Gushaka no gukwirakwiza intungamubiri bihuza uburyo udukoko dushakisha mu nkari ndetse n'imiterere y'ubuzima. Kugira ngo dukemure ikibazo cy'ibura ry'intungamubiri runaka mu byiciro bitandukanye by'ubuzima, udukoko dushobora kubona izi ntungamubiri binyuze mu kuzigaburira inyongera, urugero, binyuze mu kurya imisemburo y'inyamaswa zo mu bwoko bwa vertebrate mu buryo buzwi nka puddles. Umubu witwa Anopheles arabiani usa nkaho udafite imirire myiza, bityo ukaba ukeneye intungamubiri zo gukora imisemburo ndetse no kororoka. Intego y'ubu bushakashatsi kwari ukureba niba An. arabiensis itera uburyaryate ku nkari z'inka kugira ngo ibone intungamubiri nziza kurushaho.
Menya neza ko nta kibazo ifite. Arabiensis yakuruwe n'impumuro y'inkari nshya, imara amasaha 24, 72, n'amasaha 168, hamwe n'inka z'ingore zishaka kandi zigaburirwa amaraso (amasaha 48 nyuma y'ifunguro ry'amaraso) zapimwe muri olfactometer ya Y-tube, maze ingore zitwite zipimwa kugira ngo zibyare. Hakoreshejwe isesengura rya shimi n'iry'amashanyarazi kugira ngo hamenyekane ibintu bikora mu nkari z'inka mu byiciro byose by'imyaka ine. Uruvange rw'ibinyabutabire by'ibinyabutabire byapimwe mu igerageza rya Y-tube no mu murima. Kugira ngo harebwe inkari z'inka n'ibinyabutabire byayo by'ingenzi birimo azote nk'indyo y'inyongera ku batera malariya, ibipimo byo kurya n'imiterere y'ubuzima byarapimwe. Igipimo cy'imibu y'ingore n'ingano y'inka n'umunyu byafashwe na urea byarapimwe. Nyuma yo kurya, ingore zapimwe niba zikiriho, zihambiriye, kandi kororoka.
Shaka amaraso n'intungamubiri by'inka. Mu bushakashatsi bwa laboratwari n'ubwo mu murima, Abarabu bakuruwe n'impumuro karemano n'iy'inka nshya n'izakuze. Inka zitwite ntizitaye ku buryo inkari z'inka zikora aho zibyarira. Inka zishaka kandi zinyunyuza amaraso zifata inkari z'inka na urea kandi zigatanga izi nkari hakurikijwe amateka y'ubuzima nk'uko imiterere y'umubiri w'inka ikora kugira ngo iguruke, ikomeze kubaho, cyangwa ikororoke iba.
Kubona inkari z'inka no kuzikwirakwiza mu buryo bwo kunoza imiterere y'ubuzima bwazo. Kugaburira inkari z'inyongera bigira ingaruka ku bushobozi bw'inka mu buryo butaziguye binyuze mu kongera ubuzima bwa buri munsi n'ubwinshi bw'inka, no mu buryo butaziguye binyuze mu guhindura imikorere y'inka mu kirere, bityo bikaba bikwiye kwitabwaho mu buryo bw'ejo hazaza.
Gushaka no gukwirakwiza intungamubiri bihuza imiterere y’udukoko n’amateka y’ubuzima [1,2,3]. Udukoko dushobora guhitamo no kubona ibyo kurya no kugaburira bikurikije uko ibiryo biboneka n’ibyo intungamubiri zikenera [1, 3]. Gukwirakwiza intungamubiri biterwa n’amateka y’ubuzima kandi bishobora gutuma habaho ibisabwa bitandukanye ku bwiza bw’imirire n’ingano mu byiciro bitandukanye by’ubuzima bw’udukoko [1, 2]. Kugira ngo dusubize ibura ry’intungamubiri runaka, udukoko dushobora kubona izi ntungamubiri binyuze mu kugaburira inyongera, nko ku byondo, imyanda itandukanye n’isohoka ry’inyamaswa zo mu rutirigongo, n’inyama z’intumbi, inzira izwi ku izina ry’ibidendezi [2]. Nubwo ubwoko butandukanye bw’ibinyugunyugu n’ibihumyo byavuzwe cyane, imyobo yo kuhira nayo ibaho mu bundi buryo bw’udukoko, kandi gukurura no kurya ubwo bwoko bw’umutungo bishobora kugira ingaruka zikomeye ku buzima n’ibindi biranga ubuzima [2, 4, 5, 6] ,7]. Umubu wa malariya witwa Anopheles gambiae sensu lato (sl) ugaragara nk'umuntu mukuru 'udafite imirire myiza' [8], bityo kuhira bishobora kugira uruhare runini mu biranga amateka y’ubuzima bwawo, ariko iyi myitwarire kugeza ubu byirengagijwe. Gukoresha uburyo bwo kongera intungamubiri muri iki kintu cy'ingenzi birakwiye kwitabwaho kuko bishobora kugira ingaruka zikomeye ku ndwara.
Ingano ya azote ku mibu y’ingore ikuze ya Anopheles iragabanuka bitewe n’uko ifite karori nke zikomoka ku gihe cy’ibibyimba no kudakoresha neza ifunguro ry’amaraso [9]. Inyoni y’ingore ya Ann.gambiae sl ikunze kwishyura ibi ikongeraho ifunguro ry’inyongera ku maraso [10, 11], bityo igashyira abantu benshi mu kaga ko kwandura iyi ndwara kandi igashyira imibu mu kaga gakomeye ko kuyirya. Ubundi buryo, imibu ishobora gukoresha indyo y’inyongera ku mavuta y’inyamaswa zo mu bwonko kugira ngo ibone imvange ya azote yongera ubushobozi bwo guhangana n’impinduka no kugenda neza, nk’uko bigaragazwa n’utundi dukoko [2]. Muri urwo rwego, gukurura cyane ubwoko bumwe bw’inka bukomoka kuri An. Ubwoko bwa sl bwa Gambiya, Anopheles arabinis, inkari nshya n’izabukuru [12,13,14], birashishikaje. Anopheles arabinis ni ingenzi mu byo ikunda kandi izwiho kwishyira hamwe no kurya inka. Inkari z’inka ni umutungo ukunzwemo imvange ya azote, aho urea ingana na 50-95% bya azote yose mu nkari nshya [15, 16]. Uko inkari z'inka zigenda zikura, udukoko dukoresha iyi miterere kugira ngo tugabanye ubukana bw'ibintu birimo azote mu masaha 24 [15]. Bitewe n'ubwiyongere bwihuse bwa ammonia, bujyana no kugabanuka kwa azote y'umwimerere, udukoko duto duto twa alkalophilic (inyinshi muri zo zigira ibintu bihumanya imibu) turakura [15], dushobora kuba Ann.arabiensis y'ingore ikururwa cyane n'inkari mu masaha 24 cyangwa munsi yayo [13, 14].
Muri ubu bushakashatsi, hashakishijwe Ans isanzwe n’iy’amaraso. Mu gihe cya gonadotropin cycle yayo ya mbere, arabiensis yapimwe kugira ngo ibone imvange ya azote, harimo na urea, binyuze mu kuvanga inkari. Hanyuma, hakozwe igerageza ryinshi kugira ngo hamenyekane uburyo imibu y’ingore ikoresha iyi ntungamubiri kugira ngo irusheho kubaho, kororoka no gukomeza gushaka. Amaherezo, impumuro y’inka nshya n’izakuze yapimwe kugira ngo hamenyekane niba izi mpanuro zizewe ku imvange n’iy’amaraso. Mu gushakisha iyi ntungamubiri ishobora kuba iy’ibanze, arabiensis yavumbuye isano iri hagati y’ubusumbane bwagaragaye. Impumuro y’imvange y’ibinyabutabire by’umwimerere (VOCs) byagaragaye mu nkari zimaze amasaha 24 byasuzumwe mu gihe cy’ubutaka, byongera ibisubizo byabonetse mu gihe cya laboratwari kandi berekana ingaruka z’impumuro y’inka ku miterere itandukanye y’umubiri. Gukurura imibu. Ibisubizo byabonetse byemeza ko An. arabiensis ibona kandi igakwirakwiza imvange ya azote iboneka mu nkari z’inyamaswa zifite uruhu rw’umugongo kugira ngo igire ingaruka ku miterere y’ubuzima. Ibi bisubizo biraganirwaho mu rwego rw’ingaruka zishobora kubaho mu ndwara n’uburyo zishobora gukoreshwa mu kugenzura no kugenzura ibinyabuzima.
Inyoni zo mu bwoko bwa Anopheles arabicans (ubwoko bwa Dongola) zagumanywe ubushyuhe bwa 25 ± 2 °C, 65 ± 5% RH n'umwijima wa saa 12:12. Inyoni zashyizwe mu dusanduku twa pulasitiki (20 cm × 18 cm × 7 cm) zuzuye amazi yaciwe hanyuma zigaburirwa ibiryo by'amafi bya Tetramin® (Tetra Werke, Melle, DE). Inyoni zakusanyijwe mu bikombe bya mililitiro 30 (Nolato Hertila, Åstorp, SE) hanyuma zijyanwa mu tuzu twa Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science, Taichung, Taiwan) kugira ngo zigere ku bantu bakuru. Abantu bakuru bahawe umuti wa sucrose wa 10% ku gipimo cya kabiri kugeza ku minsi 4 nyuma yo kumera (dpe), aho abagore bashaka kwakira nyoni bahawe indyo mbere gato y'igerageza, cyangwa bicwaga inzara n'amazi yaciwe mbere y'igerageza, nk'uko byasobanuwe hano hepfo. Inyoni z'ingore zakoreshejwe mu igerageza ry'imiyoboro y'indege zaciwe inzara gusa. Amasaha 4-6 n'amazi adateguje. Kugira ngo hategurwe imibu inyunyuza amaraso kugira ngo ikorerwe isuzuma ry’umubiri nyuma yaho, abagore 4 batanzwe amaraso y’intama adateguje fibrosis (Håtunalab, Bro, SE) hakoreshejwe uburyo bwo kugaburira membrane (Hemotek Discovery Workshops, Accrington, UK). Abagore bari bahujwe cyane bahise bimurwa mu tuzu twa buri bwoko kandi bahabwa indyo itaziguye, nk'uko byasobanuwe hepfo, cyangwa 10% ya sucrose adateguje impumuro mu minsi 3 mbere y'igerageza ryasobanuwe hepfo. Abagore ba nyuma bakoreshejwe mu gusuzuma impumuro z’umubiri hanyuma bimurirwaho muri laboratwari, hanyuma banywa amazi adateguje impumuro mu gihe cy'amasaha 4-6 mbere y'igerageza.
Isuzuma ry’ibiryo ryakoreshejwe mu gupima inkari n’umunyu ku bagore bakuru b’Abarabu. Abagore bashaka kandi bagaburirwa amaraso bahawe indyo irimo inkari nshya n’izakuze 1%, urea nyinshi zitandukanye, hamwe n’ibipimo bibiri (10% sucrose n’amazi) mu gihe cy’amasaha 48. Byongeye kandi, ibara ry’ibiryo (1 mg ml-1 xylene cyanide FF; CAS 2650-17-1; Sigma-Aldrich, Stockholm, SE) ryongewe kuri iyo ndyo kandi ritangwa mu gipimo cya 4 × 4 mu miyoboro ya 250 µl microcentrifuge (Axygen Scientific, Union City, CA, US; Ishusho ya 1A) Uzuza kugeza ku nkengero (~300 µl). Kugira ngo wirinde ipiganwa hagati y’imibu n’ingaruka zishobora guterwa n’irangi, shyira imibu 10 mu isahani nini ya Petri (12 cm mu murambararo na 6 cm mu burebure; Semadeni, Ostermundigen, CH; Ishusho ya 1A) mu mwijima wose kuri 25 ± 2 cm °C na 65 ± 5% ugereranije. Ubushuhe. Aya magerageza yasubiwemo inshuro 5 kugeza ku 10. Nyuma yo kurya, imibu yashyizwe kuri -20 °C kugeza igihe isesengura rikomeje.
Shaka inkari z'inka na urea byinjizwa n'inka n'inka y'inka inywa amaraso. Mu igerageza ryo kugaburira (A), imibu y'inka yahawe indyo igizwe n'inka nshya n'izakuze, ingano zitandukanye za urea, sucrose (10%), n'amazi yaciwe (H2O). Inka z'inka zishaka inka (B) n'inka z'inka zigaburirwa amaraso (C) zabonye sucrose nyinshi kurusha izindi ndyo zose zapimwe. Menya ko inka z'inka zishaka inka zabonye inkari z'inka mu masaha 72 munsi y'inka z'amasaha 168 (B). Impuzandengo y'inka zose (± ihinduka risanzwe) igaragazwa mu gice cyo hasi. Inka z'inka zishaka inka (D, F) n'inka z'inka zinywa amaraso (E, G) bifata urea mu buryo bushingiye ku gipimo. Ingano y'impumuro (D, E) hamwe n'amazina y'inyuguti zitandukanye yari itandukanye cyane (ANOVA y'inzira imwe hakoreshejwe isesengura rya Tukey nyuma y'aho; p < 0.05). Uduce tw'amakosa tugaragaza ikosa risanzwe ry'impuzandengo (BE). Umurongo ugororotse ugaragaza umurongo wo gusubira inyuma (F, G)
Kugira ngo imibu irekure ibiryo byinjijwemo, yashyizwe ukwayo mu miyoboro ya mikorocentrifuge ya mililitiro 1.5 irimo 230 µl z'amazi yaciwe maze ingirangingo irangirika hakoreshejwe moteri ikoreshwa mu gutobora n'umugozi (VWR International, Lund, SE), hagakurikiraho gutobora kuri moteri idafite umugozi (VWR International, Lund, SE), hagakurikiraho gutobora kuri 10 krpm mu gihe cy'iminota 10. Supernatant (200 µl) yimuwe kuri microplate ya 96-well (Sigma-Aldrich) maze absorbance (λ620) ipimwa hakoreshejwe microplate reader ishingiye kuri spectrophotometer (SPECTROStar® Nano, BMG Labtech, Ortenberg, DE) nm). Ubundi buryo, imibu yasibwe muri mililitiro 1 y'amazi yaciwe, 900 µl yoherezwa muri cuvette kugira ngo ikoreshwe mu gusesengura spectrophotometric (λ 620 nm; UV 1800, Shimadzu, Kista, SE). Kugira ngo harebwe uburyo bwo kurya, hateguwe umurongo usanzwe wo gutobora kugira ngo haboneke 0.2 µl kugeza 2.4. µl ya 1 mg ml-1 xylene cyanide. Hanyuma, hakoreshejwe ubucucike bw'amabara buzwi kugira ngo hamenyekane ingano y'ibiryo buri mubu wariye.
Amakuru y’ingano yasesenguwe hakoreshejwe uburyo bumwe bwo gusesengura itandukaniro (ANOVA) hagakurikiraho igereranya rya Tukey rya post hoc pairwise (JMP Pro, v14.0.0, SAS Institute Inc., Cary, NC, US, 1989–2007). Isesengura rya Linear regression ryasobanuye ikoreshwa rya urea rishingiye ku ngano ndetse rigereranya ibisubizo hagati y’imibu ikunda gushaka imbu n’iy’imibu ikunda kunyunyuza amaraso (GraphPad Prism v8.0.0 kuri Mac, GraphPad Software, San Diego, CA, US).
Hafi 20 µl by'inkari zo muri buri tsinda ry'imyaka byashyizwe kuri Chromosorb® W/AW (10 mg 80/100 mesh, Sigma Aldrich) hanyuma bishyirwa mu tubumbe twa tin (8 mm × 5 mm). Utu tubumbe twashyizwe mu cyumba cyo gutwika cy'icyuma gipima CHNS/O (Flash 2000, Thermo Fisher Scientific, Waltham, MA, US) kugira ngo hamenyekane ingano ya azote mu nkari nshya n'izashaje hakurikijwe amabwiriza y'uruganda. Azote yose (g N l-1) yapimwe hashingiwe ku gipimo cya urea kizwi cyakoreshejwe nk'igipimo ngenderwaho.
Kugira ngo harebwe ingaruka z'imirire ku buzima bw'ingore ikeneye kandi ikamwa amaraso, imibu yashyizwe ukwayo mu masahani manini ya Petri (cm 12 z'umurambararo na cm 6 z'uburebure; Semadeni) ifite umwobo upfutseho mesh mu gipfundikizo (cm 3 z'umurambararo) hamwe n'uburyo bwo guhumeka no gutanga ibiryo. Indyo yatanzwe ako kanya nyuma ya dpe 4 kandi harimo 1% y'inkari z'inka nshya n'izashaje, ibipimo bine bya urea, n'ibipimo bibiri, 10% bya sucrose n'amazi. Buri ndyo yashyizwe kuri tampon y'amenyo (DAB Dental AB, Upplands Väsby, SE) yinjijwe mu cyuma cya 5 ml (Thermo Fisher Scientific, Gothenburg, SE), agakoresho ko kuyisukura kavanwaho, kagashyirwa hejuru y'isahani ya petri (ishusho ya 1). 1A). Hindura indyo yawe buri munsi. Komeza laboratwari nkuko byasobanuwe haruguru. Imibu yarokotse yabazwe kabiri ku munsi, mu gihe imibu yapfuye yajugunywe kugeza umubu wa nyuma upfuye (n = 40 kuri buri muti). Kurokoka kw'imibu yagaburirwaga ku ndyo zitandukanye byasesenguwe hifashishijwe uburyo bwo kubaho bwa Kaplan-Meyer na ibizamini byo kugereranya uburyo bwo kubaho hagati y’indyo (IBM SPSS Statistics 24.0.0.0).
Urusyo rukora imibu rwihariye rushingiye kuri Attisano et al.[17], rukozwe mu dupapuro twa acrylic dufite ubugari bwa mm 5 (ubugari bwa cm 10 x uburebure bwa cm 10 x uburebure bwa cm 10) tudafite udupapuro tw'imbere n'inyuma (Ishusho ya 3: hejuru). Iteraniro rizunguruka rifite umuyoboro uhagaze ukozwe mu nkingi ya chromatography ya gaze (0.25 mm id; cm 7.5 L) hamwe n'impera zifatanye n'urushinge rw'udukoko rumanitse hagati ya magnets ebyiri za neodymium ziri kuri cm 9 uvuye ahandi. Umuyoboro utambitse ukozwe mu bikoresho bimwe (cm 6.5 L) wagabanyije umuyoboro uhagaze ugakora ukuboko gufunganye n'ukuboko kwari gufite agace gato k'urupapuro rwa aluminiyumu nk'ikimenyetso cyo guhagarika urumuri.
Inka z’ingore zimaze amasaha 24 zishonje zahawe iyi ndyo yavuzwe haruguru mu gihe cy’iminota 30 mbere yo kuyifata. Imibu y’ingore igaburirwa neza, yashyizwemo ikinya ku rubura mu gihe cy’iminota 2-3, ishyirwa ku dupira tw’udukoko hakoreshejwe ishaza ry’inzuki (Joel Svenssons Vaxfabrik AB, Munka Ljungby, SE) hanyuma ihambirwa ku maboko y’imiyoboro itambitse. Flying Mill. Impinduka kuri buri ndege zanditswe n’icyuma cyandika amakuru cyihariye, hanyuma kikabikwa kandi kikagaragazwa hakoreshejwe porogaramu ya PC-Lab 2000™ (v4.01; Velleman, Gavere, BE). Uruganda rw’indege rwashyizwe mu cyumba gifite ikirere gitekanye (amasaha 12:12, urumuri: umwijima, 25 ± 2 °C, 65 ± 5% RH).
Kugira ngo harebwe imiterere y'ibikorwa byo mu ndege, intera yose yagenze (m) n'umubare wose w'ibikorwa byo mu ndege bikurikiranye byabazwe ku isaha mu gihe cy'amasaha 24. Byongeye kandi, intera mpuzandengo yagenze n'abagore ku giti cyabo yagereranyijwe mu buvuzi kandi yasesenguwe hakoreshejwe uburyo bumwe bwa ANOVA na Tukey's post hoc analysis (JMP Pro, v14.0.0, SAS Institute Inc.), aho intera mpuzandengo yafatwaga nk'ikintu gishingiye ku kintu runaka, mu gihe ubuvuzi ari ikintu cyigenga. Byongeye kandi, umubare mpuzandengo w'ibitero ubarwa mu nyongera z'iminota 10.
Kugira ngo harebwe ingaruka z'imirire ku mikorere y'imyororokere ya An.arabiensis, ingore esheshatu (4 dpe) zoherejwe mu tuzu twa Bugdorm (30 cm × 30 cm × 30 cm) nyuma yo gukusanya amaraso hanyuma zigahabwa indyo y'igerageza mu gihe cy'amasaha 48 nk'uko byasobanuwe haruguru. Hanyuma indyo yakuweho maze ibikombe byo gutera inda (30 ml; Nolato Hertila) byuzuyemo ml 20 z'amazi yaciwe bitangwa ku munsi wa gatatu mu masaha 48, bihinduka buri masaha 24. Subiramo buri gahunda y'imirire inshuro 20-50. Amagi yabazwe kandi yanditswe kuri buri kazu k'igerageza. Ingero z'amagi zakoreshejwe mu gusuzuma ingano n'uburebure bw'amagi ku giti cyayo (n ≥ 200 kuri buri ndyo) hakoreshejwe mikorosikopi ya Dialux-20 (DM1000; Ernst Leitz Wetzlar, Wetzlar, DE) ifite Kamera ya Leica (DFC) 320 R2; Leica Microsystems Ltd., DE). Amagi asigaye yashyizwe mu cyumba gicungwa n’ikirere mu buryo busanzwe bwo kororerwamo amasaha 24, kandi icyitegererezo cy’inkoko za mbere ziherutse kugaragara (n ≥ 200 kuri buri ndyo) cyapimwe, nk’uko byasobanuwe haruguru. Umubare w’amagi n’ingano y’inkoko byagereranyijwe hagati y’uburyo bwo kuvura no gukoresha uburyo bumwe bwo gusesengura bwa ANOVA na Tukey nyuma y’aho (JMP Pro, v14.0.0, SAS Institute Inc.).
Inkari zo mu mutwe zakuwe mu nka nshya (isaha imwe nyuma yo gupimwa), amasaha 24, amasaha 72 n'amasaha 168 zamaze igihe zakusanyijwe mu nka za Zebu, Arsi. Kugira ngo byorohe, inkari zakusanyijwe kare mu gitondo inka zari zikiri mu kiraro. Inkari zakusanyijwe mu nka 10 kandi mililitiro 100-200 za buri nka zashyizwe mu dufuka two gutekeshamo twa polyamide (Toppits Cofresco, Frischhalteprodukte GmbH na Co., Minden, DE) muri litiro 3 za polyamide ifite umupfundikizo mu ngoma za vinyl chloride. Inkari zo mu mutwe zakuwe mu nka zatoranijwe mu buryo butaziguye (bushya) cyangwa nyuma yo gukura ku bushyuhe bw'icyumba mu gihe cy'amasaha 24, amasaha 72 na 168, ni ukuvuga ko buri nkari yari ihagarariye buri tsinda ry'imyaka.
Ku bijyanye no gukusanya ibyuma bivangwa n'ingufu zo mu mutwe, hakoreshejwe uburyo bwo gufunga uruziga kugira ngo hanyuzwe umuyoboro wa gaze ufunguye karubone (100 ml min-1) unyuze mu gafuka ka polyamide ujya mu gasanduku k'ingufu mu gihe cy'amasaha 2.5 hakoreshejwe pompe ya diaphragm vacuum (KNF Neuberger, Freiburg, DE). Nk'uburyo bwo kugenzura, gukusanya ibyuma bivangwa n'ingufu zo mu mutwe byakozwe mu gafuka ka polyamide kari karimo ubusa. Inkingi yo gukusanya ibyuma yakozwe mu muyoboro wa Teflon (5.5 cm x 3 mm id) irimo 35 mg ya Porapak Q (50/80 mesh; Waters Associates, Milford, MA, US) hagati y'udupira tw'ikirahure. Mbere yo gukoresha, inkingi yasukwaga na ml 1 ya n-hexane (Merck, Darmstadt, DE) na ml 1 ya pentane (99.0% pure solvent GC grade, Sigma Aldrich). Ibivangwa byo mu mutwe byakuwemo 400 μl ya pentane. Ibivangwa byo mu mutwe byashyizwe hamwe hanyuma bikabikwa kuri -20°C kugeza igihe bizakoreshwa. isesengura.
Ibisubizo by'imyitwarire y'inkari zishakishwa n'umuti urya amaraso byakusanyijwe mu nkari nshya, amasaha 24, amasaha 72, n'amasaha 168, hakoreshejwe olfactometer y'ikirahure igororotse [18]. Igerageza ryakozwe mu gihe cya ZT 13-15, igihe An yari amaze igihe kinini akora akazi ko gushaka urugo. Arab [19]. Olfactometer y'ikirahure (80 cm × 9.5 cm id) yamurikiwe urumuri rutukura rwa 3 ± 1 lx ruturutse hejuru. Amakara yasukuye kandi afite ubushuhe (25 ± 2 °C, 65 ± 2% ubushuhe) yatsinze isuzuma rya bioassay kuri cm 30 s-1. Umwuka unyura mu ruhererekane rw'ibyuma bitagira umugese, bigatuma habaho urujya n'uruza rw'amazi n'imiterere y'amafu. Imashini igabanya amenyo (4 cm × 1 cm; L:D; DAB Dental AB), ihagaze ku gikoresho cya cm 5 ku mpera y'umuyaga. olfactometer, hamwe n'impinduka za stimulator buri minota 5. Mu gusesengura, 10 μl ya buri gice cy'umutwe, cyavanzwemo 1:10, cyakoreshejwe nk'igikangura. Ingano ingana ya pentane yakoreshejwe nk'igenzura. Imibu ikoresha ikoranabuhanga ryo gushaka cyangwa ikoresha amaraso yashyizwe mu dusanduku two kurekura buri masaha 2-3 mbere y'uko igerageza ritangira. Agasanduku ko kurekura kashyizwe ku ruhande rw'umuyaga wa olfactometer, maze imibu yemererwa kumenyerana n'ikirere umunota 1, hanyuma valve y'ikinyugunyugu cy'akazu irafungurwa kugira ngo irekure. Gukurura ubuvuzi cyangwa kugenzura byasesenguwe nk'umubare w'imibu yahuye n'isoko mu minota 5 nyuma yo kurekura. Buri gice cy'umutwe n'igenzura byasubiwemo nibura inshuro 30, kandi kugira ngo hirindwe ingaruka z'umunsi umwe, umubare ungana w'ubuvuzi n'igenzura byageragejwe kuri buri munsi w'igerageza. Shaka ibisubizo bivuye ku muntu n'abahawe amaraso. Itsinda ry'icyarabu n'iry'umutwe ryasesenguwe hakoreshejwe uburyo bwo gusubira inyuma bw'ibikoresho byavuzwe haruguru hagakurikiraho kugereranya ibipimo bidasanzwe (JMP Pro, v14.0.0, Ikigo cya SAS Inc.).
Uburyo An yakiraga inkari. Ibikomoka ku nkari nshya n'izakuze byasesenguwe mu byumba bya Bugdorm (30 cm × 30 cm × 30 cm; MegaView Science). Ibikombe bya pulasitiki (30 mL; Nolato Hertila) byuzuyemo mL 20 z'amazi yatetse byatanze ishingiro ryo guterana kw'inka kandi bishyirwa mu mfuruka zihanganye z'icyumba, cm 24 hagati. Ibikombe byo kuvura byahinduwe na μl 10 za buri cyumba cyo guterana kw'inka ku gipimo cya 1:10. Ingano ingana ya pentane yakoreshejwe mu guhindura igikombe cyo kugenzura. Ibikombe byo kuvura n'ibyo kugenzura byahinduwe hagati ya buri gikorwa kugira ngo harebwe ingaruka z'aho byabereye. Inka icumi zonsa amaraso zarekuwe mu byumba byo kugerageza kuri ZT 9-11 maze amagi mu bikombe abarwa nyuma y'amasaha 24. Uburyo bwo kubara igipimo cyo guterana kw'inka ni: (umubare w'amagi yashyizwe mu gikombe cyo kuvura - umubare w'amagi yashyizwe mu gikombe cyo kugenzura) / (umubare wose w'amagi yashyizwe). Buri gikorwa cyasubiwemo inshuro 8.
Isesengura rya gaze chromatographic na electron antene (GC-EAD) rya An.arabiensis y’ingore ryakozwe nk'uko byasobanuwe mbere [20]. Muri make, ibinyabutabire bishya byo mu mutwe byatandukanyijwe hakoreshejwe Agilent Technologies 6890 GC (Santa Clara, CA, US) bifite inkingi ya HP-5 (30 m × 0.25 mm id, ubugari bwa filime ya 0.25 μm, Agilent Technologies). n'inkari zishaje. Hydrogen yakoreshejwe nk'ikigero cyo kugenda hamwe n'umuvuduko w'impuzandengo wa cm 45 s-1. Buri gipimo (2 μl) cyatewe mu masegonda 30 mu buryo butagira ibice hamwe n'ubushyuhe bwo kwinjira bwa 225 °C. Ubushyuhe bw'ifuru ya GC bwashyizwe kuri 35 °C (iminota 3 ifata) kugeza kuri 300 °C (iminota 10 ifata) kuri 10 °C min-1. Mu gice cyo gutandukanya amazi ya GC, hongewemo psi 4 za azote kandi zigabanywamo 1:1 muri Gerstel 3D/2 low dead volume cross (Gerstel, Mülheim, DE) hagati y'icyuma gipima umuriro na EAD. Umuyoboro w'amazi ya GC wa EAD wanyujijwe mu murongo wo kohereza wa Gerstel ODP-2, ukurikirana ubushyuhe bw'ifuru ya GC hamwe na 5 °C, mu muyoboro w'ikirahure (10 cm × 8 mm), aho wavanzwe n'umwuka wa karuboni uyunguruye (1.5 l min-1). Antene yashyizwemo 0.5 cm uvuye aho umuyoboro usohokera. Buri mubu umwe wari ufite umwe, naho ku mibu ikeneye uwayibamo, byibuze hakozwe inshuro eshatu ku nkari z’imyaka yose.
Kumenya ibinyabutabire bikora mu matsinda y’inkari nshya n’izabukuru hakoreshejwe GC-MS; 6890 GC na 5975 MS; Agilent Technologies) kugira ngo haboneke ibisubizo by’inkoni mu isesengura rya GC-EAD, rikora mu buryo bwa electron impact ionization kuri 70 eV. GC yari ifite HP-5MS UI-coated fused silica capillary column (60 m × 0.25 mm imbere diameter, 0.25 μm film sight) hakoreshejwe helium nk'igice cyo kugenda gifite impuzandengo y'umuvuduko wa literally wa cm 35 s-1. Icyitegererezo cya 2 μl cyatewe hakoreshejwe uburyo bumwe bwo gusimbuza injector n'ubushyuhe bw'ifuru nk'ubwo gusimbuza GC-EAD. Ibinyabutabire byamenyekanye hashingiwe ku gihe cyo kubibika (Kovát index) na mass spectra ugereranije n'isomero ryakoreweho ubushakashatsi n'isomero rya NIST14 (Agilent). Ibinyabutabire byamenyekanye byemejwe hashingiwe ku bipimo nyabyo (Idosiye y'inyongera ya 1: Imbonerahamwe S2). Ku bijyanye no gupima, heptyl acetate (10 ng, 99.8% ubuziranenge bwa shimi, Aldrich) yatewe nk'igipimo cyo hanze.
Gusuzuma akamaro k'uruvange rw'impumuro ikoze mu buryo bw'ubukorano rugizwe n'ibintu bifatika byagaragaye mu nkari nshya n'izashaje kugira ngo bikurure Ans.arabiensis ishakisha kandi inyunyuza amaraso, hakoreshejwe olfactometer imwe na protocole nkuko byavuzwe haruguru. Uruvange rw'ubukorano rwagaragaje imiterere n'ibipimo by'ibintu biri mu binyabutabire bivanze mu mutwe w'inkari nshya, amasaha 24, amasaha 48, amasaha 72, n'amasaha 168 (Ishusho ya 5D-G; Dosiye y'inyongera ya 1: Imbonerahamwe S2). Kugira ngo usesengure, koresha 10 μl y'umuvuduko wa 1:100 w'uruvange rw'ubukorano, hamwe n'igipimo rusange cyo kurekura kiri hagati ya 140-2400 ng h-1, kugira ngo upime ubwiza bw'imibu ikunda n'inyunyuza amaraso. Nyuma yaho, ikizamini gikorwa ku mvange yuzuye, aho uruvange rw'ibinyabutabire by'uruvange rumwe rw'uruvange rwuzuye rukurwaho. Shaka ibisubizo ku mvange ikunda n'iy'amaraso. Uruvange rw'Abarabu na synthetic n'iy'ubukorano rwasesenguwe hakoreshejwe uburyo bwo gusubira inyuma bw'ibintu hagakurikiraho kugereranya ibidasanzwe. ibipimo (JMP Pro, v14.0.0, SAS Institute Inc.).
Kugira ngo harebwe niba inkari z'inka zishobora kuba ikimenyetso cy'aho imibu ya malariya ituye, inkari z'inka nshya n'izakuze, zakusanyijwe nkuko byasobanuwe haruguru, hanyuma amazi ashyirwa mu ndobo 3 za litiro (100 ml) zishyirwa mu mitego y'ibyatsi. (BG-HDT verisiyo; BioGents, Regensburg, DE). Imitego icumi yashyizwe muri metero 50 uvuye mu rwuri, metero 400 uvuye mu mudugudu (Silay, Etiyopiya, 5°53´24´´N, 37°29´24´´E) nta nka zihari, mu bibanza byororokeramo bihoraho no mu midugudu. Imitego itanu yashyushywe kugira ngo isa n'aho ihari, mu gihe imitego itanu yasigaye idashyushye. Buri hantu havurirwa hazunguruka nijoro mu gihe cy'amajoro atanu yose. Imibare y'imibu yafashwe mu mitego y'ibyatsi by'inka z'imyaka itandukanye yagereranyijwe hakoreshejwe uburyo bwo guhindura imiterere hakoreshejwe uburyo bwa beta binomial (JMP Pro, v14.0.0, SAS Institute Inc.).
Mu mudugudu wanduye malariya hafi y'umujyi wa Maki, mu karere ka Oromia, muri Etiyopiya (8° 11′ 08″ N, 38° 81′ 70″ E; Ishusho ya 6A). Ubushakashatsi bwakozwe hagati ya Kanama na Nzeri mbere y'uko hakoreshwa imiti ikoreshwa mu gutera imiti mu nzu buri mwaka, hamwe n'igihe cy'imvura kirekire. Inzu eshanu (20–50 uvuye ahandi) ziri ku nkengero z'umudugudu zatoranyijwe kugira ngo zikorerwe ubu bushakashatsi (Ishusho ya 6A). Ibipimo byakoreshejwe mu guhitamo amazu ni ibi: inyamaswa ntizemerewe kwinjira mu nzu, guteka mu nzu (guteka inkwi cyangwa amakara) ntibyemerewe (nibura mu gihe cy'igerageza), n'amazu afite abaturage barenga babiri, baryama mu miti idakoreshwa mu kwica udukoko. munsi y’urushundura rw’imibu rwavuwe. Icyemezo cy’amahame mbwirizamuco cyatanzwe n’Inama Ishinzwe Isuzuma ry’Amahame Mbonezamubano mu Bushakashatsi (IRB/022/2016) y’Ishami ry’Ubumenyi bw’Ubwenge Kamere (CNS-IRB), Kaminuza ya Addis Ababa, hakurikijwe amabwiriza yashyizweho n’Itangazo ry’Ishyirahamwe Mpuzamahanga ry’Abaganga rya Helsinki. Buri muyobozi w’urugo yabonetse abifashijwemo n’abakozi b’ubuvuzi. Gahunda yose yemejwe n’ubuyobozi bw’ibanze ku rwego rw’akarere n’akarere ('kebele'). Igishushanyo mbonera cy’igerageza cyakurikije igishushanyo mbonera cya kare cya 2 × 2 cy’Ikilatini, aho imvange n’igenzura byashyizwe mu mazu abiri mu ijoro rya mbere hanyuma bigasimbuza amazu mu ijoro rikurikiyeho ry’igerageza. Iyi gahunda yasubiwemo inshuro icumi. Byongeye kandi, kugira ngo harebwe imikorere y’imibu mu mazu yatoranijwe, imitego ya CDC yashyizwe mu ijoro rikurikiranye mu ntangiriro, hagati no mu mpera z’igerageza mu murima mu gihe kimwe cy’umunsi.
Uruvange rw’ibinyabutabire rurimo ibintu bitandatu bikora ku buzima rwashongeshejwe muri heptane (97.0% solvent GC grade, Sigma Aldrich) maze rurekurwa kuri 140 ng h-1 hakoreshejwe ipamba ritanga umugozi [20]. Icupa ritanga umugozi ryemerera ibintu byose gurekurwa mu rugero ruhoraho mu gihe cy’amasaha 12 y’igerageza. Heptane yakoreshejwe nk’igenzura. Aka gacupa kari kamanitswe iruhande rw’aho umutego w’urumuri rw’Ikigo gishinzwe kurwanya no gukumira indwara (CDC) winjirira (John W. Hock Company, Gainesville, FL, US; Ishusho ya 6A). Imitego yamanitswe kuri metero 0.8 - 1 uvuye hasi, hafi y’ikirenge, maze umukorerabushake aryama munsi y’urushundura rutavuwe maze akorerwa hagati ya saa 18:00 na 06:30. Imibu yafashwe n’igitsina n’imiterere yayo (itagaburirwa, ihabwa, itwite, kandi itwite [21] nyuma yaho yasuzumwe hakoreshejwe isesengura rya polymerase chain reaction (PCR) kugira ngo hamenyekane ubwoko bwagaragajwe nka A. gambiae sl. Ibice bigize uru ruganda [23]. Mu bushakashatsi bwo mu murima, gushakisha imitego mu mazu abiri byasesenguwe hakoreshejwe uburyo bwo guhuza ibintu, aho gukurura byari ibintu bihinduka bitewe n’uko ibintu bimeze, naho uburyo bwo kuvura (imvange y’ibikomoka ku bidukikije vs kugenzura) bikaba ari ingaruka zidahinduka (JMP® 14.0. 0. SAS Institute Inc.). Aha, turagaragaza agaciro ka χ2 na p bivuye mu kizamini cy’ikigereranyo cy’ingaruka.
Suzuma niba nta kibazo kirimo. Arabiensis yashoboye kubona inkari, isoko nyamukuru ya azote, urea, binyuze mu kugaburira mu buryo butaziguye, mu masaha 48 nyuma yo kuyitanga mu minsi 4 nyuma yo kugerageza (dpe) ku bagore bashaka inyange n'abanyoye amaraso (Ishusho ya 1A). Abagore bashaka inyange n'abanyoye amaraso banyoye sucrose nyinshi cyane kurusha izindi ndyo cyangwa amazi (F(5,426) = 20.15, p < 0.0001 na F(5,299) = 56.00, p < 0.0001, uko bikurikirana; Ishusho ya 1B,C). Byongeye kandi, abagore bashaka inyange bariye bike mu nkari mu masaha 72 ugereranije n'inkari mu masaha 168 (Ishusho ya 1B). Iyo bahawe indyo irimo urea, abagore bashaka inyange banyoye ingano nini ya urea kuri 2.69 mM ugereranije n'ibindi bipimo byose n'amazi, mu gihe bitagaragara ko bitandukaniye na 10% ya sucrose (F(10,813) = 15.72, p < 0.0001; Ishusho ya 1D). Ibi bitandukanye n'uko abagore barya amaraso babyitwaramo, bakunze kurya indyo nyinshi irimo urea kurusha amazi, nubwo ari munsi ya 10% ya sucrose (F(10.557) = 78.35, p < 0.0001; Ishusho ya 1).1E). Byongeye kandi, iyo ugereranya imiterere ibiri y’umubiri, abagore bafite phlebotomism bariye urea nyinshi kurusha abagore bashaka kwakira ku gipimo gito cyane, kandi aba bagore bariye urea ingano imwe ku gipimo cyo hejuru (F(1.953) = 78.82, p < 0.0001; Ishusho ya 1F, G). Nubwo kurya indyo irimo urea byasaga nkaho bifite agaciro keza (Ishusho ya 1D, E), abagore bo muri iyo mimerere yombi y’umubiri bashoboye guhindura ingano ya urea yinjijwe mu cyiciro cyose cy’ingano ya urea mu buryo bwa log-linear (Ishusho ya 1F, G). ).Mu buryo nk'ubwo, imibu isa nkaho igenzura uburyo azote yinjirira mu nkari igenzura ingano y’inkari zinjiramo, kuko ingano ya azote mu nkari igaragazwa n’ingano y’inkari zinjiramo (Ishusho ya 1B, C na B).
Kugira ngo harebwe ingaruka z'inkari na urea ku buzima bw'imibu ikeneye kandi inyunyuza amaraso, abagore bahawe inkari z'imyaka ine yose (amasaha 24, amasaha 72, n'amasaha 168 nyuma yo gukurwamo) hamwe n'ingano zitandukanye za urea, hamwe n'amazi yaciwe na sucrose 10% byakoreshejwe nk'igenzura (Ishusho ya 2A). Iri sesengura ry'ubuzima ryagaragaje ko indyo yagize ingaruka zikomeye ku buzima rusange bw'abagore bashaka utwo tunyangingo (inkari: χ2 = 108.5, df = 5, p < 0.0001; urea: χ2 = 122.8, df = 5, p < 0.0001; Ishusho ya 2B, C) n'abagore bahabwa amaraso (inkari: χ2 = 93.0, df = 5, p < 0.0001; urea: χ2 = 137.9, df = 5, p < 0.0001; Ishusho ya 2D,E). Mu bushakashatsi bwose, abagore bagaburirwa indyo y'inkari, urea, n'amazi bari bafite igipimo cyo kubaho kiri hasi cyane ugereranije n'abagore bagaburirwa indyo ya sucrose (Ishusho ya 2B-E). Abagore bashaka inkari nshya n'izamaze igihe bagaragaje igipimo cyo kubaho gitandukanye, aho abagaburirwa inkari zamaze amasaha 72 (p = 0.016) bafite amahirwe make yo kubaho (Ishusho ya 2B). Byongeye kandi, abagore bashaka inkari zamaze amasaha 135 urea bamara igihe kirekire ugereranyije n'amazi agenzura (p < 0.04) (Ishusho ya 2C). Ugereranije n'amazi, abagore bagaburirwa inkari nshya n'inkari z'amasaha 24 zamaze igihe kirekire (p = 0.001 na p = 0.012, uko bikurikirana; Ishusho ya 2D), mu gihe abagore bagaburirwa inkari z'amasaha 72 zamaze igihe kirekire ugereranyije n'abagaburirwa inkari ngufi nshya n'inkari z'amasaha 24 zamaze igihe kirekire (p < 0.0001 na p = 0.013, uko bikurikirana; Ishusho ya 2D). Iyo bagaburirwaga urea ya 135 mM, abagore bagaburirwa amaraso bamara igihe kirekire ugereranyije n'abandi bose izindi ngano za urea n'amazi (p < 0.013; Ishusho ya 2E).
Kurokoka kw'inka zo mu bwoko bwa Anopheles arabinis zirya inkari z'inka na urea. Mu isuzuma ry'ibinyabuzima (A), imibu y'inka z'inka yahawe indyo igizwe n'inka nshya n'izashaje, ingano zitandukanye za urea, sucrose (10%) n'amazi yaciwe (H2O). Kurokoka kw'inka zo mu bwoko bwa (B, C) n'izo mu bwoko bwa (D, E) byanditswe buri masaha 12 kugeza igihe inkazi zose zirya inkari (B, D) na urea (C, E), hamwe n'izigenzura, Sucrose n'amazi, byapfuye.
Intera yose hamwe n'umubare w'inshuro zagenwe mu igerageza ry'uruganda rw'indege mu gihe cy'amasaha 24 bitandukanye hagati y'imibu ikunda gushaka inyamaswa n'ikunda kunyonka amaraso, byagaragaje ko ikora buhoro muri rusange (Ishusho ya 3). Imibu ikunda gushaka inyamaswa yatangaga inkari nshya cyangwa sucrose n'amazi byagaragaje imiterere itandukanye yo kuguruka (Ishusho ya 3), aho inyamaswa z'ingore zirya inkari nshya zikora cyane mu gitondo, mu gihe izigaburirwaga amasaha 24 na 168. Imibu ikunda kurya inkari zagaragaje imiterere itandukanye yo kuguruka kandi ahanini yabaga ikora buri munsi. Imibu y'ingore yatangaga sucrose cyangwa inkari z'amasaha 72 yagaragaje imikorere mu masaha 24, mu gihe iz'ingore zatangaga amazi zagaragazaga imikorere myinshi mu gihe cy'amasaha hagati. Imibu ikunda kurya sucrose yagaragaje urwego rwo hejuru rw'imikorere nijoro no mu gitondo cya kare, mu gihe izikunda kurya inkari z'amasaha 72 zagize imiterere igabanuka mu gihe cy'amasaha 24 (Ishusho ya 3).
Imiterere y'indege y'ingore ikunda guhiga amaraso yitwa Anopheles arabinis irya inkari z'inka na urea. Mu igeragezwa ry'uruganda rwo mu kirere, imibu y'ingore yagaburirwaga inkari nshya n'izabukuru, ingano zitandukanye za urea, sucrose (10%), n'amazi yaciwe (H2O) byashyizwe ku maboko atambitse, azunguruka neza (hejuru). Ku ngore zikunda guhumeka (ibumoso) n'izikunda guhumeka amaraso (iburyo), intera yose hamwe n'umubare w'indege ku isaha kuri buri ndyo mu gihe cy'amasaha 24 byanditswe (umwirabura: imvi; urumuri: umweru). Intera mpuzandengo n'umubare mpuzandengo w'inshuro zigenda zigaragara iburyo bw'imbonerahamwe y'ibikorwa bya circadian. Utudomo tw'amakosa tugaragaza ikosa risanzwe ry'ikigereranyo. Isesengura ry'imibare reba inyandiko
Muri rusange, ibikorwa byo mu kirere by’ingore zishaka kwakira inyamaswa byakurikije icyerekezo gisa n’icy’intera yo mu kirere mu gihe cy’amasaha 24. Intera y’ikirere yagizweho ingaruka cyane n’indyo yariye (F(5, 138) = 28.27, p < 0.0001), naho ingore zishaka kwakira inyamaswa zariye inkari amasaha 72 zarutse intera ndende cyane ugereranije n’izindi ndyo zose (p < 0.0001), naho inyenzi zirya sucrose zarutse intera ndende kurusha inyenzi nshya (p = 0.022) n’inkari zimaze amasaha 24 (p = 0.022). Bitandukanye n’icyerekezo cyo mu kirere cyasobanuwe n’indyo y’inkari, inyenzi zirya urea zarutse inzobere zishaka kwakira inyamaswa zagaragaje imikorere yo mu kirere mu gihe cy’amasaha 24, zigakomera mu gice cya kabiri cy’umwijima (Ishusho ya 3). Nubwo imiterere y’ibikorwa yari isa, inyenzi zishaka kwakira inyamaswa zariye urea zongereye cyane impuzandengo y’ikirere bitewe n’ingano y’inyongera (F(5, 138) = 1310.91, p < 0.0001). abagore bagaburirwa igipimo icyo ari cyo cyose cya urea cyagurutse igihe kirekire kurusha abagore bagaburirwa amazi cyangwa sucrose (p < 0.03).
Muri rusange, imikorere y’inkari mu guhumeka amaraso yagumye ihamye kandi yagumyeho mu masaha 24 mu mirire yose, aho inkari zakomeje kwiyongera mu gice cya kabiri cy’igihe cy’umwijima ku bagore barya amazi ndetse no ku bagore barya bashya kandi bafite amasaha 24 (ishusho ya 3). Nubwo indyo y’inkari yagize ingaruka zikomeye ku ntera yo guhumeka ku bagore barya amaraso (F(5, 138) = 4.83, p = 0.0004), indyo ya urea ntiyagize ingaruka (F(5, 138) = 1.36, p = 0.24). hamwe n’izindi nkari n’indyo igenzura (bishya, p = 0.0091; amasaha 72, p = 0.0022; amasaha 168, p = 0.001; sucrose, p = 0.0017; dH2O, p = 0.036).
Ingaruka zo kugaburira inkari na urea ku bipimo byo kubyara zasuzumwe mu isuzuma ry’ibinyabuzima by’ingore (Ishusho ya 4A) kandi zasuzumwe hakurikijwe umubare w’amagi yatewe na buri ngore, ingano y’igi, n’inkoko nshya yavutse. Umubare w’amagi yatewe. Inkoko z’Abarabu zigaburirwa inkari zaratandukanye bitewe n’imirire (F(5,222) = 4.38, p = 0.0008; Ishusho ya 4B). Inkoko z’ingore zahawe inkari amasaha 24, amaraso yatewe amagi menshi cyane kurusha inkoko zigaburirwa izindi ndyo kandi zari zisa n’izahawe sucrose (Ishusho ya 4B). Mu buryo nk’ubwo, ingano y’amagi yatewe n’inkoko z’ingore zahawe inkari yatandukanaga bitewe n’imirire (F(5, 209) = 12.85, p < 0.0001), inkari z’amasaha 24 n’inkoko z’ingore zahawe sucrose zaterwaga amagi manini cyane kurusha inkoko zahawe amazi, mu gihe amagi y’inkoko z’ingore zahawe amasaha 168 y’inkari yari make cyane (Ishusho ya 4C). Byongeye kandi, imirire y’inkari yagize ingaruka zikomeye ku bunini bw’inkoko (F(5, 187) = 7.86, p < 0.0001), hamwe n'udusimba tunini cyane tuva mu magi yatewe n'ingore zonsa inkari zifite amasaha 24 na 72 ugereranyije n'udusimba twatewe n'ingore zonsa. Ingore zonsa amazi n'izonsari zimara amasaha 168 (Ishusho ya 4D).
Imikorere y’imyororokere y’ingore za Anopheles arabinis zirya inkari z’inka na urea. Imibu y’ingore yorojwe amaraso yagaburirwaga indyo irimo inkari nshya n’izakuze, ingano zitandukanye za urea, sucrose (10%), n’amazi yaciwe (H2O) mu gihe cy’amasaha 48 mbere yo gushyira mu bipimo bya bioassay no kubona substrates z’amagi. Amasaha 48 (A). Umubare w’igi (B, E), ingano y’igi (C, F) n’ingano y’ibihumyo (D, G) byagizweho ingaruka cyane n’indyo yatanzwe (inka z’inka: BD; urea: EG). Uburyo bwa buri gipimo bwapimwe hakoreshejwe amazina y’inyuguti zitandukanye bwari butandukanye cyane (ANOVA y’inzira imwe hakoreshejwe isesengura rya Tukey nyuma y’aho; p < 0.05). Uduce tw’amakosa tugaragaza ikosa risanzwe ry’ikigereranyo
Nk’igice cy’ingenzi cya azote mu nkari, urea, iyo itangwa nk’indyo ku bagore barya amaraso, yagize ingaruka zikomeye ku myororokere mu bushakashatsi bwose. Umubare w’amagi yatewe n’abagore barya urea, nyuma yo kurya amaraso, bitewe n’ingano ya urea (F(11, 360) = 4.69; p < 0.0001), abagore baryaga urea hagati ya 134 µM na 1.34 mM bateraga amagi menshi (Ishusho ya 4E). Abagore baryaga urea ya 134 µM cyangwa irenga bateraga amagi manini kurusha abagore baryaga amazi (F(10, 4245) = 36.7; p < 0.0001; Ishusho ya 4F), n’ingano y’inkari, nubwo yagizweho ingaruka n’ingano ya urea isa ku bagore babyara (F(10, 3305) = 37.9; p < 0.0001) yari ihindagurika cyane (Ishusho ya 4G).
Gukurura cyane ibikomoka ku nkari zikunda gushaka aho ziherereye. Igipimo cya arabiensis cyapimwe muri olfactometer y'ikirahure (Ishusho ya 5A) cyagizweho ingaruka cyane n'imyaka y'inkari (χ2 = 15.9, df = 4, p = 0.0032; Ishusho ya 5B). Isesengura rya nyuma ryagaragaje ko impumuro y'inkari zangiritse mu masaha 24 yateraga ubwinshi bw'ubwiza ugereranije n'ubundi buryo bwose bwo kuvura (amasaha 72: p = 0.0060, amasaha 168: p = 0.012, pentane: p = 0.00070), Uretse impumuro y'inkari nshya (p = 0.13; Ishusho ya 5B). Nubwo muri rusange imibu inyunyuza amaraso mu mpumuro y'inkari ntabwo yari itandukanye cyane (χ2 = 8.78, df = 4, p = 0.067; Ishusho ya 5C), izi nkumi zagaragaye ko zikurura cyane ibikomoka ku nkari zikunda ubwiza ugereranije n'inkari zimara amasaha 72 ugereranije kugenzura (p = 0.0066; Ishusho ya 5C).
Ibisubizo ku myitwarire ku mpumuro karemano y'inka mu gushaka inyamaswa n'amaraso bya Anopheles arabianus. Ishusho y'icyuma gipima umuyoboro w'ikirahure (A). Gukurura ibinyabutabire by'umutwe biva mu nkari nshya n'izabukuru ku mubu w'inka (B) n'umubu unyunyuza amaraso (C). Shaka uburyo Lord An acquisments yakuwe mu nkari nshya (D), amasaha 24 (E), amasaha 72 (F), n'amasaha 168 (G) y'inka zishaje biragaragara. Ibimenyetso bya elegitoroniki byo kubona antene (EAD) bigaragaza impinduka z'amashanyarazi mu gusubiza ibinyabutabire bikora mu mutwe bivuye kuri chromatograph ya gaze kandi bikabonwa n'icyuma gipima umuriro (FID). Akadomo kerekana amplitude yo gusubiza (mV) ugereranije n'igihe cyo kubika (s). Imiterere n'igipimo cyo kurekura (µg h-1) by'ibinyabutabire bikora neza biragaragara. Asterisk imwe (*) igaragaza igisubizo gihoraho cy'amplitude nke. Asterisk ebyiri (**) zigaragaza ibisubizo bidasubirwamo. Shaka ateri (H) n'umuti unyunyuza amaraso (I) An.arabiensis ifite ubwiza butandukanye ku bivange by'inkari z'inka nshya n'izashaje. Ingano y'imibu ikururwa n'amazina y'inyuguti zitandukanye yari itandukanye cyane (ANOVA y'uburyo bumwe ikoresheje isesengura rya Tukey nyuma y'aho; p < 0.05). Uduce tw'amakosa tugaragaza ikosa risanzwe ry'igipimo
Ann.arabiensis y'ingore, amasaha 72 n'amasaha 120 nyuma yo kurya amaraso, mu gihe cyo gutera intanga, nta kintu na kimwe cyagaragaye ku binyabutabire biva mu nkari nshya n'izabukuru byagaragaye ugereranije n'ibinyabutabire bya pentane (χ2 = 3.07, p > 0.05; Dosiye y'inyongera ya 1: Ishusho ya S1).
Ku bijyanye na Ann.arabiensis y’ingore, isesengura rya GC-EAD na GC-MS ryagaragaje ibintu umunani, bitandatu, bitatu na bitatu bikora ku mubiri (Ishusho ya 5D-G). Nubwo hagaragaye itandukaniro mu mubare w’ibintu byateye ingaruka z’amashanyarazi, inyinshi muri ibyo bintu zari muri buri gice cy’umutwe cy’inkari cyakusanyijwe mu nkari nshya n’izashaje. Kubwibyo, kuri buri gice cy’inkari, ibintu gusa byatanze igisubizo cy’umubiri bivuye ku ntera z’ingore ziri hejuru y’umubare w’inkende ni byo byashyizwe mu isesengura rirambuye.
Igipimo cy’irekurwa ry’ibintu bifatika mu mutwe cyariyongereye kuva kuri 29 µg h-1 mu nkari nshya kugera kuri 242 µg h-1 mu nkari zimaze amasaha 168, ahanini bitewe na p-cresol na m-formaldehyde Phenol yiyongera kimwe na phenol. Mu buryo bunyuranye, igipimo cy’irekurwa ry’ibindi bintu, nka 2-cyclohexen-1-one na decanal, cyaragabanutse uko imyaka y’inkari igenda yiyongera, ibyo bikaba byarajyanye no kugabanuka k’ubukana bw’ibimenyetso (ubwinshi) muri chromatogram (Ishusho ya 5D)-G ibumoso) hamwe n’uburyo umubiri witabira ibyo bintu (Ishusho ya 5D-G iburyo).
Muri rusange, uruvange rw’inzobere rwari rufite igipimo karemano cy’ibintu bifatika byagaragaye mu binyabutabire bishya n’ibishaje by’inkari (Ishusho ya 5D–G) kandi ntibyagaragaye ko byakuruye cyane mu gushaka inzobere (χ2 = 8.15, df = 4, p = 0.083; Ishusho ya 5H) cyangwa inzobere zinyunyuza amaraso (χ2 = 4.91, df = 4, p = 0.30; Ishusho ya 5I). Ariko, igereranya rya nyuma y’iyi miti ryagaragaje ko inzobere zishakisha inzobere zari zikurura cyane uruvange rw’inzobere mu n ...
Kugira ngo harebwe uruhare rw'ibice byihariye mu bivange by'inkari z'amasaha 24, hasuzumwe ibivange bitandatu byo gukuramo ibinyabutabire byapimwe hakurikijwe ibivange byuzuye mu gipimo cya Y-tube, aho ibinyabutabire byakuwemo. Ku mibu ishakisha aho ituye, gukuramo ibinyabutabire mu bivange byuzuye byagize ingaruka zikomeye ku buryo imyitwarire ikora (χ2 = 19.63, df = 6, p = 0.0032; Dosiye y'inyongera ya 1: Ishusho ya S2A), ibivange byose byo gukuramo byari byiza kurusha Bito kurusha bivanze neza. Mu buryo bunyuranye, gukuramo ibinyabutabire mu bivange byuzuye ntibyagize ingaruka ku buryo imyitwarire y'imibu inyunyuza amaraso (χ2 = 11.38, df = 6, p = 0.077), usibye decanal, byatumye urwego rwo hasi ugereranije n'uruvange rwuzuye Attraction (p = 0.022; Dosiye y'inyongera ya 1: Ishusho ya S2B).
Mu mudugudu wa Etiyopiya wakunze kugaragaramo malariya, uburyo inkari z'inka zikoreshwa amasaha 24 mu gukurura imibu mu mirima bwapimwe mu gihe cy'amajoro icumi (Ishusho ya 6A). Imibu 4.861 yose yafashwe kandi iramenyekana, muri yo 45.7% yari Anthropus.gambiae sl, 18.9% yari Anopheles pharoensis naho 35.4% yari Culex spp. (Idosiye y'inyongera ya 1: Imbonerahamwe S1). Anopheles arabinis ni yo yonyine iri mu bwoko bwa An.Gambian yagaragajwe hakoreshejwe isesengura rya PCR. Ugereranyije, imibu 320 yafashwe buri joro, muri icyo gihe imitego ifite imitego ivanze yafataga imibu myinshi kurusha imitego ivanze idafite uruvange (χ2(0, 3196) = 170.0, p < 0.0001). Imitego idaterwa yashyizwe kuri buri joro mu matanu yo kugenzura mu ntangiriro, hagati, no mu mpera z'igerageza. Imibare nk'iyo y'imibu yafashwe muri buri tsinda. by'imitego, bigaragaza ko nta kubogama hagati y'amazu (χ2(0, 1665) = 9 × 10-13, p > 0.05) kandi nta kugabanuka kw'abaturage mu gihe cy'ubushakashatsi. Ugereranije n'imitego yo kurwanya, umubare w'imibu yafashwe mu mitego irimo uruvange rw'ibikomoka ku bimera wiyongera cyane: gushaka umuturage (χ2(0, 2107) = 138.7, p < 0.0001), konsa amaraso vuba aha (χ2(0, 650) = 32.2, p < 0.0001) no gutwita (χ2(0, 228) = 6.27, p = 0.0123; Dosiye y'inyongera ya 1: Imbonerahamwe S1). Ibi binagaragarira mu mubare wose w'imibu yafashwe: gushaka umuturage > konka amaraso > gutwita > gutwita igice > igitsina gabo.
Isuzuma ry’imikorere y’uruvange rw’impumuro y’inkari z’inka mu masaha 24. Igerageza ryakorewe mu majyepfo yo hagati muri Etiyopiya (ikarita), hafi y’umujyi wa Maki (shyiramo), hakoreshejwe umutego woroheje wa Centers for Disease Control (CDC) (iburyo) mu mazu abiri, ufite igishushanyo mbonera cy’ikilatini kare (ishusho yo mu kirere) (A). Imitego ya CDC ikozwe mu buryo bw’impumuro ikurura kandi igafata inyamaswa z’ingore za Anopheles arabesques (B), ariko ntabwo ari Anopheles farroes (C), mu buryo butandukanye, ingaruka zishingiye ku miterere y’umubiri. Byongeye kandi, iyi mitego yafashe umubare w’imibu ya Culex yiyongera cyane. (D) Ugereranije no kugenzura. Uduti turi ibumoso tugaragaza impuzandengo y’imibu yafashwe mu duti tubiri tw’umutego uhumura (icyatsi kibisi) n’imitego yo kugenzura (ifunguye) (N = 10), mu gihe ututi turi iburyo tugaragaza impuzandengo y’umutego wo kugenzura mu duti tubiri tw’imitego yo kugenzura (ifunguye; N = 5). ). Inyenyeri zigaragaza urwego rw’uburemere bw’imibare (*p = 0.01 na ***p < 0.0001)
Ubwo bwoko butatu bwafashwe mu buryo butandukanye mu mitego irimo uruvange rw’ibikomoka ku bimera. Gushaka uwakira (χ2 (1, 1345) = 71.7, p < 0.0001), kurya amaraso (χ2 (1, 517) = 16.7, p < 0.0001) no gutwita (χ2 (1, 180) = 6.11, p = 0.0134) a .arabiensis yafatiwe mu mutego urekura uruvange rw’ibikomoka ku bimera (Ishusho ya 6B), mu gihe ingano ya An itari itandukanye. Pharoensis mu miterere itandukanye y’umubiri yabonetse (Ishusho ya 6C). Kuri Culex, umubare w’imibu ishakisha abakira ni wo wabonetse mu mitego yafatiwe mu ruvange rw’ibikomoka ku bimera (χ2 (1,1319) = 12.6, p = 0.0004; Ishusho ya 6D), ugereranije n’imitego yo kugenzura.
Imitego y’ibyambo by’inka iri hanze y’aho byashoboraga kuba biri hagati y’aho byororera n’abaturage bo mu cyaro muri Etiyopiya yakoreshejwe mu gusuzuma niba imibu ya malariya ikoresha impumuro y’inka nk’ikimenyetso cy’aho ituye. Mu gihe nta bimenyetso by’inka, ubushyuhe, kandi nta mpumuro y’inka iriho cyangwa ntayo, nta mibu yafashwe (Idosiye y’inyongera ya 1: Ishusho S3). Ariko, mu gihe hari ubushyuhe bwinshi n’impumuro y’inka, imibu ya malariya y’ingore yakururwaga ikanafatwa, nubwo yari mike, hatitawe ku myaka y’inka (χ2 (5, 25) = 2.29, p = 0.13; Dosiye y’inyongera ya 1: Ishusho S3). Mu buryo bunyuranye, abashinzwe kugenzura amazi ntibafashe imibu ya malariya ku bushyuhe bwinshi (Idosiye y’inyongera ya 1: Ishusho S3).
Imibu ya malariya ikura kandi igakwirakwiza ibintu birimo azote binyuze mu kugaburira inkari z'inka (ni ukuvuga ibiziba) kugira ngo yongere imiterere y'ubuzima, nk'uko bimeze ku tundi dukoko [2, 4, 24, 25, 26]. Inkari z'inka ni umutungo ushobora kongera gukoreshwa ujyanye n'ahantu ho kuruhukira abatera malariya, nk'aho inka ziruhukira n'ibimera birebire hafi y'amazu yo mu cyaro n'aho zibyarira. Imibu y'ingore isanga uyu mutungo binyuze mu guhumurirwa kandi ikabasha kugenzura uburyo ibintu birimo azote byinjira mu nkari, harimo na urea, igice cy'ingenzi gifite azote mu nkari [15, 16]. Bitewe n'imiterere y'umubiri w'umubu w'ingore, intungamubiri ziri mu nkari zigenewe kongera imikorere yo kuguruka no kubaho kw'imibu y'ingore ishaka kwakira, ndetse no kubaho no kubyara kw'abantu barya amaraso mu gihe cya mbere cy'ingendo ya gonadotropic. Kubwibyo, kuvanga inkari bigira uruhare runini mu mirire ku batera malariya bafunganye nk'abantu bakuru bafite imirire mibi [8], kuko biha imibu y'ingore ubushobozi bwo kubona ibintu by'ingenzi birimo azote binyuze mu kugaburira abantu bake. Iki cyavumbuwe gifite ingaruka zikomeye ku ndwara, kuko abagore bongera igihe cyo kubaho, ibikorwa n'umusaruro w'imyororokere, byose bigira ingaruka ku bushobozi bw'udukoko. Byongeye kandi, iyi myitwarire ishobora kuba intego ya gahunda zo gucunga udukoko mu gihe kizaza.