Kuwana uye kupararira kwezvinovaka muviri kunobatanidza kutsvaga zvipembenene uye hunhu hwenhoroondo yehupenyu. Kuti zvikwanise kutsiva kushomeka kwezvinovaka muviri zvakasiyana-siyana padanho rehupenyu, zvipembenene zvinogona kuwana izvi zvinovaka muviri kuburikidza nekudya zvekudya zvekuwedzera, semuenzaniso, nekudya zvinobuda mumapfupa emhuka dzinobereka mapfupa nenzira inozivikanwa semapuddles. Utunga Anopheles arabiani hunoratidzika kunge husina kudya kwakakwana uye, nekudaro, hunoda zvinovaka muviri zvemetabolism uye kubereka. Chinangwa chechidzidzo ichi chaive chekuongorora kana An. arabiensis agitation paweti yemombe kuti iwane zvinovaka muviri ichivandudza hunhu hwenhoroondo yehupenyu.
Iva nechokwadi chekuti yakachengeteka. arabiensis yakakwezvwa nekunhuhwirira kweweti yemombe itsva, yemaawa makumi maviri nemana, yemaawa makumi manomwe nemaviri, uye yemaawa zana nemakumi matanhatu nemasere, uye weti yemombe inotsvaga uye inodyiswa ropa (yemaawa makumi mana nemasere mushure mekudya kweropa) yakayerwa muY-tube olfactometer, uye weti yevakadzi vane pamuviri yakaongororwa kuti ikwanise kubereka. Ongororo yakabatana yemakemikari neelectrophysiological yakazoshandiswa kuona macompounds anoshanda mumuviri weti yemombe pamakirasi ese mana emakore. Misanganiswa yekugadzira yemakemikari anoshanda mumuviri yakaongororwa muY-tube uye mumiedzo yemunda. Kuti vaongorore weti yemombe uye urea yayo huru ine nitrogen senzira yekuwedzera yekudya kwemalaria, zviyero zvekudyisa uye hunhu hwenhoroondo yehupenyu zvakayerwa. Huwandu hweumhutu hwevakadzi uye huwandu hweweti yemombe neurea zvakayerwa. Mushure mekudyisa, weti yevakadzi yakayerwa kuti irarame, kubhururuka kwakasungirirwa, uye kubereka.
Tsvaga ropa nekudya kwemhuka yacho. Muzvidzidzo zverabhoritari nemumunda, maArabu akakwezvwa nekunhuhwirira kwechisikigo uye kwekugadzira kweweti yemombe itsva uye yakura. Hadzi dzaiva nepamuviri dzakanga dzisina hanya nemhinduro dzeweti yemombe panzvimbo dzekuberekera. Hadzi dzaitsvaga uye dzinosveta ropa dzinonyatsotora weti yemombe neurea uye dzinogovera zviwanikwa izvi zvichienderana nehupenyu hwemhuka sechikonzero chemamiriro emuviri ekubhururuka, kurarama, kana kubereka.
Anopheles arabinis kuwana uye kupararira kweweti yemombe kuti ive nehunhu hwakanaka hwenhoroondo yehupenyu. Kudyisa weti yemombe kwekuwedzera kunokanganisa kugona kwevector zvakananga nekuwedzera kurarama kwezuva nezuva uye huwandu hwevector, uye zvisina kunanga nekuchinja mashandiro ekubhururuka uye nekudaro zvinofanirwa kufungwa nezvazvo mune ramangwana.
Kuwana nekuparadzira zvinovaka muviri kunobatanidza kutsvaga zvipembenene uye hunhu hwenhoroondo yehupenyu [1,2,3]. Zvipembenene zvinokwanisa kusarudza nekuwana chikafu uye kuita kudya kunotsiva zvichienderana nekuwanikwa kwechikafu uye zvinodiwa nezvinovaka muviri [1, 3]. Kugoverwa kwezvinovaka muviri kunoenderana nemaitiro enhoroondo yehupenyu uye kunogona kutungamira kune zvinodiwa zvakasiyana zvemhando yekudya uye huwandu muzvikamu zvakasiyana zvehupenyu hwezvipembenene [1, 2]. Kuti zvitsive kushomeka kwezvinovaka muviri, zvipembenene zvinogona kuwana izvi zvinovaka muviri kuburikidza nekudya kwekuwedzera, senge pamatope, tsvina yakasiyana-siyana uye zvinobuda zvezvipenyu zvine mapfupa, uye zvakaora, maitiro anozivikanwa semadziva [2]. Kunyangwe mhando dzakasiyana dzemashavishavi nemoth dzichitsanangurwa zvakanyanya, makomba ekudiridza anowanikwawo mune dzimwe mhando dzezvipembenene, uye kukwezva nekudya marudzi aya ezviwanikwa kunogona kuve nemigumisiro yakakosha pahutano uye humwe hunhu hwenhoroondo yehupenyu [2, 4, 5, 6] ,7]. Utsi hwemalaria Anopheles gambiae sensu lato (sl) hunobuda semunhu mukuru 'asina kudya zvakakwana' [8], saka kudiridza kunogona kuita basa rakakosha muhunhu hwahwo hwenhoroondo yehupenyu, asi maitiro aya kusvika pari zvino kwaregeredzwa. Kushandiswa kwekusagadzikana senzira yekuwedzera kudya kwezvinovaka muviri muchinhu ichi chakakosha kunoda kutariswa sezvo izvi zvingave nemigumisiro yakakosha yezvirwere.
Kudya nitrogen muumhutu hwechikadzi hweAnopheles kwakaderera nekuda kwekusachengetedzwa kwemacalorie mashoma kubva padanho remakonzo uye kusashandiswa zvakanaka kwekudya kweropa [9]. Ann.gambiae sl yechikadzi inowanzo tsiva izvi nekuwedzera kudya kweropa [10, 11], zvichiita kuti vanhu vakawanda vave panjodzi yekubatwa nechirwere ichi uye kuisa umhutu panjodzi huru yekudya. Neimwe nzira, umhutu hunogona kushandisa kudya kwekuwedzera kwetsvina yemhuka dzine vertebrate kuti huwane nitrogen inosimudzira kuchinjika uye kugona kubhururuka, sezvakaratidzwa nezvimwe zvipembenene [2]. Panyaya iyi, kukwezva kwakasimba uye kwakasiyana kweimwe yemhando dzemhuri dziri muAn. Gambian sl complex, Anopheles arabinis, weti yemombe itsva uye yakwegura [12,13,14], kunonakidza. Anopheles arabinis inofarira mhuka dzinoifarira uye inozivikanwa nekubatana uye kudya mombe. Weti yemombe ipfuma yakapfuma mu nitrogen, neurea ichiverengera 50-95% yenitrogen yese muweti itsva [15, 16]. Sezvo weti yemombe ichikura, tupukanana tunoshandisa zvinhu izvi kuderedza kuomarara kwemakemikari ane nitrogen mukati memaawa makumi maviri nemana [15]. Nekuwedzera nekukurumidza kweammonia, kwakabatana nekuderera kwenitrogen yeorganic, tupukanana tunorarama mualkalophilic (tuzhinji twacho tunogadzira makemikari ane chepfu kumhutu) tunokura [15], ingave iri yemukadzi Ann.arabiensis inokwezvwa zvakanyanya neweti ine maawa makumi maviri nemana kana pasi [13, 14].
Muchidzidzo ichi, maAns anodya mombe neropa akatsvagwa. Munguva yekutanga yegonadotropin cycle, arabiensis yakaongororwa kuti iwane nitrogenous compounds, kusanganisira urea, nekusanganisa weti. Tevere, kuyedza kwakaitwa kuti vaongorore kuti umhutu hwechikadzi hunogovera sei chikafu ichi chinogona kuvandudzika kuti hurarame, kubereka uye kutsvaga zvekudya. Pakupedzisira, hwema hweweti yemombe itsva neyakura hwakaongororwa kuti vaone kana izvi zvakapa zviratidzo zvakavimbika zveAn inodya mombe neropa. Mukutsvaga kwavo chikafu ichi chinogona kuva chinovaka muviri, arabiensis akawana hukama hwemakemikari huri shure kwekukwezva kwakasiyana kwakaonekwa. Musanganiswa wekunhuhwirira kwekugadzira wemakemikari anoputika (VOCs) akaonekwa muweti yemaawa makumi maviri nemana wakaongororwazve pasi pemamiriro emunda, zvichiwedzera mhedzisiro yakawanikwa pasi pemamiriro erabhoritari uye zvichiratidza mhedzisiro yekunhuhwirira kweweti yemombe pamamiriro akasiyana emuviri. Kukwezva umhutu. Mhedzisiro yakawanikwa inosimbisa kuti An. arabiensis inowana uye inoparadzira nitrogenous compounds inowanikwa muweti yemhuka dzine vertebrate kuti ishandise hunhu hwenhoroondo yehupenyu. Mhedzisiro iyi inokurukurwa mumamiriro ezvinhu emhedzisiro inogona kuitika uye kuti inogona kushandiswa sei pakuongorora nekutonga vector.
Anopheles arabicans (Dongola strain) yakachengetedzwa pa25 ± 2 °C, 65 ± 5% RH uye 12:12 awa chiedza: rima. Makonye akarerwa mumatireyi epurasitiki (20 cm × 18 cm × 7 cm) akazadzwa nemvura yakacheneswa ndokupihwa chikafu chehove cheTetramin® (Tetra Werke, Melle, DE). Makonye akaunganidzwa mumakapu e30 ml (Nolato Hertila, Åstorp, SE) ndokuzoendeswa kumakeji eBugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) kuti akwanise kubuda. Vakuru vakapihwa 10% sucrose solution ad libitum kusvika mazuva mana mushure mekubuda (dpe), panguva iyo madzimai aitsvaga host akapihwa chikafu nguva pfupi bvunzo isati yaitwa, kana kuti akanyimwa husiku hwose nemvura yakacheneswa isati yaitwa bvunzo, sezvakatsanangurwa pazasi. Makonye akashandiswa pakuedza flight tube akanyimwa chete. Maawa mana kusvika matanhatu nemvura yakadhakwa. Kuti vagadzirire umhutu hunosveta ropa kuti vaongorore hutachiona hwemuviri hunotevera, vakadzi vana vedpe vakapihwa ropa remakwai risina fibrotic (Håtunalab, Bro, SE) vachishandisa nzira yekudyisa membrane (Hemotek Discovery Workshops, Accrington, UK). Vakadzi vakanga vazara vakazoendeswa kumakeji ega ega uye vakapihwa kudya zvakananga, sezvakatsanangurwa pazasi, kana 10% sucrose yakadhakwa kwemazuva matatu bvunzo dzisati dzaitwa dzakatsanangurwa pazasi. Vakadzi vekupedzisira vakashandiswa pakuongorora hutachiona hwemuviri hwemachubhu endege ndokuendeswa kurabhoritari, uye vakazonwa mvura yakasanganiswa yakadhakwa kwemaawa mana kusvika matanhatu bvunzo isati yaitwa.
Kuyedzwa kwekudya kwakashandiswa kuyedza kuyera weti neurea mumukadzi mukuru wechiArab. Vakadzi vanotsvaga vaenzi uye vanodyiswa ropa vakapihwa kudya kwaive ne1% weti yemombe itsva uye yakwegura yakaderedzwa, huwandu hwakasiyana hweurea, uye zvinhu zviviri zvinodzora (10% sucrose nemvura) kwemaawa makumi mana nemasere. Pamusoro pezvo, ruvara rwechikafu (1 mg ml-1 xylene cyanide FF; CAS 2650-17-1; Sigma-Aldrich, Stockholm, SE) rwakawedzerwa mukudya kwacho uye rwakapihwa mu 4 × 4 matrix mumachubhu e 250 µl microcentrifuge (Aoxygen Scientific, Union City, CA, US; Mufananidzo 1A) Zadza kusvika kumucheto (~300 µl). Kuti udzivise kukwikwidzana pakati peumhutu nemhedzisiro inogona kuitika yeruvara rwedhayi, isa umhutu gumi mudhishi hombe rePetri (12 cm mudhayamita uye 6 cm mukukwirira; Semadeni, Ostermundigen, CH; Mufananidzo 1A) murima rakakwana pa25 ± 2 cm °C uye 65 ± 5% maererano. hunyoro. Kuedza uku kwakadzokororwa ka5 kusvika ku10. Mushure mekusangana nekudya, umhutu hwakaiswa pa -20 °C kusvika pakuongorora kwakawedzerwa.
Tsvaga weti yemombe neurea zvinotorwa nemushonga wemhuka nemukadzi anosvuta ropa anonzi Anopheles arabianus. Mukuyedza kudya (A), umhutu hwechikadzi hwakapihwa chikafu chine weti yemombe itsva uye yakwegura, huwandu hwakasiyana hweurea, sucrose (10%), nemvura yakacheneswa (H2O). Humhutu hwechikadzi hunotsvaga mubatsiri (B) nehwechikadzi hunodyiswa ropa (C) hwakanwa sucrose yakawanda kupfuura chero imwe kudya yakaedzwa. Cherechedza kuti humhutu hwechikadzi hunotsvaga mubatsiri hwakanwa weti yemombe yemaawa makumi manomwe nemaviri pasi peweti yemombe yemaawa zana nemakumi matanhatu nemasere (B). Avhareji yehuwandu hwenitrogen (± standard deviation) yeweti inoratidzwa muchikamu chiri mukati. Humhutu hwechikadzi hunotsvaga mubatsiri (D, F) nehwechikadzi hunosvuta ropa (E, G) hunonwa urea zvichienderana nedosi. Avhareji yehuwandu hwemhepo inofema (D, E) ine mazita akasiyana emabhii akasiyana zvakasiyana kubva kune mumwe nemumwe (one-way ANOVA uchishandisa Tukey's post hoc analysis; p < 0.05). Error bars dzinomiririra standard error yeavean (BE). Straight dashed line inomiririra log-linear regression line (F, G)
Kuti chikafu chibudiswe, umhutu hwakaiswa muchubhu dze 1.5 ml microcentrifuge dzine 230 µl yemvura yakacheneswa uye tissue yakakanganiswa uchishandisa moteri inoraswa uye isina waya (VWR International, Lund, SE), zvichiteverwa ne centrifugation pa 10 krpm kwemaminitsi gumi. Supernatant (200 µl) yakachinjirwa kune 96-well microplate (Sigma-Aldrich) uye absorbance (λ620) yakaongororwa uchishandisa spectrophotometer-based microplate reader (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Neimwe nzira, umhutu hwakakuyiwa mu 1 ml yemvura yakacheneswa, 900 µl yayo yakachinjirwa kune cuvette kuti iongororwe ne spectrophotometric (λ 620 nm; UV 1800, Shimadzu, Kista, SE). Kuti kuyere kudya, standard curve yakagadzirwa nekusanganiswa kwe serial kuti ibudise 0.2 µl kusvika 2.4. µl ye 1 mg ml-1 xylene cyanide. Zvadaro, huwandu hwemavara eruvara rwedhayi hunozivikanwa hwakashandiswa kuona huwandu hwechikafu chakadyiwa neumhutu hwega hwega.
Ruzivo rwehuwandu rwakaongororwa uchishandisa imwe nzira yekuongorora kusiyana (ANOVA) zvichiteverwa nekuenzanisa kwaTukey's post hoc pairwise (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). Kuongororwa kweLinear regression kwakatsanangura huwandu hweurea hunoenderana nehuwandu uye kwakaenzanisa mhinduro pakati peumhutu hunotsvaga host uye hunosvuta ropa (GraphPad Prism v8.0.0 yeMac, GraphPad Software, San Diego, CA, US).
Zvinenge 20 µl zvemuenzaniso weweti kubva muboka rega rega rezera zvakasungirirwa paChromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) uye zvakaputirwa mumakapisi etin (8 mm × 5 mm). Makapisi akaiswa muimba yekupisa yeCHNS/O analyzer (Flash 2000, Thermo Fisher Scientific, Waltham, MA, US) kuti vaone huwandu hwenitrogen muweti itsva uye yakwegura zvichienderana neprotocol yemugadziri. Total nitrogen (g N l-1) yakayerwa zvichibva pahuwandu hweurea hunozivikanwa hwakashandiswa sechiyero.
Kuti vaongorore mhedzisiro yekudya pahupenyu hwemukadzi anoda rubatsiro uye anosvuta ropa, umhutu hwakaiswa mumwe nemumwe mundiro hombe dzePetri (12 cm mudhayamita uye 6 cm mukukwirira; Semadeni) neburi rakafukidzwa nemesh muvharo (3 cm mudhayamita) neKufemesa uye kupa chikafu. Kudya kwakapihwa zvakananga mushure me4 dpe uye kwaisanganisira 1% weti yemombe itsva uye yakwegura yakaderedzwa, huwandu huna hweurea, uye maviri ekudzora, 10% sucrose nemvura. Kudya kwega kwega kwakaiswa papombi pa dental tampon (DAB Dental AB, Upplands Väsby, SE) yakaiswa musirinji ye5 ml (Thermo Fisher Scientific, Gothenburg, SE), pombi yakabviswa, ndokuiswa pamusoro pedishi repetri (mufananidzo 1).1A). Chinja kudya kwako zuva nezuva. Chengetedza lab sezvakatsanangurwa pamusoro apa. Umhutu hwakapona hwakaverengwa kaviri pazuva, nepo umhutu hwakafa hwakaraswa kusvika umhutu hwekupedzisira hwafa (n = 40 pakurapwa kwega kwega). Kupona kweumhutu hwakadyiswa kudya kwakasiyana-siyana kwakaongororwa nenhamba uchishandisa Kaplan-Meyer survival curves uye bvunzo dzepamusoro-soro dzekuenzanisa kuenzanisa kugoverwa kwekupona pakati pekudya (IBM SPSS Statistics 24.0.0.0).
Chigayo chekubhururuka neumhutu chakagadzirwa naAttisano et al.[17], chakagadzirwa nemapaneru eacrylic akajeka ane ukobvu hwe5 mm (10 cm yakafara x 10 cm kureba x 10 cm kukwirira) asina mapaneru ekumberi neekumashure (Mufananidzo 3: pamusoro). Musangano wepivot une chubhu yakatwasuka yakagadzirwa nekoramu yegesi chromatography (0.25 mm id; 7.5 cm L) ine magumo akanamirwa kune tsono yezvipembenene yakasungirirwa pakati pemagineti maviri e neodymium ari kure ne9 cm. Chubhu yakatwasuka yakagadzirwa nezvinhu zvakafanana (6.5 cm L) yakapatsanura chubhu yakatwasuka kuti igadzire ruoko rwakasungwa neruoko rwaitakura chidimbu chidiki chealuminium foil sechiratidzo chinovhiringidza chiedza.
Hadzi dzaive nenzara kwemaawa makumi maviri nemana dzakapihwa chikafu chataurwa pamusoro apa kwemaminitsi makumi matatu dzisati dzadzorwa. Umhutu hwehadzi hwakadyiswa zvakakwana hwakazoiswa imwe neimwe pachando kwemaminitsi maviri kusvika matatu uye hwakanamirwa pamapini ezvipembenene newakisi (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) uye dzakazosungirirwa mumaoko emachubhu akatambanuka. Flying Mill. Kuchinja kwekufamba kwega kwega kwakanyorwa nedata logger yakagadzirwa, ndokuzochengetwa nekuratidzwa uchishandisa PC-Lab 2000™ software (v4.01; Velleman, Gavere, BE). Chigayo chekubhururuka chakaiswa muimba inodzorwa nemamiriro ekunze (12 h:12 h, chiedza: rima, 25 ± 2 °C, 65 ± 5% RH).
Kuti tifungidzire maitiro ekufamba nendege, daro rose rakafambiswa (m) uye huwandu hwese hwezviitiko zvekubhururuka zvakatevedzana zvakaverengerwa paawa mukati memaawa makumi maviri nemana. Pamusoro pezvo, avhareji yedaro rakafambiswa nemadzimai ega ega yakaenzaniswa mukurapa uye yakaongororwa uchishandisa ANOVA imwe chete uye ongororo yaTukey's post hoc (JMP Pro, v14.0.0, SAS Institute Inc.), uko avhareji yedaro yaionekwa sechinoshanduka zvichienderana ne , nepo kurapwa kuri chinhu chakazvimiririra. Pamusoro pezvo, avhareji yenhamba yekutenderera inoverengerwa mukuwedzera kwemaminetsi gumi.
Kuti vaongorore mhedzisiro yekudya pakuita kwekubereka kweAn.arabiensis, hadzi nhanhatu (4 dpe) dzakatamisirwa zvakananga kumakeji eBugdorm (30 cm × 30 cm × 30 cm) mushure mekutorwa kweropa uye dzakazopa kudya kwekuyedza kwemaawa makumi mana nemasere sezvakatsanangurwa pamusoro apa. Kudya kwakazobviswa uye makapu ekuberekesa (30 ml; Nolato Hertila) akazadzwa ne20 ml yemvura yakacheneswa akapihwa pazuva rechitatu kwemaawa makumi mana nemasere, achichinja maawa makumi maviri nemana ega ega. Dzokorora kudya kwega kwega ka20-50. Mazai akaverengwa uye akanyorerwa keji yega yega yekuyedza. Mienzaniso yemazai yakashandiswa kuongorora kusiyana kwehukuru nehurefu hwemazai ega ega (n ≥ 200 pakudya kwega) uchishandisa maikorosikopu yeDialux-20 (DM1000; Ernst Leitz Wetzlar, Wetzlar, DE) yakashongedzerwa neLeica Camera (DFC) 320 R2; Leica Microsystems Ltd., DE). Mazai akasara akachengetwa muimba inodzorwa nemamiriro ekunze pasi pemamiriro akajairwa ekurera kwemaawa makumi maviri nemana, uye sampuro shoma yemakonye ekutanga akabuda munguva pfupi yapfuura (n ≥ 200 pakudya) yakayerwa, sezvakatsanangurwa pamusoro apa. Huwandu hwemazai nehukuru hwemazai nekonye zvakaenzaniswa pakati pekurapa nekushandisa ANOVA imwe chete uye Tukey's post hoc analysis (JMP Pro, v14.0.0, SAS Institute Inc.).
Zvipuka zvinoputika zvemusoro kubva muweti itsva (awa imwe chete mushure mekutorwa kwesampuro), maawa makumi maviri nemana, maawa makumi manomwe nemaviri uye maawa zana nemakumi matanhatu nemasere zvakatorwa kubva mumienzaniso yakatorwa kubva kumombe dzeZebu, Arsi. Kuti zvive nyore, mienzaniso yeweti yakatorwa mangwanani-ngwanani mombe dzichiri mudanga. Mienzaniso yeweti yakatorwa kubva kuvanhu gumi uye 100-200 ml yemuenzaniso wega wega yakaendeswa kumabhegi ekubikira epolyamide (Toppits Cofresco, Frischhalteprodukte GmbH and Co., Minden, DE) mu3 l polyamide ine chivharo Mumadhiramu epurasitiki evinyl chloride. Zvipuka zvinoputika zvemusoro kubva mumuenzaniso wega wega weti yemombe zvakatorwa zvakananga (zvitsva) kana mushure mekuibva patembiricha yemukamuri kwemaawa makumi maviri nemana, maawa makumi manomwe nemaviri uye maawa zana nemakumi matanhatu nemasere, kureva kuti muenzaniso wega wega weti waimiririra boka rega rega rezera.
Pakuunganidza misoro yemagetsi, sisitimu yekuvhara yakashandiswa kutenderedza rwizi rwegasi rakasefetwa nekabhoni (100 ml min-1) kuburikidza nebhegi repolyamide kuenda kukoramu yemvura kwemaawa 2.5 uchishandisa pombi yemvura yediaphragm (KNF Neuberger, Freiburg, DE). Senzira yekudzora, kuunganidzwa kwenzvimbo yemhepo kwakaitwa kubva mubhegi repolyamide risina chinhu. Koramu yemvura yakagadzirwa neTeflon tube (5.5 cm x 3 mm id) ine 35 mg yePorapak Q (50/80 mesh; Waters Associates, Milford, MA, US) pakati pemagirazi emvere dzemagirazi. Usati washandisa, koramu yakasukwa ne1 ml redistilled n-hexane (Merck, Darmstadt, DE) uye 1 ml pentane (99.0% pure solvent GC grade, Sigma Aldrich). Ma adsorbed volatiles akabviswa ne400 μl yepentane. Ma Headspace akaunganidzwa akaunganidzwa ndokuzochengetwa pa -20°C kusvika ashandiswa kwenguva yakareba. ongororo.
Mhinduro dzemaitiro eAn-host-seeking uye-eating-blood extracts An.Headspace volatile extracts dzakatorwa kubva muweti itsva, maawa makumi maviri nemana, maawa makumi manomwe nemaviri, uye maawa zana nemakumi matanhatu nemasere dzakaongororwa kuti dziwane volatile extracts kubva kumumhutu weArabidopsis uchishandisa straight glass tube olfactometer [18]. Kuedza uku kwakaitwa panguva yeZT 13-15, nguva yepamusoro yeAn's home-seeking activity.Arab [19]. Glass tube olfactometer (80 cm × 9.5 cm id) yakavhenekerwa ne3 ± 1 lx yechiedza chitsvuku kubva kumusoro. Marasha akasefetwa uye akanyoroveswa nemhepo (25 ± 2 °C, 65 ± 2% relative humidity) akapfuura bioassay pa30 cm s-1. Mhepo inopfuudzwa nepakati pema screens esimbi isina tsvina, ichigadzira laminar flow uye uniform plume structure. Dental tampon dispenser (4 cm × 1 cm; L:D; DAB Dental AB), yakasungirirwa kubva pa5 cm coil pamucheto wemhepo. olfactometer, ine shanduko dzestimulator maminitsi mashanu ega ega. Pakuongorora, 10 μl yechinhu chimwe nechimwe chemusoro, chakaderedzwa 1:10, chakashandiswa sechinosimudzira. Huwandu hwakaenzana hwepentane hwakashandiswa sechinodzora. Umhutu hwemunhu mumwe nemumwe hunotsvaga host kana kuti hunosvuta ropa hwakaiswa mumakeji ekuburitsa ega maawa 2-3 kuedza kusati kwatanga. Keji yekuburitsa yakaiswa parutivi rwemhepo yeolfactometer, uye umhutu hwakabvumirwa kudzokororwa kwemaminitsi 1, uye ipapo vharuvhu yemabhururu ekeji yakavhurwa kuti iburitse. Kukwezva kurapwa kana kudzora kwakaongororwa sehuwandu hweumhutu hwakasangana nekwakabva mukati memaminitsi mashanu ekuburitsa. Chikamu chega chega chemusoro che volatile extract uye kudzora kwakadzokororwa kanenge ka30, uye kudzivirira mhedzisiro yezuva rimwe chete, nhamba imwechete yekurapa nekudzora yakaedzwa pazuva rega rega rekuyedza. Tsvaga mhinduro kubva kune host neropa-vakadyiswa Mhinduro. Arabic vs headspace sets dzakaongororwa uchishandisa nominal logistic regression ichiteverwa nekuenzanisa pairi kwe odd ratios (JMP Pro, v14.0.0, SAS Institute Inc.).
Mhinduro yekubereka yaAn. Zvimedu zvemusoro kubva muweti yemombe itsva uye yakura zvakaongororwa mumakeji eBugdorm (30 cm × 30 cm × 30 cm; MegaView Science). Makapu epurasitiki (30 mL; Nolato Hertila) akazadzwa ne20 mL yemvura yakasanganiswa zvakapa substrate yekubereka uye akaiswa mumakona akatarisana ekaji, 24 cm kubva pakati. Makapu ekurapa akagadziriswa ne10 μl yechikamu chimwe nechimwe chemusoro padilution ye1:10. Huwandu hwakaenzana hwepentane hwakashandiswa kugadzirisa kapu yekudzora. Makapu ekurapa uye ekudzora akachinjaniswa pakati pekuyedza kwega kwega kuti aone mhedzisiro yenzvimbo. Makadzi gumi akadyiswa ropa akasunungurwa mumakeji ekuyedza paZT 9-11 uye mazai mumakapu akaverengwa maawa makumi maviri nemana gare gare. Fomura yekuverenga indekisi yekubereka ndeiyi: (nhamba yemazai akaiswa mukapu yekurapa - nhamba yemazai akaiswa mukapu yekudzora) / (nhamba yese yemazai akaiswa). Kurapa kwega kwega kwakadzokororwa kasere.
Kuongororwa kwegas chromatographic uye electron antenna pattern detection (GC-EAD) yeAn.arabiensis yechikadzi kwakaitwa sezvakatsanangurwa kare [20]. Muchidimbu, fresh headspace volatile extracts dzakaparadzaniswa uchishandisa Agilent Technologies 6890 GC (Santa Clara, CA, US) ine HP-5 column (30 m × 0.25 mm id, 0.25 μm film thickness, Agilent Technologies). uye weti yekukwegura. Hydrogen yakashandiswa sechikamu chekufamba nepakati nepakati pehuwandu hwekuyerera kwe45 cm s-1. Sample yega yega (2 μl) yakabayiwa kwemasekonzi makumi matatu mu splitless mode ne inlet tembiricha ye225 °C. Tembiricha yeGC oven yakarongwa kubva pa35 °C (3 minute hold) kusvika pa300 °C (10 minute hold) pa10 °C min-1. MuGC effluent splitter, 4 psi yenitrogen yakawedzerwa ndokukamurwa 1:1 muGerstel 3D/2 low dead volume cross (Gerstel, Mülheim, DE) pakati peflame ionization detector neEAD. GC effluent capillary yeEAD yakapfuudzwa neGerstel ODP-2 transfer line, iyo inoteedzera tembiricha yeGC oven pamwe ne5 °C, mugirazi tube (10 cm × 8 mm), uko yakasanganiswa necarbon-filtered, humidified air (1.5 l min−1). Antena yakaiswa 0.5 cm kubva pakabuda chubhu. Umhutu hwega hwega hwaive neumwe chete, uye kune umhutu hwaitsvaga mudziviriri, hunenge hutatu hwakafanana hwakaitwa pasampuli dzeweti dzezera rega rega.
Kuonekwa kwemakemikari anoshanda mumuviri mumiunganidzwa yemusoro weti yemombe itsva neyakwegura uchishandisa GC yakabatana uye mass spectrometer (GC-MS; 6890 GC uye 5975 MS; Agilent Technologies) kuti ibudise mhinduro dzeantenna muGC-EAD analysis, ichishanda mu electron impact ionization mode pa70 eV. GC yaive neHP-5MS UI-coated fused silica capillary column (60 m × 0.25 mm inner diameter, 0.25 μm film thickness) ichishandisa helium se mobile phase ine avhareji linear flow rate ye35 cm s-1. Sample ye 2 μl yakapinzwa uchishandisa ma injector settings akafanana uye tembiricha ye oven seye GC-EAD analysis. Macompounds akaonekwa zvichienderana nenguva yawo yekuchengetedza (Kovát index) uye mass spectra zvichienzaniswa ne custom library ne NIST14 library (Agilent). Macompounds akaonekwa akasimbiswa nekupinzwa echokwadi standards (Additional File 1: Table S2). Kuti uwane quantification, heptyl acetate (10 ng, 99.8% makemikari akachena, Aldrich) yakabayiwa sechiyero chekunze.
Kuongorora kushanda kwemusanganiswa wekunhuhwirira kwekugadzira une mishonga inoshanda mumuviri inowanikwa muweti itsva neyekare kuti ikwezve Ans.arabiensis inotsvaga host uye inosveta ropa, uchishandisa olfactometer uye protocol yakafanana neiri pamusoro apa. Misanganiswa yekugadzira yakatevedzera kuumbwa uye huwandu hwemakemikari ari mumisanganiswa yemusoro inonyunguduka yeweti itsva, yemaawa makumi maviri nemana, yemaawa makumi mana nemasere, yemaawa makumi manomwe nemaviri, uye yemaawa zana nemakumi matanhatu nemasere (Mufananidzo 5D-G; Faira Rinowedzera 1: Tafura S2). Kuti uongorore, shandisa 10 μl ye1:100 dilution yemusanganiswa wakagadzirwa zvizere, ine huwandu hwekusunungurwa hunotangira pa140-2400 ng h-1, kuti uongorore kukwezva kwehost uye husveta ropa. Mushure meizvozvo, bvunzo inoitwa pamusanganiswa wakazara, umo musanganiswa wekubvisa wemakemikari emusanganiswa mumwe chete wemusanganiswa wakazara unobviswa. Tsvaga mhinduro kubva kuhost uye Ans inodyiswa ropa. Musanganiswa weArab vs synthetic ne subtractive wakaongororwa uchishandisa nominal logistic regression uchiteverwa nekuenzanisa pairi kune odd. zviyero (JMP Pro, v14.0.0, SAS Institute Inc.).
Kuti vaongorore kana weti yemombe ichigona kushanda senzvimbo inogara umhutu hwemalaria, weti yemombe itsva uye yakwegura, yakaunganidzwa sezvakatsanangurwa pamusoro apa, uye mvura yakaiswa mumabhaketi matatu emvura (100 ml) akasungirirwa mumisungo yekubata hove. (BG-HDT version; BioGents, Regensburg, DE). Misungo gumi yakaiswa 50 m kure mumafuro, 400 m kubva munharaunda yemusha (Silay, Ethiopia, 5°53´24´´N, 37°29´24´´E) uye pasina mombe, munzvimbo dzinoberekera mhuka nemisha. Misungo mishanu yakapiswa kuti ifananidze kuvapo kwemuputi, nepo misungo mishanu yakasiyiwa isina kupiswa. Nzvimbo yega yega yekurapa inotenderedzwa husiku hwese kwehusiku hushanu. Nhamba dzeumhutu dzakabatwa mumisungo yakachekwa neweti yemazera akasiyana dzakaenzaniswa uchishandisa logistic regression ne beta binomial distribution (JMP Pro, v14.0.0, SAS Institute Inc.).
Mumusha une malaria uri pedyo neguta reMaki, Oromia region, Ethiopia (8° 11′ 08″ N, 38° 81′ 70″ E; Mufananidzo 6A). Chidzidzo ichi chakaitwa pakati paNyamavhuvhu napakati paGunyana usati watanga kupfapfaidza mumba gore rega rega, pamwe chete nemwaka wemvura wakareba. Dzimba shanu (dzakaparadzana nemamita 20–50) dziri kunze kwemusha dzakasarudzwa kuti dziitwe chidzidzo ichi (Mufananidzo 6A). Nzira dzakashandiswa pakusarudza dzimba idzi dzaive: mhuka hadzibvumidzwe kupinda mumba, kubika mumba (kutora huni kana marasha) kwaisabvumidzwa (zvirinani panguva yekuedzwa), uye dzimba dzine vanhu vaviri chete, vanorara mumishonga isina zvipembenene. pasi pemambure eumhutu akarapwa. Mvumo yetsika yakapihwa neInstitutional Research Ethics Review Board (IRB/022/2016) yeFaculty of Natural Sciences (CNS-IRB), Addis Ababa University, zvichienderana nemirairo yakagadzwa neWorld Medical Association Declaration yeHelsinki. Mvumo kubva kumusoro wega wega wemhuri yakawanikwa nerubatsiro rwevashandi vekuwedzera hutano. Maitiro ese anotsigirwa nevatungamiriri vemunharaunda padanho redunhu newadhi ('kebele'). Dhizaini yekuedza yakatevera dhizaini ye2 × 2 Latin square, umo musanganiswa wekugadzira uye kudzora zvakapihwa dzimba dzakabatana pausiku hwekutanga uye zvakachinjaniswa pakati pedzimba pausiku hwekuedza hunotevera. Maitiro aya akadzokororwa kagumi. Pamusoro pezvo, kuti vaongorore mashandiro eumhutu mudzimba dzakasarudzwa, misungo yeCDC yakarongerwa kumhanya husiku hushanu hwakatevedzana pakutanga, pakati nepakupera kwekuyedzwa kwemunda panguva imwe chete yezuva.
Musanganiswa wekugadzira une mishonga mitanhatu inoshanda mumuviri wakanyungudutswa mu heptane (97.0% solvent GC grade, Sigma Aldrich) ndokuburitswa pa 140 ng h-1 uchishandisa donje wick dispenser [20]. Wick dispenser yakabvumira mishonga yese kuburitswa zvakaenzana mukati mekuyedza kwemaawa gumi nemaviri. Heptane yakashandiswa senzira yekudzora. Bhodhoro rakasungirirwa padyo nenzvimbo yekupinda muCenters for Disease Control and Prevention (CDC) light trap (John W. Hock Company, Gainesville, FL, US; Mufananidzo 6A). Misungo yakaturikwa 0.8 - 1 m pamusoro pevhu, pedyo netsoka yemubhedha, uye muzvipiri akarara pasi pe mosquito net isina kurapwa uye akashandiswa pakati pa18:00 na06:30. Umhutu hwakabatwa nebonde uye mamiriro emuviri (husina kudya, hwakadyiswa, hwakabata pamuviri, uye hwakabata pamuviri [21] hwakazoongororwa uchishandisa polymerase chain reaction (PCR) analysis kuti uone mhando inozivikanwa seA. gambiae sl. Nhengo dzecomplex [23]. Mukudzidza kwemumunda, trap trapping yedzimba dzakaita sepair houses yakaongororwa uchishandisa nominal logistic fit model, uko kukwezva kwaiva dependent variable uye treatment (synthetic mixture vs control) yaive fixed effect (JMP® 14.0. 0. SAS Institute Inc.). Pano, tinotaura nezve χ2 uye p-values ​​​​kubva ku likelihood ratio test.
Ongorora kana yakachengeteka. arabiensis yakakwanisa kuwana weti, iyo inonyanya kunobva nitrogen, urea, nekudyisa zvakananga, mukati memaawa makumi mana nemasere ekushandiswa kwemazuva mana mushure me (dpe) bvunzo dzekudya dzevakadzi dzinotsvaga host uye dzakapihwa ropa (Mufananidzo 1A). Vakadzi vese vanotsvaga host nevanosvuta ropa vakanwa sucrose yakawanda kupfuura chero imwe kudya kana mvura (F(5,426) = 20.15, p < 0.0001 uye F(5,299) = 56.00, p < 0.0001, zvichiteerana; Mufananidzo 1B,C). Uyezve, vakadzi vanotsvaga host vakadya zvishoma muweti mumaawa makumi manomwe nemaviri zvichienzaniswa neweti mumaawa zana nemakumi matanhatu nemasere (Mufananidzo 1B). Pavakapihwa kudya kune urea, vakadzi vanotsvaga host vakanwa huwandu hwakawanda hweurea pa2.69 mM zvichienzaniswa nezvimwe zvese zvakaunganidzwa nemvura, ukuwo zvisingasiyaniswi ne10% sucrose (F(10,813) = 15.72, p < 0.0001; Mufananidzo 1D). Izvi zvaipesana nemhinduro yevakadzi vakadyiswa ropa, avo vaiwanzo nwa zvekudya zvine urea zvakawanda kupfuura mvura, kunyangwe zviri pasi pe10% sucrose (F(10,557) = 78.35, p < 0.0001; Mufananidzo 1).1E). Uyezve, pakuenzanisa mamiriro maviri emuviri, vakadzi vane phlebotomist vakanwa urea yakawanda kupfuura vakadzi vanotsvaga host pahuwandu hwakaderera, uye vakadzi ava vakanwa huwandu hwakafanana hweurea pahuwandu hwakakwira (F(1,953) = 78.82, p < 0.0001; Mufananidzo 1F, G). Kunyange zvazvo kudya kubva mukudya kune urea kwairatidzika kunge kwakakodzera (Mufananidzo 1D,E), vakadzi vari mumamiriro ese emuviri vakakwanisa kugadzirisa huwandu hweurea inonyudzwa pamusoro pehuwandu hwese hweurea nenzira ye log-linear (Mufananidzo 1F,G). ).Saizvozvowo, umhutu hunoita sehunodzora kutorwa kwenitrogen yavo nekudzora huwandu hweweti inonyudzwa, sezvo huwandu hwenitrogen muweti huchiratidzwa nehuwandu hunonyudzwa (Mufananidzo 1B, C uye B insets).
Kuti vaongorore migumisiro yeweti neurea pakurarama kweumhutu hunotsvaga uye hunosvuta ropa, vakadzi vakadyiswa weti yezera rese ina (itsva, maawa 24, maawa 72, uye maawa 168 mushure mekuiswa) uye huwandu hwakasiyana hweurea, pamwe nemvura yakasanganiswa uye 10% sucrose zvakashandiswa senzira yekudzora (Mufananidzo 2A). Ongororo iyi yekurarama yakaratidza kuti kudya kwaive nemhedzisiro yakakura pakurarama kwese kwese kwevakadzi vanotsvaga weti (weti: χ2 = 108.5, df = 5, p < 0.0001; urea: χ2 = 122.8, df = 5, p < 0.0001; Mufananidzo 2B, C) uye vakadzi vanodyiswa ropa (weti: χ2 = 93.0, df = 5, p < 0.0001; urea: χ2 = 137.9, df = 5, p < 0.0001; Mufananidzo 2D,E).Mukuyedza kwese, vakadzi vaipiwa weti, urea, nemvura vaive nehuwandu hwakaderera hwekurarama zvichienzaniswa nevakadzi vaipiwa weti ine sucrose (Mufananidzo 2B-E). Vakadzi vaitsvaga weti itsva neyakasakara vakaratidza huwandu hwakasiyana hwekurarama, neavo vaipiwa weti yakasakara kwemaawa 72 (p = 0.016) vaine mukana wakaderera wekurarama (Mufananidzo 2B). Uyezve, vakadzi vaitsvaga weti vakapa weti ye135 mM urea vakararama kwenguva yakareba kupfuura mvura inodzorwa (p < 0.04) (Mufananidzo 2C). Zvichienzaniswa nemvura, vakadzi vaipiwa weti itsva uye weti yemaawa 24 vakararama kwenguva yakareba (p = 0.001 uye p = 0.012, zvichiteerana; Mufananidzo 2D), nepo vakadzi vaipiwa weti yemaawa 72 vakararama kwenguva yakareba kupfuura avo vaipiwa weti pfupi itsva uye weti yemaawa 24 (p < 0.0001 uye p = 0.013, zvichiteerana; Mufananidzo 2D). Pavakapa weti ye135 mM urea, vakadzi vaipiwa ropa vakararama kwenguva yakareba kupfuura vese mamwe ma "urea" nemvura (p < 0.013; Mufananidzo 2E).
Kupona kwemumhutu anosvira uye anosvira ropa anonzi Anopheles arabinis achidya weti yemombe neurea. Mukuongorora kwebioassay (A), umhutu hukadzi hwakapihwa chikafu chine weti yemombe itsva uye yakwegura, huwandu hwakasiyana hweurea, sucrose (10%) nemvura yakasanganiswa (H2O). Kupona kweumhutu hunotsvaga mumhutu (B, C) uye hunosvira ropa (D, E) kwakanyorwa maawa gumi nemaviri ega ega kusvika hunhu hwemhutu hunosvira (B, D) uye urea (C, E), uye zvinodzora, Sucrose nemvura, zvafa.
Nhanho yese uye huwandu hwemapoinzi akaonekwa mubvunzo ye "flight mill" mumaawa makumi maviri nemana zvakasiyana pakati peumhutu hunotsvaga weti uye hunosvuta ropa, izvo zvakaratidza kusashanda zvakanaka kwekubhururuka (Mufananidzo 3). Umhutu hunotsvaga weti hune weti itsva uye yakwegura kana sucrose nemvura hwakaratidza maitiro akasiyana ekubhururuka (Mufananidzo 3), nemhutu dzinodya weti itsva dzichinyanya kushanda mangwanani, nepo mhutu dzinodya weti dzine maawa makumi maviri nemana negumi nematanhatu nemasere (24) neumhutu hunodya weti hwakaratidza maitiro akasiyana ekubhururuka uye hwainyanya kushanda zuva nezuva. Umhutu hukadzi hunopa weti ye sucrose kana yemaawa makumi manomwe nemaviri hwakaratidza kushanda mukati memaawa makumi maviri nemana, nepo mhutu dzinopa mvura dzainyanya kushanda mukati menguva yepakati. Umhutu hunodya sucrose hwakaratidza huwandu hwekushanda hwakanyanya husiku uye mangwanani-ngwanani, nepo mhutu dzinodya weti dzine maawa makumi manomwe nemaviri dzakaona kudzikira kwekushanda mukati memaawa makumi maviri nemana (Mufananidzo 3).
Kubhururuka kwemhuka yemusango inosveta ropa inonzi Anopheles arabinis, inosveta ropa, ichidya weti yemombe neurea. Mubvunzo wekugadzira chigayo chekubhururuka, umhutu hwechikadzi hwakadyiswa weti yemombe itsva neyakura, huwandu hwakasiyana hweurea, sucrose (10%), nemvura yakasanganiswa (H2O) zvakasungirirwa mumaoko akatambanuka, achitenderera zvakasununguka (pamusoro). Kune mhuka dzemusango dzinosveta ropa (kuruboshwe) nedzinosveta ropa (kurudyi), daro rose nenhamba yekufamba nendege paawa yega yega yekudya mukati memaawa makumi maviri nemana zvakanyorwa (rima: grey; chiedza: chena). Avhareji daro uye avhareji yenhamba yekufamba inoratidzwa kurudyi rwegirafu yekushanda kwecircadian. Mitsetse yekukanganisa inomiririra chikanganiso chakajairwa cheavhareji. Kuongorora kwenhamba ona chinyorwa
Kazhinji, kufamba kwese kwese kwevakadzi vanotsvaga mbuva kwaitevera maitiro akafanana nekufamba kwenguva kwemaawa makumi maviri nemana. Kufamba kwenguva refu kwakakanganiswa zvakanyanya nekudya kwavakadya (F(5, 138) = 28.27, p < 0.0001), uye vakadzi vanotsvaga mbuva vakadya maawa makumi manomwe nemaviri eweti vakafamba kwenguva refu zvichienzaniswa nedzimwe nzira dzese dzekudya (p < 0.0001), uye umhutu hwakadyiswa sucrose hwakabhururuka kwenguva yakareba kupfuura umhutu hutsva (p = 0.022) uye hwakadyiswa weti maawa makumi maviri nemana (p = 0.022). Kusiyana nemaitiro ekufamba kwenguva anotsanangurwa nekudya kweweti, vakadzi vanotsvaga mbuva vakadyiswa urea vakaratidza kufamba kwenguva refu kwemaawa makumi maviri nemana, vachisvika pakakwirira mukati mehafu yechipiri yechikamu cherima (Mufananidzo 3). Kunyangwe maitiro ekushanda aive akafanana, vakadzi vanotsvaga mbuva vakadyiswa urea zvakanyanya kuwedzera kureba kwekubhururuka zvichienderana nehuwandu hwakanyudzwa (F(5, 138) = 1310.91, p < 0.0001). Kutsvaga mbuva Vakadzi vakadyiswa chero huwandu hweurea hwakabhururuka kwenguva yakareba kupfuura vakadzi vakadyiswa mvura kana sucrose (p < 0.03).
Kufamba kwese kweumhutu hunosveta ropa kwakagara kwakasimba uye kwakaramba kuripo kwemaawa makumi maviri nemana pakudya kwese, nekuwedzera kwekushanda kweweti mukati mehafu yechipiri yenguva yerima yevakadzi vanodyiswa mvura pamwe chete nevakadzi vanodyiswa vachiri vacheche uye vane maawa makumi maviri nemana ekuberekwa (mufananidzo 3). Kunyange zvazvo kudya kweweti kwakakanganisa zvakanyanya avhareji yekufamba kwese kwevakadzi vanodyiswa ropa (F(5, 138) = 4.83, p = 0.0004), kudya kweurea hakuna (F(5, 138) = 1.36, p = 0.24). Nemamwe weti uye kudya kwekudzora (kutsva, p = 0.0091; maawa makumi manomwe nemaviri, p = 0.0022; maawa gumi nematanhatu nemasere, p = 0.001; sucrose, p = 0.0017; dH2O, p = 0.036).
Mhedzisiro yekudyisa weti neurea pamatanho ekubereka yakaongororwa mukuongorora kwezvipenyu zvinoisa mazai (Mufananidzo 4A) uye yakaongororwa zvichienderana nehuwandu hwemazai anoiswa nehadzi yega yega, saizi yezai, uye mabuu ekutanga achangoburwa. Huwandu hwemazai anoiswa. Hadzi dzechiArab dzinodyiswa weti dzakasiyana zvichienderana nekudya (F(5,222) = 4.38, p = 0.0008; Mufananidzo 4B). Hadzi dzinodyiswa weti yemaawa makumi maviri nemana, kudya kweropa kunoiswa mazai akawanda kupfuura hadzi dzinodyiswa mamwe madhiri eweti uye dzakafanana nedzakadyiswa sucrose (Mufananidzo 4B). Saizvozvowo, saizi yemazai anoiswa nehadzi dzinodyiswa weti yakasiyana zvichienderana nekudya (F(5, 209) = 12.85, p < 0.0001), nehadzi dzinodyiswa weti yemaawa makumi maviri nemana uye hadzi dzinodyiswa sucrose dzinoisa mazai makuru kupfuura hadzi dzinodyiswa mvura, nepo mazai ehadzi dzinodyiswa weti kwemaawa zana nemakumi matanhatu nemasere aive madiki zvakanyanya (Mufananidzo 4C). Pamusoro pezvo, kudya kweweti kwakakanganisa zvakanyanya saizi yehonye (F(5, 187) = 7.86, p < 0.0001), ine makonye makuru zvikuru anobuda mumazai anotakurwa nehadzi dzine weti dzine makore 24 ne72 ekuberekwa kupfuura anotakurwa nemazai. Hadzi dzinodyiswa mvura uye dzinodyiswa weti kwemaawa 168 (Mufananidzo 4D).
Kubereka kweAnopheles arabinis yechikadzi ichidya weti yemombe neurea. Umhutu hwechikadzi hwakadyiswa ropa hwakapihwa zvekudya zvinosanganisira weti yemombe itsva neyekare, huwandu hwakasiyana hweurea, sucrose (10%), nemvura yakacheneswa (H2O) kwemaawa makumi mana nemasere usati waisa muongororo yebioassays uye kuwana substrates dzekuisa mazai. Maawa makumi mana nemasere (A). Nhamba yezai (B, E), saizi yezai (C, F) uye saizi yehonye (D, G) zvakakanganiswa zvakanyanya nekudya kwakapihwa (weti yemombe: BD; urea: EG). Nzira dzeparamita imwe neimwe dzakayerwa uchishandisa mazita akasiyana emavara dzaive dzakasiyana zvakanyanya kubva kune imwe neimwe (one-way ANOVA uchishandisa Tukey's post hoc analysis; p < 0.05). Error bars dzinomiririra standard error yeavhareji.
Sechikamu chikuru chenitrogen muweti, urea, kana yakapihwa sekudya kuvakadzi vanodyiswa ropa, yakakanganisa zvakanyanya parameter yekubereka mune zvese zvidzidzo. Huwandu hwemazai anokandirwa nevakadzi vanodyiswa urea, mushure mekudya muropa, zvichienderana nehuwandu hweurea (F(11, 360) = 4.69; p < 0.0001), vakadzi vanodyiswa huwandu hweurea pakati pe134 µM ne1.34 mM vanokandira mazai akawanda (Mufananidzo 4E). Vakadzi vanodyiswa huwandu hweurea hwe134 µM kana kupfuura vanokandira mazai makuru kupfuura vakadzi vanodyiswa nemvura (F(10, 4245) = 36.7; p < 0.0001; Mufananidzo 4F), uye saizi yemakonye, ​​kunyangwe ichikanganiswa nehuwandu hwakafanana hweurea mumadzimai (F(10, 3305) = 37.9; p < 0.0001) hwaive hwakasiyana-siyana (Mufananidzo 4G).
Kukwezva kwese kwese kune zvinobuda muweti yemombe zvinotsvaga host-seeking bovine. Iyo arabiensis yakaongororwa mugirazi tube olfactometer (Mufananidzo 5A) yakakanganiswa zvakanyanya nezera reweti (χ2 = 15.9, df = 4, p = 0.0032; Mufananidzo 5B). Ongororo yemashure yakaratidza kuti hwema hweweti hwakagara mumaawa makumi maviri nemana hwakakonzera mwero wakakwira wekukwezva zvichienzaniswa nemamwe marapirwo ese (maawa makumi manomwe nemaviri: p = 0.0060, maawa zana nemakumi matanhatu nemasere: p = 0.012, pentane: p = 0.00070), Kunze kwekunhuhwirira kweweti nyowani (p = 0.13; Mufananidzo 5B). Kunyangwe kukwezva kwese kweumhutu hunoyamwa ropa kune hwema hweweti kwakanga kusina kusiyana zvakanyanya (χ2 = 8.78, df = 4, p = 0.067; Mufananidzo 5C), vakadzi ava vakawanikwa vachikwezva zvakanyanya kune zvinobuda muweti yemusoro zvichienzaniswa neweti ine maawa makumi manomwe nemaviri yakakwegura zvichienzaniswa ne zvinodzora (p = 0.0066; Mufananidzo 5C).
Mhinduro dzemaitiro kune hwema hwemuto wemombe hwechisikigo uye hwekugadzira mukutsvaga Anopheles arabianus inogamuchira uye inodyiswa ropa. Chimiro chegirazi chubhu olfactometer (A). Kukwezva headspace volatile extracts kubva muweti yemombe itsva uye yakwegura kuenda kumuto wemhuka (B) uye unosveta ropa (C). Tsvaga maitiro etendekitale eLord An. Zvikamu zveHeadspace zvakaparadzaniswa kubva muweti yemombe itsva (D), 24-awa (E), 72-awa (F), uye 168-awa (G) yakwegura zvinoratidzwa. Zviratidzo zveElectron antenna detection (EAD) zvinoratidza shanduko dzemagetsi mukupindura kune bioactive compounds muheadspace yakabviswa kubva kugasi chromatograph uye yakaonekwa neflame ionization detector (FID). Chikero chebhara chinomiririra response amplitude (mV) uchienzaniswa nenguva yekuchengetedza (s). Hunhu uye mwero wekusunungurwa (µg h-1) yemakemikari anoshanda muhupenyu hunoratidzwa. Asterisk imwe chete (*) inoratidza mhinduro yakaderera yenguva dzose. Asterisk mbiri (**) dzinoratidza mhinduro dzisingadzokororwe. Tsvaga muto wemhuka (H) uye inosveta ropa (I) An.arabiensis ine kukwezva kwakasiyana kune musanganiswa wezvinhu zvakagadzirwa wekunhuhwirira kweweti yemombe itsva neyekare. Avhareji yehuwandu hweumhutu hunokwezva mazita akasiyana emavara hwakasiyana zvakanyanya kubva kune mumwe nemumwe (ANOVA yenzira imwe chete uchishandisa ongororo yaTukey's post hoc; p < 0.05). Error bars inomiririra standard error yescale.
Hanzvadzi Ann.arabiensis, maawa makumi manomwe nemaviri uye maawa zana nemakumi maviri mushure mekudya ropa, panguva yekubereka, hapana sarudzo yakaratidzwa ye headspace volatile extracts kubva muweti yemombe itsva uye yakwegura zvichienzaniswa ne pentane controls (χ2 = 3.07, p > 0.05; Faira rekuwedzera 1: Mufananidzo S1).
Kune mukadzi weAnn.arabiensis, ongororo dzeGC-EAD neGC-MS dzakaratidza makemikari masere, matanhatu, matatu uye matatu anoshanda mumuviri (Mufananidzo 5D-G). Kunyangwe musiyano muhuwandu hwemakemikari aikonzera mhinduro dzemagetsi wakaonekwa, mazhinji emakomiki aya aivepo mune yega yega headspace volatile extract yakatorwa kubva muweti nyowani uye yakwegura. Saka, pane yega yega ongororo, makemikari chete aiburitsa mhinduro yemuviri kubva kumatehwe emukadzi ari pamusoro pemuganho ndiwo akabatanidzwa mune imwe ongororo.
Mwero wekusunungurwa kwemakemikari anochengetedza muviri mumusoro wakawedzera kubva pa29 µg h-1 muweti itsva kusvika pa242 µg h-1 muweti yakacheneswa maawa 168, kunyanya nekuda kwekuwedzera kwep-cresol ne m-formaldehyde Phenol pamwe ne phenol. Kusiyana neizvi, mwero wekusunungurwa kwemamwe makemikari, akadai se 2-cyclohexen-1-one ne decanal, wakaderera nekuwedzera kwenguva yeweti, izvo zvakabatana nekuderera kwakaonekwa kwesimba rechiratidzo (kuwanda) mu chromatogram (Mufananidzo 5D)-G kuruboshwe) uye mhinduro dzemuviri kune aya makemikari (Mufananidzo 5D-G kurudyi).
Zvose, musanganiswa wekugadzira waive nechiyero chakafanana chemakemikari anoshanda mumuviri akaonekwa muzvinhu zvinoputika zvemusoro weweti mutsva newakwegura (Mufananidzo 5D–G) uye zvaiita sekuti hazvina kukwezva zvakanyanya mukutsvaga mugadziri (χ2 = 8.15, df = 4, p = 0.083; Mufananidzo 5H) kana umhutu hunosveta ropa (χ2 = 4.91, df = 4, p = 0.30; Mufananidzo 5I). Zvisinei, kuenzanisa kwemashure maviri pakati pekurapa kwakaratidza kuti umhutu hunotsvaga mugadziri hwaikwezva zvakanyanya musanganiswa wekugadzira weti yemaawa makumi maviri nemana kana tichienzanisa nepentane controls (p = 0.0086; Mufananidzo 5H).
Kuti vaongorore basa rezvikamu zvega zvega mumisanganiswa yakagadzirwa yeweti yemakore makumi maviri nemana, misanganiswa mitanhatu yekubvisa yakaongororwa zvichienderana nemisanganiswa yakazara muY-tube assay, umo mishonga yega yega yakabviswa. Kune umhutu hunotsvaga host, kubvisa mishonga yega yega kubva mumusanganiswa wakazara kwaive nemhedzisiro yakakura pamhinduro dzemaitiro (χ2 = 19.63, df = 6, p = 0.0032; Faira rekuwedzera 1: Mufananidzo S2A), misanganiswa yese yekubvisa yaive inokwezva kupfuura Diki pane yakasanganiswa zvizere. Kusiyana neizvi, kubviswa kwemakemikari ega ega kubva mumusanganiswa wakagadzirwa zvizere hakuna kukanganisa maitiro eumhutu hunosveta ropa (χ2 = 11.38, df = 6, p = 0.077), kunze kwe decanal, izvo zvakakonzera mazinga akaderera zvichienzaniswa nemusanganiswa wakazara Attraction (p = 0.022; Faira rekuwedzera 1: Mufananidzo S2B).
Mumusha unogara vanhu vane malaria muEthiopia, kushanda kwemusanganiswa weti yemombe inogadzirwa maawa makumi maviri nemana mukukwezva umhutu mumunda kwakaongororwa kwehusiku gumi (Mufananidzo 6A). Umhutu hunosvika 4,861 hwakabatwa uye hwakaonekwa, uhwo 45.7% hwaive Anthropus.gambiae sl, 18.9% hwaive Anopheles pharoensis uye 35.4% hwaive Culex spp. (Faira rekuwedzera 1: Tafura S1). Anopheles arabinis ndiyo yega nhengo yeAn.Gambian species complex yakacherechedzwa nePCR analysis. Paavhareji, umhutu hunosvika 320 hwakabatwa husiku hwega hwega, panguva iyo misungo ine zvirauro zvemusanganiswa wekugadzira yakabata umhutu hwakawanda kupfuura misungo yakabatana isina musanganiswa (χ2(0, 3196) = 170.0, p < 0.0001). Misungo isina zvirauro yakaiswa pahusiku hwega hwega hwekutonga hushanu pakutanga, pakati, uye pakupera kwekuyedza. Huwandu hwakafanana hweumhutu hwakabatwa muhuviri hwega hwega. yemisungo, zvichiratidza kuti hapana rusaruro pakati pedzimba (χ2(0, 1665) = 9 × 10-13, p > 0.05) uye hapana kudzikira kwehuwandu hwevanhu panguva yekudzidza. Zvichienzaniswa nemisungo yekudzora, huwandu hweumhutu hwakabatwa mumisungo ine musanganiswa wekugadzira hwakawedzera zvakanyanya: kutsvaga mhuka (χ2(0, 2107) = 138.7, p < 0.0001), kudya ropa munguva pfupi yapfuura (χ2(0, 650) = 32.2, p < 0.0001) uye pamuviri (χ2(0, 228) = 6.27, p = 0.0123; Faira rekuwedzera 1: Tafura S1). Izvi zvinoratidzwawo muhuwandu hweumhutu hwakabatwa: kutsvaga mhuka > kusveta ropa > pamuviri > pamuviri > pamuviri > murume.
Kuongororwa kwebasa remumunda kwekushanda kwemusanganiswa wemunhuwi wemuweti wemombe unogadzirwa kwemaawa makumi maviri nemana. Miedzo yemumunda yakaitwa kumaodzanyemba-pakati peEthiopia (mepu), pedyo neguta reMaki (isa), uchishandisa Centers for Disease Control (CDC) light trap (kurudyi) mudzimba dzakabatana, nedhizaini yechiLatin square (mufananidzo wemudenga) (A). Maphototrap eCDC akagadzirwa nemunhuwi anokwezva uye anobata Anopheles arabesques dzechikadzi (B), asi kwete Anopheles farroes (C), nenzira yakasiyana, mhedzisiro inoenderana nemamiriro emuviri. Pamusoro pezvo, misungo iyi yakabata huwandu hwakawanda hweumhutu hweCulex hunogara. (D) Zvichienzaniswa nekudzora. Misungo iri kuruboshwe inomiririra avhareji yesarudzo yeumhutu hunobatwa muzvipeya zvehunhuwi (girini) uye misungo yekudzora (yakavhurika) (N = 10), nepo misungo iri kurudyi inomiririra avhareji yesarudzo yesarudzo muzvipeya zvemisungo yekudzora (yakavhurika; N = 5). ). MaAsterisk anoratidza mazinga ekukosha kwehuwandu (*p = 0.01 uye ***p < 0.0001)
Mhando nhatu idzi dzakabatwa zvakasiyana mumisungo ine misanganiswa yakagadzirwa. Kutsvaga mudziviriri (χ2(1, 1345) = 71.7, p < 0.0001), kudya ropa (χ2(1, 517) = 16.7, p < 0.0001) uye pamuviri (χ2(1, 180) = 6.11, p = 0.0134) a .arabiensis akabatwa mumusungo achiburitsa musanganiswa wekugadzira (Mufananidzo 6B), nepo huwandu hweAn husina kusiyana. Pharoensis mumamiriro akasiyana emuviri akawanikwa (Mufananidzo 6C). Kune Culex, kuwedzera kukuru kwehuwandu hweumhutu hwakatsvaga hudziviriri kwakawanikwa mumisungo yakarukwa nemusanganiswa wekugadzira (χ2(1,1319) = 12.6, p = 0.0004; Mufananidzo 6D), zvichienzaniswa nemisungo yekudzora.
Misungo yekubata chirauro iri kunze kwenzvimbo dzinogona kuberekerwa pakati penzvimbo dzekuberekera nedzekumaruwa muEthiopia yakashandiswa kuongorora kana umhutu hwemalaria huchishandisa hwema hweweti yemombe sechiratidzo chekugara kwemhuka. Kana pasina zviratidzo zvemhuka, kupisa, uye kana paine hwema hweweti yemombe kana pasina, hapana umhutu hwakabatwa (Faira rekuwedzera 1: Mufananidzo S3). Zvisinei, kana paine tembiricha yepamusoro uye hwema hweweti yemombe, umhutu hwechikadzi hwemalaria hwakakwezvwa uye hwakabatwa, kunyangwe muhuwandu hushoma, husingatarisirwi nezera reweti (χ2(5, 25) = 2.29, p = 0.13; Faira rekuwedzera 1: Mufananidzo S3). Kusiyana neizvi, mvura haina kubata umhutu hwemalaria pakupisa kwakanyanya (Faira rekuwedzera 1: Mufananidzo S3).
Utunga hwemalaria hunotora uye hunoparadzira mishonga ine nitrogen kuburikidza nekudya weti yemombe (kureva, madziva) kuti huwedzere hunhu hwenhoroondo yehupenyu, zvakafanana nezvimwe zvipembenene [2, 4, 24, 25, 26]. Weti yemombe inzvimbo inowanikwa nyore nyore inowanikwa yakabatana nenzvimbo dzekuzororera dzezvipembenene zvemalaria, zvakaita sematanga emombe nemiti mirefu iri pedyo nemisha yekumaruwa nenzvimbo dzekuberekera vana. Utunga hwechikadzi hunowana chinhu ichi nekunhuhwirira uye hunokwanisa kudzora kutorwa kwemakemikari ane nitrogen muweti, kusanganisira urea, chinhu chikuru chine nitrogen muweti [15, 16]. Zvichienderana nemamiriro emuviri weumhutu hwechikadzi, zvinovaka muviri muweti zvinopihwa kuti zviwedzere basa rekubhururuka uye kupona kweumhutu hwechikadzi hunotsvaga muenzi, pamwe nehupenyu uye hunhu hwekubereka hwevanhu vanodyiswa ropa panguva yekutanga gonadotropic cycle. Nokudaro, kusanganiswa kweweti kunoita basa rakakosha rekudya kune zvipembenene zvemalaria zvakavharika sevanhu vakuru vasina kudya kwakakwana [8], sezvo zvichipa umhutu hwechikadzi kugona kuwana makemikari akakosha ane nitrogen nekuita kudya kune njodzi shoma. Kuwanikwa uku kune mhedzisiro yakakura yezvirwere, sezvo madzimai achiwedzera yavo tarisiro yehupenyu, chiitiko uye kubereka, zvese zvinokanganisa kugona kwevector. Uyezve, maitiro aya anogona kunge ari iwo chinangwa chezvirongwa zvekutarisira vector mune ramangwana.