Kubona intungamubiri no gukwirakwiza bihuza kurisha udukoko hamwe nubuzima bwamateka yubuzima.Kugira ngo hishyurwe ibitagenda neza mu ntungamubiri zihariye mu bihe bitandukanye byubuzima, udukoko dushobora kubona izo ntungamubiri binyuze mu kugaburira inyongera, urugero, mu kugaburira ururenda rw’intangangore mu buryo buzwi nk'ibidengeri.Umubu Anopheles arabiani bigaragara ko ari intungamubiri kandi rero, bisaba ko habaho intungamubiri.imyigaragambyo ya arabiensis kuminkari yinka kugirango ibone intungamubiri itezimbere amateka yubuzima.
Menya neza ko ifite umutekano.arabiensis yakwegewe numunuko winkari zishaje, amasaha 24, amasaha 72, n amasaha 168, kandi abategarugori bashakisha kandi bagaburirwa amaraso (ifunguro ryamasaha 48 nyuma yamaraso) bapimwe muri Y-tube olfactometer, naho abategarugori batwite basuzumaga ibizamini byimyororokere. n'ibigeragezo byo murwego
Shakisha amaraso yabakiriye nintungamubiri.Mu bushakashatsi bwa laboratoire no mu murima, abarabu bakwega impumuro karemano na sintetike yinkari zinka zishaje kandi zishaje. Abagore batwite ntibitaye ku nkari z’inka ku mbuto zororoka.
Anopheles arabinis gushaka no gukwirakwiza inkari zinka kugirango amateka yubuzima arusheho kuba meza. Kugaburira byongeye inkari zinka bigira ingaruka kubushobozi bwa vectori byongera ubuzima bwa buri munsi nubucucike bwa vector, kandi muburyo butaziguye muguhindura ibikorwa byindege bityo bigomba gutekerezwa mubyitegererezo bizaza.
Kugaburira intungamubiri no gukwirakwiza bihuza ibyatsi by’udukoko hamwe n’ibiranga amateka y’ubuzima [1,2,3] .Adukoko turashobora guhitamo no kubona ibiryo no gukora ibiryo byindishyi zishingiye ku kuboneka kw'ibiribwa n'ibikenerwa ku ntungamubiri [1, 3] .Gukwirakwiza intungamubiri biterwa n'imibereho y'amateka y'ubuzima kandi bishobora gutuma habaho ibisabwa bitandukanye ku bwiza bw'imirire ndetse no mu ntungamubiri zinyuranye ziterwa n'udukoko twinshi, nk'udukoko twinshi twangiza intungamubiri zitandukanye. no gusohora kw'intangangabo, hamwe na karrion, inzira izwi kwizina rya pisine [2] .Nubwo ubwoko butandukanye bwikinyugunyugu ninyenzi bwasobanuwe mbere na mbere, imyobo yo kuvomera nayo igaragara mubindi byateganijwe by’udukoko, kandi gukurura no kugaburira ubwo bwoko bwumutungo bishobora kugira ingaruka zikomeye kubuzima ndetse nindi mico yaranze amateka yubuzima [2, 4, 5, 6], 7]. umuntu mukuru [8], bityo kuvomera birashobora kugira uruhare runini mubiranga amateka yubuzima bwayo, ariko iyi myitwarire kugeza ubu yarirengagijwe .Gukoresha imyigaragambyo nkuburyo bwo kongera intungamubiri muri iyi modoka yingenzi bituma abantu babitaho kuko bishobora kuba bifite ingaruka zikomeye zibyorezo.
Gufata azote mu mibu ya Anopheles y’abagore bakuze ni bike kubera ububiko bwa karori nkeya buturuka ku cyiciro kinini no gukoresha nabi ifunguro ry’amaraso [9] .Umugore Ann.gambiae sl ubusanzwe arabyishyura hiyongereyeho amafunguro y’amaraso yiyongera [10, 11], bityo agashyira abantu benshi mu kaga ko kwanduza imibu. ta kubona ibinyabuzima bya azote byongera imihindagurikire y’imihindagurikire y’ikirere, nkuko bigaragazwa n’utundi dukoko [2] .Mu bijyanye n’uko, gukurura gukomeye kandi gutandukanya bumwe mu bwoko bw’abavandimwe muri An.Urwego rw’ibinyabuzima byo muri Gambiya, Anopheles arabinis, inkari z’inka zishaje kandi zishaje mu matungo azwi hamwe n’inkoko zizwi kandi zifite amikoro azwi ku nkomoko y’inka kandi zifite amoko azwi cyane. ibice, hamwe na urea bingana na 50-95% bya azote yose mu nkari nshya [15, 16] .Nkuko imyaka y’inkari y’inka, mikorobe ikoresha ubwo buryo kugira ngo igabanye ubukana bw’imyunyu ngugu mu masaha 24. amasaha cyangwa munsi yayo [13, 14].
Muri ubu bushakashatsi, hashakishijwe Ans yakiriye kandi yagaburiwe amaraso.Mu gihe cyayo cya mbere cya gonadotropine, arabiensis yasuzumwe kugira ngo haboneke ibibyimba bya azote, harimo na urea, bivanze n’inkari. Ubukurikira, hakozwe ubushakashatsi bwakozwe kugira ngo harebwe uburyo imibu y’abagore itanga ubwo buryo bw’intungamubiri kugira ngo ubuzima bwiza bwororoke kandi bushobore gutangwa neza. ubushakashatsi bwabo kuri ubwo buryo bushobora gutunga intungamubiri, arabiensis yavumbuye isano iri hagati yimiterere itandukanye igaragara.Ivangavanga impumuro nziza ya sintetike yimvange y’ibihingwa ngengabuzima (VOCs) byagaragaye mu nkari zimaze amasaha 24 byongeye gusuzumwa mu bihe by’imirima, byongera ibisubizo byabonetse mu bihe bya laboratoire kandi byerekana ingaruka z’impumuro y’inkari ku miterere itandukanye ya physiologiya.Gukurura imibu. Ibisubizo byabonetse byemeza ko An.arabiensis ibona kandi ikwirakwiza ibinyabuzima bya azote biboneka mu nkari z’intangangore kugira ngo bigire ingaruka ku mateka y’ubuzima.Ibisubizo byaganiriweho mu rwego rw’ingaruka zishobora guterwa na epidemiologiya n’uburyo byakoreshwa mu kugenzura no kugenzura virusi.
Anopheles Arabicans (Dongola strain) yagumishijwe kuri 25 ± 2 ° C, 65 ± 5% RH n’umucyo wa 12:12 h: ukwezi kwijimye. akazu (cm 30 × 30 cm × 30 cm; MegaView Science, Taichung, Tayiwani) kugirango yemere abantu bakuru.Abakuze bahawe igisubizo cya 10% sucrose ad ad libitum kugeza iminsi 4 nyuma yo kuvuka (dpe), icyo gihe abategarugori bashaka abategarugori bahabwa indyo ako kanya mbere yubushakashatsi, cyangwa bakicwa ninzara ijoro ryose hamwe namazi yatoboye mbere yubushakashatsi. kwonsa imibu kugirango bioassay ikurikiraho, 4 dpe igitsina gore yahawe amaraso yintama ya defibrotic (Håtunalab, Bro, SE) ikoresheje uburyo bwo kugaburira membrane (Hemotek Discovery Workshops, Accrington, UK) .Abagore buzuye cyane bahise bimurirwa mu kasho kamwe kandi batanga indyo yuzuye, nkuko byavuzwe haruguru, cyangwa ubushakashatsi bwakozwe na laboratoire ku munsi wa mbere. illed water ad libitum kumasaha 4-6 mbere yubushakashatsi.
Kugaburira byakoreshejwe mukugereranya inkari hamwe na urea mukuze An.Arab igitsina gore.Abagore bashakishwa kandi bagaburiwe amaraso bahawe indyo irimo inkari zinka 1 zivanze kandi zishaje, intungamubiri zitandukanye za urea, hamwe nubugenzuzi bubiri (10% sucrose namazi) kuri 48 h. Byongeye kandi, ibara ryibiryo (1 mg ml-1 xylene cyanide FF) CAS 2650-17-1; 250 µl microcentrifuge tebes (Axygen Scientific, Union City, CA, US; Igicapo 1A) Uzuza kugeza ku nkombe (~ 300 µl) .Kwirinda guhatana hagati y imibu ningaruka zishobora guterwa n’ibara ryirangi, shyira imibu 10 mu isahani nini ya Petri (cm 12 z'umurambararo na cm 6 z'uburebure; Semadeni, Ostermundigen, CH ° 1 °) kugeza ku nshuro 10. Nyuma yo guhura nimirire, imibu yashyizwe kuri -20 ° C kugeza irindi sesengura.
Shakisha inkari za bovine na urea byinjijwe na nyirarureshwa hamwe n’umugore Anopheles arabianus wonsa amaraso.Mu gihe cyo kugaburira (A), imibu y’abagore yahawe indyo igizwe ninkari z’inka zishaje kandi zishaje, ibice bitandukanye bya urea, sucrose (10%), hamwe n’amazi yatoboye (H2O). ine munsi yamasaha 168 yinkari zinka (B) .Ibisobanuro byose birimo azote yuzuye (iation gutandukana bisanzwe) yinkari bigaragarira muri inset.Abashakisha abashyitsi (D, F) hamwe no kunyunyuza amaraso (E, G) igitsina gore bafata urea muburyo buterwa na dosiye. ) .Umurongo ucagaguye ugereranya umurongo-umurongo wo gusubira inyuma (F, G)
Kurekura ibiryo byinjijwe, imibu yashyizwe ku giti cyayo mu miyoboro ya microcentrifuge ya ml 1.5 irimo 230 µl y’amazi yatoboye kandi urugingo rwahagaritswe hakoreshejwe peste na moteri idafite umugozi (VWR International, Lund, SE), hanyuma hakurikiraho centrifugation saa kumi zumugoroba mu minota 10. umusomyi wa microplate ushingiye kuri otometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm) .Ikindi kandi, imibu yashizwe muri ml 1 y'amazi yatoboye, 900 µl muri yo yimuriwe muri cuvette kugirango isesengurwe rya spropropotometometrike (λ 620 nm; µl ya 1 mg ml-1 xylene cyanide.Noneho, ubwinshi bwa optique bwibintu bizwi cyane byakoreshejwe kugirango hamenyekane ingano y'ibiryo buri mibu yatewe.
Umubare wamakuru wasesenguwe hifashishijwe isesengura ryuburyo bumwe butandukanye (ANOVA) hakurikiraho kugereranya kwa Tukey kugereranya hombi (JMP Pro, v14.0.0, SAS Institute Inc, Cary, NC, Amerika, 1989–2007) .Isesengura ryisubiramo ryasobanuye uburyo urea iterwa no kwangiza imibu (GraphPad Pris.).
Hafi ya 20 µl z'inkari z'inkari zo muri buri cyiciro cyahambirwaga kuri Chromosorb® W / AW (10 mg 80/100 mesh, Sigma Aldrich) hanyuma zigashyirwa muri tin capsules (8 mm × 5 mm) .Kapsules zinjijwe mu cyumba cyo gutwika cya CHNS / O zisesengura (Flash 2000, Thermo Fisher Scientific, Waltham, MA) g N l-1) yagereranijwe hashingiwe kumyitozo ya urea izwi nkibisanzwe.
Kugira ngo hamenyekane ingaruka z’imirire ku bashakishwa no kwakira amaraso kubaho kw’abagore, imibu yashyizwe ku giti cyayo mu byokurya binini bya Petri (cm 12 z'umurambararo na cm 6 z'uburebure; Semadeni) ifite umwobo utwikiriwe na meshi mu gifuniko (cm 3 z'umurambararo) hamwe no guhumeka no gutanga ibiryo. Amafunguro yatanzwe mu buryo butaziguye nyuma ya dpe 4 kandi harimo 1% bivanze n'amazi meza ya urea, enye zuzuye hamwe n'inkari zuzuye 10 amenyo ya tampon (DAB Dental AB, Upplands Väsby, SE) yinjijwe muri siringi ya ml 5 (Thermo Fisher Scientific, Gothenburg, SE), plunger yakuweho, hanyuma ashyirwa hejuru yisahani ya petri (ishusho 1) .1A) .Hindura imirire yawe burimunsi. Komeza imibu yapfuye umunsi wose wapfuye. = 40 kuri buri kwivuza) .Kurokoka imibu yagaburiwe ku mafunguro atandukanye yasesenguwe mu mibare hifashishijwe umurongo wo kubaho wa Kaplan-Meyer hamwe n’ibizamini byo mu rwego rwo kugereranya kugereranya ikwirakwizwa ry’imirire hagati y’imirire (Ibarurishamibare rya IBM SPSS 24.0.0.0).
Uruganda rukora imibu rusanzwe rushingiye kuri Attisano nabandi. ) yagabanyijemo umuyoboro uhagaritse kugirango ukore ukuboko gufatanye hamwe n'ukuboko kwatwaraga agace gato ka fayili ya aluminium nk'ikimenyetso kibuza urumuri.
Abagore bishwe ninzara yamasaha 24 bahawe indyo yavuzwe haruguru muminota 30 mbere yo kwifata.Imibu yagaburiwe byuzuye noneho yatewe aneste kumuntu ku rubura muminota 2-3 hanyuma igashyirwa hamwe nudukoko tw’udukoko hamwe n’ibishashara (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) hanyuma bihambirwa ku maboko ya tewolojiya yanditswemo. .
Kugira ngo ugaragaze imiterere y'ibikorwa by'indege, intera yose yagurutse (m) n'umubare rusange w'ibikorwa bikurikirana by'indege byakurikiranwe mu isaha mu gihe cy'amasaha 24. Byongeye kandi, intera mpuzandengo yatwarwaga n'abagore ku giti cyabo yagereranijwe mu buvuzi kandi isesengurwa hakoreshejwe inzira imwe ya ANOVA na Tukey isesengura ryihuse (JMP Pro, v14.0.0, SAS Institute Inc), aho intera yagereranijwe ifatwa nk'impinduka zingana, mu gihe impuzandengo yagereranijwe ni minisiteri. .
Kugirango hamenyekane ingaruka zimirire kumikorere yimyororokere ya An.arabiensis, igitsina gore esheshatu (4 dpe) cyimuriwe mu kato ka Bugdorm (30 cm × 30 cm × 30 cm) nyuma yo gukusanya amaraso hanyuma gitanga indyo yubushakashatsi kuri 48 h nkuko byasobanuwe haruguru.Ibiryo byavanyweho hanyuma ibikombe byabyaye (30 ml; Nolato Hertila) byuzuyemo amata 20 ya buri munsi ya 24. .Amagi yabazwe kandi yandikwa kuri buri kato k'ubushakashatsi. Ingero z'amagi zakoreshejwe mu gusuzuma ingano n'uburebure bw'amagi ya buri muntu (n ≥ 200 ku ndyo) ukoresheje microscope ya Dialux-20 (DM1000; Ernst Leitz Wetzlar, Wetzlar, DE) ifite Kamera ya Leica (DFC) 320 R2;Leica Microsystems Ltd, DE.
Umwanya uhindagurika wumutwe uva mumashya (1h post-sampling), 24h, 72 hr na 168 hr inkari zashaje zegeranijwe mubikusanyirizo byakuwe mu nka za Zebu, amoko ya Arsi.Kuborohereza, ingero z'inkari zegeranijwe kare mu gitondo mugihe inka zari zikiri mu kiraro. Ingero z'inkari zegeranijwe ku bantu 10 na miriyoni 100-200 za buri cyitegererezo cyimuriwe mu mifuka ya polyamide DE) muri 3 l polyamide ifite umupfundikizo Muri vinyl chloride ingoma ya plastike.Umwanya uhindagurika wa buri mwanya w'inkari za bovine wakusanyirijwe hamwe (shyashya) cyangwa nyuma yo gukura mubushyuhe bwicyumba cya 24 h, 72 h na 168 h, ni ukuvuga buri cyitegererezo cyinkari cyari gihagarariye buri cyiciro cyimyaka.
Kugirango ikusanyirizo ryumutwe ryumutwe, sisitemu ifunze-ikoreshwa kugirango ikwirakwize gaze ya gaz ikungururwa ya karubone (100 ml min-1) ikoresheje umufuka wa polyamide kugeza kuri adorption inkingi ya 2.5 h ukoresheje pompe ya vacuum ya diaphragm (KNF Neuberger, Freiburg, DE) .Nkugenzura, ikusanyirizo ryumutwe ryakozwe na tike ya polyoride ya x 5. Porapak Q. C kugeza ikoreshejwe kugirango isesengurwe.
Ibisubizo by'imyitwarire yo gushaka abashyitsi no kurya amaraso An.Ibice bivamo umuyaga uhindagurika byakusanyirijwe mu nkari nshya, 24-h, 72-h, na 168-h zasesenguwe ku bivamo imibu iva mu mibu ya Arabidopsis ikoresheje ikirahuri kiboneye cya olfactometero [18] .Ubushakashatsi bwakozwe mu gihe cya ZT 13-15 kumurikirwa na 3 ± 1 lx yumucyo utukura uva hejuru. Amakara yungurujwe kandi yujujwe (25 ± 2 ° C, 65 ± 2% ugereranije nubushuhe) yanyuze kuri bioassay kuri cm 30 s-1.Air inyuzwa murukurikirane rwicyuma cyuma kitagira umwanda, bigatuma umuyaga wa laminari uhagarara kuri cm 4 D × D; olfactometero, hamwe nimpinduka zikangura buri minota 5.Ku isesengura, 10 μl ya buri gice cyumutwe wumutwe, cyongewemo 1:10, cyakoreshejwe nkikangura.Umubare ungana wa pentane wakoreshejwe nkigenzura. Imibu yashakishaga abantu cyangwa imitsi yonsa kumaraso yashyizwe mumagambo yo kurekura buri muntu washyizwe kumurima wa olim. kuminota 1, hanyuma ikinyugunyugu kinyugunyugu cyarafunguwe kugirango kirekurwe.Igikorwa cyo kuvura cyangwa kugenzura cyasesenguwe nkigipimo cy’imibu cyahuye n’isoko mu minota 5 yo kurekurwa. Buri cyerekezo cy’umutwe uhindagurika kandi kigenzurwa byibuze inshuro 30, kandi kugira ngo wirinde ingaruka z’umunsi uwo ari wo wose, wasuzumaga amaraso yatanzwe na buri munsi. hakurikiraho kugereranya kubigereranyo bidasanzwe (JMP Pro, v14.0.0, SAS Institute Inc.).
Igisubizo cyo gutera intanga. Ibibanza byakuwe mu nkari z'inka zishaje kandi zishaje byasesenguwe mu kato ka Bugdorm (cm 30 × 30 cm × 30 cm; pentane yakoreshejwe muguhindura igikombe cyo kugenzura.Ibikombe byo kuvura no kugenzura byahinduwe hagati ya buri igeragezwa kugirango bigenzure ingaruka zumwanya. Abagore icumi bagaburiwe amaraso barekuwe mu kato k'igeragezwa kuri ZT 9-11 naho amagi mu bikombe yabazwe nyuma y'amasaha 24. Ifumbire yo kubara igipimo cyo gutera intanga ni: (umubare w'amagi yashyizwe mu gikombe cyo kuvura - umubare w'amagi yashyizweho mu gikombe cyo kugenzura).
Isesengura rya gazi ya chromatografi na electron yerekana (GC-EAD) isesengura ryumugore An.arabiensis ryakozwe nkuko byasobanuwe mbere [20] .Muri make, ibivuye mu mutwe bishya byahinduwe bitandukanijwe hakoreshejwe Agilent Technologies 6890 GC (Santa Clara, CA, Amerika) ifite ibikoresho bya HP-5 (30 m × 0,25 mm id, Ubugari bwa Agilent Technologies).n'inkari zishaje. Hydrogene yakoreshejwe nk'icyiciro kigendanwa gifite impuzandengo y'umurongo ugereranyije wa cm 45 s-1.Buri sample (2 μl) yatewe amasegonda 30 muburyo butagabanijwe hamwe n'ubushyuhe bwa inleti ya 225 ° C.Ubushyuhe bwa feri ya GC bwateguwe kuva kuri 35 ° C (gufata iminota 3) kugeza kuri 300 ° C (gufata iminota 10) kuri 10 ° C min 1. Gerstel, Mülheim, DE) hagati ya disiketi ya flame ionisation na EAD. Capillary ya GC effluent capillary ya EAD yanyujijwe kumurongo wa transfert ya Gerstel ODP-2, ikurikirana ubushyuhe bwa feri ya GC hiyongereyeho 5 ° C, mumiyoboro yikirahure (cm 10 × 8 mm), aho yari ivanze na konte imwe ya santimetero ya mitiweli (1.5 l min - 1). , no kubishakira imibu, byibuze bitatu byakorewe kuburugero rwinkari za buri myaka.
Kumenyekanisha ibinyabuzima byifashishwa mu gukusanya inkari zo mu bwoko bwa bovine zishaje kandi zishaje ukoresheje GC hamwe na mass spectrometer (GC-MS; 6890 GC na 5975 MS; Agilent Technologies) kugira ngo utange ibisubizo bya antenne mu isesengura rya GC-EAD, ukora mu buryo bwa elegitoronike ya ionisation kuri 70 eV. ugereranije impuzandengo yumurongo wa cm 35 s-1.A 2 μl sample yatewe inshinge ikoresheje igenamigambi rimwe hamwe nubushyuhe bwitanura nkubwa GC-EAD. Ibicuruzwa byagaragaye hashingiwe ku gihe cyo kubika (indangagaciro ya Kovát) hamwe na sprake ya misa ugereranije nibitabo byabigenewe hamwe nububiko bwibitabo bwa NIST14 (Agilent) .Ibikoresho byashyizwe ahagaragara byemejwe no gushiramo ibipimo bifatika (Fayili 9). Aldrich) yatewe inshinge nkibisanzwe.
Gusuzuma imikorere yimvange ya syntetique igizwe nuruvange rwa bioaktique rwerekanwe muminkari mishya kandi ishaje kugirango ikurura abashaka kwakira no kumena amaraso Ans.arabiensis, ukoresheje olfactometero imwe na protocole nkuko byavuzwe haruguru. .Ku isesengura, koresha 10 μl ya 1: 100 yo kuvanga ivangwa ryuzuye rya syntetique yuzuye, hamwe nigipimo rusange cyo kurekura kiri hagati ya 140-2400 ng h-1, kugirango usuzume igikundiro cy’imibu yonsa kandi yonsa amaraso. Nyuma yaho, ikizamini gikorerwa ku mvange zuzuye, aho kuvangavanga kuvangavanga hamwe no kuvanga amaraso hamwe nibisubizo byavanze. ression ikurikirwa no kugereranya kubigereranyo bidasanzwe (JMP Pro, v14.0.0, SAS Institute Inc.).
Kugira ngo hamenyekane niba inkari z'inka zishobora kuba nk'imiturire yakira imibu ya malariya, inkari z'inka zishaje kandi zishaje, zegeranijwe nk'uko byasobanuwe haruguru, maze amazi ashyirwa mu ndobo 3 l (100 ml) hanyuma ashyirwa mu mutego wo kuroba.(BG-HDT; nimero ya ito yafatiwe mumitego yashizwemo ninkari zimyaka itandukanye yagereranijwe ukoresheje gusubira inyuma kwa logistique hamwe no gukwirakwiza beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
Mu mudugudu w’icyorezo cya malariya hafi y’umujyi wa Maki, mu karere ka Oromia, muri Etiyopiya (8 ° 11 ′ 08 ″ N, 38 ° 81 ′ 70 ″ E; Ishusho 6A) .Ubushakashatsi bwakozwe hagati ya Kanama hagati na Nzeri hagati mbere yo gutera imiti isigaye mu ngo, hamwe n’igihe kirekire cy’imvura. inyamaswa zemewe mu nzu, nta guteka mu nzu (gushushanya inkwi cyangwa amakara) byari byemewe (byibuze mu gihe cy’igeragezwa), n’amazu afite abaturage benshi cyane, baryamye mu miti yica udukoko.munsi y’inzitiramubu ivurwa. Icyemezo cy’ibanze cyatanzwe n’ikigo gishinzwe ubushakashatsi ku myitwarire y’ubushakashatsi (IRB / 022/2016) cy’ishami ry’ubumenyi bw’ibinyabuzima (CNS-IRB), kaminuza ya Addis Abeba, hakurikijwe amabwiriza yashyizweho n’itangazo ry’ishyirahamwe ry’ubuvuzi ku isi ryatangajwe na Helsinki. igishushanyo mbonera cyakurikiranye igishushanyo mbonera cya 2 × 2 Ikilatini, aho imvange nubugenzuzi byahawe inzu zubatswe mwijoro ryambere hanyuma bikazungurana hagati yinzu mu ijoro ryakurikiyeho ryubushakashatsi.Iki gikorwa cyagarutsweho inshuro icumi. Byongeye kandi, kugereranya ibikorwa by’imibu mu mazu yatoranijwe, imitego ya CDC yashyizweho kugirango ikore amajoro atanu yikurikiranya mu ntangiriro, hagati na nyuma y’urubanza.
Uruvange rwa sintetike rurimo ibinyabuzima bitandatu bya bioaktike byashongeshejwe muri heptane (97.0% ya GC yo mu rwego rwa GC, Sigma Aldrich) hanyuma irekurwa kuri 140 ng h-1 ikoresheje imashini itanga ipamba [20] .Icyuma cya wick yemereye ibice byose kurekurwa muburyo buhoraho mugihe cyigeragezwa cyamasaha 12. Heptane yakoreshejwe nkumucungamutungo wa VC. Gainesville, FL, Amerika; Igicapo 6A) .Imitego yamanitswe hejuru ya 0.8 - 1 m hejuru yubutaka, hafi yikirenge cyigitanda, kandi umukorerabushake aryamye munsi yinzitiramubu itavuwe kandi akora hagati ya 18h00 na 06: 30.Imibu yafashwe nubusambanyi hamwe na physiologique (idakorewe, igaburirwa, itwite, kandi itwite [21] nyuma yiperereza ryakozwe na polymerase. Mu bushakashatsi bwakorewe mu murima, umutego w’amazu yombi wasesenguwe hifashishijwe urugero rwiza rwa logistique, aho gukurura byari impinduka zishingiye kandi kuvura (synthique ivanze vs kugenzura) byari ingaruka zifatika (JMP® 14.0.0. SAS Institute Inc).Hano, twatanze raporo χ2 na p-indangagaciro duhereye kubigereranyo byagereranijwe.
Suzuma niba ari umutekano.Uarabiensis yashoboye kubona inkari, izo soko ryayo nyamukuru, Urea, iburanisha kugaburira amaraso (Umutini. 56.00, p <0.0001, kubijyanye; fig. = 15.72, p <0.0001; Igishushanyo cya 1D) .Ibi byari bitandukanye nigisubizo cyigitsina gabo kitari giciro cya 10% kuruta igitsina gore cyo gushaka akazi, kandi aba bagore bo hasi, kandi aba bagore bashizwemo Urea mu buryo busa (F (1,953) = 78.82, p <0.0001;Igishushanyo 1F, G) .Mu gihe gufata ibiryo birimo urea bigaragara ko bifite agaciro keza (Igishusho 1D, E), igitsina gore muri leta zombi zifata umubiri washoboye guhindura urugero rwa urea rwinjijwe murwego rwose rwibanze rwa urea muburyo bwimbitse (Ishusho 1F, G).) .Bisa na none, imibu isa nkaho igenzura gufata azote igenga ingano yinkari zinjiye, kuko ingano ya azote mu nkari igaragarira mu mubare winjijwe (Ishusho ya 1B, C na B).
Kugira ngo hamenyekane ingaruka z'inkari na urea ku gushakisha imibu no kwonona imibu, igitsina gore cyagaburiwe inkari zifite imyaka ine yose (shyashya, 24 h, 72 h, na 168 h nyuma yo kwibizwa) hamwe n’ibice byinshi bya urea, hamwe n’amazi yatoboye hamwe na 10% ya sahrose byagize uruhare mu kugenzura (Igicapo cya 2A). 1; urea: χ2 = 122.8, df = 5, p <0.0001; Igishushanyo 2B, C) hamwe n’igitsina gore kigaburirwa amaraso (inkari: χ2 = 93.0, df = 5, p E.2c) .kumirwa n'amazi, abagore baburiwe inkari nshya hamwe ninkari zamasaha 24 zabayeho igihembo cyinkari zagabwe inshuro 72 zahagaritswe inshuro 135 zabaze inkari zirenze izindi ngingo zirenze Urea n'amazi (P <0.013; Igishusho 2e).
Kurokoka kw'abagore bakira kandi bonsa amaraso Anopheles arabinis birisha inkari z'inka na urea.Mu bioassay (A), imibu y'abagore yahawe indyo igizwe n'inkari z'inka zishaje kandi zishaje, kwibanda cyane kuri urea, sucrose (10%) n'amazi yatoboye (H2O) .Ubuzima bw'abashakishaga (B, C) hamwe n'amaraso E) yandikiwe buri gitsina (B, C) hamwe n'amaraso E). ) na urea (C, E), hamwe no kugenzura, Sucrose n'amazi, barapfuye
Intera hamwe n'umubare w'ibyiciro byagenwe mu kizamini cy'indege mu gihe cy'amasaha 24 byari bitandukanye hagati y’imibu ishakisha abashyitsi ndetse n’amaraso yonona amaraso, yerekanaga ibikorwa bike byo kuguruka muri rusange (Ishusho 3) .Imibu yashakisha abashyitsi itanga inkari nshya kandi ishaje cyangwa sucrose n'amazi yerekanaga uburyo butandukanye bwo kuguruka (Ishusho ya 3), naho abategarugori bagaburira inkari nshya bagaburiraga umuseke utandatu kandi mu gihe amasaha 168 yagaburiwe imibu. urnal.Imibu y'abagore yatangaga sucrose cyangwa inkari z'amasaha 72 yerekanaga ibikorwa mugihe cyamasaha 24, mugihe igitsina gore cyatangaga amazi cyakoraga cyane mugihe cyo hagati. Imibu yagaburiwe na sucrose yerekanaga ibikorwa byinshi murwego rwo hejuru bwijoro na kare mugitondo, mugihe abarya inkari zamasaha 72 zagabanutse cyane mubikorwa mumasaha 24 (Ishusho 3).
Imikorere y'indege ishakisha abahigi bonsa amaraso Anopheles arabinis igaburira inkari z'inka na urea.Mu kizamini cy'uruganda rw'indege, imibu y'abagore yagaburiwe inkari z'inka zishaje kandi zishaje, ibice bitandukanye bya urea, sucrose (10%), n'amazi yatoboye (H2O) byahujwe n'amasoko atambitse (yisanzuye) ku mubare w'abagore bashakishwa (hejuru). Ikiringo cyamasaha 24 cyaranditswe (umwijima: imvi; urumuri: cyera). Intera igereranije numubare ugereranije wa bouti yerekanwa iburyo bwibishushanyo mbonera byibikorwa. Utubari twibeshya twerekana ikosa risanzwe ryerekana.
Muri rusange, muri rusange ibikorwa byindege byabagore bashaka abategarugori byakurikije urugero rusa nuburebure bwindege mugihe cyamasaha 24. Intera yindege yatewe cyane nimirire yatewe (F (5, 138) = 28.27, p <0.0001), kandi abategarugori bashaka abashyitsi binjije amasaha 72 yinkari baguruka intera ndende ugereranije nandi mafunguro yose (p <0.0001), na sucrose . urea yagurutse igihe kirekire kurusha igitsina gore cyagaburiwe amazi cyangwa sucrose (p <0.03).
Muri rusange ibikorwa byindege by imibu yonsa amaraso byari bihamye kandi bikomeza amasaha arenga 24 mumirire yose, hamwe nibikorwa byinkari byiyongereye mugice cya kabiri cyigihe cyumwijima kubagore bagaburiwe kumazi kimwe nabagore bagaburiwe amasaha mashya namasaha 24 (ishusho 3) .Mu gihe indyo yinkari yagize ingaruka zikomeye bivuze intera yindege yabagore bagaburiwe namaraso (F (5, 138) = 4.83, p = 0.0004), urea. no kugenzura indyo (shyashya, p = 0.0091; amasaha 72, p = 0.0022; amasaha 168, p = 0.001; sucrose, p = 0.0017; dH2O, p = 0.036).
Ingaruka z'inkari na urea zigaburira ibipimo by'imyororokere byasuzumwe muri bioassays itera amagi (Igicapo 4A) hanyuma hakorwa iperereza ukurikije umubare w'amagi yatewe na buri gitsina gore, ingano y'igi, hamwe n'udusimba twa mbere twavutse. Umubare w'amagi yatanzwe. kandi byari bisa nabagaburiwe sucrose (Igicapo 4B) .Nubundi, ingano yamagi yashyizweho nigitsina gore yagaburiwe ninkari zitandukanye nimirire (F (5, 209) = 12.85, p <0.0001), hamwe ninkari zamasaha 24 nabagore bagaburiwe na sucrose batera amagi manini cyane ugereranije nabagore bagaburiwe amazi, mugihe amagi yabagore yagaburiwe na 168 h yinkari. 0.0001), hamwe na livine nini cyane ziva mu magi yatewe n’igitsina gore cyagaburiwe n’amasaha 24- na 72 y’amasaha kurusha amagi yatewe mu magi.
Imikorere yimyororokere yumugore Anopheles arabinis igaburira inkari zinka na urea.Imibu y’abagore yagaburiwe amaraso yagaburiwe indyo igizwe ninkari zinka zishaje kandi zishaje, kwibumbira hamwe kwa urea, sucrose (10%), namazi yatoboye (H2O) mumasaha 48 mbere yo gushyira muri bioassay no kubona amagi ya D (ingano), ingano ya B, E) byatewe cyane nimirire yatanzwe (inkari zinka: BD; urea: EG) .Ibisobanuro kuri buri kintu cyapimwe ukoresheje amazina yinyuguti zitandukanye byari bitandukanye cyane hagati yabyo (inzira imwe ANOVA ikoresheje isesengura rya post ya Tukey; p <0.05) .Ibibari byamakosa byerekana ikosa risanzwe ryibisobanuro
Nkibintu byingenzi bigize azote bigize inkari, urea, iyo bitanzwe nkibiryo byigitsina gore byigaburiwe namaraso, byagize ingaruka zikomeye kumyororokere mubushakashatsi bwose. Umubare w'amagi yatewe nigitsina gore yagaburiwe urea, nyuma yo kurya amaraso, bitewe nubushakashatsi bwa urea (F (11, 360) = 4.69; p <0.0001), igitsina gore cyagaburiraga urea hagati ya 134 µM na 1.34 mM zashyizeho amagi menshi (urema 4) amazi (F (10, 4245) = 36.7; p <0.
Muri rusange gukurura abashaka gushakisha inkari zo mu mutwe Umwanya uhindagurika.Arabiensis yasuzumwe mu kirahure cya olfactometero (ishusho 5A) yibasiwe cyane n’imyaka y’inkari (χ2 = 15.9, df = 4, p = 0.0032; Igishushanyo 5B) p = 0
Imyitwarire yimyitwarire yinkari zinka nizisanzwe mugushakisha uwakiriye kandi yagaburiwe amaraso Anopheles arabianus.Ibishushanyo mbonera byikirahure olfactometer (A) .Gukuramo imitwe ihindagurika yumutwe uva mu nkari zinka zishaje kandi zishaje kugirango zakire (B) hamwe nudusimba twamaraso (C) amasaha 24. F), hamwe n’amasaha 168 (G) inkari zinka zishaje zirerekanwa.Ibimenyetso bya elegitoroniki ya antenne (EAD) byerekana impinduka za voltage mugusubiza ibyuka bya bioaktique mumitwe yavuye kuri chromatografi ya gaze hanyuma bikagaragazwa na disiketi ya flame ionisiyoneri (FID) .Umurongo wikigereranyo ugereranya amplitude yo gusubiza (mV) ugereranije nigihe cyo kugumya (µg h) igisubizo. Inyenyeri ebyiri (**) zerekana ibisubizo bidasubirwaho. Shakisha uwakiriye (H) hamwe no kunyunyuza amaraso (I) An.arabiensis afite ubujurire butandukanye bwuruvange rwimvange rwimpumuro yinkari zinkoko zishaje kandi zishaje.p <0.05) .Ibibari byerekana amakosa yibipimo
Umugore Ann.arabiensis, 72 h na 120 h nyuma yo kurya amaraso, mugihe cyo gutera akabariro, nta cyifuzo cyerekanwe kumutwe uhindagurika uva muminkari y'inka nshya kandi ishaje ugereranije no kugenzura pentane (χ2 = 3.07, p> 0.05; Idosiye yinyongera 1: Ishusho S1).
Ku bagore Ann.arabiensis, isesengura rya GC-EAD na GC-MS ryerekanye ibice umunani, bitandatu, bitatu na bitatu bya bioactive (Igicapo 5D-G) .Nubwo byagaragaye ko itandukaniro ry’umubare w’ibintu byatangaga ibisubizo bya electrophysiologique byagaragaye, ibyinshi muri ibyo bikoresho byari muri buri gice cy’umutwe uhindagurika ukomoka ku nkari nshya kandi zashaje.
Igipimo rusange cyo kurekura ibinyabuzima bya bioaktique mu cyegeranyo cy’umutwe cyiyongereye kiva kuri 29 µg h-1 mu nkari nshya kigera kuri 242 µg h-1 mu nkari zimaze amasaha 168, bitewe ahanini na p-cresol na m-formaldehyde Phenol yiyongera kimwe na fenol. Mu buryo bunyuranye, ibipimo byo kurekura by’ibindi bikoresho, nka 2-cyclohexen-1-imwe hamwe na decan. atogramu (Igishusho 5D) -G ibumoso bwibumoso) hamwe nibisubizo bya physiologique kuri ibyo bikoresho (Igicapo 5D-G iburyo).
Muri rusange, imvange ya sintetike yari ifite igereranyo gisanzwe cyibinyabuzima byitwa bioaktique byagaragaye mubikururuka bihindagurika byumutwe winkari mushya kandi ushaje (Igicapo 5D - G) kandi nticyagaragaye nkicyifuzo gikomeye mugushakisha uwakiriye (χ2 = 8.15, df = 4, p = 0.083; igishusho 5H), kugereranya hagati yubuvuzi byerekanaga ko imibu ishakisha abashyitsi yakundaga cyane imvange ya sintetike yinkari zimyaka 24 h ugereranije no kugenzura pentane (p = 0.0086; Igicapo 5H).
Kugira ngo hamenyekane uruhare rw'ibigize buri muntu mu mvange ya sintetike y’inkari zifite imyaka 24-h, hasuzumwe imvange esheshatu zikuramo zivanze n’imvange zuzuye mu isuzuma rya Y-tube, aho zavanywemo imvange ku giti cye.Ku mibu ishakisha abashyitsi, gukuramo ibice bitandukanye bivanze bivanze byagize ingaruka zikomeye kubisubizo byimyitwarire (χ2 = 19.63, df = 6, p = 0.0032; bivanze. Mu buryo bunyuranye, kuvanaho ibice bitandukanye bivanze nubushakashatsi bwuzuye ntabwo byahinduye ibisubizo byimyitwarire y imibu yonsa amaraso (χ2 = 11.38, df = 6, p = 0.077), usibye decanal, byaviriyemo urwego rwo hasi ugereranije nuruvange rwuzuye Gukurura (p = 0.022; Idosiye yinyongera 1: Ishusho S2B).
Mu mudugudu wanduye malariya muri Etiyopiya, hasuzumwe ingaruka z’uruvange rw’inkari z’inka z’amasaha 24 mu gukurura imibu mu bihe by’umurima hasuzumwe amajoro icumi (Igishusho 6A) .Imibu yose hamwe 4.861 yarafashwe iramenyekana, muri yo 45.7% ni Anthropus.gambiae sl, 18.9% ni Anopheles. heles arabinis nimwe mubanyamuryango bonyine ba An.Gambiya yibinyabuzima byagaragajwe nisesengura rya PCR. Ugereranije, imibu 320 yafashwe nijoro, muri icyo gihe imitego hamwe nudukoko twavanze na sintetike yafashe imibu myinshi kuruta imitego yombi itavanze (χ2 (0, 3196) = 170.0, p <0.0001) yafatiwe muri buri mitego yombi, byerekana ko nta kubogama hagati yamazu (χ2 (0, 1665) = 9 × 10-13, p> 0.05) kandi nta baturage bagabanuka mugihe cy’inyigisho. Ugereranije n’imitego yo kugenzura, umubare w’imibu wafatiwe mu mitego irimo imvange ya sintetike wariyongereye cyane: abashaka gushaka (χ2 (0, 2107) = 138.7, p <0.0001) (χ2 (0, 228) = 6.27, p = 0.0123;Inyandiko yinyongera 1: Imbonerahamwe S1) .Ibi bigaragarira no mumibare yose y imibu yafashwe: uwakiriye ashaka> kumena amaraso> atwite> atwite> atwite> umugabo.
Isuzumabumenyi ryerekana umusaruro w’amasaha 24 y’uruvange rw’inkari zivanze n’inkari. Ibigeragezo byakorewe mu majyepfo ya Etiyopiya rwagati (ikarita), hafi y’umujyi wa Maki (shyiramo), hifashishijwe ikigo gishinzwe kurwanya indwara (CDC) umutego woroshye (iburyo) mu mazu yombi, ufite igishushanyo mbonera cya latine (ishusho yo mu kirere) (A). roes (C), muburyo butandukanye, ingaruka zishingiye kuri physiologique. Byongeye kandi, iyi mitego yafashe umubare wiyongereye cyane w’imibu ya Culex yakiriye.) .Ibibazo byerekana urwego rusobanura imibare (* p = 0.01 na *** p <0.0001)
Amoko atatu yafashwe mu buryo butandukanye mu mitego irimo imvange ya sintetike. Urebye uwakiriye (χ2 (1, 1345) = 71.7, p <0.0001), kugaburira amaraso (χ2 (1, 517) = 16.7, p <0.0001) no gutwita (χ2 (1, 180) = 6.11, p = 0.0134) a. haroensis mu bihe bitandukanye bya physiologique yabonetse (Igishusho 6C) .Ku Culex, gusa kwiyongera gukabije kwumubare w’imibu ushaka abashyitsi wasangaga mu mitego yatewe n’imvange ya sintetike (χ2 (1,1319) = 12.6, p = 0.0004; Ishusho 6D), ugereranije n’imitego yo kugenzura.
Imitego ya bait yakira iri hanze y’ahantu hashobora kwororerwa hagati y’ubworozi n’abaturage bo mu cyaro muri Etiyopiya yakoreshejwe mu gusuzuma niba imibu ya malariya ikoresha impumuro y’inkari z’inka nk'ahantu ho kubakira.Mu gihe hatabayeho ibimenyetso byabashitsi, ubushyuhe, kandi nta mpumuro y’inkari z’inka zigeze, nta mibu yafatwaga (Ubushuhe bw’inyama n’ubushuhe). , nubwo ari mbarwa, itagendeye kumyaka yinkari (χ2 (5, 25) = 2.29, p = 0.13; dosiye yinyongera 1: Igicapo S3) .Ibinyuranye, kugenzura amazi ntabwo byafashe imibu ya malariya mubushyuhe bwinshi (File File 1: Igicapo S3).
Umubu wa malariya ubona kandi ugakwirakwiza ibiyigize birimo azote binyuze mu kugaburira indishyi ku nkari z'inka (ni ukuvuga ibiziba) kugira ngo amateka y’ubuzima, ameze nk’utundi dukoko [2, 4, 24, 25, 26] .Inkari z'inkari ni ibikoresho byoroshye bishobora kuvugururwa bifitanye isano n’ahantu ho kuruhukira haterwa na malariya hamwe n’ibimera birebire byangiza amazu y’icyaro. gufata ibibyimba bya azote mu nkari, harimo na urea, igice kinini cya azote mu nkari [15, 16] .Bishingiye ku miterere y’imiterere y’umubu w’umugore, intungamubiri ziri mu nkari zitangwa kugira ngo ibikorwa by’indege bigabanuke kandi bikomeze kubaho imibu y’umugore ifungura amaraso mu gihe cy’imirire ya mikorobe. abantu bakuze bafite imirire mibi [8], kuko itanga imibu y'abagore ifite ubushobozi bwo kubona imiti ya azote ikomeye mu kwishora mu kugaburira ibyago bike.Iki gisubizo gifite ingaruka zikomeye z'ibyorezo, kuko igitsina gore cyongera igihe cyo kubaho, ibikorwa ndetse n’imyororokere, ibyo byose bigira ingaruka ku bushobozi bwa vector.