhov txo qis hauv cov teeb liab siv (ntau) hauv chromatogram (Fig. 5D)-G sab laug vaj huam sib luag) thiab cov lus teb physiological rau cov tebchaw (Daim duab 5D-G sab xis vaj huam sib luag).
Zuag qhia tag nrho, cov khoom sib xyaw ua ke muaj qhov zoo sib xws ntawm cov khoom siv bioactive uas pom nyob rau hauv cov tshuaj tsis hloov pauv ntawm cov zis tshiab thiab hnub nyoog lub taub hau (Fig. 5D–G) thiab tsis tshwm sim rau kev thov rov hais dua tseem ceeb hauv kev tshawb nrhiav tus tswv tsev (χ2 = 8.15, df = 4, p = 0.083; moss = 2, 4-H, Fig. = 4, p = 0.30; Fig. 5I).Txawm li cas los xij, tom qab kev sib piv ntawm kev sib piv ntawm kev kho mob pom tau tias cov yoov tshaj cum nrhiav tus tswv tsev tau ntxim nyiam rau cov khoom sib xyaw ntawm 24-h hnub nyoog tso zis piv nrog pentane tswj (p = 0.0086; Daim duab 5H).
Txhawm rau ntsuas lub luag haujlwm ntawm tus kheej cov khoom sib xyaw hauv cov khoom sib xyaw ntawm cov zis 24-h-hnub nyoog, rau qhov kev sib cais sib cais tau raug soj ntsuam los ntawm kev sib xyaw ua tiav hauv Y-tube assay, uas tus neeg sib txuas tau raug tshem tawm.Rau tus tswv tsev nrhiav yoov tshaj cum, rho tawm ib tus neeg sib xyaw los ntawm qhov sib xyaw ua tiav tau muaj txiaj ntsig zoo rau cov lus teb rau tus cwj pwm (x. cov ntaub ntawv 1: Daim duab S2A), tag nrho cov kev sib cais sib cais tau zoo nkauj dua li Me me dua qhov sib xyaw ua ke.In sib piv, kev tshem tawm ntawm ib tus neeg sib xyaw los ntawm cov khoom sib xyaw ua ke tsis cuam tshuam rau kev coj cwj pwm ntawm cov yoov tshaj cum nqus ntshav (χ2 = 11.38, df = 6, p = 0.077), nrog rau kev sib xyaw ua kom qis dua, nrog rau qhov sib txawv ntawm 0. 022; Cov Ntaub Ntawv Ntxiv 1: Daim duab S2B).
Hauv ib lub zos malaria-muaj kab mob hauv Ethiopia, qhov ua tau zoo ntawm cov khoom sib xyaw ntawm 24-teev nyuj tso zis hauv kev nyiam yoov tshaj cum nyob rau hauv thaj chaw raug soj ntsuam rau kaum hmo (Fig. 6A).Muaj tag nrho ntawm 4,861 yoov tshaj cum raug ntes thiab txheeb xyuas, ntawm 45.9% ambaepha, 8.7% ambienta. sis thiab 35.4% yog Culex spp.(Ntxiv cov ntaub ntawv 1: Table S1).Anopheles arabinis yog tib tug tswv cuab ntawm An.Gambian hom complex txheeb xyuas los ntawm PCR tsom xam.Qhov nruab nrab, 320 yoov tshaj cum raug ntes ib hmos, thaum lub sij hawm lub ntxiab nrog hluavtaws sib tov χ baits ntes tau yoov tshaj cum ntau dua 1.70 p. < 0.0001) .Cov cuab yeej uas tsis muaj cuab kav tau teeb tsa rau txhua lub tsib hmo kev tswj hwm thaum pib, nruab nrab, thiab xaus ntawm qhov kev sim.Zoo li cov yoov tshaj cum raug ntes nyob rau hauv txhua khub ntawm cov ntxiab, qhia tias tsis muaj kev cuam tshuam ntawm cov tsev (χ2(0, 1665) = 9 × 10-135, p. par > 0. Cov yoov tshaj cum ntes tau hauv cov ntxiab uas muaj cov khoom sib xyaw ua ke tau nce ntxiv: tus tswv tsev nrhiav (χ2(0, 2107) = 138.7, p < 0.0001), kev pub ntshav tsis ntev los no (χ2(0, 650) = 32.2, p < 0.0001) thiab cev xeeb tub (χ2(0.2 = 0.6; 2.2)Cov ntaub ntawv ntxiv 1: Table S1).Qhov no kuj tseem cuam tshuam txog tag nrho cov yoov tshaj cum raug ntes: tus tswv tsev nrhiav> ntshav nqus> cev xeeb tub> cev xeeb tub> txiv neej.
Kev soj ntsuam teb ntawm kev ua tau zoo ntawm 24-teev hluavtaws nyuj cov zis tsw sib xyaw.Field trials tau ua nyob rau sab qab teb-central Ethiopia (daim ntawv qhia), nyob ze lub nroog Maki (ntxig), siv Lub Chaw Tiv Thaiv Kab Mob (CDC) lub teeb ntxiab (txoj cai) hauv cov tsev ua ke, nrog rau Latin square tsim (aerial duab) dc. arabesques (B), tab sis tsis yog Anopheles farroes (C), nyob rau hauv ib tug txawv yam, ib tug physiological lub xeev-dependent nyhuv.Ntxiv rau, cov ntxiab ntes tau ntau ntau tus tswv tsev Culex yoov tshaj cum.(D) Piv nrog tswj.Cov tuav ntawm sab laug sawv cev rau nruab nrab xaiv qhov ntsuas ntawm yoov tshaj cum ntes tau nyob rau hauv khub ntawm greore odoren) thiab bait (nrog rau txoj cai) sawv cev rau qhov nruab nrab xaiv qhov ntsuas nyob rau hauv khub ntawm kev tswj cuab (qhib; N = 5).).Asterisks qhia txog qib tseem ceeb (*p = 0.01 thiab ***p < 0.0001)
Peb hom tau raug ntes sib txawv hauv cov ntxiab uas muaj cov khoom sib xyaw sib xyaw.Nrhiav tus tswv (χ2(1, 1345) = 71.7, p < 0.0001), kev pub ntshav (χ2(1, 517) = 16.7, p < 0.0001) thiab cev xeeb tub (χ2(1, 6bi4) = 1800. daig nyob rau hauv lub ntxiab tso cov khoom sib tov (Fig. 6B), thaum tus nqi ntawm An tsis txawv.Pharoensis nyob rau hauv txawv physiological xeev tau pom (Fig. 6C).Rau Culex, tsuas yog ib tug tseem ceeb nce nyob rau hauv tus naj npawb ntawm yoov tshaj cum nrhiav hosts tau pom nyob rau hauv cov ntxiab baited nrog cov hluavtaws sib tov (χ10.6 = 3.106.0,1. 6D), piv nrog cov cuab yeej tswj.
Cov cuab yeej cuab tam nyob sab nraud ntawm cov chaw muaj peev xwm ntawm cov chaw yug me nyuam thiab cov zej zog nyob deb nroog hauv Ethiopia tau siv los ntsuas seb cov yoov tshaj cum malaria siv nyuj tso zis tsw los ua tus tswv tsev cue.Thaum tsis muaj tus tswv tsev cues, cua sov, thiab nrog los yog tsis muaj nyuj tso zis tsw, tsis muaj yoov tshaj cum raug ntes nyob rau hauv 1 (Cov ntaub ntawv ntxiv, qhov kub thiab txias) tsw, poj niam malaria yoov tshaj cum tau nyiam thiab ntes, txawm tias muaj tsawg tsawg, tsis muaj hnub nyoog tso zis (χ2(5, 25) = 2.29, p = 0.13; Cov ntaub ntawv ntxiv 1: Daim duab S3 ).Tshwj xeeb, kev tswj dej tsis tau ntes cov yoov tshaj cum malaria ntawm qhov kub thiab txias (1.
Cov yoov tshaj cum malaria tau txais thiab faib cov khoom sib xyaw uas muaj nitrogen los ntawm kev them nyiaj pub rau nyuj tso zis (piv txwv li, puddles) txhawm rau txhim kho lub neej-keeb kwm zoo, zoo ib yam li lwm cov kab [2, 4, 24, 25, 26].Cow tso zis yog ib qho yooj yim muaj peev xwm txuas ntxiv txuas ntxiv mus rau qhov chaw zoo li cov kab mob loj thiab cov chaw so ntawm cov tsiaj nyeg. ral tsev thiab spawning sites.Cov poj niam yoov tshaj cum nrhiav cov peev txheej no los ntawm cov ntxhiab tsw thiab muaj peev xwm tswj tau cov tshuaj nitrogenous hauv cov zis, suav nrog urea, cov ntsiab lus nitrogenous loj hauv cov zis [15, 16].Raws li poj niam yoov tshaj cum lub cev lub cev, cov as-ham hauv cov zis yog faib rau cov poj niam uas muaj sia nyob thiab ua kom muaj sia nyob. Cov yam ntxwv ntawm kev yug me nyuam ntawm cov neeg noj ntshav thaum thawj lub voj voog gonadotropic.Yog li ntawd, cov zis sib xyaw ua lub luag haujlwm tseem ceeb rau cov kab mob malaria uas raug kaw zoo li cov neeg laus tsis muaj zaub mov noj [8], vim nws muab cov poj niam yoov tshaj cum muaj peev xwm kom tau txais cov ntsiab lus tseem ceeb nitrogenous los ntawm kev koom nrog kev noj zaub mov tsawg, qhov kev tshawb pom no muaj kev cuam tshuam loj rau cov poj niam cev xeeb tub, ua rau muaj kev pheej hmoo ntawm kev noj qab haus huv. , tag nrho cov uas cuam tshuam rau vector peev xwm. Tsis tas li ntawd, tus cwj pwm no yuav yog lub hom phiaj ntawm kev tswj hwm vector yav tom ntej.